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C H A P T E R 6

Isolation of curcumin from turmeric

Turmeric, the dried yellow rhizome of Curcuma longa, is a common oriental

spice that gives curry dishes their characteristic yellowish color and is used in

Indian cooking. The active ingredient in turmeric is curcumin, which is

approximately 0.5-6.0 % by weight of the root of turmeric. Since 1970, when

curcumin first attracted the interest of scientific medical researchers, studies of

curcumin have reported suppressive effects for the following medical conditions:

High serum cholesterol levels. Free radical damage to tissues, Diabetic cataracts,

Diabetic damage to pancreatic insulin-producing cells. Diabetic wounds,

Rheumatoid arthritis. Inflammatory bowel disease, Crohn's Disease, Psoriasis,

Cancer, Atherosclerosis, Inherited peripheral neuropathies. Liver damage by

chemicals and drugs. Microbial infections. Other four medical conditions for

which curcumin seems to hold especially exciting prospects: Alzheimer's Disease,

Parkinson's Disease, Cystic fibrosis. Ailments related to Epstein-Barr Virus.

fwww.lifelinknet.com/.../Products/PriMeric.asp] [http://jchemed. chem. wise. edu/journal/Issues/2000/Mar/abs359. html]

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Chapter VI Isolation turmeric

CurcuHiinau^s^ grouf of •phei/wUc com-fouMis LsoLated from the roots, of

Curcum-fl LoiA fl (z,Ltîberaaeae), e/ihibtt a variety of benefluaL effects oyi health

at^ 0^^ eve^^ts that hel'p ti^ -prevei^ttp^ c-ertatiA- diseases. A vast majoritij of these

studies were carried out with curcuntit^. (difeMLoLjL m.ethan€), which is a m.ajor

curcuntiiAXiid. The m.pst detailed studies usin.g curcurutin. ii^x^Lude a^ti-

it^,fLaru.m.atoru, an-tioxidant, ai^ticarcin^geiîc, an.tiviraL^ ai^ an.tiii^fsctious

activities. In addition, the wound healing and detoxifying -properties of curcumin

have also received considerable attention. As a result of extensive therapeutic

properties of curcuntin there is a need to develop newer and selective m.ethod for the

isolation of curcuntinoids from, turmêric. This chapter is a step in this direction to

im.provise the quantity of curcum.in isolation from. turm.eric roots.

kLiwôrds: Curcuw-lvuiids., Cunuma Lonâ, turm.eruc roots.


VI. 1. Introduction

Curcumin (diferuloyl methane), the natural

yellow pigment in turmeric, is isolated from the rhizomes

of the plant Curcuma longa. It constitutes about 3-4% of

the composition of turmeric. In the south and southeast

tropical Asian countries, turmeric has been used for

centuries as a spice to give the specific flavor and yellow

color to curry. Turmeric became a very important spice to

mankind when it was observed that the addition of

turmeric powder in food preparation preserved its

freshness and nutritive value. Turmeric, as an additive,

improved the palatability. Aesthetic appeal and shelf life

of perishable food items. The use of turmeric became

more popular when it was found to act as a therapeutic

agent for various illnesses. In the Ayurvedic system of

medicine, turmeric is used as a tonic and as a blood

purifier. Its role in the treatment of skin diseases and its

ability to soften rough skin resulted in the prolific use of

turmeric in topical creams and bath soaps in India. Turmeric

is also used in home remedies in the treatment of cuts,

wounds, bruises, and sprains. Its use as an anti

inflammatory and antimicrobial agent has been recognized

for more than a century. The importance of turmeric in

medicine took a new twist when it was discovered that the

dried rhizome of Curcuma longa is very rich in phenolics,

whose structures have been identified as curcuminoids

(Figure 1). Phenolics are known to possess antioxidant

properties. Free radical mediated damage to biological

systems is recognized as the initiating agent for many

diseases, such as cardiovascular diseases, cancer, and

arthritis. Turmeric and its constituents show beneficial

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effects on these diseases and on other illnesses. For

example, the low incidence of large bowel cancers in Indians

could be attributed to a high intake of natural antioxidants,

such as curcumin in the diet [1]. The anti-mutagenic and

anti-tumor effects of curcumin are most widely studied

[1]. However, in recent years, it has been shown that the

inhibition of arachidonic acid metabolism, modulation of

cellular signal transduction pathways, inhibition of

hormone, growth factor, and oncogene activity are some of

the mechanisms by which curcumin causes tumor

suppression [3]. Chemopreventive activity of curcumin is

observed when administered prior to, during, and after

carcinogen treatment as well as when it is given only

during the promotion/progression phase (starting late in

premalignant stage) of colon carcinogenesis in F 344 rats.

