Fu Xiao, Lu Rongrong, Zhao Songsong
Department of Traditional Chinese Medicine, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121000, China.
Graduate College, Liaoning Medical University, Jinzhou 121000, China.
Pharmacogn Mag. 2017 Jul-Sep;13(51):425-429. doi: 10.4103/pm.pm_141_16. Epub 2017 Jul 19.
Fuzi-Gancao herb couple is one of the commonly used herb couples involved in the traditional Chinese medicine formulations, with radix aconiti lateralis (Fuzi in Chinese) and radix glycyrrhizae (Gancao in Chinese) as a ratio of 1:1. Alkaloids and flavonoids were considered as the main active ingredients of Fuzi-Gancao herb couple. However, no analytical methods have been reported to quantitatively analyze these activity ingredients in Fuzi-Gancao herb couple simultaneously.
To develop a simple, rapid, and sensitive UHPLC-MS/MS method for simultaneous quantitation of six alkaloid and three flavonoid compounds, namely, aconitine (AC), mesaconitine (MA), hypaconitine (HA), benzoylaconitine (BAC), benzoylmesaconitine (BMA), benzoylhypaconitine (BHA), liquiritin, isoliquiritin, and licochalcone A (LCA) in Fuzi-Gancao herb couple.
The chromatographic separation was achieved using a reversed phase C18 column and a mobile phase consisted of water (0.1% formic acid in water, v/v) and acetonitrile. The detection was achieved in multiple reaction monitoring modes. The optimal mass transition ion pairs (m/z) for quantitation were 646.3/586.5 for AC, 632.4/572.5 for MA, 616.3/556.4 for HA, 604.5/572.5 for BAC, 590.4/540.4 for BMA, 574.1/542.5 for BHA, 419.4/257.1 for liquiritin, 419.4/257.1 for isoliquiritin, and 339.2/121.1 for LCA.
Good linearity was observed in validated concentration range for each analyte ( > 0.9992), and the intra- and inter-day precisions were <3.12% and 3.65%, respectively. The recovery ranged from 85.36% to 110.14%.
The results demonstrated that the method developed was reliable, rapid, and specific. Moreover, this method was successfully applied to control the quality of Fuzi-Gancao herb couple.
A simple, rapid, and sensitive UHPLC-MS/MS method for simultaneous quantitation of six alkaloid and three flavonoid compounds in Fuzi-Gancao herb couple has been developed. UHPLC-ESI-MS/MS: Ultra High Performance Liquid Chromatography-electrospray ionization-mass spectrometry; RSD: Relative standard deviations; LOD: Limit of detection; LOQ: Limit of quantification; MRM: Multiple reaction monitor; TCMs: Traditional Chinese medicines.
附子 - 甘草药对是中药方剂中常用的药对之一，由附子（中药名）和甘草（中药名）按1:1的比例组成。生物碱和黄酮类化合物被认为是附子 - 甘草药对的主要活性成分。然而，尚未见有同时定量分析附子 - 甘草药对中这些活性成分的分析方法报道。
建立一种简单、快速、灵敏的超高效液相色谱 - 串联质谱法（UHPLC-MS/MS），用于同时定量分析附子 - 甘草药对中的6种生物碱和3种黄酮类化合物，即乌头碱（AC）、中乌头碱（MA）、次乌头碱（HA）、苯甲酰乌头碱（BAC）、苯甲酰中乌头碱（BMA）、苯甲酰次乌头碱（BHA）、甘草苷、异甘草苷和甘草查尔酮A（LCA）。
采用反相C18色谱柱进行色谱分离，流动相由水（含0.1%甲酸，v/v）和乙腈组成。采用多反应监测模式进行检测。用于定量分析的最佳质荷比转换离子对（m/z）分别为：AC为646.3/586.5，MA为632.4/572.5，HA为616.3/556.4，BAC为604.5/572.5，BMA为590.4/540.4，BHA为574.1/542.5，甘草苷为419.4/257.1，异甘草苷为419.4/257.1，LCA为339.2/121.1。
各分析物在验证的浓度范围内线性良好（>0.9992），日内和日间精密度分别<3.12%和3.65%。回收率在85.36%至110.14%之间。
结果表明所建立的方法可靠、快速且具有特异性。此外，该方法成功应用于控制附子 - 甘草药对的质量。
已建立一种简单、快速、灵敏的UHPLC-MS/MS方法，用于同时定量分析附子 - 甘草药对中的6种生物碱和3种黄酮类化合物。UHPLC-ESI-MS/MS：超高效液相色谱 - 电喷雾电离 - 质谱；RSD：相对标准偏差；LOD：检测限；LOQ：定量限；MRM：多反应监测；TCMs：中药
