Traditional Salads and Soups with Wild Plants as a Source of...
13
Research Article Traditional Salads and Soups with Wild Plants as a Source of Antioxidants: A Comparative Chemical Analysis of Five Species Growing in Central Italy Valentina Savo , Francois Salomone, Elena Mattoni, Daniela Tofani , and Giulia Caneva Department of Science, Roma Tre University, Viale G. Marconi 446, Rome, Italy Correspondence should be addressed to Valentina Savo; [email protected] and Daniela Tofani; [email protected] Received 24 September 2018; Revised 30 January 2019; Accepted 18 February 2019; Published 5 March 2019 Academic Editor: G. K. Jayaprakasha Copyright © 2019 Valentina Savo et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. e interest and demand for nutraceuticals are rapidly increasing in many industrialized countries due to the emergence of health risks associated with the increased consumption of processed foods. Several wild Mediterranean plants used as traditional foods are an extraordinary source of nutraceutical substances with antioxidant properties. is study has two main aims: (1) to quantify the antioxidant properties of traditional wild food plants and (2) to determine if their use in soups (i.e., the cooking process) can alter their beneficial properties. We have evaluated the antioxidant capacity (ABTS, DPPH) and the Total Phenolic Content (Folin- Ciocalteu) of five herbaceous plants traditionally consumed in several areas of Central Italy: (A) Reichardia picroides (L.) Roth, (B) Hypochaeris radicata L., (C) Cichorium intybus L., (D) Tordylium apulum L., and (E) Helminthotheca echioides (L.) Holub. Our analyses show good levels of antioxidant capacity for all plants, with Reichardia picroides and Helminthotheca echioides having the highest levels. ere is a high correlation between the antioxidant activity and the Total Phenolic Content especially in Reichardia picroides (R 2 =0.92) and Hypochaeris radicata (R 2 =0.93). Boiling the species caused a general decrease in the antioxidant activity and polyphenols. Our study confirms the health benefits of consuming wild plants, especially raw ones in salads. It also supports the use of ethnobotanical information to study and then promote the consumption of wild food plants. 1. Introduction Wild food plants used in the traditional Mediterranean diet have received much attention in recent years for their nutraceutical properties and in particular for their content of antioxidant compounds [1–8]. Indeed, many studies have highlighted that a dietary antioxidant intake has a protec- tive effect against free radical-related pathologies, such as cardiovascular diseases [9], diabetes [10], cancer [11, 12], and neurodegenerative diseases [13]. e morbidity of these diseases has increased in the last few decades in many industrialized countries [14] and has been oſten related, among other things, to shiſts from traditional to western diets [15]. In the Mediterranean basin, ethnobotanical research has identified about 2,300 different wild plants and fungi taxa, which are still gathered and consumed as food [16]. Although in decline, the consumption of wild edible plants is still common in various areas in Italy, where they are consumed because they are considered healthy and tasty [17–19], and also because they are linked to tradition and culture [18, 20]. e traditional uses of wild food plants may contribute to the health benefits associated with the Mediterranean diet and, as a consequence, studies on their phytochemistry can validate their nutraceutical properties [5–7]. is is also supported by the fact that recent studies have highlighted that the protective effect of nutraceuticals against various diseases is linked to the association of several phytochemical molecules at low concentrations, as it occurs naturally in the diet, rather than to the ingestion of individual molecules at high concentrations, as occurs in pills of dietary supplements [21–23]. Research also supports the importance of investi- gating the antioxidant properties of the plant part that is actually consumed, rather than focusing the attention on the Hindawi Evidence-Based Complementary and Alternative Medicine Volume 2019, Article ID 6782472, 12 pages https://doi.org/10.1155/2019/6782472
Transcript of Traditional Salads and Soups with Wild Plants as a Source of...
Research ArticleTraditional Salads and Soups with Wild Plants asa Source of Antioxidants A Comparative Chemical Analysis ofFive Species Growing in Central Italy
Valentina Savo Francois Salomone Elena Mattoni Daniela Tofani and Giulia Caneva
Department of Science Roma Tre University Viale G Marconi 446 Rome Italy
Correspondence should be addressed to Valentina Savo valentinasavouniroma3itand Daniela Tofani danielatofaniuniroma3it
Received 24 September 2018 Revised 30 January 2019 Accepted 18 February 2019 Published 5 March 2019
Academic Editor G K Jayaprakasha
Copyright copy 2019 Valentina Savo et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited
The interest and demand for nutraceuticals are rapidly increasing in many industrialized countries due to the emergence of healthrisks associated with the increased consumption of processed foods Several wild Mediterranean plants used as traditional foodsare an extraordinary source of nutraceutical substances with antioxidant properties This study has two main aims (1) to quantifythe antioxidant properties of traditional wild food plants and (2) to determine if their use in soups (ie the cooking process) canalter their beneficial properties We have evaluated the antioxidant capacity (ABTS DPPH) and the Total Phenolic Content (Folin-Ciocalteu) of five herbaceous plants traditionally consumed in several areas of Central Italy (A) Reichardia picroides (L) Roth(B) Hypochaeris radicata L (C) Cichorium intybus L (D) Tordylium apulum L and (E) Helminthotheca echioides (L) Holub Ouranalyses show good levels of antioxidant capacity for all plants with Reichardia picroides and Helminthotheca echioides having thehighest levels There is a high correlation between the antioxidant activity and the Total Phenolic Content especially in Reichardiapicroides (R2=092) and Hypochaeris radicata (R2=093) Boiling the species caused a general decrease in the antioxidant activityand polyphenols Our study confirms the health benefits of consuming wild plants especially raw ones in salads It also supportsthe use of ethnobotanical information to study and then promote the consumption of wild food plants
1 Introduction
Wild food plants used in the traditional Mediterraneandiet have received much attention in recent years for theirnutraceutical properties and in particular for their contentof antioxidant compounds [1ndash8] Indeed many studies havehighlighted that a dietary antioxidant intake has a protec-tive effect against free radical-related pathologies such ascardiovascular diseases [9] diabetes [10] cancer [11 12]and neurodegenerative diseases [13] The morbidity of thesediseases has increased in the last few decades in manyindustrialized countries [14] and has been often relatedamong other things to shifts from traditional to western diets[15]
In the Mediterranean basin ethnobotanical research hasidentified about 2300 different wild plants and fungi taxawhich are still gathered and consumed as food [16] Although
in decline the consumption of wild edible plants is stillcommon in various areas in Italy where they are consumedbecause they are considered healthy and tasty [17ndash19] andalso because they are linked to tradition and culture [18 20]The traditional uses of wild food plants may contribute tothe health benefits associated with the Mediterranean dietand as a consequence studies on their phytochemistry canvalidate their nutraceutical properties [5ndash7] This is alsosupported by the fact that recent studies have highlightedthat the protective effect of nutraceuticals against variousdiseases is linked to the association of several phytochemicalmolecules at low concentrations as it occurs naturally in thediet rather than to the ingestion of individual molecules athigh concentrations as occurs in pills of dietary supplements[21ndash23] Research also supports the importance of investi-gating the antioxidant properties of the plant part that isactually consumed rather than focusing the attention on the
HindawiEvidence-Based Complementary and Alternative MedicineVolume 2019 Article ID 6782472 12 pageshttpsdoiorg10115520196782472
2 Evidence-Based Complementary and Alternative Medicine
effects of individual compounds [1] Finally although severalcompounds may contribute to the antioxidant properties incomplex systems [24 25] polyphenols are often consideredthe primary source of the antioxidant activity [26ndash30] but fewdata support a precise correlation [29 31 32]
Consequently this study is aimed at (i) evaluating andcomparing the total antioxidant capacity and the Total Phe-nolic Content in different species of wild plants traditionallyconsumed either raw or cooked in Central Italy (ii) evalu-ating the relationship between the antioxidant capacity andthe phenolic compounds contained in plant extracts to verifywhether or not the phenolic constituents are responsible forthe antioxidant activity of the species
2 Materials and Methods
21 Sampling and Plants Collection We sampled five wildplants (A) Reichardia picroides (L) Roth (Asteraceae) (B)Hypochaeris radicata L (Asteraceae) (C) Cichorium intybusL (Asteraceae) (D) Tordylium apulum L (Apiaceae) and(E) Helminthotheca echioides (L) Holub (Asteraceae) Weselected these species because they are to different extentspurposively consumed among local communities in Italybecause they are considered to have positive effects on thehealth [19 33ndash35]
We gathered four specimens of each species (20 samples)in the Tolfa Mountain area (70 Km north-west of Rome) Weselected only specimens with vigorous growth collected inareas with similar soil characteristics within an altitudinalrange of 350-400m asl growing in flat areas to eliminate theinfluence of different exposures to the sun The samples werecarefully extracted and they were carried intactly along withtheir soil to the laboratory of Roma Tre University to keepthe leaves alive until they were cut
22 Chemical Analysis
Reagents All chemicals used were of analytical grade Theused solvents and reagents were purchased from SigmaAldrich (Germany)
23 Preparation of Extracts from Crude Plants For eachsample collected 1 g of leaves was cut from the plant gentlycleaned with some paper and weighed We then put theleaves in a Falcon tube and poured 10mL of liquid nitrogeninside The leaves immediately became hard and fragile andthen they were crushed to dust We put the Falcon tube in alyophilizer (Christ Alpha 1-2 b Braun biotech internationalSavant refrigerated condensation trap RT 100) until theweight of the plant material was constant We then added5ml of methanol to the Falcon tube We mixed the solutionusing an ultrasound apparatus (Sonica Soltec 2002MH) for60 minutes at 30∘C Afterwards the solution was filteredin an Eppendorf test tube (final volume 10ml) put in anitrogen atmosphere and left in the freezer (-20∘C) untilanalysis We performed three analyses for each plant species(namely DPPH analysis ABTS analysis and Total PhenolicContent)
24 Preparation of Extracts from Cooked Plants We collected1 g of leaves from each plant put the leaves in a smallbeaker containing 10ml of boiling water and left them tocook for 5 minutes After that the leaves were recoveredand gently dried over a clean piece of paper We then putthe leaves in a Falcon tube and poured 10mL of liquidnitrogen inside and continued the procedure as previouslydescribed for the crude material (ie leaves were frozen withliquid nitrogen crushed to dust lyophilized to remove thewater and extracted with 5ml of methanol in an ultrasoundapparatus for 60minutes at 30∘C the solutionwas filtered andput in nitrogen atmosphere and then placed in the freezer)We performed three analyses for each plant species (namelyDPPH analysis ABTS analysis and Total Phenolic Content)
25 DPPH Analysis We performed a DPPH analysis ofthe samples with some adjustments following the methoddescribed by Brand-Williams et al [36] We prepared a 75120583M solution of DPPH in methanol Plant extracts werediluted and analyzed at three different final concentrationsranging from 15 to 5mgml We added 50 120583l of each samplesolution to 0950ml of the DPPH solution and left themin the dark After 30 minutes we measured the absorbanceof the samples at 517 nm using the Shimadzu UV-2401 PCspectrophotometer We used 50 120583L of pure ethanol as acontrol We repeated four measurements for each plantsample
Subsequently we plotted the percentages of DPPH inhi-bition vs the antioxidant concentrations and elaboratedlinear regressions using the Graphpad Prism 41 program(httpwwwgraphadcom) From each graph we extrapo-lated IC50 values as the concentration of the sample thathalves the DPPH radical absorbance Plants with a lower IC50value contained higher levels of antioxidants We performedstatistical analyses applying Studentrsquos t-test and ANOVA asanalyses of variance for the IC50 values We also calculatedthe Antiradical Activity (ARA) as the inverse of IC50 Wecalculated all values including relative errors through thepropagation of uncertainty
26 ABTS Analysis To measure the antioxidant capacityof all samples we followed the method of Pellegrini et al[37] with some adjustments We prepared the ABTS radicalcation solution mixing 10ml of 70mM aqueous solution ofABTS with 10ml of 228mM aqueous solution of K2S2O8 anddiluting it to a 25ml final volume (ABTS+∙ solution) Thenwe left the solution at room temperature overnight Beforethe analysis we diluted the ABTS+∙ solution with ethanolto reach an absorbance of 070plusmn020 In each analysis weadded 10 120583l of plant extracts to 1ml of the diluted ABTS+∙solution All plant extracts were analyzed in ethanol (02of water) at room temperature using three different finalconcentrations ranging from 01 to 10mgml We measuredthe extent of colour fading after 3 minutes at 120582=734 nmusing a Shimadzu UV-2401 PC spectrophotometer Weperformed four measurements for each concentration Wealso run solvent blanks and we used Trolox (6-Hydroxy-2578-tetramethylchroman-2-carboxylic acid) as a reference
Evidence-Based Complementary and Alternative Medicine 3
antioxidant (Trolox is the hydrophilic derivative of alpha-tocopherol)
