秦　优，陈　林，梁雪娟,3，糜亚男，肖　娟，张水寒,3*（1.湖南中医药大学，湖南长沙41008；.湖南省中医药研究院中药研究所，湖南长沙410013；3.湖南省011数字中医药协同创新中心，湖南长沙41008）



雪峰虫草高效液相色谱指纹图谱研究

秦优1,2，陈林2，梁雪娟2,3，糜亚男1,2，肖娟2，张水寒1,2,3*
（1.湖南中医药大学，湖南长沙410208；2.湖南省中医药研究院中药研究所，湖南长沙410013；3.湖南省2011数字中医药协同创新中心，湖南长沙410208）

〔摘要〕目的建立雪峰虫草药材的高效液相色谱指纹图谱，为评价其质量提供依据。方法采用高效液相色谱法，色谱柱为月旭AQ-C18（4.6 mm×250 mm，5 μm）；检测波长260 nm；以甲醇-0.1%甲酸水溶液为流动相进行梯度洗脱；体积流量0.5 mL/min；柱温30℃；进样量20 μL。结果建立了11批雪峰虫草药材高效液相色谱指纹图谱，确定共有峰15个，采用对照品结合HPLC-Q-TOF-MS/MS技术对7个特征峰进行了指认，并采用相似度分析、主成分分析、聚类分析的方法对指纹图谱进行了分析。结论建立的雪峰虫草高效液相指纹图谱方法简便、可靠且重复性好，为雪峰虫草的质量控制提供了有效的手段。

〔关键词〕雪峰虫草；高效液相色谱；指纹图谱；质量控制；聚类分析度；相似度分析

HPLC Fingerprint of Ophiocordyceps xuefengensis

QIN You1,2, CHEN Lin2, LIANG Xuejuan2,3, MI Yanan1，2, XIAO Juan2, ZHANG Shuihan1,2,3*
（1.Hunan University of Chinese Medicine, Changsha, Hunan 410208, China; 2.Hunan Academy of Chinese Medicine, Changsha, Hunan 410013, China;3.2011 Collaboeation and Innovation Center for Digital Chinese Medicine in Hunan, Changsha, Hunan 410208, China）

〔Abstract〕Objective To establish HPLC fingerprint for Ophiocordyceps xuefengensis and provide conference for quality evaluation. Methods The HPLC method was used with the condition that the column was Yuexu AQ-C18（4.6 mm×250 mm, 5 μm）; The detection wavelength was set at 260 nm; The mobile phase was eluted with gradient by mixture of methanol and water（0.1%formic acid）; The flow rate was 0.5 mL/min; The column temperature was 30℃; The injection was 20 μL. Results The HPLC fingerprint for Ophiocordyceps xuefengensis collected from eleven different sources was set up. Fifteen common peaks werefound, seven of them had been identified by reference substances and HPLC -Q -TOF -MS/MS. The fingerprint was analyzed with methods of cluster analysis, principal component analysis and similarity computation. Conclusion The method of HPLC fingerprint for Ophiocordyceps xuefengensis is simple, rapid and can be used for identification and evaluation of Ophiocordyceps xuefengensis.

〔Keywords〕Ophiocordyceps xuefengensis; HPLC; fingerprints; quality control; cluster analysis; simillarity analysis

雪峰虫草为线虫草菌科真菌雪峰虫草菌Ophiocordyceps xuefengensis sp. nov寄生在蝙蝠蛾科昆虫幼虫上的子座和幼虫尸体的干燥复合体或充满菌丝的干燥幼虫尸体（菌核体），为线虫草菌科线虫草菌属的一个新种，经鉴定是目前同属真菌中与冬虫夏草亲缘关系最近的一个种[1]，因其产于雪峰山脉而得名。主要成分是腺苷、虫草素、D-甘露醇、多糖、氨基酸等[2]。……