黃芩藥材及其製劑Nested PCR-DNA定序鑑定方法之建立

Establish of Nested PCR-DNA Sequencing Method for the Identification of Scutellariae Radix and Its Chinese Medicine Preparations

呂康祖(Kang-Tsu Lu) ； 謝詠筌(Yung-Chuan Hsieh) ； 劉芳淑(Fang-Su Liu) ； 羅吉方(Chi-Fang Lo) ； 林哲輝(Jer-Huei Lin)

《食品藥物研究年報》 1期 (2010/11) Pp. 264-270

https://doi.org/10.6945/ARFDR.201011.0264

摘要

黃芩為Scutellaria baicalensis Georgi之乾燥根，為中藥常用藥村，然而因為生長環境不同，藥材外觀差異大，難以確認是否為正確藥材，因此，研究中將應用Nested PCR-DNA定序方法鑑別黃芩藥材的真偽，並建立製劑中黃芩藥材的檢驗鑑別方法。利用Gen Bank中ITS(internal transcribed spacer)序列資料設計引子組合，用以進行Nested PCR，再分析比對檢體經擴增後的ITS序列作為鑑定依據。結果顯示，黃芩藥材69件藥材檢體中，出現2種DNA序列，其中11件的序列較其他58件鑑定為Scutellaria baicalensis的序列多出1個鹽基。收集黃芩製劑檢體46件為24種方劑，在不同藥材組合的方劑中，可用建立之Nested PCR-DNA定序方法確認黃芩藥材基原，而不受其他藥材成分的干擾，確認本方法的專一性。

關鍵字

黃芩 ； Nested PCR ； DNA定序 ； 鑑定

並列摘要

Scutellariae Radix, the dried root of Scutellaria baicalensis Georgi, is widely used in Chinese medicine preparations. Due to various growth environments, Scutellariae Radix is difficult to authenticate by its externals. In this study, a nested polymerase chain reaction (nested PCR) followed by DNA sequencing method was developed and employed to identify the authentic Scutellariae Radix and to detect its presence in Chinese medicine preparations. Using the sequence of the internal transcribed spacer (ITS) obtained from GenBank, we designed the primer sets for nested PCR to amplify the ITS fragments of samples. The revealed sequences were compared with GenBank databases. Fifty eight out of 69 raw material samples tested were identified as S. baicalensis, while 11 samples had sequences with one more base than that of S. baicalensis. Authentic Scutellariae Radix in 46 preparation samples of 24 formulas could be identified specifically by the established nested PCR-DNA sequencing methods.

並列關鍵字

Scutellariae Radix ； ITS ； nested PCR ； DNA sequencing