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Extraction and isolation of turmerone from turmeric

Article  in  Electronic Journal of Environmental, Agricultural and Food Chemistry · August 2013

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ISSN: 1579-4377

EXTRACTION AND ISOLATION OF TURMERONE FROM TURMERIC

Surwase V.S.a, Laddha K.S.a, Kale R.V.b, Syed Imran Hashmi*c and Lokhande S.M.b
a
Pharmaceutical Sciences and Technology Division, Institute of Chemical Technology, University of Mumbai,
Matunga (E), Mumbai-400019 (MS) India
b
Assistant Professor, Food Technology Division, Department of Technology, Shivaj University, Kolhapur-416004
(MS) India
c
Research Associate, Department of Food Trade & Business Management, College of Food Technology, M.A.U.,
Parbhani (MS) India
*imran.foodresearch@gmail.com

ABSTRACT

Turmerone is principle flavouring compound of turmeric (Curcuma longa L.). The objective of
the research work was to isolate turmerone from turmeric oil and its characterization. Turmerone
was extracted from turmeric oil. It was further purified with activated charcoal or preparative
TLC. Turmerone shows violet spot at Rf of 0.72 with vanillin-sulfuric acid on heating. A UV
spectrum of the isolated compound shows two peaks of almost same intensity at 233.5 nm and
236 nm. IR spectra values in cm-1 were found to be 2988.7 and 2936.8 (for aromatic C=C
stretching), 1735.4 and 1446.2 (for C=O Stretching), 939.0, 847.4 (for –CH bending). GC
spectra of isolated compound shows the first peaks at retention time of 7.227 min. with area
99.2% and second peak at retention time of 9.667 min. with area 0.8%. GC-MS spectra of the
isolated compound in positive ionization mode showed molecular ion peaks at m/z: 217.2 and
219.2 which correspond to molecular weight of ar-turmerone and turmerone.

KEYWORDS
Turmerone, isolation, characterization, Turmeric.
Hashmi, S. I. et al. EJEAFChe, 10(5), 2011 [2173-2179]
 

INTRODUCTION

Variety of chemical constituents occurs widely in nature. Volatile constituent from the plant
sources plays important role in the various fields like fragrance, food, pharmaceutical and
aromatherapy (Jae et al., 2006). Turmeric (Curcuma longa L.) is an important tropical spice
commercially traded for its aroma and colouring properties. It forms an essential ingredient of
curry powders and is extensively used in traditional medicine as a carminative, stomachic,
anthelmintic, laxative and as a cure for liver disorders (Neena, 2009). Active compounds in
turmeric are typically classified as non-volatile or volatile. Major non-volatile curcuminoids are
curcumin, demethoxycurcumin and bisdemethoxycurcumin (Hastak et al., 1993). The turmeric
volatile oil is yellowish, stiff and commonly evolves a few slightly aromatic flavors (Paranee et
al., 2009). The major compounds identified in turmeric volatile oil are ar-turmerone, turmerone,
ar-curcumene, zingiberene, α-phellandrene, curlone, 1, 8-cineol and some other sesquiterpenes.
Turmerones are sesquiterpenoid cyclic ketones accounts for 40-50% of volatile oil
(Govindarajan and Stahl, 1980). Turmerones are the major constituents present in oil and possess
many biological activities such as antivenom activity, antiplatelet property, anticancer agent,
anti-oxidant activity etc. Turmerone and ar-turmerone are known to be the character impact
compounds of turmeric contributing to the dry turmeric aroma. These compounds together with
1:8 cineol that imparts a camphory note has been reported to be responsible for the top note of
the spice. Turmerone is used in flavour and fragrance concentrates where spicy-woody note is
required (Khanna, 1999). Therefore considering the use of turmeric oil and turmerone, the
research work was undertaken to study the properties and isolate constituent from it. The
methodology adapted for the same include collection and identification of raw material,
standardization of raw material, extraction and isolation of turmerone from turmeric oil and
characterization of isolated turmerone by TLC, UV-spectroscopy, GC, IR, and GC-MS.

