蛴螬脂溶性成分的研究

冯 皓，陈瑞兵，张 磊（第二军医大学药学院药用植物学教研室，上海 200433）

·论著·

蛴螬脂溶性成分的研究

冯 皓，陈瑞兵，张 磊（第二军医大学药学院药用植物学教研室，上海 200433）

目的研究蛴螬提取物中的脂溶性成分。方法采用乙酸乙酯回流提取蛴螬中的脂溶性成分，对其进行气相色谱-质谱法（GC-MS）分析，并对主要脂溶性成分进行定性分析。结果从蛴螬脂溶性成分中鉴定出33个化合物，十五酸、Z-11-十六烯酸、十六酸、十七酸、十八烯酸和十八酸为其主要的脂溶性成分。结论本研究为进一步研究动物药和确认蛴螬的有效成分奠定了基础。

蛴螬；鳃金龟；乙酸乙酯；气相色谱-质谱

1 Introduction

In China，the subfamily Melolonthidae is represented in the literature about 500species［1］.Among them，four species larvae of Holotrichia diomphalia Bates，Holotrichia oblila Fald，Holotrichia sauteri Moser and Holotrichia parallela Motschulsky are traditionally used in China to treat arthrolithiasis，tetanus，erysipelas and superficial infection［2-4］.

The chemical composition of themedicinal animal has few been studied in detail.We reported previously the composition on the petroleum ether extract of H.diomphalia using GC-MS［5］，however，liposoluble constituent extracted from H.diomphalia has not been studied.Through the investigation of H.diomphalia，we discovered that the ethylacetate extract of H.diomphalia contained a host of liposoluble constituent.Thus，the extract was identified by GC-MS and we firstly reported the result on GC-MS determination of liposoluble constituentofethylacetate extractof H.diomphalia.

2 Materials andmethods

2.1 Materials Medicalmaterials of H.diomphalia were purchased from local market and identified by Professor Zhang Hanming，Second M ilitary Medical University，Shanghai，China.All standard compounds were purchased from Sigma（St.Louis，MO，USA）.Ultrapure water was produced by a Milli-Q Reagent Water System（M illipore，MA，USA）.All experimentswere carried out on a Thermo Focus DSQ gas chromatograph and interfaced to a Thermo Focus DSQ mass selective detector.

2.2 Ext ractmethods Dried（50℃）and chopped H.diomphalia powder was refluxed three times（3heach time）with 75%ethanol.After filtration，the clear supernatant was then concentrated at 60℃.Concentrated ethanol extract was partitioned between water and petroleum ether（60-90℃）.After removing petroleum ether fraction，the aqueous layer was partitioned again with acetic ether.The extract of acetic ether was evaporated and the residue was used for the following experiment.

2.3 GC-MS condition GC-MS analysis was carried out on a Thermo Focus DSQ gas chromatograph fitted w ith a fused silica HP-5MS capillary column（30m× 0.25mm×0.25μm）.The oven temperaturewas programmed from 50–300°C at15°C／min.Helium was used asa carrier gasata flow rate of 1.0m l／min.The gas chromatograph was coupled to a Thermo Focus DSQ mass selective detector.The MS operating parameters were：ionization voltage，70eV；ion source temperature，250℃.Identification of components of the petroleum ether extract was based on retention times（Rt）of relative fat acid and computermatching with the NIST98.Library，as well as comparison of the fragmentation patterns of the mass spectra w ith those reported in the literatures［6，7］.

3 Results

In total，as shown in the Table 1，33compounds were identified in ethylacetate extract of H.diomphalia through GC-MS.Pentadecylic acid，Z-11-Hexadecenoic acid，hexadecanoic acid，heptadecanoic acid，oleic acid and octadecanoic acid weremain components of the ethylacetate extract.The retention time（Rt）and relative percentage content（%）of compoundswere listed in Table 1.

4 Discussion

In China，H.diomphalia，asa traditionalmedicinal animal，has been used to treat arthrolithiasis and infection，but the chemical composition and active compounds has notbeen identified and studied.Here，based on the previous research of the composition on the petroleum ether extract of H.diomphalia，this time we focused on the liposoluble constituent using GC-MSand 33compoundswere identified.Thiswork will contribute to the study of active compounds in H.diomphalia andmedicinalanimals.

Table 1 Composition of the ethylacetate extractof H.diomphalia

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［收稿日期］2015-01-23［修回日期］2015-04-02［本文编辑］顾文华

Liposoluble constituent of ethylacetate extract of Holotrichia diomphalia larvae

FENG Hao，CHEN Ruibing，ZHANG Lei（Departmentof Pharmaceutical Botany，Schoolof Pharmacy，Second M ilitary Medical University，Shanghai200433，China）

ObjectiveTo analyze the chemical components of the liposoluble constituent of Holotrichia diomphalia larvae.MethodsEssential liposoluble compoundswere extracted from H.diomphalia through ethylacetate.GC-MSanalysiswas used to identify itsmain components.Results33peakswere separated and 33compoundswere identified from ethylacetate-extracted H.diomphalia.The main compounds were pentadecylic acid，Z-11-Hexadecenoic acid，hexadecanoic acid，heptadecanoic acid，oleic acid and octadecanoic acid.ConclusionThiswork w ill contribute to the study ofmedicinal animals and confirmation of active compounds in H.diom phalia.

Holotrichia diom phalia；Melolonthidae；ethylacetate；GC-MS

R931.74

A

1006-0111（2015）06-0533-03

10.3969／j.issn.1006-0111.2015.06.014

2015-01-29

2015-09-15［本文编辑］李睿旻

冯 皓，博士.研究方向：新药研发.E-mail：fenghao＠sphchina.com

张 磊，硕士生导师.研究方向：药用植物次生代谢工程.Tel：（021）81871307；E-mail：leizhang100＠163.com