GC-MS Analysis of Chemical Constituents of Volatile Oil from Euonymus fortune Produced in Different Areas of Guangxi

Shuang LIANG Senhua LU Jianjun HUANG Xiucai WU Lihong HE

Abstract The chemical constituents of the essential oil of Euonymus fortune from different areas were compared and analyzed by GC-MS. The results showed that 25 chromatographic peaks were separated and 13 chemical constituents were identified in Binyang County， Nanning City， accounting for 97.25% of the total volatile oil. Twenty nine chromatographic peaks were separated from Rong'an County， Liuzhou City， and 14 chemical constituents were identified， accounting for 89.91% of the total volatile oil. Thirty two chromatographic peaks were separated from Mengshan County， Wuzhou City， and 12 chemical constituents were identified， accounting for 92.16% of the total volatile oil. Twenty eight chromatographic peaks were separated from Rong County， Yulin City， and 11 chemical constituents were identified， accounting for 84.79% of the total volatile oil. Five of the chemical constituents of the volatile oil from the three places of origin are common to the four. They are phytol， palmitic acid， n-nonane， cumene， hexahydrofarnesyl acetone. It can be seen that the main components of volatile oil from different producing areas are different.

Key words Euonymus fortune; Volatile oils; GC-MS; Guangxi

Fugangteng is the dry aboveground part of Euonymus fortune， a plant of Celastraceae. It is a commonly used medicinal material for the Zhuang and Yao ethnic groups in Guangxi. It is pungent and neutral in nature and flavor. It has the effects of promoting qi and blood circulation， relaxing meridians and collaterals， nourishing liver and kidney， and stopping bleeding， etc.， and can be used to treat lumbar muscle strain， rheumatic arthralgia， metrorrhagia， hemoptysis， irregular menstruation， traumatic fractures， traumatic bleeding， etc.[1-2]. Clinically， E. fortune can be used to treat neurasthenia， chronic heart failure， chronic prostatitis， chronic pharyngitis， and angina pectoris[3-10]. The main chemical components of E. fortune include flavonoids， pentacyclic triterpenes， sugar alcohols， phenolic acids and steroids[11-18]. E. fortune has hemostatic[19-21]， anti-myocardial hypoxia， cardiovascular and cerebrovascular-protecting[22-26]， anti-aging and immunity-improving[26-27]， myocardial hypoxia-correcting[28-31]， and insecticidal effects[32]. At present， there are few comparative studies on the types and relative contents of the volatile oil of E. fortune from different places. Therefore， in this study， the volatile oil of E. fortune from 4 different producing areas in Binyang County of Nanning City， Rong'an County of Liuzhou City， Mengshan County of Wuzhou City， and Rong County of Yulin City was extracted by steam distillation and determined for composition by GC-MS， and the common chemical components of the volatile oil from E. fortune produced in different areas were analyzed and compared， so as to explore the influence of producing area on the composition and content of the volatile oil of E. fortune. This study provides a theoretical basis for the clinical application， quality control and resource development and utilization of this medicinal material.

Materials and Methods

Materials

The materials were identified as the whole plant of E. fortune of Euonymus in Celastraceae by Professor Teng of Guangxi University of Chinese Medicine. The specific information is shown in Table 1.

Instrument and Reagents

Agilent 7890A-5975C gas chromatography-mass spectrometer （Agilent Technologies Inc.）; DB-1MS quartz capillary column （30 m×0.25 mm×0.25 μm）; volatile oil extractor; ML3002 electronic analytical balance[Mettler-Toledo Instruments （Shanghai） Co.， Ltd].

Anhydrous sodium sulfate （analytical purity， Guangdong Guanghua Technology Co.， Ltd.）; ethyl acetate （chromatographic purity， Sinopharm Chemical Reagent Co.， Ltd.）; pure water.

Methods

Essential oil extraction

A certain amount of E. fortune medicinal powder （150 g） that had been crushed to 20 meshes was weighed， added in a 2 000 ml round-bottomed flask， and added with distilled water 10 times the amount of powder. After soaking the medicinal powder for 1 h， the flask was connected to the volatile oil extractor and reflux condenser， followed by steam distillation according to the method in the appendix of the 2015 edition of Chinese Pharmacopoeia （without xylene） until the distillate contained no oil droplets （about 6 h）， obtaining a light yellow oil with a special fragrance， which was dissolved in ethyl acetate and collected. The distillate was dried with anhydrous sodium sulfate， sealed and stored in a refrigerator for later use.

