LOVELY PROFESSIONAL UNIVERSITY

PROJECT NAME:

“Biochemical Analysis of Active ingredient’s in Asparagus

Racemosus (Shatavari) and its various uses”

NAME OF STUDENTS:Neeraj Kumar Yadav

BATCH:2008

YEAR OF PASSING:2012

SECTION:B1RO7

REGISTRATION NO.:10807700

ROLL NO.:RB1RO7B27

ABSTRACT

Asparagus Racemosus is an important medicinal plant of tropical and subtropical India. Its

medicinal usage has been reported in the Indian and British Pharmacopoeias and in traditional

systems of medicine such as Ayurveda, Unani and Siddha. Asparagus racemosus is one of the

important medicinal plants found in India, China and other parts of the world. This plant is

known to produce steroidal saponins called Shatavarins. In the present investigation,

alkaloid profiling of Asparagus racemosus plant samples, including roots, leaves and callus,

were performed which were collected from different localities. The Crude alkaloid fraction CAF

of samples suggest that roots are rich in alkaloid content as compared to leaves and callus. The

maximum CAF was obtained from the in vitro regenerated roots . The TLC analysis of all the

samples, including roots, leaves and callus, showed that roots are rich in saponins and they also

contain other alkaloids besides saponins. Callus tissues also contain saponins, but in the leave

samples alkaloids could not be estimated. However, saponins was detected in all the root and leaf

samples by TLC. The alkaloid profiling through TLC detected a number of alkaloids from all the

plant samples. The highest saponins content was obtained from the in vitro regenerated roots

and the least from the leaves.

Contents:

Introduction

Literature Review:

Geographical Distribution

Chemical Constituents

Cultivation

Recent Research

Extraction And Isolation

Materials And Methods

Alkaloid Confirmation

Estimation Of Alkaloids:

Results

Discussion

Pharmacological Applications Of Asparagus Racemosus

Conclusion

Uses & Benefits Of Asparagus

Reference:

Introduction

The World Health Organization (2003) has estimated that 80% of the population of developing

countries being unable to afford pharmaceutical drugs rely on traditional medicines, mainly plant

based, to sustain their primary health care needs. India is one of the most medico-culturally

diverse countries in the world where the medicinal plant sector is part of a time-honoured

tradition that is respected even today. Here, the main traditional systems of medicine include

Ayurveda, Unani and Siddha. The earliest mention of the use of plants in medicine is found in

the Rigveda which was written between 4500 and 1600 BC. It is however in Ayurveda that the

specific properties of plants and their use as medicinal drugs has been dealt with in great detail.

‘Ayurveda’ literally translated means science of life . Ananthacharya (1939) in defining this

system of medicine said Ayurveda scrutinizes the subtle process of life, studies its nature, ways

and conditions of development and deduces there-from a universal course of conduct for man’s

guidance in life . Ayurveda has eight divisions dealing with different aspects of the art of

healing. These include kaya cikitsa (internal medicine), salya tantra (surgery), salakya tantra

(treatment of diseases of the head and neck region), agada tantra (toxicology), bhuta vidya

(management of mental ailments), bala tantra (pedi-atrics), rasayana tantra (rejuvenation therapy

and geriatrics) and vajikarana tantra (science of aphrodisiacs). Around 1250 plants are presently

used in various Ayurvedic formulations. Asparagus Racemosus Willd. is one such important

medicinal plant which is regarded as a ‘rasayana’ (plant drugs promoting general well being by

increasing cellular vitality and resistance) in the Ayurvedic system of medicine . Asparagus

Racemosus is an important medicinal plant of tropical and subtropical India. Its medicinal usage

has been reported in the Indian and British Pharmacopoeias and in traditional systems of

medicine such as Ayurveda, Unani and Siddha. Asparagus racemosus is one of the important

medicinal plants found in India, China and other parts of the world. This plant is known to

produce steroidal saponins called Saponinss. The hydro-alcoholic and aqueous extracts of

Asparagus racemosus were subjected to different chemical tests for the detection of

phytoconstituents such as carbohydrates, glycosides, alkaloids, proteins, amino acids,

tannins, phenolics, saponins, flavonoids, triterpenoids, steroids, fixed oils, gums and

mucilages.

Literature Review:

Cytotoxic, antioxidant, tyrosinase inhibitory, antimicrobial activities of the crude ethanol extract

of dry powdered roots of Asparagus racemosus (Liliaceae) were investigated. The root of A.

racemosus is a potential broad spectrum antibiotic. TLC and HPLC finger printing showed the

presence of steroids-terpenes, alkaloids and flavonoids.

Potduang et.al ,2008

Asparagus species (family Liliaceae) are medicinal plants of temperate Himalayas. They

possess a variety of biological properties, such as being antioxidants, immunostimulants, anti-

inflammatory, antihepatotoxic, antibacterial, antioxytocic, and reproductive agents. The article

briefly reviews the isolated chemical constituents and the biological activities of the plant

species. The structural formula of isolated compounds and their distribution in the species

studied are also given.

Negi et.al,2010

Reverse pharmacology approach to examine the plants for drug development is a viable

approach. To fully validate this approach, further clinical trials are needed to examine their

potential. It is anticipated that this approach will not be as expensive as currently used and the

compounds/drugs isolated will be safe. Also one must question why using a single chemical

compound is preferred as a drug as compare to extracts from the whole plants. Benefits of a

single chemical entity may be in convenience to understand its molecular mechanism. However,

it may not be beneficial to the patient when examined, in part, due to the possibility of

development of resistance to a single chemical entity. It is possible that when whole plant extract

or combination of plant extracts are used, it may exhibit improved bioavailability and lower

toxicity, as compared to single chemical entity.

