冬虫夏草水提物HPLC指纹图谱及模式识别分析

HPLC Fingerprint and Pattern Recognition Analysis of Aqueous Extracts from Ophiocordyceps sinensis

目的:通过建立不同产地冬虫夏草的高效液相色谱(HPLC)指纹图谱,评价不同产地冬虫夏草质量差异,比较冬虫夏草与冬虫夏草菌丝体质量差异,为冬虫夏草药材的质量鉴别提供科学依据。方法:采用HPLC法,以甲醇-0.3%乙酸溶液为流动相进行梯度洗脱,检测波长260 nm,对10批冬虫夏草和9批冬虫夏草菌丝体进行指纹图谱研究。通过相似度评价结合主成分分析、聚类分析对冬虫夏草指纹图谱进行综合评价。结果:建立了冬虫夏草HPLC指纹图谱,标定18个共有峰,指认出其中5个共有峰,分别为胞苷、尿苷、肌苷、鸟苷和腺苷;方法的精密度、稳定性和重复性的峰面积RSD分别小于2.99%、3.04%和3.16%,符合指纹图谱分析要求。10批冬虫夏草相似度均大于0.92;冬虫夏草与冬虫夏草菌丝体相似度为0.563～0.693,通过指纹图谱特征可实现有效区分,表明二者化学成分存在较大差异。主成分分析和聚类分析将样品分为3类,冬虫夏草单独为1类,该结果与相似度分析结果基本一致。结论:建立的HPLC图谱结合主成分分析和聚类分析,可以快速进行冬虫夏草的真伪鉴别,为冬虫夏草质量控制提供理论参考。

Objective： HPLC fingerprints of Ophiocordyceps sinensis from different areas were established to provide a scientific basis for quality control of O.sinensis and cultivated mycelium.Method： Chromatographic fingerprints of 10 batches of O.sinensis and 9 batches of cultivated mycelium were established by HPLC with 0.3% acetic acid and methanol as mobile phase for gradient elution; the detection wavelength was set at 260 nm. The fingerprints were analyzed by using similarity evaluation,principal component analysis（ PCA） and cluster analysis（ CA）. Results： The reference HPLC fingerprint of O. sinensis was established with 18 common peaks,and 5 of common peaks were identified,including cytidine,uridine,inosine,guanosine and adenosine.The peak area RSD of precision,stability and repeatability of the method were less than 2.99%,3.04% and 3.16%,respectively,which conformed to the requirements of fingerprint test. The similarity of 10 batches O. sinensis was greater than 0.92. There were some differences between O.sinensis and cultivated mycelium,with the similarity value from 0.563 to 0.693.The results of PCA and CA were consistent with similarity analysis,which all samples were classified into 3 groups and 10 batches of O.sinensis belonged to 1 group.Conclusion： The HPLC fingerprint combined with PCA and CA methods could be quickly used to identify genuine O.sinensis,and provide scientific basis to evaluate the quality of O.sinensis.