鲜冬虫夏草的理化鉴别  

Physico-chemical Identification of Fresh Cordyceps Sinensis

作　　者：李文庆 陈铃 周建桥 温宝庆 梅全喜 李春红 李文佳 钱正明 Li Wenqing;Chen Ling;Zhou Jianqiao;Wen Baoqing;Mei Quanxi;Li Chunhong;Li Wenjia;Qian Zhengming(Key Laboratory of State Administration of Traditional Chinese Medicine,Sunshine Lake Pharma Co. Ltd. ,Guangdong Dongguan 523850,China;Shenzhen Bao'an Authentic TCM Therapy Hospital)

机构地区：[1]广东东阳光药业有限公司国家中医药管理局重点研究室,广东东莞523850 [2]深圳市宝安纯中医治疗医院

出　　处：《中国药师》2019年第5期962-965,共4页China Pharmacist

基　　金：东莞市院士工作站资助项目(编号:DGYSZ-2018-03);国家重点研发计划(编号:2018YFC1706101)

摘　　要：目的:对鲜冬虫夏草进行理化鉴别研究。方法:采用聚合酶链式反应法(PCR)、蛋白质电泳法、TLC法、HPLC法。PCR法基于内转录间隔区2(ITS2)序列设计引物进行鉴定;蛋白质电泳法通过超声提取蛋白质,聚丙烯酰胺电泳检测; TLC法采用4%磷酸氢二钠溶液浸渍处理后的硅胶GF254薄层板,以异丙醇-乙酸乙酯-甲醇-水-浓氨试液(5∶3∶1∶1∶0. 1)为展开剂,紫外光灯(254 nm)下检视; HPLC法以Welch,Ultimate AQ-C_(18)(150 mm×4. 6 mm,5μm)为色谱柱;以0. 04 mol·L^(-1)磷酸二氢钾溶液-甲醇为流动相,梯度洗脱;柱温为30℃;流速为1 ml·min^(-1);检测波长为260 nm。结果:PCR法可鉴别除发酵冬虫夏草菌粉外的7种常见混淆品;蛋白质电泳法可鉴别8种常见混淆品,TLC法可鉴别除蝉花、戴氏虫草外的6种常见混淆品,HPLC法可鉴别除蝉花外的7种常见混淆品。结论:该研究可为鲜冬虫夏草的药材鉴定和质量标准制定提供参考。Objective:To research physico-chemical identification of fresh Cordyceps sinensis.Methods:Polymerase chain reaction(PCR),protein electrophoresis,TLC and HPLC discrimination were adopted in the present study.The primers used in the PCR experiment were based on the ITS2 sequence;In the protein electrophoresis experiment,ultrosonic extraction and polyacrylamide gel electrophoresis separation were employed;In the TLC analysis,the plate was GF254 plate soaked in 4%disodium hydrogen phosphate solution,the developing system was isopropano-acetic ether-methanol-water-strong ammonia solution(5∶3∶1∶1∶0.1),and the plate was inspected under 254 nm;In the HPLC analysis,an Ultimate AQ-C18(150 mm×4.6 mm,5μm)column was gradiently eluted by0.04 mol·L^-1 monopotassium phosphate-methanol,the column temperature was 30℃,the flow rate was 1 ml·min^-1,and the detection wavelength was 260 nm.Results:PCR could identify 7 common confusing substances except for Hirsutella sinensis;Protein electrophoresis could identify 8 common confusing substances;TLC could identify 6 common confusing substances except for Cordyceps ciecadae and Cordyceps tasi;HPLC could identify 7 common confusing substances except for Cordyceps ciecadae.Conclusion:The research is useful for the identification and quality control of fresh Cordyceps sinensis.

关 键 词：鲜冬虫夏草 理化鉴别 聚合酶链式反应 蛋白质电泳 薄层色谱法 高效液相色谱法 

分 类 号：R282.5[医药卫生—中药学]