Ndjoko K, Wolfender J L, Hostettmann K
Institut de Pharmacognosie et Phytochimie, BEP, Université de Lausanne, Switzerland.
J Chromatogr B Biomed Sci Appl. 2000 Jul 21;744(2):249-55. doi: 10.1016/s0378-4347(00)00316-9.
Ginkgolic acids (GAs) are toxic phenolic compounds present in the fruits and leaves of Ginkgo biloba L. (Ginkgoacae). Their maximum level in phytopharmaceuticals containing ginkgo extracts has been recently restricted to 5 microg/g by the Commission E of the former Federal German Health Authority. In order to detect ginkgolic acids at these low levels, a sensitive and selective analytical method, based on liquid chromatography-electrospray mass spectrometry (LC-ES-MS) has been developed. The three main phenolic acids (1-3) of the chloroform fruit extract were isolated and used as standards for quantification. In the LC-ES-MS negative ion mode, calibration curves with good linearities (r=0.9973, n=6) were obtained in the range of 0.5-10 microg/g for compounds 1, 2 and between 0.1 and 7.5 microg/g (r=0.9949, n=6) for ginkgolic acid 3. The detection limits at a SIN ratio of 3 were 0.1 (3) and 0.25 microg/g (1, 2). Recoveries were around 101% at 5 microg/g for the substances detected in the leaf extracts. Good precision was achieved with relative standard deviations of less than 4% (n=6). The optimised method was applied to verify whether the amount of gingkolic acids was below 5 microg/g in a standardised leaf extract which is a constituent of a phytopreparation.
银杏酸(GAs)是银杏科银杏属植物银杏果实和叶子中存在的有毒酚类化合物。前联邦德国卫生局的E委员会最近将含银杏提取物的植物药中银杏酸的最高含量限制为5微克/克。为了检测这些低含量的银杏酸,已开发出一种基于液相色谱 - 电喷雾质谱(LC - ES - MS)的灵敏且选择性的分析方法。氯仿果实提取物中的三种主要酚酸(1 - 3)被分离出来并用作定量标准品。在LC - ES - MS负离子模式下,化合物1、2在0.5 - 10微克/克范围内获得了线性良好的校准曲线(r = 0.9973,n = 6),银杏酸3在0.1至7.5微克/克范围内(r = 0.9949,n = 6)。在SIN比为3时,检测限分别为0.1(3)和0.25微克/克(1、2)。叶提取物中检测到的物质在5微克/克时的回收率约为101%。相对标准偏差小于4%(n = 6),实现了良好的精密度。应用优化后的方法来验证一种标准化叶提取物(一种植物制剂的成分)中银杏酸的含量是否低于5微克/克。