Aggarwal et.al,2011

Asparagus racemosus is an herbaceous perennial plant. It is the most important rasayana herb in

Ayurvedic medicine which grows in low forest areas throughout India. Their medicinal

usages have been reported in indigenous systems of medicine, Indian and British

Pharmacopoeias. In Ayurveda, A. racemosus has been used extensively as an adaptogen to

increase the non,specific resistance of organisms against a variety of stress. A. racemosus plant

contain steroidal saponins, isoflavones, asparagamine and polysaccharides, which play a

major role in treatment of diarrhoea and dysentery. Roots and rhizomes of A. racemosus has

potent antioxidant, antitussive, antidyspepsia, antiulcer and anticancer activity. A. racemosus is

also useful as immunostimulant, galactogogue. The present article gives the detailed exploration

of phytochemistry, ethnopharmacology and bioactivity of A. racemosus, in an attempt to give a

direction for further research.

Vandana garg et.al,2008

Asparagus racemosus is mainly known for its phytoestrogenic properties. With an increasing

realization that hormone replacement therapy with synthetic oestrogens is neither as safe nor as

effective as previously envisaged, the interest in plant-derived oestrogens has increased

tremendously making Asparagus racemosus particularly important. The plant has been shown to

aid in the treatment of neurodegenerative disorders and in alcohol abstinence-induced

withdrawal symptoms. In Ayurveda, Asparagus racemosus has been described as a rasayana herb

and has been used extensively as an adaptogen to increase the non-specific resistance of

organisms against a variety of stresses. Besides use in the treatment of diarrhoea and dysentery,

the plant also has potent antioxidant, immunostimulant, anti-dyspepsia and antitussive effects.

Due to its multiple uses, the demand for Asparagus racemosus is constantly on the rise; however,

the supply is rather erratic and inadequate. Destructive harvesting, combined with habitat

destruction in the form of deforestation has aggravated the problem. The plant is now considered

‘endangered’ in its natural habitat. Therefore, the need for conservat ion of this plant is crucial.

This article aims to evaluate the biological activities, pharmacological applications and clinical

studies of Asparagus racemosus in an attempt to provide a direction for further research.

Keeping in mind the fact that it is the active principle that imparts medicinal value to a plant;

consistency in quality and quantity needs to be maintained to ensure uniform drug efficacy. Also,

deliberate or inadvertent adulteration needs to be dealt with at an early stage. To overcome these

prevalent problems, the availability of genetically superior and uniform planting material is

essential. This can be obtained by a combination of various biotechnological tools involving

chemoprofiling, tissue culture and use of molecular markers. Along with the application of these

methods, proper agro-techniques and adequate marketing opportunities would encourage

cultivation of Asparagus racemosus and thereby contribute to its conservation. There are also

several gaps in the existing literature with regard to the pharmacological actions of Asparagus

racemosus . These include an incomplete understanding about the interaction/synergy between

Asparagus racemosus and other plant constituents in polyherbal formulations; lack of

information regarding the mode of action of the various constituents of Asparagus racemosus ,

etc. Consequently, we have suggested a ‘systems biology’ approach that includes metabolite

profiling, metabolic fingerprinting, metabolite target analysis and metabonomics to enable

further research.

Nishritha Bopana ,et.al,2007

Authors report that Asparagus racemosus has an antimicrobial activity against common

pathogens, but when it is combined with Antibacterial drug like Roxythromycin, Cefixime

and Levofloxacin the combinations help in - inhibiting growth of Staphylococcus aureus,

Staphylococcus epidermis, Escherichia coli and Bacillus subtilis. However, these

combinations have powerful effect against bacteria with less side- effects. The medicinal

importance of the Asparagus racemosus in the prevention of aerobic and anaerobic bacterial

infections is obvious considering the growing number of these developing resistance

organisms to conventional antibiotics. Phytochemical Analysis of the Asparagus racemosus

helps to find out the presence of chemical constituents in the plant extract. In the present paper,

the status on the above mentioned combinations has been discussed.

M.R. PRAJAPATI,et.al,2011

Geographical distribution :

Asparagus Racemosus is distributed throughout tropical Africa, Java, Australia, India, Srilanka

and Southern parts of China. In India, It is found in tropical, sub tropical regions and in

Himalayas upto 1000 to 1500 m.

Chemical Constituents:

The chemical ingredients in the Shatavari plant, including steroidal saponins, isoflavones,

asparagamine (an alkaloid substance similar to aspirin), and polysaccharides, make this plant a

natural chemical source.The following active constituents are present is Shatavari plant:

Steroidal saponins, known as shatavarins I-IV. Shatavarin I is the major glycoside with 3 glucose

and rhamnose moieties attached to sarsasapogenin

Isoflavones including 8-methoxy-5,6,4'- trihydroxyisoflavone 7-O-beta-D-glucopyranoside.

Asparagamine, a polycyclic alkaloid

Racemosol, a cyclic hydrocarbon (9,10- dihydrophenanthrene), Polysaccharides, mucilage

Cultivation

Soils: The plant prefers light (sandy), medium (loamy) and heavy (clay) soils and requires well-

drained soil. Black, well drained and fertile soil is good for cultivation. But can be cultivated in

loose and medium black soil.