We calculated the dose-response curves as the percentageof absorbance decrease ( ABTS inhibition) against theamount of antioxidant concentration for each plant collectedWe performed linear regressions and extrapolated slopes ofthe dose-response relationship using the Graphpad Prism41 program (httpwwwgraphadcom) The level of signif-icance for linear regressions was p lt0005 for all datasetsfor either the crude or the cooked samples We reportedthe antioxidant capacities as Trolox Equivalent AntioxidantCapacity (TEAC) defined as the concentration (mmoll)of Trolox having the equivalent antioxidant capacity of 1kgfwl solution of the plant extract under investigation Wecalculated the average TEAC values for each group of plantsandwe performed statistical analyses applying Studentrsquos t-testand ANOVA as analyses of variance of the TEAC values
27 Total Phenolic Content (TPC) The Folin-Ciocalteureagent assay [38] was used to determine the Total PhenolicContent (TPC) As a first step we diluted 25ml of Folin-Ciocalteu commercial reagent to 25ml with deionized waterobtaining a new solution (solution A) We mixed 010ml ofeach plant sample with 075ml of solution A and let to restfor three minutes at 25∘C before adding 075ml of a saturatedsodium carbonate solution We let the new mixed solutionrest for another 120minutes beforemeasuring the absorbanceat 725 nm Analyses were performed in quadruplicate foreach plant collected We used Gallic acid as a standard forthe calibration curve The Total Phenolic Content (TPC) wasexpressed as Gallic Acid Equivalents (GAE) ie the mg ofGallic acid corresponding to the polyphenols present in l gof dry plant material As for the other analyses we calculatedthe average TPC value for each plant and performed Studentrsquost-test and analysis of variance (ANOVA)
28 Correlation between Antioxidant Capacity and TotalPhenolic Content We correlated the antioxidant capacity andTotal Phenolic Content Specifically we used TPC and Anti-radical Activity (ARA) to calculate the antioxidant capacityas a function of the presence of phenolic compounds inthe plants We analyzed the data as a whole and then asdisaggregated sets for the four plant samples of each speciesWe elaborated linear regressions and determined slopes usingthe Graphpad Prism 41 program (httpwwwgraphadcom)
29 Statistical Analyses and Literature Search All data wereexpressed as mean plusmn standard error (SE) To test differencesamongDPPHABTS andTBCwe performed a series of one-way analyses of variance (ANOVA) and BonferronirsquosMultipleComparison Test Statistical analyses were performed usingthe Graphpad Prism 41 program (httpwwwgraphadcom)
We carried out a literature search on the antioxidantcapacity and phenolic content of the selected plants We usedcommon scientific literature search engines and databases(ie Google Scholar Pubmed and Science Direct) using askeywords the scientific names of the plants and the names ofthe various analytical tests Subsequently when possible we
compared the antioxidant activity and polyphenol contentswith literature data and possible reasons behind significantdifferences in values were discussed
3 Results and Discussion
31 Ethnobotanical Sampling and Plants Collection In Italythe gathering and use of wild edible species even if it isdecreasing and practiced mainly by older people is stillwidespread throughout the entire country mostly in ruralareasThe plants we selected are commonly consumed raw orcooked in Central Italy to prepare various traditional dishes[17 18 39] The Tolfa area where we gathered the samplesis considered very important from an ethnobotanical pointof view [40 41] In the area the leaves of these plants arecommonly consumed fresh in salads or cooked in a tasty soupcalled ldquoAcquacottardquo For this reason we sampled and ana-lyzed four plants from each of the five wild species Reichardiapicroides (L) Roth (Asteraceae) (A) Hypochaeris radicataL (Asteraceae) (B) Cichorium intybus L (Asteraceae) (C)Tordylium apulum L (Apiaceae) (D) and Helminthothecaechioides (L) Holub (Asteraceae) (E) Furthermore threeplants Hypochaeris radicata (BC) Cichorium intybus (CC)and Helminthotheca echioides (EC) were also analyzed aftercooking In Italy all five plants are traditionally used besidesas food also for their medicinal properties ie they are usedto treat heart problems infections and diabetes and as adepurative [7 17 42] These plants have a wide distributionin Italy and can grow in many different habitats mostlyarid and ruderal [43] As such their cultivation could bepromoted in marginal and arid lands or abandoned fields[44 45]
32 DPPH Assay The DPPH radical scavenging assay is oneof the most extensively used methods for estimating theantioxidant efficacy of molecules and plant samples [46] Wereported graphically the extent of the antioxidant capacitythat was determined as the amount of antioxidant that halvesthe DPPH radical concentration (IC50) of the four samples ofeach species in Figures S1 and S2 of Supporting Information(SI) In Figure 1 and Tables 1 and 2 we showed the obtainedIC50 average values for the five fresh plants (A-E) and thethree cooked plants (BC CC and EC) All the analyzedplants showed antioxidant properties In Table 1 it is possibleto observe that in the fresh samples IC50 ranges from269plusmn005mgml for Hypochoeris radicata (B1) to the higher057plusmn002mgml for the antioxidant capacity of Reichardiapicroides (A3) The cooking caused a general lowering ofthe antioxidant activity in all samples In this case IC50ranged from 31plusmn03mgml of Cichorium intybus (CC3) to173plusmn007mgml ofHelminthotheca echioides (EC4) (Table 2)The comparison of fresh Cichorium intybus IC50 values withthose reported in literature was in some cases difficult aspublished data dealt with antioxidant analyses of differentparts of the plant (02-1mgml for roots [47]) or cultivatedvarieties (57 nmol Troloxmg of fresh weight for red chicory[48]) Some works instead reported lower values of IC50for wild plants of Cichorium intybus (111mgml [6]) The
4 Evidence-Based Complementary and Alternative Medicine
A B BC C CC D E EC
DPPH
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
raw plantscooked plants
0
1
2
IC50
(mg
ml)
3
4
Figure 1 DPPH assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as IC50 ie the inhibition concentration that halves the DPPH radical activity Lower IC50 valuesindicate higher antioxidant capacity Statistical analyses of all average data were performed using ANOVA
literature on the antioxidant capacity of cooked plants wasinstead quite scarce (ie [49])
Statistical analyses performed applying Studentrsquos t-test tothe samples gave a level of significance of p lt 0005 for all thefresh samples and p lt 005 for the cooked samples (blue barsin Figures S1 and S2)The average IC50 value for each group ofcrude samples showed a level of significance of p lt 0001 andof p lt 005 for the cooked samples (Figure 1) The StatisticalAnalysis of Variation (ANOVA) of IC50 values showed asignificant difference between the fresh plant species the IC50value of A and E proved to be similar to each other butsignificantly different from B C and D Instead the B Cand D species showed IC50 values that were not statisticallydifferent from each other (Figure S1) For the cooked plantsthe differences among the IC50 values of all the samples werenot statistically significant (Figure S2)
33 ABTS Assay The ABTS assay also supported the antiox-idant properties of the selected plants This method ofanalysis is generally more sensitive than the DPPH assayto phenolic and flavonoid contents [50] In Figure 2 weshowed the relative average values expressed as TroloxEquivalent Antioxidant Capacity (TEAC) for each specieseither crude or cooked and their relative errors calculatedwith the propagation of uncertainty (see also Tables 1and 2) The TEAC values of crude samples ranged fromthe lower 37plusmn01mmolkgfw of Cichorium intybus (C4)to 154plusmn05mmolkgfw of Helminthotheca echioides (E1)
(Table 1) while for the cooked leaves TEAC values spannedfrom 179plusmn006mmolkgfw of Hypochaeris radicata (BC1)to 416plusmn01mmolkgfw of Helminthotheca echioides (EC4)(Table 2) As in the DPPH analysis the lower TEAC valuesof the cooked samples could be explained by the instabilityof phenolic antioxidants at high temperatures The averageTEAC value of 49plusmn1mmolkgfw obtained from the analysisof the four samples of the crudeCichorium intybus is coherentwith literature data (see also [51] for the wild specimens)Since Cichorium intybus is generally considered as havinga significant level of antioxidant capacity [51] the higherTEAC values obtained for all the other examined species(crude samples) supported their good antioxidant capac-ity Nevertheless the average TEAC value for each speciesshowed a high standard deviation (Figures S3 and S4) Otherstatistical analyses instead confirmed the robustness of thedata and attested the antioxidant capacity of the plants Thelevel of significance for the Studentrsquos t-test was p lt 001 forthe crude samples and p lt 005 for the cooked samplesThe Analysis of Variation (ANOVA) revealed that significantdifferences in TEAC values only exist between E and othercrude plant samples In fact Helminthotheca echioides (E)showed an average TEAC value of 12plusmn2mmolkgfw morethan 25 times higher than that of Cichorium intybus (Fig-ure 2) supporting its good antioxidant capacity All the otherTEAC values of crude (A B C and D) or cooked (BC CCand EC) samples were not statistically different from eachother
Evidence-Based Complementary and Alternative Medicine 5
Table1Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
crud
eplants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Reich
ardiapicroides
A1086plusmn003
117plusmn004
112plusmn03
200plusmn07
A210
6plusmn003
094plusmn003
62plusmn02
138plusmn05
A3057plusmn002
176plusmn005
64plusmn02
30plusmn1
A4075plusmn007
133plusmn012
62plusmn02
258plusmn09
Hypochaerisradicata
B1269plusmn005
037plusmn001
45plusmn01
122plusmn04
B219
0plusmn004
053plusmn001
56plusmn02
184plusmn07
B313
9plusmn007
072plusmn004
88plusmn03
224plusmn08
B4210plusmn007
048plusmn002
59plusmn02
140plusmn05
Cichorium
intybu
s
C1223plusmn002
045plusmn000
39plusmn01
152plusmn06
C216
9plusmn008
059plusmn003
61plusmn
02
224plusmn08
C317
6plusmn003
057plusmn001
58plusmn02
142plusmn05
C4230plusmn003
044plusmn001
37plusmn01
105plusmn04
Tordylium
apulum
D1
200plusmn005
050plusmn001
61plusmn
02
138plusmn05
D2
179plusmn001
056plusmn000
80plusmn03
178plusmn07
D3
191plusmn
012
052plusmn003
61plusmn
02
219plusmn08
D4
203plusmn006
049plusmn001
60plusmn02
124plusmn05
Helm
inthotheca
echioides
E1091plusmn001
109plusmn001
154plusmn05
33plusmn1
E2116plusmn004
086plusmn003
107plusmn03
198plusmn07
E3071plusmn002
142plusmn004
121plusmn04
276plusmn10
E414
7plusmn007
068plusmn003
116plusmn04
202plusmn07
6 Evidence-Based Complementary and Alternative Medicine
Table2Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
cooked
plants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Hypochaerisradicata
BC1
248plusmn005
040plusmn001
179plusmn006
81plusmn
03
BC2
22plusmn02
045plusmn004
220plusmn007
118plusmn04
BC3
20plusmn03
050plusmn008
32plusmn01
85plusmn05
BC4
264plusmn001
038plusmn000
207plusmn006
92plusmn03
Cichorium
intybu
s
CC1
270plusmn004
037plusmn001
268plusmn008
129plusmn05
CC2
237plusmn006
042plusmn001
253plusmn008
97plusmn04
CC3
31plusmn
03
032plusmn003
205plusmn006
84plusmn03
CC4
23plusmn01
043plusmn002
38plusmn01
122plusmn04
Helm
inthotheca
echioides
EC1
30plusmn04
033plusmn004
35plusmn01
114plusmn04
EC2
18plusmn01
056plusmn004
291plusmn009
132plusmn07
EC3
25plusmn01
040plusmn002
295plusmn009
80plusmn03
EC4
173plusmn007
058plusmn002
42plusmn01
128plusmn05
Evidence-Based Complementary and Alternative Medicine 7
raw plantscooked plants
ABTS
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
0
3
6
9
12
15
TEAC
(mm
olk
g)
Figure 2 ABTS assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as TEAC ie mmol of Trolox equivalent per kg of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA
34 Total Phenolic Content The average Total Phenolic Con-tent (TPC) of the plant extracts is presented in Figure 3 whilethe TPC values for all the samples are provided in the FiguresS5 and S6The crude plant extracts showed a polyphenol levelranging from 105plusmn04mg GAEgdw in Cichorium intybus(C4) to 33plusmn1mg GAEgdw in Helminthotheca echioides (E1)(Table 1) On the other hand cooked leaves displayed alower TPC ranging from 80plusmn03mg GAEgdw (EC3) to132plusmn07 (EC2) mg GAEgdw both measured in samplesof Helminthotheca echioides Literature data about crudeextracts of Cichorium intybus indicated a lower pheno-lic content (066mg GAEgdw [6] or higher (226plusmn10mgGAEgdw [52]) than the mean value of the four crude plantextracts analyzed (16plusmn5mg GAEgdw) This could be dueeither to the intrinsic differences in the growing environ-ment of the plant (ie soil exposure to solar radiationand hydric supply [52]) or to different sample preparationsthat could have safeguarded polyphenols from degrada-tion
TPC values and their relative errors calculated throughthe propagation of uncertainty are reported in Tables 1 and 2The level of significance is p lt 005 for all data The ANOVAtest for the average TPC values confirmed a low variationin phenolic concentrations among the various plants eithercrude or cooked these concentrations were not statisticallydifferent from each other except to somemarginal differencesbetween E and B or C and D (crude samples)
35 Correlation between Antioxidant Capacity and TotalPhenolic Content The Antiradical activity (ARA) as theinverse of IC50 is reported in Table 1 andwas used to correlateTPC with the antioxidant capacity of crude samples (Figures4 and 5) In Figure 4 we showed the graph plotting thevalues of ARA and TPC of all the crude plant samples understudy The graph showed a linear correlation (plt00001)with a slope of 0047plusmn0009 and -01plusmn02 as 119910 intercept Asregards Figure 4 the value of R2 (059) suggests a good linearcorrelation between the two variables (see [6]) In Figure 5 weshowed the same linear correlations between ARA and TPCbut with data disaggregated for the four plant samples of eachspecies ForReichardia picroidesA (R2=092) andHypochaerisradicata B (R2=093) the antioxidant capacity proved to belinearly dependent on TPC while a weaker correlation wasobserved with Cichorium intybus (C) Tordylium apulum (D)and Helminthotheca echioides (E) The slope of each curvecorrelates the antioxidant capacity with the polyphenolscontained in the plant the higher the value of the slope thehigher the antioxidant capacity of the polyphenols in thesamples FurthermoreHypochaeris radicata (B) exhibited anantioxidant activity completely dependent on polyphenoliccompounds as shown by119910 intercepts tending towards zero inthe absence of polyphenolic compounds (x=0) whereas theantioxidant capacity of Reichardia picroides (A) Cichoriumintybus (C) Tordylium apulum (D) and Helminthothecaechioides (E) is probably due to other kinds of antioxidants
