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Xhmka PACTHTE^tHoro CMPba. 2010. №1. C. 161-163.
UDC 547.913
INVESTIGATION OF KAZAKHSTANI FLORA. I. GC/MS ANALYSIS OF THYMUS MARSCHALLIANUS WILLD ESSENTIAL OIL OBTAINED BY SUPERCRITICAL CO2 EXTRACTION
© E.M. Suleimenov1, S. Machmudah2, M.Yu. Ishmuratova3, M. Sasaki2, M. Goto2
department of Chemistry, Munaitpassov 5, L.N. Gumilyov Eurasian National University, Astana, 010008 (Kazakhstan). E-mail: [email protected] 2Department of Applied Chemistry and Biochemistry, Kurokami 2-39-1, Kumamoto University, Kumamoto, 860-8555 (Japan)
3Zhezkazgan Botanic Garden, Zhezkazgan, 100600 (Kazakhstan)
The composition of extracts of Thymus marschallianus Willd (Labiatae) from two regions of Kazakhstan, obtained by supercritical C02 extraction was investigated by GC/MS.
Keywords: Thymus marschallianus (Labiatae), supercritical fluid extraction (SCFE), GC/MS
Acknowledgments - the authors thank the Matsumae International Foundation (Japan) for a research grant. Introduction
T. marschallianus Willd. (Labiatae) is quite spread in an average strip of Kazakhstan, and its essential oil represents practical interest as a source of preparations of antibacterial [1] and antioxidative actions [2-3].
Chemical compound of T. marschallianus was studied insufficiently. A phenolic glycosides [4], rosmarinic acid [5] and essential oil were found in T. marschallianus. Essential oil composition was investigated by GC from various regions of the world and are quite different [6-12]. So, for example, in the essential oil from Hungary the basic component is thymol 32,99% [6]. In oil from Russia the basic components are borneol - 30,0, a-pinene - 10,7 and sabinene -9,9%, while phenolic compounds are - p-cymene - 8,1, carvacrole - 3,1, thymol - 5,5% [7]. In essential oil from Kazakhstan the main components are p-cymene, thymol and y-terpinene, with the small maintenance of p-bisabolene, a-pinene and terpineole [8]. In essential oil of the thyme growing on Caucasus geraniol was found as main component -30,28% [9]. Detailed research of essential oil of T. marschallianus from China obtained by water distillation was carried out by GC/MS [10]. Thymol - 32,87, y-terpinene - 22,41, carvol - 8,02 and p-cymene - 7,72% were found as main components of this type of essential oil of T. marschallianus. It is necessary to note, that research of essential oil of T. marschallianus obtained by method C02 extraction haven’t been investigated before except of T. serpyllum [11].
Experimental part
Plant material. Plants were collected by authors in Zhezkazgan Botanic (A) garden and in vicinities of Astana (B) (Kazakhstan Republic) and deposited in Zhezkazgan Botanic garden (Voucher code is 2008.07.06.01.04).
SCFE. Cooler system - Iwaki Asahi Glass cooling unit CLU-33; Heater of extractor - Eyela WF0-400; Pump -Jasco PU-1586 Intelligent Prep. Pump; Automatic Back Pressure regulator - Jasco BP-2080 plus; Gas meter - Wet gas Meter Sinagawa, model W-NK-1A; Extractor - Thar designs, Inc. USA, Vessel 25 mL V. Supercritical extraction was carried out at a pressure of 15 MPa, flow rate of C02 - 3 ml/min and temperature of extractor at 40 °C (Figure).
* Author for correspondence.
We collected extracts within an hour. The yield of essential oil is from 2,26 to 1,55% for raw material A and B respectively. The obtained extracts were dissolved in 1 ml of ethanol, cooled overnight at - 20 °C, filtered and analyzed by
gc/ms.