Climate- crop responses well to tropical and hot climate..

Irrigation: The tamarind is adapted to semiarid regions of the tropics and can withstand drought

conditions quite well. They require minimum irrigation so avoid over-watering..

Fertilization: one ploughing, three harrowings and then apply 20-25 tonns of farm yard manure.

Harvest: 1) raised beds -1x3 m in the month of May or june.

2) Seed –one kg for one hectare area.

3) Apply 50 gram urea in the bed after 20-25 days.

Seedlings become ready within 6-8 weeks for transplantation in the main field.

Transplanting- 1) Size of pit-45x45x45

2) spacing-row to row-1.5m and plant to plant-1.0m

3) Fill the pits with 20-30 gram lindane or carbaryl and 5 kgs of FYM

4) Time of transplanting – july-august

5) provide the crop with 50 gms of 15:5:15(suphala) per plant when it starts with good growth.

6) Carry out timely weeding operations. Generally shatavari crop does not affect with pest and

diseases.

Harvesting- 1) first harvesting – 1.5-2 years after transplanting, which continues for 10-15 years.

Male and female plants must be grown if seed is required.

Recent research

Michael Thomsen (2002) has done extensive research on this plant. He analysed the adaptogenic,

diuretic, antitussive (suppresses cough), antibacterial, immunological, digestive, antioxytocic,

hormonal, galactogogue (increase in female milk production) properties, toxicity and

cytoprotective effect of this plant on human body. He says that this plant acts as adaptogen,

antitussive, antioxidant, antibacterial, immunomodulator, digestive, cytoprotective,

galactogogue, anti-oxytocic (preventing the stimulation of the involuntary muscles of the

uterus), antispasmodic, antidiarrhoeal and sexual tonic in human body.

Materials And Methods

Extraction And Isolation

Purification or isolation of alkaloids from a plant is always difficult process because an alkaloids

bearing plant generally contains a complex mixture of several alkaloids with glycoside organic

acid also complicate the process. Following steps are involved in isolation process.

Extraction: -

Total alkaloid contents were estimated from different plant parts of Asparagus racemosus, (e.g.,

callus, leaves and roots) collected from different habitats as well as in vitro regenerated plants.

Hundred gram of powdered dry samples of Asparagus racemosus were soaked in 10.0 mL

methanol and left for 30 min. After 30 min, the soaked plant material was filtered. The residue

obtained after filtration is further dissolved in 5.0 mL methanol and filtered after 10 min, the

same step is repeated once again and the final filtrate is collected in 50 mL conical flask. The

extract was evaporated to dryness in the soxhlet evaporator. The crude extract was dissolved in

100 mL of 0.01 M HCl. The pH of filtered solutions were adjusted to 6.0 with 0.01 M NaOH.

The crude extracts obtained were used for TLC analysis. The crude extract obtained, was

concentrated to dryness to yield Crude Alkaloid Fraction (CAF). Flow Chart of extraction

Powdered Drug/ Macerated Plant

Light Petroleum ether.

Filtration.

Filtrate Plant residue

Evaporate 1. CH3OH (72 hrs. extraction)

2. Filtration

Fat 3Evaporate

Crude Plant extract

1. Dissolve Water

2. Acidify to pH-2

3. Steam distillation

4.Filter

Filtrate

Ether

Ether soluble Acid Solution

Evaporate Ether + NaOH

Basic Material

Aqueous residue Ether solution

Evaporate.

(Crude Alkaloids)

Alkaloid confirmation

Presence of alkaloids confirmed by doing following tests:

Mayer’s reagent [( solution A i.e Hgcl2 + d water) +solution B i.e KI + d water )] Created by

dissolving 1.358 grams of HgCl2 in 60 milliliters of water, and pouring the solution into a

solution of 5 grams of KI in 10 milliliters of water. Gave Cream coloured precipitates.

Wagner’s reagent (Iodine and KI in Distilled water). Prepared by dissolving 2g of iodine and 6g

of KI in 100ml of water. Specimen with Wagner's Reagent gave reddish brown precipitates.

Estimation of alkaloids:

TLC

Extractive values

(1) Alcohol soluble extractive value

Accurately weighed 5 gm coarse and air dried drug material was macerated with 100ml

ethanol (99%) in a stoppered flask for 24 hrs. with frequent shaking for 6 hrs. It was then filtered

rapidly through filter paper taking precautions to prevent excessive loss of ethanol. The volume

was made up to 100ml with ethanol. The residue was evaporated in a flat bottom shallow dish,

dried at 105 0C, weighed and kept in a desiccators. Average extractive value in

percentage w/w (on dry basis) was calculated with reference to air dried drug (Table-2).

Water soluble extractive value

5 gm coarse and air dried drug material was macerated with water in a stoppered flask for 24

hrs. with frequent shaking for first 6 hrs . The extract was filtered rapidly through filter paper

taking precaution to prevent excessive loss of solvent. The residue was evaporated in a flat

bottom shallow dish, dried at 105 0C weighed and kept in a desiccators. Average extractive

value in Percentage w/w (on dry weight basis) was calculated with reference to air dried

drug (Table-2).