8 Evidence-Based Complementary and Alternative Medicine
raw plantscooked plants
Total Phenolic Content
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
00
05
10
15
20
25
30
35
GA
E (m
ggd
w)
Figure 3 Total Phenolic Content of the raw and cooked plant extracts determined using the Folin-Ciocalteu assayThe averages data of all raw(A-E) and cooked (BC CC and EC) species are presented as Gallic Acid Equivalents (GAE) ie mg of Gallic acid corresponding to thepolyphenols contained per gram of dry weight (gdw) Statistical analyses of all average data were performed using ANOVA
20
15
10
05
00
ARA
(mlm
g)
00 100 200 300 400 500
GAE (mggdw)
Figure 4 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content (TPC) for all data of the crude plant extractsy = 0047x - 014 R2 = 059 p lt00001 Linear regressions and best fit values were calculated using the Graphpad Prism 41 program(httpwwwgraphadcom)
(ie carotenoids) as shown by 119910 intercepts different fromzero at x=0
Our results show that the examined wild food plants havea good antioxidant activity (see [53]) although with some
differences among plants and a certain variability amongsamples of the same species Among the analyzed plantsHelminthotheca echioides (E) and Reichardia picroides (A)have shown the best performances Helminthotheca echioides
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
2 Evidence-Based Complementary and Alternative Medicine
effects of individual compounds [1] Finally although severalcompounds may contribute to the antioxidant properties incomplex systems [24 25] polyphenols are often consideredthe primary source of the antioxidant activity [26ndash30] but fewdata support a precise correlation [29 31 32]
Consequently this study is aimed at (i) evaluating andcomparing the total antioxidant capacity and the Total Phe-nolic Content in different species of wild plants traditionallyconsumed either raw or cooked in Central Italy (ii) evalu-ating the relationship between the antioxidant capacity andthe phenolic compounds contained in plant extracts to verifywhether or not the phenolic constituents are responsible forthe antioxidant activity of the species
2 Materials and Methods
21 Sampling and Plants Collection We sampled five wildplants (A) Reichardia picroides (L) Roth (Asteraceae) (B)Hypochaeris radicata L (Asteraceae) (C) Cichorium intybusL (Asteraceae) (D) Tordylium apulum L (Apiaceae) and(E) Helminthotheca echioides (L) Holub (Asteraceae) Weselected these species because they are to different extentspurposively consumed among local communities in Italybecause they are considered to have positive effects on thehealth [19 33ndash35]
We gathered four specimens of each species (20 samples)in the Tolfa Mountain area (70 Km north-west of Rome) Weselected only specimens with vigorous growth collected inareas with similar soil characteristics within an altitudinalrange of 350-400m asl growing in flat areas to eliminate theinfluence of different exposures to the sun The samples werecarefully extracted and they were carried intactly along withtheir soil to the laboratory of Roma Tre University to keepthe leaves alive until they were cut
22 Chemical Analysis
Reagents All chemicals used were of analytical grade Theused solvents and reagents were purchased from SigmaAldrich (Germany)
23 Preparation of Extracts from Crude Plants For eachsample collected 1 g of leaves was cut from the plant gentlycleaned with some paper and weighed We then put theleaves in a Falcon tube and poured 10mL of liquid nitrogeninside The leaves immediately became hard and fragile andthen they were crushed to dust We put the Falcon tube in alyophilizer (Christ Alpha 1-2 b Braun biotech internationalSavant refrigerated condensation trap RT 100) until theweight of the plant material was constant We then added5ml of methanol to the Falcon tube We mixed the solutionusing an ultrasound apparatus (Sonica Soltec 2002MH) for60 minutes at 30∘C Afterwards the solution was filteredin an Eppendorf test tube (final volume 10ml) put in anitrogen atmosphere and left in the freezer (-20∘C) untilanalysis We performed three analyses for each plant species(namely DPPH analysis ABTS analysis and Total PhenolicContent)
24 Preparation of Extracts from Cooked Plants We collected1 g of leaves from each plant put the leaves in a smallbeaker containing 10ml of boiling water and left them tocook for 5 minutes After that the leaves were recoveredand gently dried over a clean piece of paper We then putthe leaves in a Falcon tube and poured 10mL of liquidnitrogen inside and continued the procedure as previouslydescribed for the crude material (ie leaves were frozen withliquid nitrogen crushed to dust lyophilized to remove thewater and extracted with 5ml of methanol in an ultrasoundapparatus for 60minutes at 30∘C the solutionwas filtered andput in nitrogen atmosphere and then placed in the freezer)We performed three analyses for each plant species (namelyDPPH analysis ABTS analysis and Total Phenolic Content)
25 DPPH Analysis We performed a DPPH analysis ofthe samples with some adjustments following the methoddescribed by Brand-Williams et al [36] We prepared a 75120583M solution of DPPH in methanol Plant extracts werediluted and analyzed at three different final concentrationsranging from 15 to 5mgml We added 50 120583l of each samplesolution to 0950ml of the DPPH solution and left themin the dark After 30 minutes we measured the absorbanceof the samples at 517 nm using the Shimadzu UV-2401 PCspectrophotometer We used 50 120583L of pure ethanol as acontrol We repeated four measurements for each plantsample
Subsequently we plotted the percentages of DPPH inhi-bition vs the antioxidant concentrations and elaboratedlinear regressions using the Graphpad Prism 41 program(httpwwwgraphadcom) From each graph we extrapo-lated IC50 values as the concentration of the sample thathalves the DPPH radical absorbance Plants with a lower IC50value contained higher levels of antioxidants We performedstatistical analyses applying Studentrsquos t-test and ANOVA asanalyses of variance for the IC50 values We also calculatedthe Antiradical Activity (ARA) as the inverse of IC50 Wecalculated all values including relative errors through thepropagation of uncertainty
26 ABTS Analysis To measure the antioxidant capacityof all samples we followed the method of Pellegrini et al[37] with some adjustments We prepared the ABTS radicalcation solution mixing 10ml of 70mM aqueous solution ofABTS with 10ml of 228mM aqueous solution of K2S2O8 anddiluting it to a 25ml final volume (ABTS+∙ solution) Thenwe left the solution at room temperature overnight Beforethe analysis we diluted the ABTS+∙ solution with ethanolto reach an absorbance of 070plusmn020 In each analysis weadded 10 120583l of plant extracts to 1ml of the diluted ABTS+∙solution All plant extracts were analyzed in ethanol (02of water) at room temperature using three different finalconcentrations ranging from 01 to 10mgml We measuredthe extent of colour fading after 3 minutes at 120582=734 nmusing a Shimadzu UV-2401 PC spectrophotometer Weperformed four measurements for each concentration Wealso run solvent blanks and we used Trolox (6-Hydroxy-2578-tetramethylchroman-2-carboxylic acid) as a reference
Evidence-Based Complementary and Alternative Medicine 3
antioxidant (Trolox is the hydrophilic derivative of alpha-tocopherol)
We calculated the dose-response curves as the percentageof absorbance decrease ( ABTS inhibition) against theamount of antioxidant concentration for each plant collectedWe performed linear regressions and extrapolated slopes ofthe dose-response relationship using the Graphpad Prism41 program (httpwwwgraphadcom) The level of signif-icance for linear regressions was p lt0005 for all datasetsfor either the crude or the cooked samples We reportedthe antioxidant capacities as Trolox Equivalent AntioxidantCapacity (TEAC) defined as the concentration (mmoll)of Trolox having the equivalent antioxidant capacity of 1kgfwl solution of the plant extract under investigation Wecalculated the average TEAC values for each group of plantsandwe performed statistical analyses applying Studentrsquos t-testand ANOVA as analyses of variance of the TEAC values
27 Total Phenolic Content (TPC) The Folin-Ciocalteureagent assay [38] was used to determine the Total PhenolicContent (TPC) As a first step we diluted 25ml of Folin-Ciocalteu commercial reagent to 25ml with deionized waterobtaining a new solution (solution A) We mixed 010ml ofeach plant sample with 075ml of solution A and let to restfor three minutes at 25∘C before adding 075ml of a saturatedsodium carbonate solution We let the new mixed solutionrest for another 120minutes beforemeasuring the absorbanceat 725 nm Analyses were performed in quadruplicate foreach plant collected We used Gallic acid as a standard forthe calibration curve The Total Phenolic Content (TPC) wasexpressed as Gallic Acid Equivalents (GAE) ie the mg ofGallic acid corresponding to the polyphenols present in l gof dry plant material As for the other analyses we calculatedthe average TPC value for each plant and performed Studentrsquost-test and analysis of variance (ANOVA)
28 Correlation between Antioxidant Capacity and TotalPhenolic Content We correlated the antioxidant capacity andTotal Phenolic Content Specifically we used TPC and Anti-radical Activity (ARA) to calculate the antioxidant capacityas a function of the presence of phenolic compounds inthe plants We analyzed the data as a whole and then asdisaggregated sets for the four plant samples of each speciesWe elaborated linear regressions and determined slopes usingthe Graphpad Prism 41 program (httpwwwgraphadcom)
29 Statistical Analyses and Literature Search All data wereexpressed as mean plusmn standard error (SE) To test differencesamongDPPHABTS andTBCwe performed a series of one-way analyses of variance (ANOVA) and BonferronirsquosMultipleComparison Test Statistical analyses were performed usingthe Graphpad Prism 41 program (httpwwwgraphadcom)
We carried out a literature search on the antioxidantcapacity and phenolic content of the selected plants We usedcommon scientific literature search engines and databases(ie Google Scholar Pubmed and Science Direct) using askeywords the scientific names of the plants and the names ofthe various analytical tests Subsequently when possible we
compared the antioxidant activity and polyphenol contentswith literature data and possible reasons behind significantdifferences in values were discussed
3 Results and Discussion
31 Ethnobotanical Sampling and Plants Collection In Italythe gathering and use of wild edible species even if it isdecreasing and practiced mainly by older people is stillwidespread throughout the entire country mostly in ruralareasThe plants we selected are commonly consumed raw orcooked in Central Italy to prepare various traditional dishes[17 18 39] The Tolfa area where we gathered the samplesis considered very important from an ethnobotanical pointof view [40 41] In the area the leaves of these plants arecommonly consumed fresh in salads or cooked in a tasty soupcalled ldquoAcquacottardquo For this reason we sampled and ana-lyzed four plants from each of the five wild species Reichardiapicroides (L) Roth (Asteraceae) (A) Hypochaeris radicataL (Asteraceae) (B) Cichorium intybus L (Asteraceae) (C)Tordylium apulum L (Apiaceae) (D) and Helminthothecaechioides (L) Holub (Asteraceae) (E) Furthermore threeplants Hypochaeris radicata (BC) Cichorium intybus (CC)and Helminthotheca echioides (EC) were also analyzed aftercooking In Italy all five plants are traditionally used besidesas food also for their medicinal properties ie they are usedto treat heart problems infections and diabetes and as adepurative [7 17 42] These plants have a wide distributionin Italy and can grow in many different habitats mostlyarid and ruderal [43] As such their cultivation could bepromoted in marginal and arid lands or abandoned fields[44 45]
32 DPPH Assay The DPPH radical scavenging assay is oneof the most extensively used methods for estimating theantioxidant efficacy of molecules and plant samples [46] Wereported graphically the extent of the antioxidant capacitythat was determined as the amount of antioxidant that halvesthe DPPH radical concentration (IC50) of the four samples ofeach species in Figures S1 and S2 of Supporting Information(SI) In Figure 1 and Tables 1 and 2 we showed the obtainedIC50 average values for the five fresh plants (A-E) and thethree cooked plants (BC CC and EC) All the analyzedplants showed antioxidant properties In Table 1 it is possibleto observe that in the fresh samples IC50 ranges from269plusmn005mgml for Hypochoeris radicata (B1) to the higher057plusmn002mgml for the antioxidant capacity of Reichardiapicroides (A3) The cooking caused a general lowering ofthe antioxidant activity in all samples In this case IC50ranged from 31plusmn03mgml of Cichorium intybus (CC3) to173plusmn007mgml ofHelminthotheca echioides (EC4) (Table 2)The comparison of fresh Cichorium intybus IC50 values withthose reported in literature was in some cases difficult aspublished data dealt with antioxidant analyses of differentparts of the plant (02-1mgml for roots [47]) or cultivatedvarieties (57 nmol Troloxmg of fresh weight for red chicory[48]) Some works instead reported lower values of IC50for wild plants of Cichorium intybus (111mgml [6]) The
4 Evidence-Based Complementary and Alternative Medicine
A B BC C CC D E EC
DPPH
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
raw plantscooked plants
0
1
2
IC50
(mg
ml)
3
4
Figure 1 DPPH assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as IC50 ie the inhibition concentration that halves the DPPH radical activity Lower IC50 valuesindicate higher antioxidant capacity Statistical analyses of all average data were performed using ANOVA
literature on the antioxidant capacity of cooked plants wasinstead quite scarce (ie [49])
