82 International Journal of Natural Products Research 2014; 4(3): 82-87

ISSN: 2249-0353

Original Article

Synthesis and characterization of bioactive Curcumin derived from

selected turmeric plants in India

Muhamed haneefa.M1, Jayandran.M

1, Anand.B

1, Balasubramanian.V

2, Muthu mariappan.S

3

1Department of Chemistry,Mahendra Engineering College, Namakkal-637503, India.

2 Dean & HOD, Department of Chemistry, AMET University, Chennai, India.

3Departmentof Chemistry, V.O.Chidambaram College, Tuticorin-628008, India.

1 Corresponding author: Email: honey79101@gmail.com, Tel: +91-9965409669

Email: jayandranchem@gmail.com, Tel: +91-9443899072

Received 22 July 2014; Accepted 08 August 2014

Abstract

This paper reports an investigation of the bioactive "Curcumin" present in the crude plant extracts of 4 selected turmeric

plants i.e. BSR-01, BSR-02, CL-101, CL-219. Curcumin is a significant spice that is extracted from the turmeric rhizomes

(Curcuma longa L). It has several types of biological and pharmacological activities, including anticancer, anti-

inflammatory and antioxidant properties, etc., Based on the various papers and reports about curcumin importance, we

have shown more interest to deal the isolation process of curcumin from turmeric in the easy and fast manner with high

recovery. This investigation was carried out to determine and compare the quantitative amounts of curcumin that are

present in 4 different varieties of turmeric. The extraction of the herb curcumin from turmeric was attempted by using a

"Soxhlet" solvent extraction technique with 95% ethanol as a solvent, then the quantification of curcumin in turmeric was

normally based on spectrophotometric measurement. The presence of curcumin was confirmed by UV-Visible and

elemental analytical techniques. Morphology studies (SEM) and XRD crystal studies were also investigated.

© 2014 Universal Research Publications. All rights reserved

Key words: Turmeric rhizomes, curcuma longa L, curcumin, soxhlet apparatus, solvent extraction

1. Introduction

Turmeric is known as golden spice of India which

comes from the root Curcuma longa L., a member of the

ginger family (Zingaberaceae). It has the chemical structure

(1,7-bis (4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-

dione) and is water insoluble. Its bright yellow pigment is

used as a food coloring agent. Tumeric has been used as a

spice for a food preservative and a dye for a long period.

The medicinal properties of this spice have been slowly

revealing themselves over the centuries [1-4].

Curcumin is known for its antioxidant [5], anti-

inflammatory [6], antiviral, antibacterial [7], antifungal [8],

and cancer chemopreventive actions [9-10]. Curcumin

exhibiting hypolipidemic [11–12] activities and also been

studied extensively as a chemopreventive agent in several

cancers. Additionally it has been suggested that curcumin

may contribute in part to the lower rate of colorectal cancer

in Asian countries compared to rates in other countries [13–

15].

The turmeric rhizome contains a variety of

pigments among which curcumin is a major pigment. The

curcuminoids are polyphenols and are responsible for the

yellow color of turmeric and which has been shown to have

a wide range of therapeutic effects [16]. The polyphenols

are connected by two α,β-unsaturated carbonyl groups. The

two carbonyl groups form a diketone. The diketones form

stable enols or are easily deprotonated and form enolates,

while the α,β-unsaturated carbonyl is a good Michael

acceptor and undergoes nucleophilic addition. Curcumin

can be used for boron quantification in the curcumin

method. It reacts with boric acid forming a red colored

compound, known as rosocyanine [17]. Western scientists

first isolated the curcumin molecule in 1815, obtained its

crystalline form in 1870 and determined its overall

structure in 1910 [18].

Curcumin is a liposoluble compound and can be

easily dissolved into organic solvent such as methanol,

ethanol, and acetone. However, poor water solubility often

limits its biomedical uses using aqueous systems [19].