Determination of total phenolics by spectral analysis

Phenolic substances all absorb UV light, and all of them have some absorbance at 280 nm. This

property can be used to determine phenolics by spectral analysis. One problem with this method is that

each class of phenolic substances has a different absorptivity (extinction coefficient, e) at 280 nm.

Thus, the results cannot be related to any specific standard and are reported directly in absorbance units

(AU).

Materials

Sample.

Filter membrane.

Cuvettes, transparent at 280 nm (e.g., quartz )

Spectrophotometer, set to 280 nm

1. Filter a sample or blank (deionized or distilled water) with a PTFE filter membrane or

other material to achieve clarity.Nylon or other membranes that absorb phenolics should not

be used. Membranes can be tested for phenolic absorption by comparing absorbance after

single and double filtration.

2. Transfer an appropriate volume of sample to a quartz cuvette and measure absorbance at

280 nm in a spectrophotometer. If absorbance is not within the acceptable precision of

the spectrophotometer (usually A < 2 AU), dilute sample as necessary and repeat.

3. Subtract absorbance of blank, and correct absorbance to original concentration and a 1-cm

cuvette path length. Subtract 4 AU to report final value.

For instance, if a sample is diluted ten-fold with water and a reading of 0.85 AU is

observed with a 2-mm cell, the correction would be as follows:

Total phenol = [A280 ×DF × (1 cm/b)] – 4

where DF is the dilution factor, b is the cell path length, and 4 is an arbitrary correction for non

phenolic absorbance.result in Table 3

4. Phytochemical Screening

The fresh bark was collected and dried in shade and r educed to coarse powder. The powdered

material was extracted with Petroleum ether, Chloroform, Ethanol and water in Soxhlet

apparatus. The extract was filtered hot and solvent removed by distillation under reduced

pressure . The percentage yield was calculated and the extract was further subjected to

Phytochemical tests for Alkaloids, Glycosides, Flavonoids ,Carbohydrates ,Tannins (Table-4).

5. Results and Discussion

Extractive values of alcohol and water extracts are given in Table-2. Phytochemical screening

shows the presence of carbohydrates, glycosides, Tannins and absence of alkaloids and

flavanoids Table-4. Rf value given for pet. Ether inTable-5, chloroform inTable-6, ethanol

Table-7 and water Table-8.

The Ficus arnottiana Miq, bark was defatted with Petroleum ether then extracted with

Chloroform and after that with ethanol solvent and Water in soxhlet apparatus. Moisture content

is zero in pet ether & chloroform, ethanol extract and 1% in aqueous extract found out by

Karl Fischer Reagent. Extractive values of absolute alcohol is 18% and water is 22%. The total

Ash value of the bark is 38. Phytochemical Screening shows presence of carbohydrate in pet

ether and chloroform extract extracts while absence in ethanol and aqueous extract, fats

and oils are positive in chloroform extract absence in other extract Steroids, Glycosides,

Saponins and alkaloids are present strongly in ethanol extract and chloroform extract while

marginally in aqueous extract. These are not found in pet ether extract phenolic, tannin, resin

compounds are strong l positive in ethanol extract. Thin Layer Chromatography of

Pet.Ether, chloroform, ethanol & aqueous extracts have been performed in different

solvent system of varying degree of polarity using silicagel G of TLC grade. In pet. ether and

aqueous extract only one compounds are extracted while in Chloroform extract four

compounds have been extracted and ethanol extract three compound have been extracted. This

shows that chemical constituents of Ficus arnottiana Miq. is extractable in semi polar solvent

ethanol extract.

Table- 4 Phytochemical screening of Ficus arnottiana Miq. :

ALKALOIDS

S.No. CHEMICAL TEST PET.ETHER1 CHLOROFORM ETHANOL WATER

A Dragendorff’s test - ++ + -

B Mayer’s test - ++ + -

C Wagner’s test + - - +

Key: strongly present ++

Present +

Absent -

Rf value determination: Rf value calculated using different solvent system for

different extracts of Ficus arnottiana Miq.

by TLC.

Table 5 Rf Values for Petrolieum ether extract by TLC:

S.No. Solvent Solvent front No. of spots Spot Rf Value

System height (cm) height (cm)

1. Chloroform:Methanol (7:3) 5.5 1 5.4 0.98

2. Methanol:Benzene (5:5) 5.2 - - -

3. Benzene:Ethyl acetate (9:1) 5 - - -

4. Benzene:Ethyl acetate (5:5) 5.9 1 5.7 0.96

5. n-Butenol:Acetic acid (5:4) 5.5 1 5.3 0.96

Table-6 Rf Values for Chloroform extract by TLC

S. No Solvent Solvent front No.of Spot Rf Value

system height (cm) spots height (cm)

1. Chloroform:Methanol (7:3) 5.4 4 1.5, 3.6, 3.7, 5.3, 0.27, 0.66, 0.68, 0.98

2. Methanol:Benzene (5:5) 5.7 2 4.2, 5.6 0.73, 0.98

3. Benzene:Ethyl acetate (9:1) 5.7 3 4.3, 2.8, 2.3 0.75, 0.49, 0.40

4. Benzene:Ethyl acetate (5:5) 5.6 2 1.1, 2.1 0.19, 0.37

5. n-Butenol:Acetic acid (5:4) 6.0 1 5.1 0.85

Table-7 Rf Values for Ethanolic extract by TLC

S. No Solvent Solvent front No.of Spot Rf Value

system height (cm) spots height (cm)