Statistical analyses performed applying Studentrsquos t-test tothe samples gave a level of significance of p lt 0005 for all thefresh samples and p lt 005 for the cooked samples (blue barsin Figures S1 and S2)The average IC50 value for each group ofcrude samples showed a level of significance of p lt 0001 andof p lt 005 for the cooked samples (Figure 1) The StatisticalAnalysis of Variation (ANOVA) of IC50 values showed asignificant difference between the fresh plant species the IC50value of A and E proved to be similar to each other butsignificantly different from B C and D Instead the B Cand D species showed IC50 values that were not statisticallydifferent from each other (Figure S1) For the cooked plantsthe differences among the IC50 values of all the samples werenot statistically significant (Figure S2)
33 ABTS Assay The ABTS assay also supported the antiox-idant properties of the selected plants This method ofanalysis is generally more sensitive than the DPPH assayto phenolic and flavonoid contents [50] In Figure 2 weshowed the relative average values expressed as TroloxEquivalent Antioxidant Capacity (TEAC) for each specieseither crude or cooked and their relative errors calculatedwith the propagation of uncertainty (see also Tables 1and 2) The TEAC values of crude samples ranged fromthe lower 37plusmn01mmolkgfw of Cichorium intybus (C4)to 154plusmn05mmolkgfw of Helminthotheca echioides (E1)
(Table 1) while for the cooked leaves TEAC values spannedfrom 179plusmn006mmolkgfw of Hypochaeris radicata (BC1)to 416plusmn01mmolkgfw of Helminthotheca echioides (EC4)(Table 2) As in the DPPH analysis the lower TEAC valuesof the cooked samples could be explained by the instabilityof phenolic antioxidants at high temperatures The averageTEAC value of 49plusmn1mmolkgfw obtained from the analysisof the four samples of the crudeCichorium intybus is coherentwith literature data (see also [51] for the wild specimens)Since Cichorium intybus is generally considered as havinga significant level of antioxidant capacity [51] the higherTEAC values obtained for all the other examined species(crude samples) supported their good antioxidant capac-ity Nevertheless the average TEAC value for each speciesshowed a high standard deviation (Figures S3 and S4) Otherstatistical analyses instead confirmed the robustness of thedata and attested the antioxidant capacity of the plants Thelevel of significance for the Studentrsquos t-test was p lt 001 forthe crude samples and p lt 005 for the cooked samplesThe Analysis of Variation (ANOVA) revealed that significantdifferences in TEAC values only exist between E and othercrude plant samples In fact Helminthotheca echioides (E)showed an average TEAC value of 12plusmn2mmolkgfw morethan 25 times higher than that of Cichorium intybus (Fig-ure 2) supporting its good antioxidant capacity All the otherTEAC values of crude (A B C and D) or cooked (BC CCand EC) samples were not statistically different from eachother
Evidence-Based Complementary and Alternative Medicine 5
Table1Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
crud
eplants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Reich
ardiapicroides
A1086plusmn003
117plusmn004
112plusmn03
200plusmn07
A210
6plusmn003
094plusmn003
62plusmn02
138plusmn05
A3057plusmn002
176plusmn005
64plusmn02
30plusmn1
A4075plusmn007
133plusmn012
62plusmn02
258plusmn09
Hypochaerisradicata
B1269plusmn005
037plusmn001
45plusmn01
122plusmn04
B219
0plusmn004
053plusmn001
56plusmn02
184plusmn07
B313
9plusmn007
072plusmn004
88plusmn03
224plusmn08
B4210plusmn007
048plusmn002
59plusmn02
140plusmn05
Cichorium
intybu
s
C1223plusmn002
045plusmn000
39plusmn01
152plusmn06
C216
9plusmn008
059plusmn003
61plusmn
02
224plusmn08
C317
6plusmn003
057plusmn001
58plusmn02
142plusmn05
C4230plusmn003
044plusmn001
37plusmn01
105plusmn04
Tordylium
apulum
D1
200plusmn005
050plusmn001
61plusmn
02
138plusmn05
D2
179plusmn001
056plusmn000
80plusmn03
178plusmn07
D3
191plusmn
012
052plusmn003
61plusmn
02
219plusmn08
D4
203plusmn006
049plusmn001
60plusmn02
124plusmn05
Helm
inthotheca
echioides
E1091plusmn001
109plusmn001
154plusmn05
33plusmn1
E2116plusmn004
086plusmn003
107plusmn03
198plusmn07
E3071plusmn002
142plusmn004
121plusmn04
276plusmn10
E414
7plusmn007
068plusmn003
116plusmn04
202plusmn07
6 Evidence-Based Complementary and Alternative Medicine
Table2Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
cooked
plants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Hypochaerisradicata
BC1
248plusmn005
040plusmn001
179plusmn006
81plusmn
03
BC2
22plusmn02
045plusmn004
220plusmn007
118plusmn04
BC3
20plusmn03
050plusmn008
32plusmn01
85plusmn05
BC4
264plusmn001
038plusmn000
207plusmn006
92plusmn03
Cichorium
intybu
s
CC1
270plusmn004
037plusmn001
268plusmn008
129plusmn05
CC2
237plusmn006
042plusmn001
253plusmn008
97plusmn04
CC3
31plusmn
03
032plusmn003
205plusmn006
84plusmn03
CC4
23plusmn01
043plusmn002
38plusmn01
122plusmn04
Helm
inthotheca
echioides
EC1
30plusmn04
033plusmn004
35plusmn01
114plusmn04
EC2
18plusmn01
056plusmn004
291plusmn009
132plusmn07
EC3
25plusmn01
040plusmn002
295plusmn009
80plusmn03
EC4
173plusmn007
058plusmn002
42plusmn01
128plusmn05
Evidence-Based Complementary and Alternative Medicine 7
raw plantscooked plants
ABTS
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
0
3
6
9
12
15
TEAC
(mm
olk
g)
Figure 2 ABTS assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as TEAC ie mmol of Trolox equivalent per kg of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA
34 Total Phenolic Content The average Total Phenolic Con-tent (TPC) of the plant extracts is presented in Figure 3 whilethe TPC values for all the samples are provided in the FiguresS5 and S6The crude plant extracts showed a polyphenol levelranging from 105plusmn04mg GAEgdw in Cichorium intybus(C4) to 33plusmn1mg GAEgdw in Helminthotheca echioides (E1)(Table 1) On the other hand cooked leaves displayed alower TPC ranging from 80plusmn03mg GAEgdw (EC3) to132plusmn07 (EC2) mg GAEgdw both measured in samplesof Helminthotheca echioides Literature data about crudeextracts of Cichorium intybus indicated a lower pheno-lic content (066mg GAEgdw [6] or higher (226plusmn10mgGAEgdw [52]) than the mean value of the four crude plantextracts analyzed (16plusmn5mg GAEgdw) This could be dueeither to the intrinsic differences in the growing environ-ment of the plant (ie soil exposure to solar radiationand hydric supply [52]) or to different sample preparationsthat could have safeguarded polyphenols from degrada-tion
TPC values and their relative errors calculated throughthe propagation of uncertainty are reported in Tables 1 and 2The level of significance is p lt 005 for all data The ANOVAtest for the average TPC values confirmed a low variationin phenolic concentrations among the various plants eithercrude or cooked these concentrations were not statisticallydifferent from each other except to somemarginal differencesbetween E and B or C and D (crude samples)
35 Correlation between Antioxidant Capacity and TotalPhenolic Content The Antiradical activity (ARA) as theinverse of IC50 is reported in Table 1 andwas used to correlateTPC with the antioxidant capacity of crude samples (Figures4 and 5) In Figure 4 we showed the graph plotting thevalues of ARA and TPC of all the crude plant samples understudy The graph showed a linear correlation (plt00001)with a slope of 0047plusmn0009 and -01plusmn02 as 119910 intercept Asregards Figure 4 the value of R2 (059) suggests a good linearcorrelation between the two variables (see [6]) In Figure 5 weshowed the same linear correlations between ARA and TPCbut with data disaggregated for the four plant samples of eachspecies ForReichardia picroidesA (R2=092) andHypochaerisradicata B (R2=093) the antioxidant capacity proved to belinearly dependent on TPC while a weaker correlation wasobserved with Cichorium intybus (C) Tordylium apulum (D)and Helminthotheca echioides (E) The slope of each curvecorrelates the antioxidant capacity with the polyphenolscontained in the plant the higher the value of the slope thehigher the antioxidant capacity of the polyphenols in thesamples FurthermoreHypochaeris radicata (B) exhibited anantioxidant activity completely dependent on polyphenoliccompounds as shown by119910 intercepts tending towards zero inthe absence of polyphenolic compounds (x=0) whereas theantioxidant capacity of Reichardia picroides (A) Cichoriumintybus (C) Tordylium apulum (D) and Helminthothecaechioides (E) is probably due to other kinds of antioxidants
8 Evidence-Based Complementary and Alternative Medicine
raw plantscooked plants
Total Phenolic Content
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
00
05
10
15
20
25
30
35
GA
E (m
ggd
w)
Figure 3 Total Phenolic Content of the raw and cooked plant extracts determined using the Folin-Ciocalteu assayThe averages data of all raw(A-E) and cooked (BC CC and EC) species are presented as Gallic Acid Equivalents (GAE) ie mg of Gallic acid corresponding to thepolyphenols contained per gram of dry weight (gdw) Statistical analyses of all average data were performed using ANOVA
20
15
10
05
00
ARA
(mlm
g)
00 100 200 300 400 500
GAE (mggdw)
Figure 4 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content (TPC) for all data of the crude plant extractsy = 0047x - 014 R2 = 059 p lt00001 Linear regressions and best fit values were calculated using the Graphpad Prism 41 program(httpwwwgraphadcom)
(ie carotenoids) as shown by 119910 intercepts different fromzero at x=0
Our results show that the examined wild food plants havea good antioxidant activity (see [53]) although with some
differences among plants and a certain variability amongsamples of the same species Among the analyzed plantsHelminthotheca echioides (E) and Reichardia picroides (A)have shown the best performances Helminthotheca echioides
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
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Disease Markers
Hindawiwwwhindawicom Volume 2018
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Hindawiwwwhindawicom Volume 2013
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Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
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Computational and Mathematical Methods in Medicine
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
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Evidence-Based Complementary and Alternative Medicine 3
antioxidant (Trolox is the hydrophilic derivative of alpha-tocopherol)
We calculated the dose-response curves as the percentageof absorbance decrease ( ABTS inhibition) against theamount of antioxidant concentration for each plant collectedWe performed linear regressions and extrapolated slopes ofthe dose-response relationship using the Graphpad Prism41 program (httpwwwgraphadcom) The level of signif-icance for linear regressions was p lt0005 for all datasetsfor either the crude or the cooked samples We reportedthe antioxidant capacities as Trolox Equivalent AntioxidantCapacity (TEAC) defined as the concentration (mmoll)of Trolox having the equivalent antioxidant capacity of 1kgfwl solution of the plant extract under investigation Wecalculated the average TEAC values for each group of plantsandwe performed statistical analyses applying Studentrsquos t-testand ANOVA as analyses of variance of the TEAC values
27 Total Phenolic Content (TPC) The Folin-Ciocalteureagent assay [38] was used to determine the Total PhenolicContent (TPC) As a first step we diluted 25ml of Folin-Ciocalteu commercial reagent to 25ml with deionized waterobtaining a new solution (solution A) We mixed 010ml ofeach plant sample with 075ml of solution A and let to restfor three minutes at 25∘C before adding 075ml of a saturatedsodium carbonate solution We let the new mixed solutionrest for another 120minutes beforemeasuring the absorbanceat 725 nm Analyses were performed in quadruplicate foreach plant collected We used Gallic acid as a standard forthe calibration curve The Total Phenolic Content (TPC) wasexpressed as Gallic Acid Equivalents (GAE) ie the mg ofGallic acid corresponding to the polyphenols present in l gof dry plant material As for the other analyses we calculatedthe average TPC value for each plant and performed Studentrsquost-test and analysis of variance (ANOVA)
28 Correlation between Antioxidant Capacity and TotalPhenolic Content We correlated the antioxidant capacity andTotal Phenolic Content Specifically we used TPC and Anti-radical Activity (ARA) to calculate the antioxidant capacityas a function of the presence of phenolic compounds inthe plants We analyzed the data as a whole and then asdisaggregated sets for the four plant samples of each speciesWe elaborated linear regressions and determined slopes usingthe Graphpad Prism 41 program (httpwwwgraphadcom)
29 Statistical Analyses and Literature Search All data wereexpressed as mean plusmn standard error (SE) To test differencesamongDPPHABTS andTBCwe performed a series of one-way analyses of variance (ANOVA) and BonferronirsquosMultipleComparison Test Statistical analyses were performed usingthe Graphpad Prism 41 program (httpwwwgraphadcom)
We carried out a literature search on the antioxidantcapacity and phenolic content of the selected plants We usedcommon scientific literature search engines and databases(ie Google Scholar Pubmed and Science Direct) using askeywords the scientific names of the plants and the names ofthe various analytical tests Subsequently when possible we
compared the antioxidant activity and polyphenol contentswith literature data and possible reasons behind significantdifferences in values were discussed
3 Results and Discussion
31 Ethnobotanical Sampling and Plants Collection In Italythe gathering and use of wild edible species even if it isdecreasing and practiced mainly by older people is stillwidespread throughout the entire country mostly in ruralareasThe plants we selected are commonly consumed raw orcooked in Central Italy to prepare various traditional dishes[17 18 39] The Tolfa area where we gathered the samplesis considered very important from an ethnobotanical pointof view [40 41] In the area the leaves of these plants arecommonly consumed fresh in salads or cooked in a tasty soupcalled ldquoAcquacottardquo For this reason we sampled and ana-lyzed four plants from each of the five wild species Reichardiapicroides (L) Roth (Asteraceae) (A) Hypochaeris radicataL (Asteraceae) (B) Cichorium intybus L (Asteraceae) (C)Tordylium apulum L (Apiaceae) (D) and Helminthothecaechioides (L) Holub (Asteraceae) (E) Furthermore threeplants Hypochaeris radicata (BC) Cichorium intybus (CC)and Helminthotheca echioides (EC) were also analyzed aftercooking In Italy all five plants are traditionally used besidesas food also for their medicinal properties ie they are usedto treat heart problems infections and diabetes and as adepurative [7 17 42] These plants have a wide distributionin Italy and can grow in many different habitats mostlyarid and ruderal [43] As such their cultivation could bepromoted in marginal and arid lands or abandoned fields[44 45]