Curcumin was synthesized from turmeric by various

methods such as solvent extraction, high performance

liquid chromatography (HPLC) and supercritical carbon

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Universal Research Publications. All rights reserved

83 International Journal of Natural Products Research 2014; 4(3): 82-87

dioxide extraction method. The solvents used are acetone,

dichloromethane, 1,2-dichloroethane, methanol, ethanol,

isopropanol and light petroleum (hexanes). The reported

curcumin content percentage has been varied from 3.5 to

9.0% in different commercially available turmeric samples

[20]. There are approximately 30 varieties have been

recognized in the type of curcuma. Several studies have

shown that soil factors, including nutrients and level of

acidity as well as the genus diversity, may affect the

content of curcumin in plants that are the source of turmeric

[21–22].

Based on the above discussions the aim of this

study was designed to extract and quantitate the curcumin

from four different types of turmeric samples to find the

high yield curcumin variety. The synthesis was performed

by using soxhlet extraction method and 95% ethanol was

used as a solvent. Depending upon the amount of curcumin

present in these four turmeric varieties the properties and

functions of curcumin is varied. The turmeric herbs

(samples) were collecteded from various places in

Tamilnadu (India).

2. Materials and methods

The experiment was carried out by solvent

extraction to extract the curcumin from turmeric. Turmeric

samples of four varieties such as CL-101, CL-219, BSR-01,

BSR-02 were obtained from Coimbatore, Salem, Erode and

Madhurai in Tamilnadu respectively. The solvent used 95%

Ethyl alcohol and Acetone were purchased from E.Merck

(India) Ltd. The standard curcumin powder was ordered

from HPLC India. All reagents were of analytical grade and

used as received.

2.1. Synthesis of curcumin

2.1.1. Processing care

One kilogram of fresh turmeric rhizomes from

each plot (comprising 30% mother rhizomes and 70%

primary and secondary rhizomes) were boiled in pure water

for 45-60 minutes till the rhizomes became soft and emitted

a typical turmeric odour . After boiling, the rhizomes were

dried under sun light to attain 8% moisture content. The

recovery of dry turmeric rhizomes then cleaned, crushed

and powdered [23-24]

2.1.2. Plant extraction

In the present work, curcumin was quantitatively

extracted in soxhlet apparatus (invented in 1879 by Franz

von Soxhlet) from turmeric by using 95% ethanol as a

solvent and the curcumin content was estimated as per the

method of Manjunath et al.-1991. The dried turmeric

powder below 300 mesh (IS- 2446, 1963) were taken in a

soxlet apparatus at the rate of 5 g was refluxed with 250 ml

of 95% ethanol for 2 hours and 30 minutes. The extract was

cooled and filtered quantitatively into a 100 ml volumetric

flask; the residue was then transferred to the filter, washed

thoroughly and volume was made up to 100 ml with Ethyl

alcohol. 5 ml of this filtered extract was pipetted out into a

100 ml volumetric flask and volume made up using

ethanol. It was mixed well and the absorbance of this

solution was measured at 425 nm against alcohol blank.

From the absorbance value the curcumin percentage was

calculated. The above ethanolic residual extract was

evaporated and dried then recrystalized by 95% ethanol

[25-26]. This was used for further analyses.

2.2.3. Standard solution

The 925 mg of standard curcumin powder was

taken in a 100 ml volumetric flask which was dissolved in

alcohol after adding small quantity of acetone and the

volume was made up to 100 ml. Again 1 ml of this solution

was transferred into a 100 ml volumetric flask and volume

was made up with alcohol. This standard solution

(containing 0.0025g/1ml) was read at 425 nm against

alcohol blank in spectrophotometer and the curcumin

content obtained by this method was determined and

expressed as percent.

2.2. Characterization

The UV-Visible absorption spectra of the samples were

measured on a Shimadzu UV-Vis V-530A

spectrophotometer in the range of 425nm. Elemental

analyses were carried out with Elementar Vario EL III

series used to collect the micro analytical data (C, H and N)

and compared with the calculated theoritical values.

An X-ray measurement of the prepared solids

were carried out using a Panalytical X’Pert Powder

X’Celerator Diffractometer (Netherland). The patterns were

run with Cu Kα radiation at 40 kV and 30 mA with

scanning speed in the range of 10 ο

to 80 ο

2θ of 2ο min

-1.

