UFLC-MS/MS法同时测定黄芪桂枝五物汤中4个活性成分的含量

Simultaneous Determination of Four Active Components in Huangqi Guizhi Wuwu Decoction by UFLC-MS/MS

本文建立了UFLC-MS/MS法同时测定黄芪桂枝五物汤中4个活性成分(毛蕊异黄酮苷、桂皮酸、芍药苷和芍药内酯苷)的含量。采用Shim-Pack XR-ODS柱,0.1%甲酸水溶液(A)-乙腈(B)流动相,梯度洗脱,流速为0.4 mL/min。采用3200 QTRAP三重四极杆串联质谱仪,电喷雾电离源,负离子检测,多重反应检测(MRM)。毛蕊异黄酮苷、桂皮酸、芍药苷和芍药内酯苷分别在0.2~10.0μg/mL(r=0.999 9)、0.5~25.0μg/mL(r=0.999 8)、2.0~100.0μg/mL(r=0.999 6)、0.5~25.0μg/mL(r=0.999 5)呈良好的线性关系;平均回收率(n=9)分别为98.5%、99.0%、98.7%和98.8%。6批黄芪桂枝五物汤样品含量测定结果:毛蕊异黄酮苷0.143~0.153 mg/mL、桂皮酸0.109~0.123 mg/mL、芍药苷2.881~2.990 mg/mL、芍药内酯苷0.436~0.446 mg/mL。结果表明该方法快速、准确,可用于黄芪桂枝五物汤的质量标准研究,同时为其在兽药领域开发提供参考。

In this study, an UFLC-MS/MS method for simultaneous determination of four active constituents(calycosin-7-O-β-D-glucoside, cinnamic acid, paeoniflorin and albiflorin) in Huangqi Guizhi Wuwu decoction was developed. Chromatographic separation was performed on Shim-Pack XR-ODS column. The mobile phase consisted of 0.1% formic acid(A) and acetonitrile(B) in water at a flow rate of 0.4 mL/min. The four analytes were determined with gradient elution. Detection was performed on 3200 QTRAP mass spectrometry equipped with electro-spray ionization source in negative ionization mode. Quantification was performed using multiple reactions monitoring(MRM) mode. The linear ranges were 0.2-10.0 μg/mL(r=0.999 9) for calycosin-7-O-β-D-glucoside, 0.5-25.0 μg/mL(r=0.999 8) for cinnamic acid, 2.0-100.0 μg/mL(r=0.999 6) for paeoniflorin, 0.5-25.0 μg/mL(r=0.999 5) for albiflorin. The average recovery(n=9) of the four components were 98.5%, 99.0%,98.7% and 98.8%,respectively. The content ranges of four components in six batches were 0.143-0.153 mg/mL for calycosin-7-O-β-D-glucoside, 0.109-0.123 mg/mL for cinnamic acid, 2.881-2.990 mg/mL for paeoniflorin and 0.436-0.446 mg/mL for albiflorin. The results showed that the method was fast and accurate, could be used for the quality standard study of Huangqi Guizhi Wuwu decoction, and has potential for adoption in the field of veterinary medicinal development.