32 DPPH Assay The DPPH radical scavenging assay is oneof the most extensively used methods for estimating theantioxidant efficacy of molecules and plant samples [46] Wereported graphically the extent of the antioxidant capacitythat was determined as the amount of antioxidant that halvesthe DPPH radical concentration (IC50) of the four samples ofeach species in Figures S1 and S2 of Supporting Information(SI) In Figure 1 and Tables 1 and 2 we showed the obtainedIC50 average values for the five fresh plants (A-E) and thethree cooked plants (BC CC and EC) All the analyzedplants showed antioxidant properties In Table 1 it is possibleto observe that in the fresh samples IC50 ranges from269plusmn005mgml for Hypochoeris radicata (B1) to the higher057plusmn002mgml for the antioxidant capacity of Reichardiapicroides (A3) The cooking caused a general lowering ofthe antioxidant activity in all samples In this case IC50ranged from 31plusmn03mgml of Cichorium intybus (CC3) to173plusmn007mgml ofHelminthotheca echioides (EC4) (Table 2)The comparison of fresh Cichorium intybus IC50 values withthose reported in literature was in some cases difficult aspublished data dealt with antioxidant analyses of differentparts of the plant (02-1mgml for roots [47]) or cultivatedvarieties (57 nmol Troloxmg of fresh weight for red chicory[48]) Some works instead reported lower values of IC50for wild plants of Cichorium intybus (111mgml [6]) The
4 Evidence-Based Complementary and Alternative Medicine
A B BC C CC D E EC
DPPH
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
raw plantscooked plants
0
1
2
IC50
(mg
ml)
3
4
Figure 1 DPPH assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as IC50 ie the inhibition concentration that halves the DPPH radical activity Lower IC50 valuesindicate higher antioxidant capacity Statistical analyses of all average data were performed using ANOVA
literature on the antioxidant capacity of cooked plants wasinstead quite scarce (ie [49])
Statistical analyses performed applying Studentrsquos t-test tothe samples gave a level of significance of p lt 0005 for all thefresh samples and p lt 005 for the cooked samples (blue barsin Figures S1 and S2)The average IC50 value for each group ofcrude samples showed a level of significance of p lt 0001 andof p lt 005 for the cooked samples (Figure 1) The StatisticalAnalysis of Variation (ANOVA) of IC50 values showed asignificant difference between the fresh plant species the IC50value of A and E proved to be similar to each other butsignificantly different from B C and D Instead the B Cand D species showed IC50 values that were not statisticallydifferent from each other (Figure S1) For the cooked plantsthe differences among the IC50 values of all the samples werenot statistically significant (Figure S2)
33 ABTS Assay The ABTS assay also supported the antiox-idant properties of the selected plants This method ofanalysis is generally more sensitive than the DPPH assayto phenolic and flavonoid contents [50] In Figure 2 weshowed the relative average values expressed as TroloxEquivalent Antioxidant Capacity (TEAC) for each specieseither crude or cooked and their relative errors calculatedwith the propagation of uncertainty (see also Tables 1and 2) The TEAC values of crude samples ranged fromthe lower 37plusmn01mmolkgfw of Cichorium intybus (C4)to 154plusmn05mmolkgfw of Helminthotheca echioides (E1)
(Table 1) while for the cooked leaves TEAC values spannedfrom 179plusmn006mmolkgfw of Hypochaeris radicata (BC1)to 416plusmn01mmolkgfw of Helminthotheca echioides (EC4)(Table 2) As in the DPPH analysis the lower TEAC valuesof the cooked samples could be explained by the instabilityof phenolic antioxidants at high temperatures The averageTEAC value of 49plusmn1mmolkgfw obtained from the analysisof the four samples of the crudeCichorium intybus is coherentwith literature data (see also [51] for the wild specimens)Since Cichorium intybus is generally considered as havinga significant level of antioxidant capacity [51] the higherTEAC values obtained for all the other examined species(crude samples) supported their good antioxidant capac-ity Nevertheless the average TEAC value for each speciesshowed a high standard deviation (Figures S3 and S4) Otherstatistical analyses instead confirmed the robustness of thedata and attested the antioxidant capacity of the plants Thelevel of significance for the Studentrsquos t-test was p lt 001 forthe crude samples and p lt 005 for the cooked samplesThe Analysis of Variation (ANOVA) revealed that significantdifferences in TEAC values only exist between E and othercrude plant samples In fact Helminthotheca echioides (E)showed an average TEAC value of 12plusmn2mmolkgfw morethan 25 times higher than that of Cichorium intybus (Fig-ure 2) supporting its good antioxidant capacity All the otherTEAC values of crude (A B C and D) or cooked (BC CCand EC) samples were not statistically different from eachother
Evidence-Based Complementary and Alternative Medicine 5
Table1Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
crud
eplants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Reich
ardiapicroides
A1086plusmn003
117plusmn004
112plusmn03
200plusmn07
A210
6plusmn003
094plusmn003
62plusmn02
138plusmn05
A3057plusmn002
176plusmn005
64plusmn02
30plusmn1
A4075plusmn007
133plusmn012
62plusmn02
258plusmn09
Hypochaerisradicata
B1269plusmn005
037plusmn001
45plusmn01
122plusmn04
B219
0plusmn004
053plusmn001
56plusmn02
184plusmn07
B313
9plusmn007
072plusmn004
88plusmn03
224plusmn08
B4210plusmn007
048plusmn002
59plusmn02
140plusmn05
Cichorium
intybu
s
C1223plusmn002
045plusmn000
39plusmn01
152plusmn06
C216
9plusmn008
059plusmn003
61plusmn
02
224plusmn08
C317
6plusmn003
057plusmn001
58plusmn02
142plusmn05
C4230plusmn003
044plusmn001
37plusmn01
105plusmn04
Tordylium
apulum
D1
200plusmn005
050plusmn001
61plusmn
02
138plusmn05
D2
179plusmn001
056plusmn000
80plusmn03
178plusmn07
D3
191plusmn
012
052plusmn003
61plusmn
02
219plusmn08
D4
203plusmn006
049plusmn001
60plusmn02
124plusmn05
Helm
inthotheca
echioides
E1091plusmn001
109plusmn001
154plusmn05
33plusmn1
E2116plusmn004
086plusmn003
107plusmn03
198plusmn07
E3071plusmn002
142plusmn004
121plusmn04
276plusmn10
E414
7plusmn007
068plusmn003
116plusmn04
202plusmn07
6 Evidence-Based Complementary and Alternative Medicine
Table2Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
cooked
plants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Hypochaerisradicata
BC1
248plusmn005
040plusmn001
179plusmn006
81plusmn
03
BC2
22plusmn02
045plusmn004
220plusmn007
118plusmn04
BC3
20plusmn03
050plusmn008
32plusmn01
85plusmn05
BC4
264plusmn001
038plusmn000
207plusmn006
92plusmn03
Cichorium
intybu
s
CC1
270plusmn004
037plusmn001
268plusmn008
129plusmn05
CC2
237plusmn006
042plusmn001
253plusmn008
97plusmn04
CC3
31plusmn
03
032plusmn003
205plusmn006
84plusmn03
CC4
23plusmn01
043plusmn002
38plusmn01
122plusmn04
Helm
inthotheca
echioides
EC1
30plusmn04
033plusmn004
35plusmn01
114plusmn04
EC2
18plusmn01
056plusmn004
291plusmn009
132plusmn07
EC3
25plusmn01
040plusmn002
295plusmn009
80plusmn03
EC4
173plusmn007
058plusmn002
42plusmn01
128plusmn05
Evidence-Based Complementary and Alternative Medicine 7
raw plantscooked plants
ABTS
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
0
3
6
9
12
15
TEAC
(mm
olk
g)
Figure 2 ABTS assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as TEAC ie mmol of Trolox equivalent per kg of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA
34 Total Phenolic Content The average Total Phenolic Con-tent (TPC) of the plant extracts is presented in Figure 3 whilethe TPC values for all the samples are provided in the FiguresS5 and S6The crude plant extracts showed a polyphenol levelranging from 105plusmn04mg GAEgdw in Cichorium intybus(C4) to 33plusmn1mg GAEgdw in Helminthotheca echioides (E1)(Table 1) On the other hand cooked leaves displayed alower TPC ranging from 80plusmn03mg GAEgdw (EC3) to132plusmn07 (EC2) mg GAEgdw both measured in samplesof Helminthotheca echioides Literature data about crudeextracts of Cichorium intybus indicated a lower pheno-lic content (066mg GAEgdw [6] or higher (226plusmn10mgGAEgdw [52]) than the mean value of the four crude plantextracts analyzed (16plusmn5mg GAEgdw) This could be dueeither to the intrinsic differences in the growing environ-ment of the plant (ie soil exposure to solar radiationand hydric supply [52]) or to different sample preparationsthat could have safeguarded polyphenols from degrada-tion
TPC values and their relative errors calculated throughthe propagation of uncertainty are reported in Tables 1 and 2The level of significance is p lt 005 for all data The ANOVAtest for the average TPC values confirmed a low variationin phenolic concentrations among the various plants eithercrude or cooked these concentrations were not statisticallydifferent from each other except to somemarginal differencesbetween E and B or C and D (crude samples)
35 Correlation between Antioxidant Capacity and TotalPhenolic Content The Antiradical activity (ARA) as theinverse of IC50 is reported in Table 1 andwas used to correlateTPC with the antioxidant capacity of crude samples (Figures4 and 5) In Figure 4 we showed the graph plotting thevalues of ARA and TPC of all the crude plant samples understudy The graph showed a linear correlation (plt00001)with a slope of 0047plusmn0009 and -01plusmn02 as 119910 intercept Asregards Figure 4 the value of R2 (059) suggests a good linearcorrelation between the two variables (see [6]) In Figure 5 weshowed the same linear correlations between ARA and TPCbut with data disaggregated for the four plant samples of eachspecies ForReichardia picroidesA (R2=092) andHypochaerisradicata B (R2=093) the antioxidant capacity proved to belinearly dependent on TPC while a weaker correlation wasobserved with Cichorium intybus (C) Tordylium apulum (D)and Helminthotheca echioides (E) The slope of each curvecorrelates the antioxidant capacity with the polyphenolscontained in the plant the higher the value of the slope thehigher the antioxidant capacity of the polyphenols in thesamples FurthermoreHypochaeris radicata (B) exhibited anantioxidant activity completely dependent on polyphenoliccompounds as shown by119910 intercepts tending towards zero inthe absence of polyphenolic compounds (x=0) whereas theantioxidant capacity of Reichardia picroides (A) Cichoriumintybus (C) Tordylium apulum (D) and Helminthothecaechioides (E) is probably due to other kinds of antioxidants
8 Evidence-Based Complementary and Alternative Medicine
raw plantscooked plants
Total Phenolic Content
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
00
05
10
15
20
25
30
35
GA
E (m
ggd
w)
Figure 3 Total Phenolic Content of the raw and cooked plant extracts determined using the Folin-Ciocalteu assayThe averages data of all raw(A-E) and cooked (BC CC and EC) species are presented as Gallic Acid Equivalents (GAE) ie mg of Gallic acid corresponding to thepolyphenols contained per gram of dry weight (gdw) Statistical analyses of all average data were performed using ANOVA
20
15
10
05
00
ARA
(mlm
g)
00 100 200 300 400 500
GAE (mggdw)
Figure 4 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content (TPC) for all data of the crude plant extractsy = 0047x - 014 R2 = 059 p lt00001 Linear regressions and best fit values were calculated using the Graphpad Prism 41 program(httpwwwgraphadcom)
(ie carotenoids) as shown by 119910 intercepts different fromzero at x=0
Our results show that the examined wild food plants havea good antioxidant activity (see [53]) although with some
differences among plants and a certain variability amongsamples of the same species Among the analyzed plantsHelminthotheca echioides (E) and Reichardia picroides (A)have shown the best performances Helminthotheca echioides
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
4 Evidence-Based Complementary and Alternative Medicine
A B BC C CC D E EC
DPPH
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
raw plantscooked plants
0
1
2
IC50
(mg
ml)
3
4
Figure 1 DPPH assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as IC50 ie the inhibition concentration that halves the DPPH radical activity Lower IC50 valuesindicate higher antioxidant capacity Statistical analyses of all average data were performed using ANOVA
literature on the antioxidant capacity of cooked plants wasinstead quite scarce (ie [49])
Statistical analyses performed applying Studentrsquos t-test tothe samples gave a level of significance of p lt 0005 for all thefresh samples and p lt 005 for the cooked samples (blue barsin Figures S1 and S2)The average IC50 value for each group ofcrude samples showed a level of significance of p lt 0001 andof p lt 005 for the cooked samples (Figure 1) The StatisticalAnalysis of Variation (ANOVA) of IC50 values showed asignificant difference between the fresh plant species the IC50value of A and E proved to be similar to each other butsignificantly different from B C and D Instead the B Cand D species showed IC50 values that were not statisticallydifferent from each other (Figure S1) For the cooked plantsthe differences among the IC50 values of all the samples werenot statistically significant (Figure S2)
33 ABTS Assay The ABTS assay also supported the antiox-idant properties of the selected plants This method ofanalysis is generally more sensitive than the DPPH assayto phenolic and flavonoid contents [50] In Figure 2 weshowed the relative average values expressed as TroloxEquivalent Antioxidant Capacity (TEAC) for each specieseither crude or cooked and their relative errors calculatedwith the propagation of uncertainty (see also Tables 1and 2) The TEAC values of crude samples ranged fromthe lower 37plusmn01mmolkgfw of Cichorium intybus (C4)to 154plusmn05mmolkgfw of Helminthotheca echioides (E1)