1. Chloroform:Methanol (7:3) 5.7 2 0.9, 5.2 0.15

2. Methanol:Benzene (5:5) 6 1 5.9 0.98

3. Benzene:Ethyl acetate (9:1) 5.8 - - -

4. Benzene:Ethyl acetate (5:5) 6.0 3 1.5, 2.1,2.5 0.25, 0.35, 0.41

5. n-Butenol:Acetic acid (5:4) 6.0 1 4.2 0.7

Table-8 Rf Values for Aqueous extract by TLC

S. No Solvent Solvent front No.of Spot Rf Value

system height (cm) spots height (cm)

1. Chloroform:Methanol (7:3) 5.7 1 1.3 0.22

2. Methanol:Benzene (5:5) 5.6 - - -

3. Benzene:Ethyl acetate (9:1) 5.7 - - -

4. Benzene:Ethyl acetate (5:5) 5.5 - - -

5. n-Butenol:Acetic acid (5:4) 6.0 1 5.2 0.86

Fig.1 Pet. Ether extract.

Fig.2 Chloroform extract.

Fig.3 Ethanolic extract.

Fig.4 Aqueous extract.

Table: 2 Extractive values of bark of Ficus arnottiana Miq.

S/No. Solvent used Average extractive value in % w/w on dry weight basis

1. Ethanol 18

2. (Absolute) Water 22

Table: 3 Phenolics content values of bark of Ficus arnottiana Miq.

S.No. Extract Phenolics content (AU)

1 Pet.ether Absence

2 Chloroform 451

3 Ethanol 263.5

4 Aqueous 418.5

S/No. Solvent used Average extractive value in % w/w on dry weight basis

RESULTS

The synthesis of Alkaloid is confermed.

Alkaloid profiling by thin layer chromatography (TLC):

TLC investigation showed the presence of steroids-terpenes, alkaloids and flavonoids (Table 2,

Figure)

Table 1: Rf values of chief constituents detected on TLC of 3 different extracts from the root

powder of A. racemosus

Zone Steroids-terpenes Alkaloids Flavonoids

Rf

value

visible colour

Rf

value

visible colour

Rf

value

UV-365nm

fluorescence

1

2

3

4

5

6

7

8

9

10

11

12

13

13-15

15-17

19-21

25-27

29-30

33-35

39-42

46-51

55-57

58-62

62-67

67-69

86-88

yellow

sky blue

grayish brown

yellow brown

brown

yellow

light gray

grayish purple

violet

violet

yellow

grayish blue

grayish yellow

55-57

orange-brown

0-3

52-56

sky blue

blue-green

Discussion

The CAF were reported to be higher in the roots as compared to the callus, regenerated in MS

medium which is in accordance to the earlier findings Alkaloid analysis by TLC showed that

saponins is the major alkaloid in the roots and present in all the three root samples as well as in

the callus tissues. Leaves of all the plants were found to be devoid of alkaloids or present in very

low amount, which is not detectable by TLC. The earlier reports also suggested that saponins is

the major component of roots in Asparagus racemosus. Zoning patterns of steroids-terpenes,

alkaloids and flavonoids on the TLC fingerprints were specific enough to be used for the

identification of A. racemosus root powder. The alkaloid profiling of different plant samples

envisage that the production of secondary metabolites by plants depends greatly on the

physiological and developmental stages of the plants. The synthetic capacity of de-differentiated

tissue often differs substantially from that of fully differentiated tissues, both qualitatively and

quantitatively, because of differences in the enzyme profiles which regulate the organ-specific

expression of the biosynthetic genes. The differentiated cultures often show biochemical and

genetic stability and hence, offers a predictable and high-productivity system which does not

require extensive optimization. The biosynthesis usually occurs in an organ in a tissue -specific

manner and is often temporally restricted during the development

Pharmacological applications of Asparagus Racemosus

Asparagus Racemosus has been used in Ayurveda as a galactagogue, aphrodisiac, anodyne,

diuretic, antispasmodic and nervine tonic since time immemorial. The plant finds use in about 64

ayurvedic formula-tions which include traditional formulations such as ‘Shatavari kalpa’,

‘Phalaghrita’, ‘Vishnu taila’, etc.. Abana® (containing 10 mg Satavari root extract per tablet),

Diabecon®(containing 20 mg Satavari root extract per tablet), EveCare®(containing 32 mg

Satavari root extract per 5 ml syrup), Geriforte®(containing 20 mg Satavari root powder per

tablet), Himplasia®(containing 80 mg Satavari root powder per tablet), Lukol®(containing 40

mg Satavari root extract per tablet) and Menosan®(containing 110 mg Satavari root extract per

tablet) are some formulations containing Asparagus Racemosus developed by Himalaya Herbal

Healthcare, India (Table 1).

Fig. 1. Active principles of Asparagus

Racemosus (I) Shatavarin, (II) Sarsasa-pogenin, (III) Racemosol and (IV) Asparagamine.