The crystallite size of crystalline phases in the

investigated solid was based on X-ray diffraction line

broadening and calculated by using Scherrer equation.

where d is the average crystallite size of the phase under

investigation, B is the Scherrer constant (0.89), λ is the

wavelength of X-ray beam used, β is the full-width half

maximim (FWHM) of diffraction and θ is the Bragg's

angle.

Scanning electron microscopy (SEM) images were

recorded by using a JEOL Model JSM - 6390LV scanning

electron microscope equipped with an energy-dispersive

spectrum (EDS) capability.

3. Results and Discussion

3.1. UV-vis spectrum of curcumin

One of the most convenient techniques for

charecterization of curcumin compound is UV-vis

spectroscopy. Curcumin was quantitatively extracted by

refluxing the material in alcohol and was estimated

spectrometrically using Shimadzu UV-Vis V-530A

spectrophotometer in the range of 300-600 nm wavelength.

The well established curcumin structure is represented as

follows (Figure 1). Curcumin exhibits strong broad

absorption peak at around 425 nm. This can be due either to

an n→π* transition or to a combination of π→π* and

n→π* transitions. Therefore curcumin content was

estimated spectrophotometrically in the range of around

425 nm for all turmeric extracts (Figures.1a-1d).

Figure. 1. Structure of curcumin

84 International Journal of Natural Products Research 2014; 4(3): 82-87

Figure. 1 a. UV-vis spectrum of CL-101 curcumin

Figure. 1 b. UV-vis spectrum of CL-219 curcumin

Figure. 1 c. UV-vis spectrum of BSR-01 curcumin

Figure. 1 d. UV-vis spectrum of BSR-02 curcumin

Table 1. Turmeric varieties and its curcumin amount.

S.No Turmeric variety

(curcumin) Weight (g) Absorbance Curcumin yield (%)

1 CL-101 5.014 2.4839 8.59%

2 CL-219 5.008 1.1448 4.25%

3 BSR-01 5.005 2.3128 9.24%

4 BSR-02 5.007 1.2594 4.68%

3.1.1. Estimation of curcumin content In this work four type of turmeric varieties CL-

101, CL-219, BSR-01, BSR-02 were used to determine the

content of curcumin. The extracted Curcumin was

quantitatively estimated spectrophotometrically and it

wasconverted into percentage of curcumin and expressed

by using the following formula. The concentration of

curcumin in turmeric samples were determined in g/100 g

converted into percentage. The results obtained were

tabulated for further analysis (Table 1).

After drying soxhlet extract were weighed and

weight percentage of curcumin were calculated those are

shown in table.1. Maximum concentration of curcumin was

obtained in ethanol extract in the form of dark black orange

colour. From the result, the percentage has been estimated

to be between 4.25% and 9.24% in these 4 different

turmeric samples. Curcumin concentrations of both BSR-

01 and CL-101 extracts reached around 9% (w/w),

significantly higher than those in other turmeric extracts

(Table 1).

Since among the four varieties of turmeric

samples found that the two turmeric varieties, BSR1 and

CL101 having the highest percentage (9.24% and 8.59%)

of curcumin which were collected from erode and covai

respectively. From this experiment we have come to the

conclusion that the soxlet extraction method by using 95%

ethanol was the most efficient in extracting curcumin from

the turmeric rhizomes into extracts.

3.2. Elemental analysis

The recrystallized powdered curcumin were stable at room

-0.1

2.5

1

2

300 600400 500

Abs

Wavelength [nm]

-0.1

1.2

0

0.5

1

300 600400 500

Abs

Wavelength [nm]

-0.1

2.4

1

2

300 600400 500

Abs

Wavelength [nm]

0

1.3

0.5

1

300 600400 500

Abs

Wavelength [nm]

85 International Journal of Natural Products Research 2014; 4(3): 82-87

Table 2. Elemental analysis data of Curcumin

Curcumin Experimental value Theoritical value

C H N C H N

C21H20O6 69.43 5.20 - 68.47 5.47 -

Table 3. Average crystallite size of Curcumin varieties

Turmeric variety (Curcumin) Average size, K= 0.9λ/βcosθ

CL-101 57 nm

CL-219 70 nm

BSR-01 40 nm

BSR-02 63 nm

temperature and are non-hygroscopic. The analytical data

of curcumin was obtained from a Elementar Vario EL III

series analyzer as in the table 2. The molecular formula of

curcumin C21H20O6. There is no disagreement between the

theoritical and experimental values.