Curcumin is also a powerftil inhibitor of the proliferation of

several tumor cells, as well as an anti-inflammatory agent.

It exhibits anti-clastogenic, anti-ftingal and anti-viral

properties. However, the lack of information regarding the

mechanisms of action of curcumin has precluded its clinical

use in western countries. Several recent studies have given

some insight into the molecular basis for the action of

curcumin at the cellular level. This review looks at some of

these insights into the cellular processes, molecular, and/or

biochemical mechanisms that are influenced by curcumin

[4-9].

"aCO^ O o

H O - ^ 7 - C H = C H — C - C H j - C - C H = C H - ^ ^ O H

Curcumul

HfifK O O .

H O - ^ ^ C H = C H - C - C H j - C - C H = C H - ^ y - O H

Cuicumin 11 ( demeUioxycutGuiDm )

"° \ / ' "~ CH—C—CHj—C—CH: = C H - ^ V o H

Curcumin III ( biademethoxycufcumin )

Figure VI.1

Structure of some natural curcuminoids

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VI.1.1. The biological source ofcurcumin

Curcumin genus in the plant family of Zingiberacea,

is the biological source for curcuminoids, including

curcumin. Curcuma longa, the yellow tuberous root that is

referred to as turmeric, was taken from India to Southeast

Asia, China, North Australia, West Indies, and South

America. Subsequently, its cultivation spread to many

African countries. India, however, remains the largest

producer of tiormeric in the world, with a figure of 4,87,000

metric tones in production, of which 27 ,750 metric tones

are exported. The yellow pigmented fraction of Curcuma

longa contains curcuminoids, which are chemically related

to its principal ingredient curcumin. The three main

curcuminoids isolated from turmeric are curcumin,

demethoxy curcumin, and bisdemethoxy curcumin (Figure

I). Curcuminoids are present in 3-5% of turmeric.

Curcumin is the important active ingredient responsible for

the biological activity of turmeric. Curcumin, C2H20O6

(m.p. 1 84°C), or diferuloyl methane was first isolated in

1815. The crystalline form of curcumin was obtained in

1910, and Lampe solved its structure in 1913. It is insoluble

in water, but soluble in ethanol and acetone [4-9].

VI.1.2. Biological activities of curcumin

VI. 1.2.1. Anti-injlammatrory property

Inflammation is a necessary process for fighting

infections. It results from a series of complex reactions,

triggered by the host immunological response. Uncontrolled

inflammatory responses may lead to undesirable effects,

such as tissue damage. Many of the diseases, such as

Peptide in turmeric Besides the phenolics, turmeric is also a source of a water-soluble peptide with antioxidant properties. This compound has been identified as turmerin, a heat-stable noncyclic peptide which has 40 amino acid residues, and is resistant to the proteolytic action of the enzymes trypsin and pepsin. Turmerin, which is found in turmeric at a concentration of 0.1 percent, has been shown in some experiments to be a more potent antioxidant than curcuminoids or butylated hydroxyanisole (BHA). Turmerin is rich in methionine, the sulfur-containing amino acid, and a known antioxidant, which may in part explain the strong antioxidant properties of this compound.

[Turmeric and the healing curcuminoids. Muhammed Majeed, Ph.D., Vladimir Badmaev, M.D., Ph.D.and Frank Murray]

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rheumatoid arthritis, are the result of sustained production of

inflammatory mediators causing physical damage to joints.

Many inflammatory mediators have been implicated in these

complex reactions, some of which are modulated by

curcumin [10-14].

VI.1.2.2. Antioxidant properties

The discovery of the antioxidant properties of

curcumin explains many of its wide ranging

pharmacological activities. Curcumin is an effective

antioxidant and scavenges superoxide radicals, hydrogen

peroxide, and nitric oxide from activated macrophages. It

inhibits the inducible nitric oxide synthase activity in

macrophages. Human keratinocytes are protected from

Xanthinexanthine oxidase injury by virtue of the antioxidant

property of curcumin. Oral administration of 30mg/kg body

weight of curcumin in rats for 10 days reduces the iron-

induced hepatic damage by lowering lipid peroxidation.

