目的 建立特异性引物扩增鉴定金嗓清音丸中僵蚕成分.方法 根据家蚕、白僵菌、绿僵菌线粒体全基因序列差异设计特异性引物PM2、PB2、PA2,采用十六烷基三甲基溴化铵法、酚仿抽提法提取纯化总DNA,并以此为模板进行特异性引物扩增和方法学考察,产物以2％琼脂糖凝胶电泳检测.结果 家蚕、白僵菌、绿僵菌DNA经PCR扩增后,分别在82、94、144 bp处产生条带,并且未观察到非特异性扩增条带.PM2、PB2、PA2对相应DNA的检测灵敏度分别为0.10、0.10、0.010 ng/μL.来自同一厂家的6批样品中同时检出家蚕、白僵菌DNA,但未检出绿僵菌DNA,而另外2批样品仅检出家蚕DNA.结论 该方法方便可靠,专属性强,有助于更好地监控金嗓清音丸等含僵蚕中药成方制剂的质量,可为其临床用药的安全性和有效性提供保障,并为其他中成药中动物药的专属性鉴定提供参考.

目的 基于特异性引物扩增建立中成药中僵蛹成分的鉴定技术.方法 根据家蚕、白僵菌和绿僵菌的全基因组序列差异设计了特异性引物PM、PB和PA,采用CTAB法、三氯甲烷抽提法从供试品中提取与纯化DNA,并进行聚合酶链式反应(PCR),扩增产物以2％琼脂糖凝胶电泳检测.结果 家蚕、白僵菌及绿僵菌的DNA经扩增后分别在61、94、144 bp处产生条带,且引物间无交叉反应,体现出很强的专属性,家蚕、白僵菌和绿僵菌DNA的检出限分别为1、0.1、1 ng.在6批天蚕片和天蚕胶囊中,均检测出家蚕和白僵菌,未检测出绿僵菌.结论 该方法准确快速,可用于鉴定天蚕片和天蚕胶囊中的僵蛹成分.

AIM To establish a UPLC method for the simultaneous content determination of glycosides,rutinoside,myricitrin,hyperoside,isoquercitrin,guaijaverin,astragalin,quercitrin and afzelin in the shoots of Toona sinensis Roemer.METHODS The analysis of 60％ methanol extract of T.sinensis shoots was performed on a 35 ℃ thermostatic Waters XBridge Shield RP18 column (2.1 mm × 150 mm,1.7 μm),with the mobile phase comprising of acetonitrile-water flowing at 0.35 mL/min in a gradient elution manner,and the detection wavelength was set at 350 nm.RESULTS Eight flavonol glycosides showed good linear relationships within their own ranges (r≥ 0.9992),whose average recoveries were 98.3％-103.5％ with the RSDs of 0.46％-3.36％.CONCLUSION This accurate,stable and reproducible method can be used for the quality control of T.sinensis shoots.