Extraction and purification of curcuminoids from

Turmeric (curcuma longa L.)

S.J.Kulkarni, K.N. Maske, M.P.Budre & R.P.Mahajan

Dept. Of Biotechnology, Sinhgad college of Engineering, Pune. India.
E-Mail : sitsjk@yahoo.com, kalpanamaske@gmail.com, mona_prithvi@yahoo.com & Reshmamahajan30@gmail.com

Abstract - Turmeric comes from the root of the Curcuma longa plant and has a tough brown skin and a deep orange flesh. Turmeric
has long been used as a powerful anti-inflammatory in both the Chinese and Indian systems of medicine. Turmeric was traditionally
called "Indian saffron" because of its deep yellow-orange color and has been used throughout history as a condiment, healing remedy
and textile dye. Turmeric is rich in curcuminoids. Curcuminoids vary in chemical structures, Physico-chemical characteristics. The
present work reports on extraction method using Soxhlet extractor. Isolation and purification of curcuminoids was carried out by
column chromatography. The quantification of curcumin in maximum resultant extract (by methanol) was performed using pre
validated HPLC methodology. Percentage yield of curcumin by HPLC was 12.39% .extracted curcuminoids were subjected to
spectrophotometer to check it’s percentage amount in extracted sample. Different solvent were used for extraction, among them
methanol showed maximum yield of each curcuminoids. Separation of curcuminoids were tested in TLC chloroform: methanol at
95:5 showed RF value at 0.67, 0.6, 0.506 as curcumin, dimethoxycurcumin,bis demethoxycurcumin respectively. the methanol
extract was subjected to silica gel column chromatography with chloroform: methanol at increasing polarity followed by TLC to
check purity of extracted curcumin.

I. INTRODUCTION The antioxidant property of curcumin can prevent

Turmeric is commonly known for its medicinal
values in the Indian traditional systems of medicine.
Turmeric has been used traditionally in “ayurvedic
medicine” as an antiseptic, wound healing, and anti
inflammatory compounds. Curcumin,
dimethyoxycurcumin and bis demethoxycurcumin is a
dietary photochemical obtained from dried rhizomes of
the turmeric plant (curcuma Longa), Curcumin is a main
coloring substance in Curcuma longa and two related
compounds, demethoxycurcumin (DMC) and
bisdemethoxycurcumin (BDMC), are altogether known
as curcuminoid. The value of the turmeric products is
based on their curcuminoid content. Quantitative
estimation of curcuminoids can be carried out
photometrically based on its absorbance at 420 nm .The
principal colouring components of curcumin exhibit a
keto-enol tautomerism and antioxidative properties.
rancidity of foods and provide foodstuffs containing less
oxidized fat or free radicals. The powerful anti-
oxidation property of curcumin has an important role in
keeping curry for a long time without it turning rancid.
Curcuminoids are poorly soluble in the hydrocarbon
solvents. Curcumin is an oil soluble pigment, practically
insoluble in water at acidic and neutral pH, soluble in
alkali. Preparations of water-soluble curcumin by
incorporation into various surfactant micellar systems
(acetone, methanol, and ethanol) have been reported [1].
It is stable at high temperatures and in acids, but
unstable in alkaline conditions and in the presence of
light.
Curcumin is widely used to colour many foods
[2]. Curcumin is listed for use in dairy products, fats,
oils and fat emulsions, edible ices, fruit and vegetable
products, confectionery, cereal products, bakery wares,
meat and meat products, fish and fish products, eggs and
eggs products, spices, soups, sauces and protein
products, Curcumin, the most active curcuminoid found
in turmeric, has been shown to possess a multitude of
beneficial effects in the treatment of cancers,
cardiovascular disease, and inflammation
[3]. A daily dose of 2 grams of Curcuma domestic
extract was found to provide pain relief that was
equivalent to ibuprofen for the relief of pain associated
with osteoarthritis of the knee[3]. Commercial capsules
of curcumin contain piperine, a compound found in

International Journal of Pharmacology and Pharmaceutical Technology (IJPPT), ISSN: 2277 – 3436, Volume-1, Issue- 2, 2017

