基于UPLC-MS/MS法的栝楼桂枝汤使药甘草配伍研究

Study on compatibility of licorice by gualou guizhi decoction based on UPLC-MS/MS

目的建立超高效液相色谱串联质谱法(UPLC-MS/MS)同时测定栝楼桂枝汤中5种甘草活性成分的含量,研究栝楼桂枝汤不同配伍对甘草活性成分含量的影响。方法甘草与栝楼桂枝汤中其他味药按照不同配伍组别分成9组,采用Waters CORTECS C_(18)色谱柱(2. 1 mm×100 mm,1. 6μm),乙腈-0. 1%甲酸水进行梯度洗脱,5种成分在多反应监测模式(MRM)下检测,甘草酸m/z 821. 45/351. 2;芹糖异甘草苷m/z 549. 16/255. 06;芹糖甘草苷m/z549. 16/255. 06;甘草苷m/z 417. 1/255. 3;异甘草苷m/z 417. 1/255. 3,分别对不同配伍组的含量进行比较分析。结果甘草中5种活性成分在6 min内均完成色谱分离与检测,同时栝楼桂枝汤佐药配伍组和君药配伍组对使药甘草5种活性成分含量有影响,此外,发现大枣、生姜配伍改变了甘草黄酮活性成分的溶出、君药天花粉配伍改变了甘草酸的溶出。结论所建立的栝楼桂枝汤甘草活性成分测定方法简便、快速,可应用于甘草配伍研究,能更好地探寻甘草的复方配伍机制,为其临床合理用药提供参考。

Objective UPLC-MS/MS method was established to determine the contents of five active ingredients of licorice in Gualou Guizhi Decoction.The effects of different compatibility of Gualou Guizhi Decoction and licorice on the content of active ingredients of licorice were studied.Methods The other components of Gualou Guizhi Decoction were divided into 9 groups according to their compatibility with licorice.Waters CORTECS C18 column(2.1 mm×100 mm,1.6μm)was used.Gradient elution was carried out with acetonitrile-0.1%formic acid water,and 5 components were detected under multireaction detection mode(MRM).Glycyrrhizic acid m/z 821.45/351.2;Celery isoglycyrrhizin m/z 549.16/255.06;Apigenoside m/z 549.16/255.06;Glycyrrhizin m/z 417.1/255.3;Isoglycoside m/z 417.1/255.3.The contents of different compatibility groups were compared and analyzed.Results The chromatographic separation and detection of five active ingredients in licorice were completed within 6 minutes.At the same time,the adjuvant compatibility group of Gualou Guizhi Decoction and the monarch compatibility group had influence on the content of five active ingredients in licorice.In addition,it was also found that jujube and ginger changed the dissolution of Flavonoids from licorice,and trichosanthin compatibility changed the dissolution of glycyrrhizic acid.Conclusion Based on UPLC-MS/MS method,the determination of active ingredients of licorice in Gualou Guizhi Decoction is simple and rapid,which can be applied to the study of licorice compatibility,better explore the compatibility mechanism of licorice compound,and provide reference for clinical rational drug use.