MATERIALS AND METHODS

Collection and identification: Turmeric powder was purchased from local market, Mumbai. It  
was identified by physicochemical and sensory characteristics.
Standardization: Standardization of turmeric was done on the basis of its physico-chemical
analysis. Moisture, protein, crude fibre and ash content of turmeric powder was determined by
standard method (AOAC, 1995), carbohydrate and crude fiber content was determined by
methods illustrated by Sadasivam and Manuckam (1996). While water and alcohol soluble
extractives and volatile oil was determined by method described in Indian Pharmacopeia (2007).
Extraction and isolation of turmerone: 1kg of turmeric powder (Curcuma longa L.) was
extracted with 4 l of petroleum ether (b.p.60-80oC) using soxhlet assembly for 12 h. The extract
obtained was then concentrated by using distillation. The solid material from the soxhlet
assembly was extracted twice more with petroleum ether and the concentrated filtrates or
extracts were combined. This extract was nothing but turmeric oil. Turmeric oil was then
fractioned between petroleum ether and methanol using separating funnel. Repeated
fractionation was carried out with methanol. Petroleum ether fraction was then treated with
activated charcoal to remove any impurities present. This fraction was further purified by
preparative TLC. Thus, repeated fractionation and charcoal treatment resulted in isolation of pure
turmerone.

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Hashmi, S. I. et al. EJEAFChe, 10(5), 2011 [2173-2179]
 

Extraction method
Extraction of turmeric powder with petroleum ether using soxhlet assembly (12h)

Concentration of extract by distillation

Turmeric oil

Fractionation of turmeric oil with petroleum ether and methanol

Repeated fractionation with methanol result in isolation of turmerone

It was further purified by activated charcoal and preparative TLC


Chemical evaluation of isolated oil viz. saponification value, acid value, ester value, peroxide
value and acetyl value were carried out and test conducted by Standard methods (Anon, 2007).
Physical properties: Physical properties extractive value, refractive index, optical rotation,
melting point, viscosity, specific gravity, density etc. are determined by standard methods
(AOAC, 1995).
Characterization of Isolated Compound
Think Layer Chromatography of volatile oil of turmeric and isolated compounds was performed
by the Ascending technique. The process parameters includes Silica gel 60F254 pre-coated TLC
plate (Merck) as Adsorbent, Toluene: Ethyl acetate (93:7) as Chromatography solvent. Vanillin-
sulphuric acid was used as spraying agent after the length 5.0 cm length.
Gas Chromatographic analysis of Turmeric oil and its column fractions we prepared with
chloroform solvent and were analyzed using Chemito Ceres 800 Plus chromatography equipped
with FID detector, using OV-17 packed column (3.0m x 18") using nitrogen as carrier gas with
the carrier gas flow rate of 3.6 bars. Injector port temperature was 280oC; detector temperature
was 290oC with the holding time of 20 min at 290oC.
UV/visible spectrum were obtained in ethanol on Jasco V-530 UV/visible
spectrophotometer and Mass spectrum was recorded on Micromass, Q-TOF MS ES+.
GC-MS analysis turmeric oil and its column fractions were analyzed using a Shimadzu
17A-GC chromatograph equipped with a QP-5000 (Quadrapole) Mass Spectrometer. The sample
was diluted 25 times with chloroform and 1 µl was injected. A fused silica column SPBTM-1 (30
m x 0.32 mm film thickness 0.25 µm) coated with polydimethylsiloxane was used. Injector port
temperature was 280oC; detector temperature was 290oC and oven temperature was maintained
at 60oC for 3 min and then increased to 290oC 2oC/ min at which temperature of the column was
maintained for 5 min; helium was the carrier gas at a flow rate of 1 ml/min; split ratio was 1:25;
ionization voltage, 70 eV.

RESULTS AND DISCUSSION

Physicochemical properties of turmeric powder: The turmeric powder before isolation of


bioactive compound was subjected to physico-chemical analysis. The obtained results are
depicted in Table-1. The results revealed that powder was sufficiently dried with the lower
moisture content of 4.37 per cent. The values for protein, carbohydrates, crude fibre and ash

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Hashmi, S. I. et al. EJEAFChe, 10(5), 2011 [2173-2179]
 

content were comparable with earlier reports (Kapoor, 1990). The water extractive of turmeric
powder found to be higher that alcohol extract.
Table-1: Physical evaluation of Turmeric Powder
Sr. No. Tests Results
1 Moisture content 4.37%
2 Carbohydrate content 51.29%
3 Protein content 10.65%
4 Volatile oil content in turmeric powder 4.5%
5 Crude fibre content 2.5%
6 Ash value 5.26%
7 Extractive value Water soluble extractive 12.40%
Alcohol soluble extractive 4.60%

Chemical properties of turmeric oil: Quality of oil in terms of chemical properties like saponification
value, acid value, ester value, acetyl value, peroxide value, refractive index and specific gravity were
determined as depicted in Table-2. The results revealed that the chemical properties of oil extracted by
using ether are comparable with other commercially available turmeric oil.