Effect on neurodegenerative disorders

In Alzheimer’s and Parkinson’s diseases, excitotoxicity and oxidative stress are the major

mechanisms of neuronal cell death. Therefore, to combat neurodegenerative disorders, there is a

need for a compound that can retard or reverse this neuronal damage. Asparagus Racemosus is a

well-known nervine tonic in the Ayurvedic system of medicine. Parihar and Hemnani (2004)

conducted a study to investigate the potential of methanolic extract of Asparagus Racemosus

roots against kainic acid (KA)-induced hippocampal and striatal neuronal damage in mice. Intra-

hippocampal and intra-striatal injections of KA to anes-thetized mice resulted in the production

of excitotoxic lesions in the brain. After KA injection, impairment of hippocampus and striatal

regions of brain was observed accompanied by increased lipid peroxidation, increased protein

carbonyl content, decreased glutathione peroxidase (GPx) activity and reduced glutathione

(GSH) content. GSH is an important antioxidant which acts as a nucleophilic scavenger of toxic

compounds and as a substrate in the GPx-mediated destruction of hydroperox-ides which would

otherwise accumulate to toxic levels in brain tissues. The mice treated with Asparagus

Racemosus extract showed an enhancement in GPx activity and GSH content, and reduction in

membranal lipid peroxidation and protein carbonyl. They concluded that the plant extract plays

the role of an antiox-idant by attenuating free radical induced oxidative damage. ‘EuMil’, a

polyherbal formulation containing the stan-dardized extracts of Withania somnifera , Ocimum

sanctum , Asparagus Racemosus and Emblica officinalis was evaluated for its anti-stress activity

in rats (Bhattacharya et al., 2002). Chronic electroshock stress for 14 days was found to increase

the rat brain tribulin activity and decrease the monoamine neuro-transmitter levels. ‘EuMil’

treatment normalized the perturbed nor-adrenalin, dopamine and 5-hydroxytryptamine

concentra-tions and also attenuated the tribulin activity. ‘Mentat’, a herbal psychotropic

preparation containing Asparagus Racemosus has been found to be effective in the treat-ment of

alcohol abstinence induced withdrawal symptoms such as tremors, convulsions, hallucinations

and anxiety in ethanol administered rats (Kulkarni and Verma, 1993) due to its anticon-vulsant

and anxiogenic action. However, it is unlikely that these are the only reasons for its de-addiction

potential and therefore can be examined further. Neurological and psychiatric disorders together

account for more chronic suffering than all other disorders combined (Cowan and Kandel, 2001).

Treating these problems however, remains a challenging field in medical science. Keeping in

mind the encouraging leads and the limited data regarding the use of Asparagus Racemosus in

treating neurological disorders; more studies need to be conducted to fully exploit the potential

of Satavari in this area.

Anti-diarrhoeal effects

Diarrhoea has long been recognized as one of the most important health problems faced globally

particularly by the population of developing countries. Each year diarrhoea is esti-mated to kill

about 2.2 million people globally, a majority of whom are infants and children below the age of

5 years (WHO, 2005). Nanal et al. (1974) found Satavari to be extremely effective in the

treatment of Atisar (diarrhoea), Pravahika (dysentery) and Pittaj shool (gastritis) as described in

Ayurvedic texts such as Sushruta Samhita and Sharangdhar Samhita . Ethanol and aque-ous

extracts of Asparagus Racemosus roots exhibited significant anti-diarrhoeal activity against

castor oil induced diarrhoea in rats demonstrating an activity similar to loperamide .The release

of ricinoleic acid from castor oil results in inflammation and irritation of the intestinal mucosa

caus-ing the release of prostaglandins which stimulate motility and secretion. It is well known

that ‘prostaglandin E’ causes diar-rhoea in experimental animals and human beings. Therefore,

the action of this extract can be attributed to the inhibition of prostaglandin biosynthesis which in

turn inhibits gastro-intestinal motility and secretion. Since the Asparagus Racemosus root extract

is composed of saponins, alkaloids, flavonoids, sterols and terpenes; further analysis is needed to

identify the exact phytoconstituent(s) that imparts the anti-diarrhoeal action.

Anti-dyspepsia effects

Asparagus Racemosus also finds use in Ayurveda in the treat-ment of dyspepsia. The plant was

found to have an effect comparable to a modern allopathic drug metoclopramide which is a

dopamine antagonist used in dyspepsia to reduce gastric emptying time. In this study, 2 g

powdered root of Asparagus Racemosus was compared to a standard treatment of

metoclopramide (10 mg tablet) in eight normal healthy male vol-unteers, and the gastric

emptying halftime was observed. There was no statistically significant difference between the

actions of Asparagus Racemosus and metoclopramide. They hypoth-esized that Satavari might

be a mild dopamine agonist. This isolated study merely supports the use of Satavari in traditional

Ayurvedic medicine as an anti-dyspeptic drug. It does not elabo-rate its mechanism of action

which can be an avenue for further research.

Cardio protective effects

Increase in serum lipid levels especially cholesterol along with the generation of reactive oxygen

species are the major reasons for the development of coronary artery disease and atherosclerosis.

‘Abana’, a herbo-mineral formulation contain-ing 10 mg Asparagus Racemosus extract per

tablet, was found to have significant hypocholesterolaemic effect in rats and there-fore

demonstrated a potential for use as a cardio-protective agent . They found that the total choles-

terol, phospholipids and triglyceride levels were significantly lower (37–45%) as against the

control. Since ‘Abana’ is a poly-herbal formulation, further research needs to be conducted on

the exact role that the Asparagus Racemosus component plays in the hypolipidaemic action.