GC/MS. Component composition of essential oil was analyzed by the method of GC/MS using a Hewlett Packard GCD system. An HP-5 column (60 m x 0,25 mm id.) was used with 1 mL/min Helium as carrier gas. GC oven temperature was held at 60 °C for 10 min and programmed to 220 °C at a rate of 4 °C/min, and then held isothermal for 10 min. Split flow was adjusted at 50 mL/min. The injector temperature was at 250 °C. Mass spectra were recorded at 70 eV. Mass range was m/z 35-425. Percentage values of each compound were automatically calculated from peak areas of the total ion chromatogram. Library search was carried out using Wiley GC/MS Library. Composition of CO2 extract of T. marschallianus given in the Table.
Scheme of supercritical CO2 extraction
Composition of CO2 extract of Thymus marschallianus
RT Library Area% RT Library Area%
A B A B
4,86 a-Thujene** Tr - 22,77 p-Cymen-7-ol Tr -
5,07 a-Pinene Tr - 23,03 Thymol 42,47 70,91
5,58 Camphene Tr - 23,29 Carvacrol 2,28 -
6,71 P-Pinene Tr - 24,89 Limonene 13,17 -
7,50 Sabinene Tr - 25,70 Copaene Tr -
8,87 o-Cymene Tr - 27,13 Caryophyllene 0,92 2,65
9,39 p-Cymene*** 15,71 6,39 27,61 Methyl a-hydroxybutyrate 0,48 -
9,76 1,8-Cineole 0,85 - 27,76 Aromadendrene Tr -
11,71 y-Terpinen 1,32 4,78 29,14 y-Muurolene Tr -
12,21 3-Carene 0,62 - 29,79 P-Selinene Tr -
13,54 p-Cymenene Tr - 30,08 P-Bisabolene 2,72 3,98
16,29 Camphor Tr - 30,50 P-Cadinene Tr Tr
17,38 Camphene 3,21 - 31,03 Carveol Tr -
17,95 a-Phellandrene 0,83 - 31,79 Carvone Tr -
18,41 3,4-Dimethylstyrene 0,50 - 32,06 Unknown 1**** 0,70 -
18,61 m-Mentha-6,8-diene 7,00 - 32,20 Spathulenol 0,86 -
20,58 1-Isopropyl-2-methoxy-4- methylbenzene 0,52 - 34,84 Guaiazulene+Cadalin Tr -
21,13 Ocimene 5,45 - 42,22 Dibutyl phthalate Tr -
22,52 m-Mentha-1,8-diene Tr - 56,62 Diisooctyl ester of phthalic acid - 11,29
TOTAL 99,61 99,99
A - Essential oil from T. marschallianus from raw material collected in Zhezkazgan (Karaganda region). B - Essential oil from T. marschallianus from raw material collected in vicinities of Astana. ** - Tr- components less then 0.1%; *** - basic components in bold; **** - Unknown 1: EIMS, 70 eV, m/z (% rel. int.): 256(3), 235(1), 220(1), 202(40), 187(32), 177(11), 165(6), 159(100), 145(46), 131(75), 117(44), 105(72), 91(81), 79(48), 67(24), 55(27).
Results and discussion
As shown in table, in the essential oil of T. marschallianus from the raw materials collected in Zhezkazgan 38 components were found, main of them are thymol - 42,47, p-cymene - 15,71, limonene - 13,17, m-mentha-6,8-diene -7,00 and camphene - 3,21%, and among 7 found out components from raw materials collected in vicinities of Astana the main components are thymol - 70,91, diisooctyl ester of phthalic acid - 11,29, p-cymene - 6,39, y-terpinen -4,78% and p-bisabolene - 3,98%. It is necessary to note the high maintenance of thymol in the last essential oil.
Conclusions
Thus, for the first time we investigated the component composition of essential oil of T. marschallianus (Labi-atae) from two regions of Kazakhstan obtained by supercritical C02 extraction method.
References
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Received July 21, 2009
Revised November 9, 2009
