應用ITS序列鑑定麻黃藥材及其製劑

Identification of Ephedrae Herba and Its Preparations by PCR-amplified ITS Region Sequencing Method

李蕙君(Hui-Chun Lee) ； 謝詠筌(Yung-Chuan Hsieh) ； 呂康祖(Kang-Tsu Lu) ； 蔡佳芬(Chia-Fen Tsai) ； 陳惠芳(Hwei-Fang Cheng)

《食品藥物研究年報》 6期 (2015/12) Pp. 215-221

摘要

麻黃基原為麻黃科Ephedraceae植物草麻黃Ephedra sinica Stapf、中麻黃 Ephedraintermedia Schrenk et C. A. Mey.及木賊麻黃Ephedra equisetina Bge.之乾燥草質莖，本研究以ITS (internal transcribed spacer)片段序列作為基因標記，設計對Ephedra屬專一之引子組進行Nested PCR-DNA定序，建立藥材及製劑之麻黃基原鑑別方法。收集食品藥物管理署標本室收藏及查驗登記留樣合計31件藥材，實驗結果呈現2種不同序列，其中27件藥材檢體與草麻黃Ephedra sinica序列完全符合，而另外4件檢體與Ephedra equisetina有99%的序列相似度，應為木賊麻黃。在製劑鑑定方法的建立上，麻黃製劑檢體共有18件，其鑑定結果均為E. sinica。結果顯示，本研究建立以NestedPCR-DNA定序方法在藥材或多種不同複方製劑中均可專一地鑑別麻黃基原。

關鍵字

麻黃 ； Nested PCR-DNA定序

並列摘要

Ephedrae Herba consists of the dried stem or aerial part of three specific Ephedrae species, namely Ephedra sinica Stapf, Ephedra intermedia Schrenk et C. A. Mey. and Ephedra equisetina Bge. This study aimed on the development of a nested polymerase chain reaction (Nested PCR) and DNA sequencing method for the identification of genuine Ephedrae Herba. The concept of this method was to amplify internal transcribed spacer (ITS) regions by Nested PCR and then the PCR products were analyzed by auto-sequencer. The established method was applied to a survey of 31 herbs and 18 Chinese medicine preparations. The results showed that 27 and 4 herbal samples were identified as Ephedra sinica and Ephedra equisetina, respectively. The 18 Chinese medicine preparations were found all belong to species of Ephedra sinica. This method is selective for the identification of Ephedrae Herba in forms of herb and its Chinese medicine preparations.

並列關鍵字

ephedrae herba ； ITS ； nested PCR and DNA sequencing