Asparagus Racemosus has also been investigated for the reduction of cholesterol levels in

hypercholesteremic rats by Visavadiya and Narasimhacharya (2005). They found that Asparagus

Racemosus root powder supplements decreased lipid peroxidation and caused a dose-dependent

reduction in lipid profiles. The total lipids, total cholesterol and triglycerides in plasma and liver

as well as plasma LDL (low-density lipopro-tein) and VLDL (very low-density lipoprotein)-

cholesterol decreased by more than 30%. Though it can be hypothesized that the

hypercholesteremia is alleviated by decreasing exogenous cholesterol absorption and increasing

conversion of endogenous cholesterol to bile acid; more research needs to be conducted to

comprehend the mechanism of action responsible for this action.

Anti-bacterial effects

In an isolated study, different concentrations of the methanol extract of the roots of Asparagus

Racemosus have also shown considerable antibacterial efficacy under in vitro con-ditions against

Escherichia coli , Shigella dysenteriae , Shigella sonnei, Shigella flexneri , Vibrio cholerae ,

Salmonella typhi , Salmonella typhimurium , Pseudomonas putida , Bacillus sub-tilis and

Staphylococcus aureus (Mandal et al., 2000b). The antibacterial effect of Asparagus Racemosus

may also be play-ing a secondary role in its action with respect to other functions of the plant as

well and therefore needs to be studied in greater detail.

Immunoadjuvant effects

The immunoadjuvant potential of Asparagus Racemosus was studied in experimental animals

immunized with diphtheria, tetanus, and pertussis (DTP) vaccine. After challenge, animals

treated daily with Asparagus racemo-sus aqueous root extract (100 mg/kg body weight) showed

a significant increase (p = 0.0052) in antibody titres to Bordetella pertussis as against the

untreated animals. Reduced mortality coupled with overall improved health status was observed

in treated animals and this indicated the development of a protec-tive immune response. Extracts

and formulations prepared from Asparagus race-mosus exhibited various

immunopharmacological actions such as increases in white cell counts, haemagglutinating and

haemolytic antibody titres in cyclophosphamide (CP)-treated mouse ascitic. CP is widely used in

the treatment of a variety of malignant and non-malignant immunopathological disorders and has

several side effects such as leucopenia, anaemia, etc. Since macrophages play an important role

in the development of intraperitoneal adhesions, the modulation of macrophage activity would

pro-vide a new approach for the prevention and management of post-operative adhesions.

Asparagus Racemosus being reported to be an immunomodulator and immunostimulant,

significantly decreased the adhesion scores by increasing macrophage phago-cytosis by more

than 50% in experimental animals treated with the plant extract. It was demonstrated that a

combination of Asparagus Racemosus , Withania som-nifera and Tinospora cordifolia extracts

protected mice against CP induced neutropenia. Pre-treatment for 15 days with these drugs

produced a striking leucocytosis with a predominant neu-trophilia. The leucopenia and

specifically neutropenia induced by CP was significantly reduced by the plant extract. The total

WBC and absolute neutrophil counts following the treatment with Asparagus Racemosus had

risen very high (18800 ± 2001, 13366 ± 501.96, respectively) such that the percentage fall after

CP administration (63.77% total counts, 58.13% neu-trophil counts) was greater than that in the

control group (43.78%, 25.18%, respectively) (Dhuley, 1997). In addition, it was found that

Asparagus Racemosus , Withania somnifera and Tinospora cordifolia along with Picrorhiza

kurrooa significantly inhibited carcinogen ochratoxin-induced suppression of chemo-tactic

activity and production of interleukin-1 and TNF-by macrophages (Thatte et al., 1987). The role

of Asparagus Racemosus as an immunoadjuvant in traditional therapy is well documented and

therefore it can be applied to evade the toxic side effects of synthetic chemothera-peutic drugs

without compromising on its anti-tumour activity. Interestingly, in Ayurvedic medicine, AIDS is

thought to be a disease of decreased ‘ojas’, defined as the essential energy of the body. Satavari

is said to aid in the formation of ‘ojas’ and has been used in immune therapy (Canadian AIDS

Treatment N. Bopana, S. Saxena / Journal of Ethnopharmacology 110 (2007) 1–15 11

Information Exchange, 2005). It is in situations like these that the function of Asparagus

Racemosus as an immunoadjuvant can be scrutinized for use in adjuvant therapy in the

management of HIV.

Antitussive effects

In yet another isolated report the methanol extract of Aspara-gus racemosus roots showed

significant antitussive activity on sulphur dioxide induced cough in mice with the cough

inhibition being comparable to that of 10–20 mg/kg of codeine phosphate (Mandal et al., 2000a).

This action has not been well documented and can be worked upon further.

Challenges in conservation and sustainable use of Asparagus Racemosus

Due to its multiple uses, the demand for Asparagus racemo-sus is constantly on the rise; however

the supply is rather erratic and inadequate. Destructive harvesting combined with habitat

destruction in the form of deforestation adds to the magnitude of the problem. All of this has

resulted in the drastic shrinkage of its population. In nature, the species is propagated through

seeds in March–April. Apart from this method, Aspara-gus racemosus can also be propagated

vegetatively but this is a very slow and laborious technique. Irrespective of the mode of

propagation the plant is ready for harvesting only by the third year. Hence, this is not an effective

solution to meet the growing demand for this plant. Considering the escalating demands of the

market for a continuous and uniform supply of the plant material, and the increasing depletion of

the forest resource base, cultivation of the plant rather than collection from wild will be an

effective strategy. However, the basis for a sound conservation strategy would lie not only in

increasing the area under cultivation, but also in attaining greater productivity to ensure

reasonably higher financial returns to the growers.

