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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]
Chapter VI Isolation turmeric
CurcuHiinau^s^ grouf of •phei/wUc com-fouMis LsoLated from the roots, of
Curcum-fl LoiA fl (z,Lt^iberaaeae), 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^gei^ic, 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, turm^eric. This chapter is a step in this direction to
im.provise the quantity of curcum.in isolation from. turm.eric roots.
kLiw^ords: Curcuw-lvuiids., Cunuma Lon^a, turm.eruc roots.
Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
[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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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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Chapter VI Isolation turmeric
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2. Anto, R.J., George. J., Babu, K.V., Rajasekharan. K.N. and Kuttan. R. (1996). 3. Antimutagenic and anticarcinogenic activity of natural and synthetic curcuminoids. Mutat. Res.,
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PART
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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