Protection from radiation by dietary curcumin administered

to mice is also attributed to the antioxidant property of

curcumin. Curcumin protects renal cells and neural glial

cells from oxidative stress. Interestingly, curcumin not only

exhibits antioxidative and free radical scavenging

properties, but also enhances the activities of other

antioxidants, such as superoxide dismutase, catalase, and

glutathione peroxidase. Lipid peroxidation is lower in liver,

kidney, spleen, and brain microsomes from retinol deficient

rats that are fed with 0.1% dietary curcumin for three weeks.

Another mechanism by which curcumin protects against

oxidative stress in endothelial cells is by the induction of

heme oxygenase-1 [15-18].

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VI.L2.3. Effect ofcurcumin on lymphocytes

Mucosal CD4 (+) T cells and B cells increase in

animals treated with curcumin, suggesting that it modulates

lymphocyte-mediated immune functions [19]. Dietary

curcumin increases antibody response in rats in vivo. In vitro,

curcumin enhances IgM production in rat spleen

lymphocytes. Han et al. studied the ability of curcumin to

modulate proliferative responses of normal splenic and

transformed B-lymphocytes. These observations indicate

that curcumin arrests growth and induced apoptosis of B

cell lymphomas more effectively than normal B

lymphocytes. In BKS-2 B lymphoma cells, the inhibitory

effects of curcumin appear to be mediated by the down-

regulation of survival genes (egr-1, c-myc, bcl-XL and NF-

KB), as well as the tumor suppressor gene p53. On the other

hand, curcumin may be an effective adjunct in the

prevention of post-transplant lymphoproliferative disorder in

patients undergoing therapy with cyclosporine. A, because

curcumin blocks the B-cell immortalization by EBV, which

is promoted by oxidative stress induced by cyclosporin A

[20-21].

VI. 1.2.4. Effect ofcurcumin on platelet aggregation

Curcumin inhibits platelet-activating factor (PAF),

ADR arachidonic acid (AA), epinephrine, and collagen

mediated platelet aggregation. However, at lower doses

(20-25|JM), curcumin inhibits only PAF and AA mediated

platelet aggregation and not those mediated by other agonists.

Pretreatment of platelets with curcumin resulted in the

inhibition of platelet aggregation induced by calcium

ionophore A 23187, but not by the PKC activator, PMA

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[22]. Curcumin also inhibited thromboxane A2 (TXA2)

formation by platelets. These observations suggest that

curcumin mediated preferential inhibition of PAF- and AA-

induced platelet aggregation involves inhibitory effects on

TXA2 synthesis and Ca ^ signaling [23].

VI. 1.2.5. Effects of curcumin on detoxification

mechanisms curcumin, cell cycle, and

apoptosis

Curcumin is an apoptotic agent. While a low

concentration of curcumin is known to arrest cell

proliferation in the G0-GI/G2/S phase, a high concentration

of curcumin induces apoptosis in rat A7r5 cells [24].

Several hallmarks of apoptosis. including DNA laddering,

chromatin condensation and fragmentation, and an

apoptosis specific cleavage of 28S and 18S ribosomal

RNA were observed after treatment of immortalized

mouse embryo fibroblast NIH 3T3, erb B2 oncogene-

transformed NIH 3T3, mouse sarcoma SI80, human colon

cancer cell HT-29, human kidney cancer cell 293, and

human hepatocellular carcinoma Hep 02 cells with

curcumin [25]. Curcumin-induced apoptosis in human basal

cells was shown to be dependent on a p53-signaling path

way. Curcumin was shovra to accumulate in plasma

membrane, endoplasmic reticulum, and nuclear envelope

and to produce apoptosis-like changes in plasma mem

branes in rat thymocytes The sequence and extent of primary

events during apoptosis induced by curcumin have been

compared with those occurring during dexamethasone-

induced apoptosis in rat thymocytes (Curcumin-treated cells

National Cancer Institute research

update

The National Cancer Institute is currently developing curcumin as a drug for the treatment of cancer. The rationale behind this effort is based on the combination of potential chemopreventive mechanisms, the validated biological activity in preclinical studies and its proven safety to humans.