(Table 1) while for the cooked leaves TEAC values spannedfrom 179plusmn006mmolkgfw of Hypochaeris radicata (BC1)to 416plusmn01mmolkgfw of Helminthotheca echioides (EC4)(Table 2) As in the DPPH analysis the lower TEAC valuesof the cooked samples could be explained by the instabilityof phenolic antioxidants at high temperatures The averageTEAC value of 49plusmn1mmolkgfw obtained from the analysisof the four samples of the crudeCichorium intybus is coherentwith literature data (see also [51] for the wild specimens)Since Cichorium intybus is generally considered as havinga significant level of antioxidant capacity [51] the higherTEAC values obtained for all the other examined species(crude samples) supported their good antioxidant capac-ity Nevertheless the average TEAC value for each speciesshowed a high standard deviation (Figures S3 and S4) Otherstatistical analyses instead confirmed the robustness of thedata and attested the antioxidant capacity of the plants Thelevel of significance for the Studentrsquos t-test was p lt 001 forthe crude samples and p lt 005 for the cooked samplesThe Analysis of Variation (ANOVA) revealed that significantdifferences in TEAC values only exist between E and othercrude plant samples In fact Helminthotheca echioides (E)showed an average TEAC value of 12plusmn2mmolkgfw morethan 25 times higher than that of Cichorium intybus (Fig-ure 2) supporting its good antioxidant capacity All the otherTEAC values of crude (A B C and D) or cooked (BC CCand EC) samples were not statistically different from eachother
Evidence-Based Complementary and Alternative Medicine 5
Table1Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
crud
eplants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Reich
ardiapicroides
A1086plusmn003
117plusmn004
112plusmn03
200plusmn07
A210
6plusmn003
094plusmn003
62plusmn02
138plusmn05
A3057plusmn002
176plusmn005
64plusmn02
30plusmn1
A4075plusmn007
133plusmn012
62plusmn02
258plusmn09
Hypochaerisradicata
B1269plusmn005
037plusmn001
45plusmn01
122plusmn04
B219
0plusmn004
053plusmn001
56plusmn02
184plusmn07
B313
9plusmn007
072plusmn004
88plusmn03
224plusmn08
B4210plusmn007
048plusmn002
59plusmn02
140plusmn05
Cichorium
intybu
s
C1223plusmn002
045plusmn000
39plusmn01
152plusmn06
C216
9plusmn008
059plusmn003
61plusmn
02
224plusmn08
C317
6plusmn003
057plusmn001
58plusmn02
142plusmn05
C4230plusmn003
044plusmn001
37plusmn01
105plusmn04
Tordylium
apulum
D1
200plusmn005
050plusmn001
61plusmn
02
138plusmn05
D2
179plusmn001
056plusmn000
80plusmn03
178plusmn07
D3
191plusmn
012
052plusmn003
61plusmn
02
219plusmn08
D4
203plusmn006
049plusmn001
60plusmn02
124plusmn05
Helm
inthotheca
echioides
E1091plusmn001
109plusmn001
154plusmn05
33plusmn1
E2116plusmn004
086plusmn003
107plusmn03
198plusmn07
E3071plusmn002
142plusmn004
121plusmn04
276plusmn10
E414
7plusmn007
068plusmn003
116plusmn04
202plusmn07
6 Evidence-Based Complementary and Alternative Medicine
Table2Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
cooked
plants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Hypochaerisradicata
BC1
248plusmn005
040plusmn001
179plusmn006
81plusmn
03
BC2
22plusmn02
045plusmn004
220plusmn007
118plusmn04
BC3
20plusmn03
050plusmn008
32plusmn01
85plusmn05
BC4
264plusmn001
038plusmn000
207plusmn006
92plusmn03
Cichorium
intybu
s
CC1
270plusmn004
037plusmn001
268plusmn008
129plusmn05
CC2
237plusmn006
042plusmn001
253plusmn008
97plusmn04
CC3
31plusmn
03
032plusmn003
205plusmn006
84plusmn03
CC4
23plusmn01
043plusmn002
38plusmn01
122plusmn04
Helm
inthotheca
echioides
EC1
30plusmn04
033plusmn004
35plusmn01
114plusmn04
EC2
18plusmn01
056plusmn004
291plusmn009
132plusmn07
EC3
25plusmn01
040plusmn002
295plusmn009
80plusmn03
EC4
173plusmn007
058plusmn002
42plusmn01
128plusmn05
Evidence-Based Complementary and Alternative Medicine 7
raw plantscooked plants
ABTS
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
0
3
6
9
12
15
TEAC
(mm
olk
g)
Figure 2 ABTS assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as TEAC ie mmol of Trolox equivalent per kg of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA
34 Total Phenolic Content The average Total Phenolic Con-tent (TPC) of the plant extracts is presented in Figure 3 whilethe TPC values for all the samples are provided in the FiguresS5 and S6The crude plant extracts showed a polyphenol levelranging from 105plusmn04mg GAEgdw in Cichorium intybus(C4) to 33plusmn1mg GAEgdw in Helminthotheca echioides (E1)(Table 1) On the other hand cooked leaves displayed alower TPC ranging from 80plusmn03mg GAEgdw (EC3) to132plusmn07 (EC2) mg GAEgdw both measured in samplesof Helminthotheca echioides Literature data about crudeextracts of Cichorium intybus indicated a lower pheno-lic content (066mg GAEgdw [6] or higher (226plusmn10mgGAEgdw [52]) than the mean value of the four crude plantextracts analyzed (16plusmn5mg GAEgdw) This could be dueeither to the intrinsic differences in the growing environ-ment of the plant (ie soil exposure to solar radiationand hydric supply [52]) or to different sample preparationsthat could have safeguarded polyphenols from degrada-tion
TPC values and their relative errors calculated throughthe propagation of uncertainty are reported in Tables 1 and 2The level of significance is p lt 005 for all data The ANOVAtest for the average TPC values confirmed a low variationin phenolic concentrations among the various plants eithercrude or cooked these concentrations were not statisticallydifferent from each other except to somemarginal differencesbetween E and B or C and D (crude samples)
35 Correlation between Antioxidant Capacity and TotalPhenolic Content The Antiradical activity (ARA) as theinverse of IC50 is reported in Table 1 andwas used to correlateTPC with the antioxidant capacity of crude samples (Figures4 and 5) In Figure 4 we showed the graph plotting thevalues of ARA and TPC of all the crude plant samples understudy The graph showed a linear correlation (plt00001)with a slope of 0047plusmn0009 and -01plusmn02 as 119910 intercept Asregards Figure 4 the value of R2 (059) suggests a good linearcorrelation between the two variables (see [6]) In Figure 5 weshowed the same linear correlations between ARA and TPCbut with data disaggregated for the four plant samples of eachspecies ForReichardia picroidesA (R2=092) andHypochaerisradicata B (R2=093) the antioxidant capacity proved to belinearly dependent on TPC while a weaker correlation wasobserved with Cichorium intybus (C) Tordylium apulum (D)and Helminthotheca echioides (E) The slope of each curvecorrelates the antioxidant capacity with the polyphenolscontained in the plant the higher the value of the slope thehigher the antioxidant capacity of the polyphenols in thesamples FurthermoreHypochaeris radicata (B) exhibited anantioxidant activity completely dependent on polyphenoliccompounds as shown by119910 intercepts tending towards zero inthe absence of polyphenolic compounds (x=0) whereas theantioxidant capacity of Reichardia picroides (A) Cichoriumintybus (C) Tordylium apulum (D) and Helminthothecaechioides (E) is probably due to other kinds of antioxidants
8 Evidence-Based Complementary and Alternative Medicine
raw plantscooked plants
Total Phenolic Content
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
00
05
10
15
20
25
30
35
GA
E (m
ggd
w)
Figure 3 Total Phenolic Content of the raw and cooked plant extracts determined using the Folin-Ciocalteu assayThe averages data of all raw(A-E) and cooked (BC CC and EC) species are presented as Gallic Acid Equivalents (GAE) ie mg of Gallic acid corresponding to thepolyphenols contained per gram of dry weight (gdw) Statistical analyses of all average data were performed using ANOVA
20
15
10
05
00
ARA
(mlm
g)
00 100 200 300 400 500
GAE (mggdw)
Figure 4 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content (TPC) for all data of the crude plant extractsy = 0047x - 014 R2 = 059 p lt00001 Linear regressions and best fit values were calculated using the Graphpad Prism 41 program(httpwwwgraphadcom)
(ie carotenoids) as shown by 119910 intercepts different fromzero at x=0
Our results show that the examined wild food plants havea good antioxidant activity (see [53]) although with some
differences among plants and a certain variability amongsamples of the same species Among the analyzed plantsHelminthotheca echioides (E) and Reichardia picroides (A)have shown the best performances Helminthotheca echioides
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
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Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 5
Table1Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
crud
eplants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Reich
ardiapicroides
A1086plusmn003
117plusmn004
112plusmn03
200plusmn07
A210
6plusmn003
094plusmn003
62plusmn02
138plusmn05
A3057plusmn002
176plusmn005
64plusmn02
30plusmn1
A4075plusmn007
133plusmn012
62plusmn02
258plusmn09
Hypochaerisradicata
B1269plusmn005
037plusmn001
45plusmn01
122plusmn04
B219
0plusmn004
053plusmn001
56plusmn02
184plusmn07
B313
9plusmn007
072plusmn004
88plusmn03
224plusmn08
B4210plusmn007
048plusmn002
59plusmn02
140plusmn05
Cichorium
intybu
s
C1223plusmn002
045plusmn000
39plusmn01
152plusmn06
C216
9plusmn008
059plusmn003
61plusmn
02
224plusmn08
C317
6plusmn003
057plusmn001
58plusmn02
142plusmn05
C4230plusmn003
044plusmn001
37plusmn01
105plusmn04
Tordylium
apulum
D1
200plusmn005
050plusmn001
61plusmn
02
138plusmn05
D2
179plusmn001
056plusmn000
80plusmn03
178plusmn07
D3
191plusmn
012
052plusmn003
61plusmn
02
219plusmn08
D4
203plusmn006
049plusmn001
60plusmn02
124plusmn05
Helm
inthotheca
echioides
E1091plusmn001
109plusmn001
154plusmn05
33plusmn1
E2116plusmn004
086plusmn003
107plusmn03
198plusmn07
E3071plusmn002
142plusmn004
121plusmn04
276plusmn10
E414
7plusmn007
068plusmn003
116plusmn04
202plusmn07
6 Evidence-Based Complementary and Alternative Medicine
Table2Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
cooked
plants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Hypochaerisradicata
BC1
248plusmn005
040plusmn001
179plusmn006
81plusmn
03
BC2
22plusmn02
045plusmn004
220plusmn007
118plusmn04
BC3
20plusmn03
050plusmn008
32plusmn01
85plusmn05
BC4
264plusmn001
038plusmn000
207plusmn006
92plusmn03
Cichorium
intybu
s
CC1
270plusmn004
037plusmn001
268plusmn008
129plusmn05
CC2
237plusmn006
042plusmn001
253plusmn008
97plusmn04
CC3
31plusmn
03
032plusmn003
205plusmn006
84plusmn03
CC4
23plusmn01
043plusmn002
38plusmn01
122plusmn04
Helm
inthotheca
echioides
EC1
30plusmn04
033plusmn004
35plusmn01
114plusmn04
EC2
18plusmn01
056plusmn004
291plusmn009
132plusmn07
EC3
25plusmn01
040plusmn002
295plusmn009
80plusmn03
EC4
173plusmn007
058plusmn002
42plusmn01
128plusmn05
Evidence-Based Complementary and Alternative Medicine 7
raw plantscooked plants
ABTS
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
0
3
6
9
12
15
TEAC
(mm
olk
g)
Figure 2 ABTS assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as TEAC ie mmol of Trolox equivalent per kg of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA
34 Total Phenolic Content The average Total Phenolic Con-tent (TPC) of the plant extracts is presented in Figure 3 whilethe TPC values for all the samples are provided in the FiguresS5 and S6The crude plant extracts showed a polyphenol levelranging from 105plusmn04mg GAEgdw in Cichorium intybus(C4) to 33plusmn1mg GAEgdw in Helminthotheca echioides (E1)(Table 1) On the other hand cooked leaves displayed alower TPC ranging from 80plusmn03mg GAEgdw (EC3) to132plusmn07 (EC2) mg GAEgdw both measured in samplesof Helminthotheca echioides Literature data about crudeextracts of Cichorium intybus indicated a lower pheno-lic content (066mg GAEgdw [6] or higher (226plusmn10mgGAEgdw [52]) than the mean value of the four crude plantextracts analyzed (16plusmn5mg GAEgdw) This could be dueeither to the intrinsic differences in the growing environ-ment of the plant (ie soil exposure to solar radiationand hydric supply [52]) or to different sample preparationsthat could have safeguarded polyphenols from degrada-tion
TPC values and their relative errors calculated throughthe propagation of uncertainty are reported in Tables 1 and 2The level of significance is p lt 005 for all data The ANOVAtest for the average TPC values confirmed a low variationin phenolic concentrations among the various plants eithercrude or cooked these concentrations were not statisticallydifferent from each other except to somemarginal differencesbetween E and B or C and D (crude samples)
35 Correlation between Antioxidant Capacity and TotalPhenolic Content The Antiradical activity (ARA) as theinverse of IC50 is reported in Table 1 andwas used to correlateTPC with the antioxidant capacity of crude samples (Figures4 and 5) In Figure 4 we showed the graph plotting thevalues of ARA and TPC of all the crude plant samples understudy The graph showed a linear correlation (plt00001)with a slope of 0047plusmn0009 and -01plusmn02 as 119910 intercept Asregards Figure 4 the value of R2 (059) suggests a good linearcorrelation between the two variables (see [6]) In Figure 5 weshowed the same linear correlations between ARA and TPCbut with data disaggregated for the four plant samples of eachspecies ForReichardia picroidesA (R2=092) andHypochaerisradicata B (R2=093) the antioxidant capacity proved to belinearly dependent on TPC while a weaker correlation wasobserved with Cichorium intybus (C) Tordylium apulum (D)and Helminthotheca echioides (E) The slope of each curvecorrelates the antioxidant capacity with the polyphenolscontained in the plant the higher the value of the slope thehigher the antioxidant capacity of the polyphenols in thesamples FurthermoreHypochaeris radicata (B) exhibited anantioxidant activity completely dependent on polyphenoliccompounds as shown by119910 intercepts tending towards zero inthe absence of polyphenolic compounds (x=0) whereas theantioxidant capacity of Reichardia picroides (A) Cichoriumintybus (C) Tordylium apulum (D) and Helminthothecaechioides (E) is probably due to other kinds of antioxidants
8 Evidence-Based Complementary and Alternative Medicine
raw plantscooked plants
Total Phenolic Content
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
00
05
10
15
20
25
30
35
GA
E (m
ggd
w)
Figure 3 Total Phenolic Content of the raw and cooked plant extracts determined using the Folin-Ciocalteu assayThe averages data of all raw(A-E) and cooked (BC CC and EC) species are presented as Gallic Acid Equivalents (GAE) ie mg of Gallic acid corresponding to thepolyphenols contained per gram of dry weight (gdw) Statistical analyses of all average data were performed using ANOVA
20
15
10
05
00
ARA
(mlm
g)
00 100 200 300 400 500
GAE (mggdw)
Figure 4 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content (TPC) for all data of the crude plant extractsy = 0047x - 014 R2 = 059 p lt00001 Linear regressions and best fit values were calculated using the Graphpad Prism 41 program(httpwwwgraphadcom)
(ie carotenoids) as shown by 119910 intercepts different fromzero at x=0
Our results show that the examined wild food plants havea good antioxidant activity (see [53]) although with some