Table-2: Chemical evaluation of turmeric oil


Sr. No. Tests Results
1 Volatile oil content in turmeric oil 16.8%
2 Specific gravity 0.92342
2 Refractive index of 1.5154-1.5160
3 Saponification value 36.465
4 Acid value 3.4782
5 Ester value 32.9868
6 Acetyl value 26.87
7 Peroxide value 4.2851
Characterization of isolated turmerone by TLC, UV, IR, GC and GC-MS: Characterization of isolated
compound was carried out with the analytical techniques like TLC, UV, IR, GC and GC-MS.
Table-3: Characterization data of isolated compound
Sr. No. Parameters Observations
1 Colour Light yellow
2 Odour Dry, woody, spicy
3 TLC RF value 0.72
4 UV λmax (ethanol) 233.5 nm and 236 nm
5 Infrared spectroscopy (cm-1) 2988.7, 2936.8, 1735.4, 1446.2, 1374.0, 939.0, 847.4
6 Mass spectrometry m/z: 217.2 and 219.2 [M+1]
 

Fig-1: TLC of Turmeric oil and isolated compound

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Hashmi, S. I. et al. EJEAFChe, 10(5), 2011 [2173-2179]
 

Thin layer chromatography is an important tool for quick identification of required constituents in
drug or in extract. The thin layer chromatography (TLC) was performed for presence of turmerone in
turmeric oil. Mobile phase comprising of toluene:ethyl acetate (93:7). Comparative TLC of turmeric oil
and turmerone was done. Turmerone shows violet spot at Rf of 0.72 with vanillin-sulfuric acid on
heating. This spot matched exactly with the reported Rf in same mobile phase.
The UV absorption maxima (λ max) of the isolated compound were recorded using ethanol as
solvent. A UV spectrum of the isolated compound shows two peaks of almost same intensity at 233.5 nm
and 236 nm.
IR spectra values in cm-1 were found to be 2988.7 and 2936.8 (for aromatic C=C stretching),
1735.4 and 1446.2 (for C=O Stretching), 939.0, 847.4 (for –CH bending).
Gas chromatography was carried out using OV-17 packed column with FID detector. The sample
was diluted using chloroform and 10 µl was injected. The flow of N2 gas was maintained at 3.6 bars.
Injector temperature was kept at 2800C and at detector temperature 2900C it was hold for 200C.There was
rise in temperature from 1000C to 2900C with 250C/min. GC spectra of isolated compound shows the first
peaks at retention time of 7.227 min. with area 99.2% and second peak at retention time of 9.667 min.
with area 0.8% (Table-4).

Table-4: Gas chromatographic studies of turmeric isolated compound


Compound GC Studies
Retention time (min) Area (%) Height (%)
Turmerone 7.227 99.2 99.4
Ar-turmerone 9.677 00.8 00.6

GC-MS spectra of the isolated compound in positive ionization mode showed molecular ion
peaks at m/z: 217.2 and 219.2 which correspond to molecular weight of ar-turmerone and turmerone. On
the basis of the data obtained and the reported values, the isolated compound was identified and
confirmed to be turmerone. The purity of the isolated compound was determined by GC.

Fig-2: UV-spectra of isolated compound Fig-3: IR spectra of isolated compound

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Hashmi, S. I. et al. EJEAFChe, 10(5), 2011 [2173-2179]
 

Fig-4: GC-MS Mass spectra of isolated compound

Fig-5: Peak fragmentation of isolated compound


CONCLUSION

Thus based on the results of the tests carried out and from spectral studies, the observed data was
found to match well with that of reported data for turmerone and the isolated compound was
identified as turmerone.

REFERENCES
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