Uses & Benefits of Asparagus

I. The roots are useful in nervous disorders, dyspepsia, tumors, scalding of urine, throat

infections, tuberculosis, cough bronchitis and general debility.

II. It helps with nervousness, pain, restless sleep, disturbing dreams and people with weak

emotional and physical heart.

III. The herb is useful for treating anorexia, insomnia, hyperactive children and people who

are under-weight.

IV. Asparagus is considered as a rejuvenating female tonic for overall health and vitality.

V. Satawari is used for treating sexual debility, infertility in both the sexes, and menopausal

symptoms and increases milk secretion during lactation.

VI. The herb is useful in pregnancy for threatened abortion.

VII. It is useful for the treatment of ulcerative disorders of stomach and Parinama Sula,

clinical entity akin to the duodenal ulcer diseases.

VIII. The paste of fresh leaves is used to apply on the burning sensation of the skin in

smallpox and bullae.

IX. The fresh juice of the roots, mixed with honey, helps in reducing the burning sensation

pain in tumors, due to pitta.

X. Asparagus proves to be an effective demulcent for the dry and inflamed membranes of

the lungs, stomach, kidneys and sexual organs.

XI. The herb is also an extremely nutritious tonic for women from menarche to menopause.

XII. Since it increases the urinary output, it is beneficial in urinary stones and Dysurea.

XIII. Asparagus is anabolic to uterus and thus, helpful in uterine hypoplasia in young girls.

XIV. It improves uterine growth, mitigates dysmenorrheal and menorrhagia, augments fertility

and imparts anabolic properties.

XV. The herb works as a rejuvenative to improve eyesight, when consumed for a prolonged

duration.

XVI. When taken with a cup of saffron milk, asparagus is good for post-menopausal women.

XVII. It curbs the intensity of the bronchospasms and decreases the frequency of paroxysms.

XVIII. Conclusion

XIX. The pharmacological studies conducted on Asparagus race-mosus indicate the immense

potential of this plant in the treatment of conditions such as menopausal symptoms,

neurode-generative disorders, diarrhoea, dyspepsia, etc. However, gaps in the studies

conducted are apparent which need to be bridged in order to exploit the full medicinal

potential of Asparagus Racemosus . Since most drugs containing Satavari that are

available in the market are in the form of polyherbal formulations, it is difficult to

attribute a particular medicinal action as being solely due to the Asparagus Racemosus

component of the drug. Aside from possible synergistic effects existing among the plant

extract constituents, another likelihood could be the formation of ‘pro-drugs’ which are

defined as compounds activated in the body after the administration of a particular

medicineWhile most of the research has been in vivo which has helped to validate the

applicability on the human system; in vitro stud-ies would have facilitated a better

understanding of the mode of action of Asparagus Racemosus . Due to the non-

availability of commercial Shatavarin standards, most studies first involve the extraction

and purification of the active principle to be used as a reference standard which makes the

process more cumber-some. The availability of authentic metabolite standards would not

only hasten secondary metabolite assays but also make the results more reliable and

reproducible. All of these issues can be addressed by adopting a ‘systems biology’

approach wherein a very large number of com-ponents (at the levels of genome,

transcriptome, proteome or metabolome as well as physiological parameters such as

blood pressure, etc.) are catalogued and statistical methods are used to infer correlations

to understand mechanisms of action. A key technology in systems biology is

‘metabolomics’. Metabolomics refers to the exhaustive profiling (identifica-tion, analysis

and quantification) of the ‘metabolome’ which includes all the metabolites in a cell. It is a

complex inter-disciplinary field of research that requires a combination of bioscience,

analytical chemistry, organic chemistry, chemomet-rics and informatics. Strategies for

metabolomics analysis can include the metabolite profil-ing of Asparagus Racemosus by

identification and quantification of pre-defined metabolites by chromatographic

separations (by high performance liquid chromatography, gas chromatogra-phy, and

electrophoresis) followed by spectroscopic detection (by mass spectrometry or nuclear

magnetic-resonance spec-trometry). Metabolic fingerprinting should be done for sample

classification purposes to ensure purity and avoid deliberate or inadvertent adulteration

with material from other Aspara-gus species.

Reference:

Buppachart Potduang, Maneerat Meeploy, Rattanasiri Giwanon, Yaowaluck Benmart, Montree

Kaewduang and Winai Supatanakul,2008 “BIOLOGICAL ACTIVITIES OF ASPARAGUS

RACEMOSUS” , Pharmaceuticals and Natural Products Department ,Thailand Institute of

Scientific and Technological Research.

J. S. Negi, P. Singh, G. P. Joshi, M. S. Rawat, and V. K. Bisht,2010, “Chemical constituents of

Asparagus”, Department of Chemistry, HNB Garhwal University, Srinagar, Garhwal, and Herbal

Research and Development Institute, Gopeshwar, Uttarakhand, India

Bharat B. Aggarwal,* Sahdeo Prasad, Simone Reuter, Ramaswamy Kannappan, Vivek R. Yadev,

Byoungduck Park, Ji Hye Kim, Subash C. Gupta, Kanokkarn Phromnoi, Chitra Sundaram,

Seema Prasad, Madan M. Chaturvedi, and Bokyung Sung,2011, “Identification of Novel Anti-

inflammatory Agents from Ayurvedic Medicine for Prevention of Chronic Diseases” ,