differences among plants and a certain variability amongsamples of the same species Among the analyzed plantsHelminthotheca echioides (E) and Reichardia picroides (A)have shown the best performances Helminthotheca echioides
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
6 Evidence-Based Complementary and Alternative Medicine
Table2Summarytablew
ithallthe
results
obtained
from
theD
PPHA
BTSandFo
lin-C
iocalteuassays
onthee
xtractso
fthe
cooked
plants
Plant
Sample
DPP
HIC50
ARA
ABT
STE
ACFO
LINGAE
(mgml)
(mlm
g)(m
molkgfw)
(mggdw
)
Hypochaerisradicata
BC1
248plusmn005
040plusmn001
179plusmn006
81plusmn
03
BC2
22plusmn02
045plusmn004
220plusmn007
118plusmn04
BC3
20plusmn03
050plusmn008
32plusmn01
85plusmn05
BC4
264plusmn001
038plusmn000
207plusmn006
92plusmn03
Cichorium
intybu
s
CC1
270plusmn004
037plusmn001
268plusmn008
129plusmn05
CC2
237plusmn006
042plusmn001
253plusmn008
97plusmn04
CC3
31plusmn
03
032plusmn003
205plusmn006
84plusmn03
CC4
23plusmn01
043plusmn002
38plusmn01
122plusmn04
Helm
inthotheca
echioides
EC1
30plusmn04
033plusmn004
35plusmn01
114plusmn04
EC2
18plusmn01
056plusmn004
291plusmn009
132plusmn07
EC3
25plusmn01
040plusmn002
295plusmn009
80plusmn03
EC4
173plusmn007
058plusmn002
42plusmn01
128plusmn05
Evidence-Based Complementary and Alternative Medicine 7
raw plantscooked plants
ABTS
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
0
3
6
9
12
15
TEAC
(mm
olk
g)
Figure 2 ABTS assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as TEAC ie mmol of Trolox equivalent per kg of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA
34 Total Phenolic Content The average Total Phenolic Con-tent (TPC) of the plant extracts is presented in Figure 3 whilethe TPC values for all the samples are provided in the FiguresS5 and S6The crude plant extracts showed a polyphenol levelranging from 105plusmn04mg GAEgdw in Cichorium intybus(C4) to 33plusmn1mg GAEgdw in Helminthotheca echioides (E1)(Table 1) On the other hand cooked leaves displayed alower TPC ranging from 80plusmn03mg GAEgdw (EC3) to132plusmn07 (EC2) mg GAEgdw both measured in samplesof Helminthotheca echioides Literature data about crudeextracts of Cichorium intybus indicated a lower pheno-lic content (066mg GAEgdw [6] or higher (226plusmn10mgGAEgdw [52]) than the mean value of the four crude plantextracts analyzed (16plusmn5mg GAEgdw) This could be dueeither to the intrinsic differences in the growing environ-ment of the plant (ie soil exposure to solar radiationand hydric supply [52]) or to different sample preparationsthat could have safeguarded polyphenols from degrada-tion
TPC values and their relative errors calculated throughthe propagation of uncertainty are reported in Tables 1 and 2The level of significance is p lt 005 for all data The ANOVAtest for the average TPC values confirmed a low variationin phenolic concentrations among the various plants eithercrude or cooked these concentrations were not statisticallydifferent from each other except to somemarginal differencesbetween E and B or C and D (crude samples)
35 Correlation between Antioxidant Capacity and TotalPhenolic Content The Antiradical activity (ARA) as theinverse of IC50 is reported in Table 1 andwas used to correlateTPC with the antioxidant capacity of crude samples (Figures4 and 5) In Figure 4 we showed the graph plotting thevalues of ARA and TPC of all the crude plant samples understudy The graph showed a linear correlation (plt00001)with a slope of 0047plusmn0009 and -01plusmn02 as 119910 intercept Asregards Figure 4 the value of R2 (059) suggests a good linearcorrelation between the two variables (see [6]) In Figure 5 weshowed the same linear correlations between ARA and TPCbut with data disaggregated for the four plant samples of eachspecies ForReichardia picroidesA (R2=092) andHypochaerisradicata B (R2=093) the antioxidant capacity proved to belinearly dependent on TPC while a weaker correlation wasobserved with Cichorium intybus (C) Tordylium apulum (D)and Helminthotheca echioides (E) The slope of each curvecorrelates the antioxidant capacity with the polyphenolscontained in the plant the higher the value of the slope thehigher the antioxidant capacity of the polyphenols in thesamples FurthermoreHypochaeris radicata (B) exhibited anantioxidant activity completely dependent on polyphenoliccompounds as shown by119910 intercepts tending towards zero inthe absence of polyphenolic compounds (x=0) whereas theantioxidant capacity of Reichardia picroides (A) Cichoriumintybus (C) Tordylium apulum (D) and Helminthothecaechioides (E) is probably due to other kinds of antioxidants
8 Evidence-Based Complementary and Alternative Medicine
raw plantscooked plants
Total Phenolic Content
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
00
05
10
15
20
25
30
35
GA
E (m
ggd
w)
Figure 3 Total Phenolic Content of the raw and cooked plant extracts determined using the Folin-Ciocalteu assayThe averages data of all raw(A-E) and cooked (BC CC and EC) species are presented as Gallic Acid Equivalents (GAE) ie mg of Gallic acid corresponding to thepolyphenols contained per gram of dry weight (gdw) Statistical analyses of all average data were performed using ANOVA
20
15
10
05
00
ARA
(mlm
g)
00 100 200 300 400 500
GAE (mggdw)
Figure 4 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content (TPC) for all data of the crude plant extractsy = 0047x - 014 R2 = 059 p lt00001 Linear regressions and best fit values were calculated using the Graphpad Prism 41 program(httpwwwgraphadcom)
(ie carotenoids) as shown by 119910 intercepts different fromzero at x=0
Our results show that the examined wild food plants havea good antioxidant activity (see [53]) although with some
differences among plants and a certain variability amongsamples of the same species Among the analyzed plantsHelminthotheca echioides (E) and Reichardia picroides (A)have shown the best performances Helminthotheca echioides
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 7
raw plantscooked plants
ABTS
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
0
3
6
9
12
15
TEAC
(mm
olk
g)
Figure 2 ABTS assay of the raw and cooked plant extractsThe average antioxidant capacity for four plants of each species either raw (A-E)or cooked (BC CC and EC) is presented as TEAC ie mmol of Trolox equivalent per kg of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA
34 Total Phenolic Content The average Total Phenolic Con-tent (TPC) of the plant extracts is presented in Figure 3 whilethe TPC values for all the samples are provided in the FiguresS5 and S6The crude plant extracts showed a polyphenol levelranging from 105plusmn04mg GAEgdw in Cichorium intybus(C4) to 33plusmn1mg GAEgdw in Helminthotheca echioides (E1)(Table 1) On the other hand cooked leaves displayed alower TPC ranging from 80plusmn03mg GAEgdw (EC3) to132plusmn07 (EC2) mg GAEgdw both measured in samplesof Helminthotheca echioides Literature data about crudeextracts of Cichorium intybus indicated a lower pheno-lic content (066mg GAEgdw [6] or higher (226plusmn10mgGAEgdw [52]) than the mean value of the four crude plantextracts analyzed (16plusmn5mg GAEgdw) This could be dueeither to the intrinsic differences in the growing environ-ment of the plant (ie soil exposure to solar radiationand hydric supply [52]) or to different sample preparationsthat could have safeguarded polyphenols from degrada-tion
TPC values and their relative errors calculated throughthe propagation of uncertainty are reported in Tables 1 and 2The level of significance is p lt 005 for all data The ANOVAtest for the average TPC values confirmed a low variationin phenolic concentrations among the various plants eithercrude or cooked these concentrations were not statisticallydifferent from each other except to somemarginal differencesbetween E and B or C and D (crude samples)
35 Correlation between Antioxidant Capacity and TotalPhenolic Content The Antiradical activity (ARA) as theinverse of IC50 is reported in Table 1 andwas used to correlateTPC with the antioxidant capacity of crude samples (Figures4 and 5) In Figure 4 we showed the graph plotting thevalues of ARA and TPC of all the crude plant samples understudy The graph showed a linear correlation (plt00001)with a slope of 0047plusmn0009 and -01plusmn02 as 119910 intercept Asregards Figure 4 the value of R2 (059) suggests a good linearcorrelation between the two variables (see [6]) In Figure 5 weshowed the same linear correlations between ARA and TPCbut with data disaggregated for the four plant samples of eachspecies ForReichardia picroidesA (R2=092) andHypochaerisradicata B (R2=093) the antioxidant capacity proved to belinearly dependent on TPC while a weaker correlation wasobserved with Cichorium intybus (C) Tordylium apulum (D)and Helminthotheca echioides (E) The slope of each curvecorrelates the antioxidant capacity with the polyphenolscontained in the plant the higher the value of the slope thehigher the antioxidant capacity of the polyphenols in thesamples FurthermoreHypochaeris radicata (B) exhibited anantioxidant activity completely dependent on polyphenoliccompounds as shown by119910 intercepts tending towards zero inthe absence of polyphenolic compounds (x=0) whereas theantioxidant capacity of Reichardia picroides (A) Cichoriumintybus (C) Tordylium apulum (D) and Helminthothecaechioides (E) is probably due to other kinds of antioxidants
8 Evidence-Based Complementary and Alternative Medicine
raw plantscooked plants
Total Phenolic Content
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
00
05
10
15
20
25
30
35
GA
E (m
ggd
w)
Figure 3 Total Phenolic Content of the raw and cooked plant extracts determined using the Folin-Ciocalteu assayThe averages data of all raw(A-E) and cooked (BC CC and EC) species are presented as Gallic Acid Equivalents (GAE) ie mg of Gallic acid corresponding to thepolyphenols contained per gram of dry weight (gdw) Statistical analyses of all average data were performed using ANOVA
20
15
10
05
00
ARA
(mlm
g)
00 100 200 300 400 500
GAE (mggdw)
Figure 4 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content (TPC) for all data of the crude plant extractsy = 0047x - 014 R2 = 059 p lt00001 Linear regressions and best fit values were calculated using the Graphpad Prism 41 program(httpwwwgraphadcom)
(ie carotenoids) as shown by 119910 intercepts different fromzero at x=0
Our results show that the examined wild food plants havea good antioxidant activity (see [53]) although with some
differences among plants and a certain variability amongsamples of the same species Among the analyzed plantsHelminthotheca echioides (E) and Reichardia picroides (A)have shown the best performances Helminthotheca echioides
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
8 Evidence-Based Complementary and Alternative Medicine
raw plantscooked plants
Total Phenolic Content
A B BC C CC D E EC
Reichardia picroides Hypochaeris radicata Cichorium intybus Tordylium apulum Helminthotheca echioides
00
05
10
15
20
25
30
35
GA
E (m
ggd
w)
Figure 3 Total Phenolic Content of the raw and cooked plant extracts determined using the Folin-Ciocalteu assayThe averages data of all raw(A-E) and cooked (BC CC and EC) species are presented as Gallic Acid Equivalents (GAE) ie mg of Gallic acid corresponding to thepolyphenols contained per gram of dry weight (gdw) Statistical analyses of all average data were performed using ANOVA
20
15
10
05
00
ARA
(mlm
g)
00 100 200 300 400 500
GAE (mggdw)
Figure 4 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content (TPC) for all data of the crude plant extractsy = 0047x - 014 R2 = 059 p lt00001 Linear regressions and best fit values were calculated using the Graphpad Prism 41 program(httpwwwgraphadcom)
(ie carotenoids) as shown by 119910 intercepts different fromzero at x=0
Our results show that the examined wild food plants havea good antioxidant activity (see [53]) although with some
differences among plants and a certain variability amongsamples of the same species Among the analyzed plantsHelminthotheca echioides (E) and Reichardia picroides (A)have shown the best performances Helminthotheca echioides
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
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Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
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Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 9
GAE (mggdw)
25
20
15
10
05
00
ARA
(mlm
g)
0 5 10 15 20 25 30 35 40 45 50
ABC
DE
Figure 5 Correlation between the Antiradical Activity (ARA) and Total Phenolic Content for each raw species (A-E) A Reichardia picroides(X) y = 0047x +025 R2 = 092 B Hypochaeris radicata (◼) y = 0031x +0009 R2 = 093 C Cichorium intybus (Δ) y = 0011x +034 R2 =052 D Tordylium apulum (x) y = 0004x +045 R2 = 033 and E Helminthotheca echioides (∙) y = 003x+025 R2 = 045 A (plt 005) B(plt 005) C (plt 01) D (plt 03) and E (plt 01) Linear regressions and best fit values were calculated using the Grahpad Prism 4 program(httpwwwgraphadcom)
(E) exhibited the highest antioxidant capacity using eitherthe ABTS or the DPPH assays coupled with the highestpolyphenolic content
The antioxidant capacity and the phenolic content are cor-related in the plants under study A strong correlation existsin Reichardia picroides A (R2=092) andHypochaeris radicataB (R2=093) instead in Cichorium intybus (C) Tordyliumapulum (D) andHelminthotheca echioides (E) the correlationdecreases due to the likely presence of nonpolyphenolicantioxidants Similarly Dalar et al [54] found a correlationbetween total phenolics and antioxidant capacity in theirexperiments but they also detected a significant level ofother redox-active compounds besides phenolics In anotherstudy instead the contribution of phenolic components tothe antioxidant capacity was found to be at only 58 [55]
4 Conclusions
Our study demonstrated that the examined wild food plantsconsumed in traditional recipes in Central Italy are indeedrich in antioxidant compounds Ethnobotanical researchmight guide the study and then the revitalization of ahealthyMediterranean diet that incorporates wild plants withantioxidant capacity We also verified that this antioxidantcapacity is mainly correlated with the presence of phenoliccompounds although further analysis (ie the identificationof the individual constituents of the mixture by HPLCor LC-MS) would better determine the presence of otherantioxidant compounds We were also able to assess thatthe way food is consumed (raw or cooked) can significantly
alter the health benefits of the ingested plants In view of thegrowing demand for food rich in natural antioxidants by theglobalmarket the use of ethnobotanical information to studyand then promote new agronomic products from wild foodplants could constitute an important way to produce incomein many rural regions