A Phase I trial assessing the safety and pharmacokinetics, studies on patients with colon polyps (in view of the inhibitory effect of curcumin on the formation of arachidonic acid metabolites) and a Phase II chemoprevention trial on patients with oral leukoplakia are included in these plans.

[http://www.curcuminoid s.com/nci.htm]

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exhibit typical features of apoptotic cell death, including

shrinkage, transient phosphatidylserine exposure, increased

membrane permeability and decrease in mitochondrial

membrane potential. The level of anti-apoptotic protein

Bcl-2 was decreased in the presence of curcumin [26].

VI. 1.2.6. Clinical evaluation of curcumin as an anti-

carcinogenic agent in humans

Because of their safety and the fact that they are not

perceived as 'medicine,' food-derived products are of great

interest in the development of chemopreventive agents,

which may have a widespread long-term use in populations

at normal risk. Curcumin is one such diet-derived agent that

is being clinically evaluated as a chemopreventive agent for

major cancer targets, including the breast, prostate, colon,

and lung [27]. For developing such agents, the National

Cancer Institute (NCI) has advocated co-development of a

single or a few putative active compounds (including

curcumin) that are contained in the food-derived agent. The

active compounds provide mechanistic and pharmacologic

data that may be used to characterize the chemopreventive

potential of the extract, and these compounds may be useful

as chemopreventives in higher risk subjects (patients wdth

precancers or previous cancers). Other critical aspects for

developing the food-derived products are careful analysis

and definition of the extract to ensure reproducibility

(eg., growth conditions, chromatographic characteristics, or

composition) and basic science studies to confirm

epidemiologic findings associating the food product vsdth

cancer prevention [28].

Curcumin for Cancer

In clinical studies, a topical ointment containing five percent curcumin applied to cancerous growth on the skin in 62 patients was found to reduce foul smell, itching, pain and the discharge of fluid from the lesion (wound) in a significant majority of the patients.8 The foul odor was considerably reduced in more than 90 percent of the patients; pain and itching subsided in 50 percent; and fluid discharge was reduced in 70 percent of the cases. Turmeric extracts or alone in combination with betel leaf extract was effective against tumors induced by a powerful carcinogen (a nitrosamine derivative) in the mouth mucosa of hamsters. Curcumin also inhibited the action of another potent carcinogen (a nitro quinoline derivative) in inducing tumors in the mouth mucosa of rats. In a clinical study, the effectiveness of curcumin was tested in patients with oral cancer. One hundred patients were given 500 mg of curcumin three times a day for 30 days. A significant number of patients improved on the curcumin regimen and responded with dramatic clinical improvement within 15 days, while others responded more gradually throughout the 30day treatment period.

[Turmeric and the healing curcuminoids. Muhammed Majeed, Ph.D., Vladimir Badmaev, M.D., Ph.D.and Frank Murray]

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VLl.2.7, Wound-healing properties of curcumin

Tissue repair and wound healing are complex

processes that involve inflammation, granulation, and tissue

remodeling. Interactions of different cells, extracellular

matrix proteins, and their receptors are involved in wound

healing and are mediated by cytokines and growth factors.

Curcimiin enhances cutaneous wound healing in rats and

guinea pigs by increasing the formation of granulation

tissue, biosynthesis of extracellular matrix proteins, and

TGF-pi in wounds [29]. Curcumin also accelerated wound

healing in streptozotocin-induced diabetic swiss albino rats

and genetically diabetic (C57/KsJdb+/db+) mice by

increasing the formation of granulation tissue, faster

re-epitheliatization, and increased collagenization [30].

Systemic treatment with curcumin after local muscle injury

leads to faster restoration of normal tissue architecture, as

well as an increased expression of biochemical markers

associated with muscle regeneration [31].