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 Extraction and purification of curcuminoids from Turmeric (curcuma longa L.)
pepper which aids absorption of curcumin into the blood
stream. Curcumin might be potentially useful in some
kidney diseases by preventing renal inflammation.[ 12]
The most conventional method for extraction of
curcumin has been Soxhlet extraction with heating time
ranging as long as up to 12 h. the Soxhlet extraction
process is a time consuming, laborious and makes use of
bulk amount of organic solvents. as the heating process
continues for long hours, the approach possibly involves
high risk of thermal decomposition of target molecules
.Soxhlet extraction method operate through cell
permeation followed by solubilizing the active
constituents by the extracting solvent. Curcumin present
inside the oleoresin cells which in turn is covered by
tightly packed cork cells probably makes the entry route
for the solvent and time consuming. A number of
studies are undertaken to separate curcuminoid pigments
by thin layer chromatography (TLC), column
chromatography. HPLC method was sensitive, precise,
and accurate for detection and quantification of
curcuminoids in the extract of rhizomes curcuma longa
II. MATERIALS AND METHODS
Curcuma longa (turmeric) collected from Satara
dist (Rajapuri). All solvents /chemicals used were of
AR/HPLC grade and obtained from SRL. The reference
curcumin purchased from HIGHMEDIA Company in
India.
METHODS
1. Extraction of curcuminoids
Fresh rhizomes were cleaned, washed with
deionsed water, sliced and dried in the sun for one week
and again. Dried at 50˚c in a hot air oven for six hours.
Dried rhizomes were cut in small pieces, powdered by
electronic mill. Six gm of sample were taken into a
thimble and placed in a Soxhlet apparatus, were set up
with various solvent from non polar to polar. 250 ml of
solvent was added and extracted according to their
boiling point for seven hours. The solvents used were
chloroform (B.P. =61˚c).ethyl acetate (B.P. =77˚c),
methanol (B.P. =65˚c) and acetone (B.P. =56.53˚c).
After completion of extraction the dark brown
extract was then cooled, concentrated using rotary
evaporator get a crude dried extract which was black
orange in colour. Each raw sample of turmeric was
extracted by the same method and yield was calculated.
2. Estimation of curcuminoids: by
sphectrophotometric analysis Procedure
i) Preparation of standard: 1mg of pure curcumin
was dissolved into methanol and water such that
concentration was 60µg/ml, got reading at 420 nm. They
are shown in graph no (3). Refer this graph as standard.
International Journal of Pharmacology and Pharmaceutical Technology (IJPPT), ISSN: 2277 – 3436, Volume-1, Issue- 2, 2017
82
ii) Sphectrophotometric analysis :
To find out concentration of extracted sample by
using spectrophotometer, 1 mg of sample were mixed
with methanol and water same as standard solution, OD
was taken at 420 nm. Because all three components of
curcuminoids has λmax at 420 nm.
iii) Separation of curcuminoids by TLC:
Methanol extracts were tested on TLC for presence
different curcuminoids. The TLC pre-coated silica gel.
Plate were developed using glass beaker , which was
pre-saturated with mobile phase for 20 min and each
plate was developed up to a height of about 6.8 cm.
chloroform : methanol mobile phase was used with
composition 95: 5. After development, plates were
removed and dried. Spots were analyzed.
III. COLUMN CHROMATOGRAPHY
A. Sample preparation
Six gram of fine powdered rhizomes were subjected
to Soxhlet extraction and solvent used were methanol
for seven hours. The extract was concentrated in rotary
evaporator .this crude curcuminoids mixture contained
curcumin, demethoxycurcumin and bis
demethoxycurcumin,
B. Silica gel Column chromatography
Methanol extract was subjected to Column
chromatography in silica gel glass column. About 1gm
of crude curcuminoids were mixed with 1 ml methanol
and loaded on to the column (34×1.5cm) and eluted with
chloroform: Methanol followed by methanol with
increasing polarity. All the collected fractions were
subjected to TLC and detected as yellow Spots.
IV. RESULT AND DISCUSSION:
After drying Soxhlet extract were weighted and
weight percentage of curcuminoids were calculated
those are shown in table (no.1). Maximum concentration
of curcuminoids was obtained in methanol extract in the
form of dark black orange colour .
Concentrations of extracted dried sample were
analyzed on spectrophotometer at 420 nm with respect
to standard graph (fig 3). We got regration values such
as 0.998, 0.997, 0.986, and 0.991 for methanol (with
conc. 60µg/ml), acetone (100µg/ml), ethyl acetate
(200µg/ml) and chloroform (100µg/ml) respectively.
Sample was run on TLC (fig.4), we got three
different spots of C, DMC, BDMC. RF values for these
spots were calculated and found to be similar that of
reported values [10] (Table no.2) . According to the RF
values curcumin was analyzed by running standard
 Extraction and purification of curcuminoids from Turmeric (curcuma longa L.)

curcumin along with the sample. The HPLC profile of .Table 1.weight % extract of curcuminoids
extracted crude curcuminoids showed curcumin and its
analogous DMC and BDMC to present in chromatogram Solvent % wt. extract Dry wt. (from 6
(fig 2B.) Spiking in methanol extract was compared to gm turmeric)
that of standard curcumin HPLC profile (fig 2A) Acetone 4.6% 0.28gm
Chloroform 4.3% 0.26gm
Screening of solvent for extraction Methanol 5.6% 0.34gm
Different solvent with varying polarity were used Ethyl acetate 4.5% 0.27gm
for extraction of curcuminoids from turmeric rhizomes
Total weight % extract of curcuminoid was determined
.various solvents used were chloroform , ethyl acetate,
as describe in the text.
methanol , acetone. After concentrating each extract
total yield were determined and percentage yield of Table 2.Separation of curcuminoids by TLC
curcuminoids in the extract were analyzed by
Sr TLC Mobile Ratio RF values
spectrophotometer at 420nm. Extract was also subjected phase
to the TLC and got RF values as 0.67, 0.6, 0.506 for C, no C DMC BDMC
DMC, BDMC respectively. According to this, curcumin 1 Chloroform: 19:1 0.67 0.6 0.506
was found to be the major compound followed by DMC methanol
and BDMC. Hence methanol used as extracting solvent 2 Chloroform: 19:1 0.75 0.55 0.27
for curcumin extraction. methanol
(ref no .10)
Each plate was developed to a height of about 6.8cm
C=curcumin, DMC=demethoxycurcumin,
BDMC=bisdemethoxycurcumin

1 y = 0.128x
R² = 0.998
0.8
Absorbance 420nm)

0.6

Fig 2(A). HPLC profile of standard curcumin (By TUV 0.4

India private limited.) 0.2
0
0 2 4 6 µg/ml 8
[Curcumin]

Fig. 3. Standard calibration curve of curcumin

Figure 2(B): HPLC profile of purified curcuminoids.

Curcumin, Demethoxycurcumin, .and
Bisdemethoxycurcumin Shows single peak at the
retention time of 9.213, 8.533, 7.915 min Respectively.
(By TUV India private limited.)
Fig.4 TLC result of methanol extract

International Journal of Pharmacology and Pharmaceutical Technology (IJPPT), ISSN: 2277 – 3436, Volume-1, Issue- 2, 2017
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 Extraction and purification of curcuminoids from Turmeric (curcuma longa L.)

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