HPLC指纹图谱在鉴别冬虫夏草上的应用

Identification of Cordyceps sinensis by HPLC Fingerprint

[目的]建立冬虫夏草HPLC指纹图谱,为其真伪鉴别提供有效方法,并定量测定其6种共有核苷类成分(尿嘧啶、尿苷、胸腺嘧啶、腺嘌呤、腺苷、虫草素)的质量。[方法]采用HPLC,色谱柱Waters Symmetry RP-C18(250×4.6 mm,5μm),以乙酸缓冲盐(p H 6.5,A)-甲醇(B)为流动相,梯度洗脱,检测波长260 nm,柱温25℃,流速0.8 m L/min。应用中药色谱指纹图谱相似度计算软件对指纹图谱进行相似度评价、聚类分析。[结果]所建立的方法重复性和精密度良好,建立的冬虫夏草指纹图谱包含24个共有峰,各峰分离良好,确认出尿嘧啶、尿苷、胸腺嘧啶、腺嘌呤、腺苷、虫草素6个共有成分。考察了29批西藏冬虫夏草,3批西藏以外地区冬虫夏草,2批蛹虫草,1批假虫草的指纹图谱与冬虫夏草指纹对照图谱的相似度。通过相似度分析,能很好的区分冬虫夏草、蛹虫草和假虫草。[结论]所建立的方法简单可靠,可做为冬虫夏草真伪鉴定的有效实验方法和依据。

[Objective] To establish HPLC fingerprints for indentification of Cordyceps sinensis in Tibet.[Methods] The HPLC analysis was performed on Waters Symmetry RP-C18 column（250×4.6 mm, 5 μm）and detected at 260 nm. The mobile phase was consisted of methnol and acetate buffer（p H 6.5） using a gradient elution. The column temperature was 25℃, and the flow rate was 0.8 m L/min. The similarity of fingerprints was evaluated by the fingerprint similarity calculation software, then, duster analysis was studied. [Results] The precision and repeatability of the method were good as the relative standard deviation（RSD） of intrabatch was less than 5%. The fingerprints of 29 batches of Cordyceps sinensis in Tibet, 3 Cordyceps sinensis from Sichuan and Qinghai, 2 Cordyceps militaris, and 1 fake Cordyceps sinensis were compared in characteristic components and similarity. The results demonstrated that Cordyceps sinensis was differentiated from the Cordyceps militaris and fake Cordyceps sinensis.[Conclusion] The method was simple, feasible and reproducible, and can be offered as technical basis for the identification of Cordyceps sinensis.