VI. 1.2.8. Curcumin and diabetes

Feeding diabetic rats curcumin improved their

metabolic status [32], Diabetic rats maintained on a 0.5%

curcumin diet for 8 weeks excreted comparatively lower

amounts of albumin, urea, creatinine, inorganic

phosphorus, sodivmi, and potassium. On the other hand,

glucose excretion or the fasting sugar level was unaffected

by dietary curcumin and also the body weights were not

improved to any significant extent. Diabetic rats fed a

curcimiin diet had a lower relative liver weight at the end of

the study, compared to other diabetic groups. Diabetic rats

According to ayurveda, diabetes is a metabolic kapha type of disorder in which diminished functioning ofagni leads to a tendency toward high blood sugar. Ayurvedic practitioners attack diabetes using a multiprong approach. First, they address diet modification, eliminating sugar and simple carbohydrates, and emphasizing complex carbohydrates. Protein is limited, since excessive intake can damage the kidneys. Fat is also limited because there is often a deficiency of pancreatic enzymes, making fat digestion difficult

The most important herbs for all doshas are shilajit, gudmar turmeric, neem, amalaki, guggul, and arjuna. Turmeric with aloe vera gel (1 to 3 gms./.035 to . 1 oz) is best used during the early stages of diabetes for regulating pancreas and liver functions.

[http://www.holisticonline. com/Remedies/Diabetes/di abetes_ayurveda.htm]

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fed a curcumin diet also showed lowered lipid peroxidation

in plasma and urine. The extent of lipid peroxidation,

however, was still higher in cholesterol fed diabetic groups,

compared to diabetic rats fed with control diet. The

mechanism by which curcumin improves this situation is

probably by virtue of its hypocholesterolemic influence

[33], antioxidant nature and free radical scavenging

property [32].

VI. 1.2.9. Antiviral properties of curcumin

In vitro, curcumin (0.32 mg/ml) moderately

inhibited the activity of human simplex virus-2. Curcumin

provided significant protection in a mouse model of

intravaginal human simplex virus-2 challenge [34]. Cur

cumin is also highly effective in inhibiting Type I Human

Immunodeficiency Virus, (HIV) long terminal repeat

directed gene expression, and viral replication. Curcumin

inhibited p24 antigen production in cells either acutely or

chronically infected with HIV-1. However, curcumin failed

to inhibit the HIV-I multiplication in acutely infected MT-4

cells. Nevertheless, curcumin specifically inhibited the

enzymatic reactions associated with HIV-I integrase but not

other viral (HI V-I reverse transcriptase) and cellular (UNA

polymerase II) nucleic acid processing enzymes [35,36].

Curcumin was isolated as early as 1815. Daube in

1870 obtained it in crystalline form [37], To our

knowledge, no information has been reported in the

literature about the process of isolating pure curcuminoids

from turmeric. Therefore, the objective of the present study

was to develop a new method for the isolation of curcumin

from turmeric.

Curcumin against HIV

The importance of studies of the turmeric derivatives as potential anti-HlV biological response modifiers is obvious. The World Health Organization reported in 1994 that some 17 million people have been infected with HIV worldwide. The vaccine solution to prevent HlV infections is still a remote possibility, particularly in view of the fact that HIV mutates very rapidly. This means that developing resistance through vaccination to one form of HIV may be rendered futile when a new form of HIV evolves. It is noteworthy that some forms of HIV became resistant to the specific antiviral mechanism offered by one of the few useful anti-HIV agents, azidothymidine (AZT) Turmeric derivatives, on the other hand, may offer in the battle against HIV a nonspecific mechanism or mechanisms that may regulate the immunological response in the direction of natural resistance of the organism to being infected by the virus. Interestingly, the nonspecific or bioprotectant nature of curcuminoids may be studied as an independent therapy modality against the HIV infection, as well as an add-on to specific antiviral drugs, possibly diminishing their inherent toxicity.

[Turmeric and the healing curcuminoids Muhammed Majeed, Ph D , Vladimir Badmaev, M D , Ph D and Frank Murray]

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VI.2. Materials and methods

VI.2.1. Chemicals

Methanol, ethanol (HPLC grade), isopropyl alcohol,

n-butyl alcohol, iso butyl alcohol, secondary butyl alcohol,

iso amyl alcohol (Analytical grade) (BDH, India ltd).

VI. 2.2. Procedure

Turmeric rhizomes were powdered and sieved

through 30 mesh size sieve to obtain sample of uniform

particle size. The resulting powder was extracted with

acetone using cold percolation process in which the solvent

uniformly seeps through the particle bed (sample powder),

allowing the efficient diffusion of the soluble from the

powder into the solvent after the contact time of 90 min. the

solvent ratio was 5 volumes calculated on the dry weight of

powdered turmeric rhizome. The extraction procedure was

repeated for 7 times and the individual extracts combined

before concentrating. The extracts were filtered and

concentrated by distillation under vacuum at temperature

less than50°C to produce turmeric oleioresin.