3.3. XRD analysis

Crystalline compounds give characteristic X-ray

diffractogram. The crystallite size of material, packing and

morphology tested using XRD spectrometer with Cu source

on the basis of powder diffraction method. Quantitative

analysis of Xray powder diffraction technique is a

measurement of a series of d spacing, the interplanar

spacings from the position of the diffraction peaks. The

diffraction angle is a recorded in terms of 2θ and all 2θ

values are readily converted to d-values expressed in

angstroms units for a given wave length of X rays. The

sample was rotated during the data collection to reduce

orientation effects, and the data was recorded using a

curved photosensitive detector. The X ray was measured in

the range of 2θ=10 to 80ο at steps of (100) at ambient

temperature. The crystal structures of Curcumin for four

turmeric varieties were shown in the Figure 2a-2d.

Figure. 2 a. XRD pattern of CL-101 curcumin

Figure. 2 b. XRD pattern of CL-219 curcumin

Figure. 2 c. XRD pattern of BSR-01 Curcumin

Figure. 2 d. XRD pattern of BSR-02 Curcumin

X ray diffraction studies of curcumin were

investigated from the angle of 10 0 to 80

0. The intensity vs

angle (2θ in degrees) was plotted which showed the

decrease in intensities and broadened slightly while moving

further in all curcumin varieties due to size effect. The

crystallite size for those above 4 varieties of turmeric

curcumin was determined by Scherrer formula. The

average crystallite size obtained is as in the table 3.

3.4. SEM analysis Morphology of synthesized curcumin for all the

four turmeric varieties were charaterized by SEM analysis.

The samples were placed in an evacuated chamber and

scanned in a controlled pattern by an electron beam.

Interaction of the electron beam with the specimen

produces a variety of physical phenomenon that detected,

were used to form images and provide information about

the specimens. The SEM images of CL-101, CL-219, BSR-

Position [°2Theta] (Copper (Cu))

20 30 40 50 60 70

Counts

0

500

1000

CL101-C

Position [°2Theta] (Copper (Cu))

20 30 40 50 60 70

Counts

0

500

1000

1500

CL219-C

Position [°2Theta] (Copper (Cu))

20 30 40 50 60 70

Counts

0

200

400

600 BSRI-C

Position [°2Theta] (Copper (Cu))

20 30 40 50 60 70

Counts

0

200

400

600

BSRII-C

86 International Journal of Natural Products Research 2014; 4(3): 82-87

Figure. 3 a. SEM image of CL-101 curcumin

Figure. 3 b. SEM image of CL-219 curcumin

Figure. 3 c. SEM image of BSR-01 curcumin

Figure. 3 d. SEM image of BSR-02 curcumin

01 and BSR-02 are as follows (Figure 3a-3d). SEM images

of those compounds had shown the cubic, spherical and

some elongated morphology of material.

4. Conclusion

It is observed that all varieties are statistically

significant from each other in respect of curcumin

extraction. Among the four turmeric varieties CL-101

collected from Erode (92%) is statistically superior over all.

The variety CL-219 collected fom Salem (41%) is

statistically inferior in curcumin extraction. Based on the

reported results, it may be concluded that to extract

maximum curcumin percentage it is strongly recommended

that it is better to go for BSR-01 variety (Erode) and then

CL-101 variety (Covai) by using soxhlet extraction method

with 95% ethanol solvent. This result will be very much

useful in future researches such as pharmacokinetic studies

and chemoprevention investigations related with the

curcumin.

Acknowledgements

We gratefully acknowledge Department of

Physics, Manonmaniam Sundaranar University,

Tirunelveli, India for providing XRD analysis facilityand

Department of Chemistry, V.O.Chidambaram College,

Tuticorin, India for providing UV analysis facility. We also

87 International Journal of Natural Products Research 2014; 4(3): 82-87

thank SAIF, Cochin for SEM analysis and Elemental

analytical facilities. Moreover we are thankful to Mahendra

Engineering College and AMET University for their

support to do this work.

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Source of support: Nil; Conflict of interest: None declared