Yang Li, Wang Yun, Wang Longxing, Xiao Hongbin, Wang Zhengtao, Hu Zhibi
Key Laboratory of Standardization of Chinese Medicines of Ministry of Education, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Zhangjiang Hi-tech Park, Shanghai 201203, China.
J Chromatogr A. 2009 Mar 13;1216(11):2098-103. doi: 10.1016/j.chroma.2008.06.043. Epub 2008 Jul 4.
Longdan Xiegan Decoction (LXD) is a formulated preparation composed of 10 ingredient herbs, with iridoids as the main bioactive components. In this study, a rapid, simple and reliable method of simultaneous determination of four iridoid glycosides in LXD using high-performance liquid chromatography (HPLC) coupled with electrospray ionization mass spectrometry (MS) was first developed and validated. The four iridoid glycosides references were isolated from LXD extract and purified using a preparative HPLC chromatography. The sample preparation for quantification comprised of a simple ultrasonic extraction and the satisfactory chromatographic separation of the four structurally similar iridoid glycosides was effected in less than three minutes on a CAPCELL PAK C(18) MGII column (3 microm, 100 mm x 2.0 mm), using an elution system of 10% methanol and their concentrations in different batches of LXD and ingredient herbs were simultaneously determined by HPLC-MS/MS using a multiple reaction monitoring (MRM) mode. The method was validated with respect to the overall intra- and inter-day variation (RSD less than 8%) and the limits of quantification for the four iridoid glycosides were 35, 20, 37 and 33 ng/mL, respectively.
龙胆泻肝汤（LXD）是一种由10味中药组成的配方制剂，以环烯醚萜类化合物为主要生物活性成分。本研究首次建立并验证了一种快速、简便且可靠的高效液相色谱（HPLC）-电喷雾电离质谱（MS）联用同时测定LXD中4种环烯醚萜苷的方法。从LXD提取物中分离出4种环烯醚萜苷对照品，并采用制备型HPLC色谱法进行纯化。定量分析的样品制备采用简单的超声提取法，在CAPCELL PAK C(18) MGII柱（3μm，100mm×2.0mm）上，使用10%甲醇洗脱系统，不到3分钟即可实现4种结构相似的环烯醚萜苷的良好色谱分离，并采用多反应监测（MRM）模式通过HPLC-MS/MS同时测定不同批次LXD及其药材中它们的含量。该方法在日内和日间总体变异方面得到验证（相对标准偏差小于8%），4种环烯醚萜苷的定量限分别为35、20、37和33 ng/mL。