A detailed study was carried out for isolation of

curcuminoids from turmeric oleioresin extracted as above.

In 100ml beaker 20 g turmeric oleioresin and 20g solvent

were added, mixed well and kept aside for 48 hrs at room

temperature for curcumioids precipitation. The precipitated

curcuminoid crystals were purified by washing several

times with the solvent.

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VI.2.3. HPLC method for quantitative estimation of

curcuminoids

The HPLC method described below was used to

estimate the curcuminoids. The area under the peaks is

measured for quantitative analysis of the sample.

Chromatographic System

HPLC Pump

HPLC Detector SPD 10 A

Syringe

Injector

Column

LC 10 AD - Shimadzu Make

Shimadzu Make

Hamilton 100 ml Syringe

77251 Rheodyne Injector

250 X 4.6 mm SS column

containing amino packing 5

micron particle size (NEC

Phenomenex column is

suitable).

Instrument Conditions

Mobile Phase

Injection size

Flow rate

Detector

Wavelength

Alcohol and methanol are mixed in

the ratio 60:40 and the mixture is

degassed and filtered.

20 ml

1 ml per minute

UV

254 nm

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Preparation of standards

Standard Curcumin: 25 mg of standard curcumin is

accurately weighed into a 25 ml volumetric flask and

dissolved in methanol with warming, if necessary and the

volume made up wdth methanol, followed by mixing.

Standard Bismethoxycurcumin and standard

Demethoxycurcumin: In each case, 10 mg of the compound

is accurately weighed into a 25 ml volumetric flask,

dissolved and diluted to volume with methanol.

Preparation of Sample

25 mg of the sample is accurately weighed into a 25

ml volumetric flask, dissolved and diluted to volume with

methanol.

VI.2.4. Procedure

Each standard preparation and sample preparation is

injected, in duplicate, separately into the chromatograph.

The response of the sample preparation in terms of areas

under the three major peaks corresponding to BDMC, DMC

and curcumin and the area under the major peaks for the

respective standards is measured.

The content of each curcuminoid is calculated as follows:

Content of Curcumin

Sample area of curcumin x standard weight x standard purity

Standard area of curcumin x sample weight

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Content of Bisdemethoxycurcumin

Sample area of BDMC X Standard weight X Standard purity

Standard area of BDMC X Sample weight

Content of Demethoxy Curcumin

Sample area of DMC X Standard weight X Standard purity

Standard area of DMC X Sample weight

io4 L . 1 . . / ^ . .

The chromatograms obtained for the standards

are shown

o o o •^ ^ "t C

VI. 2.5. Effect of solvents

20 gm of turmeric oleioresin was taken in different

beakers and 20 gm of different solvents were added and

mixed well. The mixture was kept for 48 h at room

temperature for curcuminoids precipitation.

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VI.2.6. Effect of time

20 gm of turmeric oleioresin and 20 gm n-butyl

alcohol was added and curcumin precipitation was studied

at different time intervals.

VI.2.7. Effect of Temperature

20 gm of turmeric oleioresin and 20 gm of n- butyl

alcohol was added and incubated at different temperatures

for two days.

VI.2.8. Effect of volume of n-butyl alcohol for the

precipitation of curcuminoids

Different ratio of n- butyl alcohol was added to the

turmeric oleoresin and incubated at room temperature for

two days and yield was calculated.

^7.2.9. Effect of ageing of oleoresin

Ageing of the oleioresin was studied, after

standardizing all other parameters like effect of solvent,

temperature, solvent concentration and time.

After extraction of oleioresin fi'om turmeric

rhizomes, oleioresin was kept for ageing to improve the

isolation of curcuminoids. Every day 20 gm of this

oleioresin was taken in a beaker and to this 20 gm of n-

butyl alcohol was added and dissolved well and kept aside

for two days at room temperature for curcuminoids

precipitation.

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VI.3. Results and discussion

95.05 94.50 o^ 7A „., ^^ 88.90 • • • • _ 90.07 92.00

Effect of different alcohols for the precipitation of curcuminoids

1. Methanol 2. Ethanol 3. isopropanol 4. n-butanol

n Yield • Purity

5. iso butanol 6. sec butanol 7. iso amyl alcohol

95.77 Effect of Time on precipitation

of curcuminoids

1. 2. 3. 4. 5.