Abbreviations
DPPH 22-diphenyl-1-picrylhydrazyl radicalANOVA Analysis of varianceARA Antiradical activityABTS 221015840-azinobis(3-ethylbenzothiazoline-6-
sulfonic acid) diammoniumsalt
TEAC Trolox equivalent antioxidant capacityTrolox 6-Hydroxy-2578-tetramethylchroman-2-
carboxylicacid
TPC Total phenolic contentGAE Gallic acid equivalentskgfw Kilograms fresh weightgdw Grams dry weightSE Standard error
Data Availability
The data used to support the findings of this study areincluded within the article and the supplementary informa-tion files
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
10 Evidence-Based Complementary and Alternative Medicine
Conflicts of Interest
The authors declare no financial conflicts of interest
Acknowledgments
The authors would like to thank Paolo Maria Guarrera forproviding useful comments and suggestions
Supplementary Materials
Figure S1 DPPH assay of the crude plant extracts Dataof each plant sample and averages of all species (A-E) arepresented as IC50 ie the inhibition concentration that halvesthe DPPH radical activity Lower IC50 values indicate higherantioxidant capacity Statistical analyses of all average datawere performed using ANOVA Values for A and E resultednot statistically different from each other but different fromall other values (p lt 001) except E versus B (plt005)Figure S2 DPPH assay of the cooked plant extracts Data ofplant samples and averages of species (BC CC and EC)are presented as IC50 ie the inhibition concentration thathalves the DPPH radical activity Lower IC50 values indicatehigher antioxidant capacity Statistical analyses of all averagedata were performed using ANOVA Values resulted notstatistically different from each other Figure S3 ABTS assayof the crude plant extracts Data of each plant sample andaverages of all species (A-E) are presented as TEAC iemmoles of Trolox equivalent per kilogram of fresh weight(kgfw) Statistical analyses of all average data were performedusing ANOVA All average data resulted not statisticallydifferent from each other except for E that was statisticallydifferent from A (plt 005) B (plt 001) C (plt 0001) and D(plt 001) Figure S4 ABTS assay of the cooked plant extractsData of plant samples and averages of species (BC CC andEC) are presented as TEAC ie mmoles of Trolox equivalentper kilogram of fresh weight (kgfw) Statistical analyses of allaverage data were performed using ANOVA All average dataresulted not statistically different from each other Figure S5Total Phenolic Content of the crude plant extracts determinedusing the Folin-Ciocalteu assayData of each plant sample andaverages of all species (A-E) are presented as mg of Gallicacid per gram of dry weight (gdw) Statistical analyses ofall average data were performed using ANOVA All averagedata resulted not statistically different from each other exceptto some marginal differences between E and B or C andD Figure S6 Total Phenolic Content of the cooked plantextracts determined using the Folin-Ciocalteu assay Data ofplant samples and averages of species (BC CC and EC) arepresented as mg of Gallic acid per gram of dry weight (gdw)Statistical analyses of all average data were performed usingANOVA All average data resulted not statistically differentfrom each other (Supplementary Materials)
References
[1] The local Food-Nutraceuticals Consortium ldquoUnderstandinglocal Mediterranean diets a multidisciplinary pharmacological
and ethnobotanical approachrdquo Pharmacological Research vol52 pp 353ndash366 2005
[2] S Salvatore N Pellegrini O V Brenna et al ldquoAntioxidantcharacterization of some Sicilian edible wild greensrdquo Journal ofAgricultural and Food Chemistry vol 53 no 24 pp 9465ndash94712005
[3] M Heinrich W E Muller and C Galli Local MediterraneanFood plants Karger Basel Switzerland 2006
[4] D Martins L Barros A M Carvalho and I C F R FerreiraldquoNutritional and in vitro antioxidant properties of edible wildgreens in Iberian Peninsula traditional dietrdquo Food Chemistryvol 125 no 2 pp 488ndash494 2011
[5] P Morales A M Carvalho M C Sanchez-Mata M CamaraM Molina and I C F R Ferreira ldquoTocopherol compositionand antioxidant activity of Spanish wild vegetablesrdquo GeneticResources and Crop Evolution vol 59 no 5 pp 851ndash863 2012
[6] P Morales I C F R Ferreira A M Carvalho et al ldquoMediter-ranean non-cultivated vegetables as dietary sources of com-pounds with antioxidant and biological activityrdquo LWT - FoodScience and Technology vol 55 no 1 pp 389ndash396 2014
[7] A Ranfa A Maurizi B Romano and M Bodesmo ldquoTheimportance of traditional uses and nutraceutical aspects ofsome edible wild plants in human nutrition the case of Umbria(central Italy)rdquo Plant Biosystems vol 148 no 2 pp 297ndash3062014
[8] L Barros P Morales A M Carvalho and I C F R FerreiraldquoAntioxidant potential of wild plant foodsrdquo in MediterraneanWild Edible Plants Ethnobotany and Food Composition TablesM C Sanchez-Mata and J Tardıo Eds pp 209ndash232 SpringerNew York NY USA 2016
[9] M G L Hertog E J M Feskens and D Kromhout ldquoAntioxi-dant flavonols and coronary heart disease riskrdquoThe Lancet vol349 no 9053 p 699 1997
[10] J Montonen P Knekt R Jarvinen and A Reunanen ldquoDietaryantioxidant intake and risk of type 2 diabetesrdquo Diabetes Carevol 27 no 2 pp 362ndash366 2004
[11] A Trichopoulou P Lagiou H Kuper and D TrichopoulosldquoCancer and Mediterranean dietary traditionsrdquo Cancer Epi-demiology Biomarkers amp Prevention vol 9 no 9 pp 869ndash8732000
[12] C La Vecchia ldquoAssociation between Mediterranean dietarypatterns and cancer riskrdquo Nutrition Reviews vol 67 no 1 ppS126ndashS129 2009
[13] Y Gilgun-Sherki E Melamed and D Offen ldquoThe role ofoxidative stress in the pathogenesis of multiple sclerosis theneed for effective antioxidant therapyrdquo Journal of Neurology vol251 no 3 pp 261ndash268 2004
[14] R J Koene A E Prizment A Blaes and S H Konety ldquoSharedrisk factors in cardiovascular disease and cancerrdquo Circulationvol 133 no 11 pp 1104ndash1114 2016
[15] D Stuckler and M Nestle ldquoBig food food systems and globalhealthrdquo PLoS Medicine vol 9 no 6 p e1001242 2012
[16] A C Hadjichambis D Paraskeva-Hadjichambi A Della etal ldquoWild and semi-domesticated food plant consumption inseven circum-Mediterranean areasrdquo International Journal ofFood Sciences and Nutrition vol 59 no 5 pp 383ndash414 2008
[17] P M Guarrera Usi e Tradizioni Della Flora Italiana Aracneeditrice Rome Italy 2006
[18] M Ghirardini M Carli N del Vecchio et al ldquoThe importanceof a taste A comparative study on wild food plant consumptionin twenty-one local communities in Italyrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 3 no 1 p 22 2007
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 11
[19] P M Guarrera and V Savo ldquoPerceived health properties of wildand cultivated food plants in local and popular traditions ofItaly a reviewrdquo Journal of Ethnopharmacology vol 146 no 3pp 659ndash680 2013
[20] PMGuarrera andV Savo ldquoWild food plants used in traditionalvegetable mixtures in Italyrdquo Journal of Ethnopharmacology vol185 pp 202ndash234 2016
[21] R H Liu ldquoHealth benefits of fruit and vegetables are from addi-tive and synergistic combinations of phytochemicalsrdquoAmericanJournal of Clinical Nutrition vol 78 no 3 pp 517Sndash520S 2003
[22] R G Mehta G Murillo R Naithani and X Peng ldquoCancerchemoprevention by natural products how far have we comerdquoPharmaceutical Research vol 27 no 6 pp 950ndash961 2010
[23] R H Liu ldquoHealth benefits of phytochemicals in whole foodsrdquoin Nutritional Health Strategies For Disease Prevention N JTemple TWilson and D R Jacobs Eds pp 293ndash310 SpringerScience New York NY USA 2012
[24] A P Simopoulos ldquoOmega-3 fatty acids and antioxidants inedible wild plantsrdquo Biological Research vol 37 no 2 pp 263ndash277 2004
[25] M S Brewer ldquoNatural antioxidants sources compoundsmechanisms of action and potential applicationsrdquo Comprehen-sive Reviews in Food Science and Food Safety vol 10 no 4 pp221ndash247 2011
[26] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001
[27] M Kapiszewska E Soltys F Visioli A Cierniak and G ZajacldquoThe protective ability of the Mediterranean plant extractsagainst the oxidative DNA damage The role of the radical oxy-gen species and the polyphenol contentrdquo Journal of Physiologyand Pharmacology Journal vol 56 pp 183ndash197 2005
[28] D Heimler L Isolani P Vignolini S Tombelli and A RomanildquoPolyphenol content and antioxidative activity in some speciesof freshly consumed saladsrdquo Journal of Agricultural and FoodChemistry vol 55 no 5 pp 1724ndash1729 2007
[29] F Conforti S Sosa M Marrelli et al ldquoThe protective abilityof Mediterranean dietary plants against the oxidative damagethe role of radical oxygen species in inflammation and thepolyphenol flavonoid and sterol contentsrdquo Food Chemistry vol112 no 3 pp 587ndash594 2009
[30] P Vanzani M Rossetto V de Marco L E Sacchetti M GPaoletti and A Rigo ldquoWild mediterranean plants as traditionalfood a valuable source of antioxidantsrdquo Journal of Food Sciencevol 76 no 1 pp C46ndashC51 2011
[31] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005
[32] I Hinneburg H J Damien Dorman and R Hiltunen ldquoAntiox-idant activities of extracts from selected culinary herbs andspicesrdquo Food Chemistry vol 97 no 1 pp 122ndash129 2006
[33] P M Guarrera ldquoFood medicine and minor nourishment in thefolk traditions of Central Italy (Marche Abruzzo and Latium)rdquoFitoterapia vol 74 no 6 pp 515ndash544 2003
[34] S Nebel and M Heinrich ldquoThe use of wild edible plants in theGraecanic area in Calabria Southern Italyrdquo in Ethnobotany ofthe New Europe M Pardo-de-Sanatayana A Pieroni and R KPuri Eds pp 172ndash188 Berghahn NY USA 2010
[35] A Ranfa F Orlandi A Maurizi and M Bodesmo ldquoEthnob-otanical knowledge and nutritional properties of two edible
wild plants from Central Italy Tordylium apulum L andUrospermum dalechampii (L) FW Schmidrdquo Journal of AppliedBotany and Food Quality vol 88 pp 249ndash254 2015
[36] W Brand-Williams M E Cuvelier and C Berset ldquoUse of a freeradical method to evaluate antioxidant activityrdquo LWT - FoodScience and Technology vol 28 no 1 pp 25ndash30 1995
[37] N Pellegrini F Visioli S Buratti and F Brighenti ldquoDirectanalysis of total antioxidant activity of olive oil and studieson the influence of heatingrdquo Journal of Agricultural and FoodChemistry vol 49 no 5 pp 2532ndash2538 2001
[38] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungstic acid reagentsrdquoAmeri-can Journal of Enology and Viticulture vol 16 pp 144ndash158 1965
[39] A Pieroni V Janiak C M Durr S Ludeke E Trachsel andM Heinrich ldquoIn vitro antioxidant activity of non-cultivatedvegetables of ethnic Albanians in southern Italyrdquo PhytotherapyResearch vol 16 no 5 pp 467ndash473 2002
[40] P M Guarrera Il Patrimonio Etnobotanico Del Lazio LePiante Del Lazio NellrsquoUso Terapeutico Alimentare DomesticoReligioso EMagico Etnobotanica Laziale E Della Media PenisolaItaliana A Confronto Regione Lazio Assessorato Alla CulturaE Dipartimento Di Biologia Vegetale Universita La SapienzaRome Italy 1994
[41] P M Guarrera V Savo and G Caneva ldquoTraditonal usesof plants in the Tolfa-Cerite-Manziate area (central Italy)rdquoEthnobiology Letters vol 6 no 1 pp 119ndash161 2015
[42] L Cornara A La Rocca S Marsili and M G MariottildquoTraditional uses of plants in the Eastern Riviera (LiguriaItaly)rdquo Journal of Ethnopharmacology vol 125 no 1 pp 16ndash302009
[43] S Pignatti Flora drsquoItalia Edagricole Bologna Italy 1982[44] M Spina M Cuccioloni L Sparapani et al ldquoComparative
evaluation of flavonoid content in assessing quality of wild andcultivated vegetables for human consumptionrdquo Journal of theScience of Food andAgriculture vol 88 no 2 pp 294ndash304 2008
[45] DHeimler L Isolani P Vignolini andA Romani ldquoPolyphenolcontent and antiradical activity of Cichorium intybus L frombiodynamic and conventional farmingrdquo Food Chemistry vol114 no 3 pp 765ndash770 2009
[46] S B Kedare and R P Singh ldquoGenesis and development ofDPPHmethod of antioxidant assayrdquo Journal of Food Science andTechnology vol 48 no 4 pp 412ndash422 2011
[47] H Liu QWang Y Liu G Chen and J Cui ldquoAntimicrobial andantioxidant activities of cichorium intybus root extract usingorthogonal matrix designrdquo Journal of Food Science vol 78 no2 pp M258ndashM263 2013
[48] V Lavelli ldquoAntioxidant activity of minimally processed redchicory (Cichorium intybus L) evaluated in xanthine oxidase-myeloperoxidase- and diaphorase-catalyzed reactionsrdquo Journalof Agricultural and Food Chemistry vol 56 no 16 pp 7194ndash7200 2008
[49] F Boari M Cefola F Di Gioia B Pace F Serio and V CantoreldquoEffect of cooking methods on antioxidant activity and nitratecontent of selected wild Mediterranean plantsrdquo InternationalJournal of Food Sciences and Nutrition vol 64 no 7 pp 870ndash876 2013
[50] A Floegel D-O Kim S-J Chung S I Koo and O K ChunldquoComparison of ABTSDPPH assays to measure antioxidantcapacity in popular antioxidant-rich US foodsrdquo Journal of FoodComposition and Analysis vol 24 no 7 pp 1043ndash1048 2011
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
12 Evidence-Based Complementary and Alternative Medicine
[51] A Montefusco G Semitaio P P Marrese et al ldquoAntioxidantsin varieties of chicory (Cichorium intybus L) and wild poppy(Papaver rhoeas L) of Southern Italyrdquo Journal of Chemistry vol2015 Article ID 923142 8 pages 2015
[52] A Dalar and I Konczak ldquoCichorium intybus from eastern ana-tolia phenolic composition antioxidant and enzyme inhibitoryactivitiesrdquo Industrial Crops and Products vol 60 pp 79ndash852014
[53] N Pellegrini M Serafini B Colombi et al ldquoTotal antioxidantcapacity of plant foods beverages and oils consumed in Italyassessed by three different in vitro assaysrdquo Journal of Nutritionvol 133 no 9 pp 2812ndash2819 2003
[54] A Dalar Y Uzun M Turker M Mukemre and I KonczakldquoHealth attributes of ethnic vegetables consumed in the EasternAnatolia region of Turkey antioxidant and enzyme-inhibitorypropertiesrdquo Journal of Ethnic Foods vol 3 no 2 pp 142ndash1492016
[55] D Fraisse C Felgines O Texier and L J Lamaison ldquoCaffeoylderivatives major antioxidant compounds of some wild herbsof theAsteraceae familyrdquo Journal of Food and Nutrition Sciencesvol 2 no 3 pp 181ndash192 2011
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