Sample treatment

A certain amount of the ethyl acetate solution of volatile oil （1 ml） was added in a centrifuge tube， which was then added with an appropriate amount of anhydrous sodium sulfate for dehydration. Centrifugation was next performed in a centrifuge at 1 000 r/min for 20 min， obtaining the supernatant as the test solution.

GC-MS conditions

Chromatographic condition： Quartz capillary column DB-1MS capillary column （30 m×0.25 mm×0.25 μm）; temperature programming： starting with a temperature at 50 ℃， which was held for 1 min and increased at 5 ℃/min to 250 ℃， which was held for 5 min; carrier gas： He （≥99.99%）; column flow rate： 1.0 ml/min; injection port temperature： 250 ℃; split ratio： 20∶1; injection volume： 1 μl; solvent delay： 3.0 min.

MS conditions： EI source; electron energy： 70 eV; ion source temperature： 220 ℃; quadrupole temperature： 150 ℃; mass spectrometer interface temperature： 280 ℃; full scan （Scan） acquisition mode， with a mass range of 50-500 amu and a scan interval of 0.2 s.

Measurement

The samples under "Sample treatment" was diluted with ethyl acetate， subjected to water removal with anhydrous sodium sulfate， and filtered with 0.45 μm microporous membrane. The treated sample was analyzed according to chromatographic conditions under "GC-MS conditions" and determined for the total ion current of E. fortune volatile oil. The chemical component represented by each chromatographic peak was determined by searching through the Nist.08 mass spectrometry data system.

Data processing

The chromatographic and mass spectrum information obtained from the GC-MS analysis results were subjected to automatic search and analysis by the data processing system and its memory library （Nist.08）， and the relative percentage of each chemical component in the volatile oil was determined by the peak area normalization method.

Results and Analysis

GC-MS results

The volatile oil of E. fortune extracted from the 4 producing areas was analyzed by GC-MS according to the above test conditions. After the total ion current was measured， it was searched with the mass spectrum database （Nist 02 and Wiley 275）， and the relative content of each component in each sample was determined by the peak area normalization method. The total ion current chromatograms of the volatile oil are shown in Fig. 1， Fig. 2， Fig. 3 and Fig. 4， and the composition analysis results are shown in Table 2， Table 3， Table 4， and Table 5. The E. fortune produced in Binyang County， Nanning， showed 25 chromatographic peaks separated， from which 13 chemical components were identified， accounting for 97.25% of the total volatile oil. The main components were phytol （74.49%）， palmitic acid （14.29%）， cumene （2.90%）， nonane （1.36%）， etc. The E. fortune produced in Rong'an County， Liuzhou， Guangxi was separated with 29 chromatographic peaks， from which 14 chemical components were identified， accounting for 89.91% of the total volatile oil. Its main components were palmitic acid （44.36%）， phytol （19.98%）， a-linolenic acid （14.38%）， etc. Thirty two chromatographic peaks were isolated from the E. fortune produced in Mengshan County， Wuzhou， Guangxi， and 12 chemical components were identified， accounting for 92.16% of the total volatile oil. The main components were phytol （72.83%）， palmitic acid （7.92%）， cumene （2.89%）， nonane （1.25%）， etc. From the E. fortune produced in Rong County， Yulin， Guangxi， 28 chromatographic peaks were separated， and 11 chemical components were identified， accounting for 84.79% of the total volatile oil. The main components were phytol （53.35%）， palmitic acid （19.31%）， cumene （3.42%）， nonane （1.59%）， etc. Among the chemical components of the volatile oil from the leaves of E. fortune leaves in the four producing areas， there were five common components， namely phytol， palmitic acid， nonane， cumene， and hexahydrofarnesylacetone （Table 6）.

Conclusions and Discussion

The chemical composition of the volatile oil of E. fortune from different producing areas had certain differences in types and contents， which might be caused by environmental differences. The differences in the types and contents of chemical components and characteristic components of E. fortune may affect the pharmacological effects and other aspects. The results of this study provide a scientific basis for the further development and utilization of E. fortune.

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