12 hr 24 hr 48 hr 72 hr 96 hr

n Yield • Purity

Time in Hours

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T3

s.

93,77

17 69

BB?3:

I Effect of Temperature

on precipitation of curcuminoids

1. 2. 3.

Yield

25°C oo°c 20''C

[Purity

100 , 92.84 94.50 95.10

17 56

Percentage of n-butynol added

Percentage of n-butanol and Oleioresin for the

precipitation of curcuminoids

1.75% 2.100% 3. 150%

Yield Purity

6 8 10 12 14 16

Number of days of ageing

20 22

Effect of Ageing of turmeric oleioresin for

the precipitation of curcuminoids

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From the above observations n-butyl alcohol was

selected as the best solvent for the precipitation on the basis

of yield and purity of curcuminoids as shown in graph 1.

Further, the work was carried out to ascertain the time

required for precipitation and it was found that 48 hrs were

required to get good results (graph 2). Similarly, the

procedure was repeated at different temperatures ranging

from -20°C to room temperature (23±3°C), maximum yield

and purity of the precipitated curcuminoids were found at

room temperature as shown in graph 3. From graph 4, it

was clear that the ratio of oleoresin and n-butyl alcohol

plays a key role in precipitating the curcuminoids; the ratio

(1:1) gave good results.

VI.4. Conclusion

The turmeric spice has been used for many

centuries mainly as a food additive, primarily because of its

golden yellow color. The medicinal properties of this spice

were recognized in Indian folklore medicine and in

Ayurveda, which is an ancient Indian traditional system of

medicine. It was used as a tonic for improving health and in

various combinations for the treatment of diseases such as

common cold. The major break through in realizing the

medicinal value of turmeric came with the isolation of

phenolics called "curcuminoids" , of which curcumin is the

major constituent. Even though a large number of studies

unequivocally identified the numerous pharmaceutical

actions of curcuminoids, its acceptance as a wonder

compound is slowly forthcoming. The objective of the

present investigation was to provide a novel method for

extracting curcuminoids from turmeric rhizome by an

appropriate solvent for obtaining high yield and purity of

curcuminoids.

Health in focus

Turmeric is used in ancient Hindu medicine as a treatment for sprains and swelling. While the therapeutic use of this treasured spice has been commonplace throughout history, emerging medical research has begun to elucidate curcumin's beneficial effects for a range of diseases and conditions. Much of the recent science has focused on its effects against cancer, both therapeutically and prophylactically. Curcumin's potential apparently stems from its ability to suppress the proliferation of a wide variety of tumor cells and to inhibit harmful molecules and enzymes, as well as its antioxidant and antiinflammatory properties. Some studies have even suggested that curcumin can inhibit cancer metastasis.

In research studies, curcumin has shown potential activity against cataract formation, liver injury and the resultant damage from heart attack and stroke. More important, curcumin's antiinflammatory effects and apparent effectiveness in keeping Alzheimer's disease at bay are attracting the notice of more and more medical researchers. [http://chinese-school.netfirms.com/review s/curcumin-alzheimers.html]

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PART

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—- 'i-;!i'rafe4i-s

Analytical studies

The properties of every substance from the air we breathe to the vast range of materials and products which we use in our private lives are directly or indirectly a function of chemical composition.

These properties are influenced for better or worse, by the presence or absence of one or more chemical species, sometimes at concentration levels which are barely detectable. Consider the contamination of a foodstuff with a toxic metal or compound, or the presence of even a few milligrams of asbestos per cubic meter in the air we breathe.

It is obvious therefore,e that chemical composition in many instances decides the value of products and materials which form the basis of national and international trade. As a consequence, it is not surprising that there are more government regulations, standard specifications and trading agreements associated with chemical composition than all other properties put together. It is here that analytical chemistry plays its role.

Analytical chemistry is the science concerned with the systematic identification or characterization of established chemical species and their determination to known degrees of certainty at any level of concentration and in any matrix in which they may occur\www.uUe/~childsD/CinA/cina4/TOC5 Analvtical.htm

http://www.uUe/~childsD/CinA/cina4/TOC5