Wang Ruwei, Kobayashi Yuta, Lin Yu, Rauwald Hans Wilhelm, Yao Jianbiao, Fang Ling, Qiao Hongxiang, Kuchta Kenny
Zhejiang CONBA Pharmaceutical & Drug Research Development Corporation, Hangzhou, China.
Faculty of Medicine, Shimane University, Izumo, Japan.
Planta Med. 2015 Jan;81(1):71-8. doi: 10.1055/s-0034-1383312. Epub 2014 Dec 17.
An HPLC quantification method for ginkgolic acid derivatives in Ginkgo biloba leaf extracts was developed. Using 13 : 0 ginkgolic acid as a marker compound, the relative correlation factors of the four other ginkgolic acid derivatives - namely, 15 : 0 ginkgolic acid, 15 : 1 ginkgolic acid, 17 : 1 ginkgolic acid, and 17 : 2 ginkgolic acid - to 13 : 0 ginkgolic acid were determined by HPLC and subsequently used for calculating their contents in ten hydro-ethanolic refined extract samples. In other words, the content of 13 : 0 ginkgolic acid in the extracts was determined using the isolated compound as an external standard. Subsequently the now known concentration of this compound functioned as an internal standard for the quantification of the other four ginkgolic acid derivatives via the described correlation factors. This HPLC method was validated by two independent control measurements, one with an external standard for every individual compound and one based on the present method with the single marker compound alone. The results did not differ significantly in any of the 10 tested extract samples. The protocol presented here thus not only uses the same reference substance for G. biloba extracts as the current Chinese Pharmacopoeia method but also incorporates the advantages of the current European Pharmacopoeia approach. It is simple, reproducible, and can be used to determine the total contents of ginkgolic acid derivatives in G. biloba leaf extracts.
建立了一种用于测定银杏叶提取物中银杏酸衍生物的高效液相色谱定量方法。以13:0银杏酸作为标记化合物，通过高效液相色谱法测定了其他四种银杏酸衍生物（即15:0银杏酸、15:1银杏酸、17:1银杏酸和17:2银杏酸）与13:0银杏酸的相对校正因子，并随后用于计算其在10个水乙醇精制提取物样品中的含量。换句话说，提取物中13:0银杏酸的含量是使用分离得到的化合物作为外标进行测定的。随后，该化合物已知的浓度通过所述校正因子作为内标用于其他四种银杏酸衍生物的定量分析。该高效液相色谱方法通过两项独立的对照测量进行了验证，一项是对每种单独化合物使用外标，另一项是基于仅使用单一标记化合物的本方法。在10个测试提取物样品中的任何一个中，结果均无显著差异。因此，本文提出的方案不仅使用与现行《中国药典》方法相同的对照品用于银杏提取物，还结合了现行《欧洲药典》方法的优点。它简单、可重复，可用于测定银杏叶提取物中银杏酸衍生物的总含量。
