UFLC-MS/MS法同时测定黄芪建中汤中6个活性成分的含量

Simultaneous determination of six active components in Huangqi Jianzhong decoction by UFLC-MS/MS

目的:建立一种同时测定黄芪建中汤中6个活性成分(毛蕊异黄酮-7-O-β-D-吡喃葡萄糖苷、桂皮酸、芍药苷、芍药内酯苷、甘草苷和甘草酸)含量的超快速液相色谱-串联质谱法。方法:采用Shim-Pack XR-ODS色谱柱(75 mm×3.0 mm,2.2μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0~6.0 min,12%B→25%B;6.0~7.0 min,25%B→45%B;7.0~11.0 min,45%B→60%B;11.0~13.0 min,60%B→70%B),平衡时间5 min,流速0.4 mL·min^(-1),柱温30℃,进样量5μL;采用电喷雾电离源(ESI源),负离子模式检测,多反应监测(MRM)。结果:毛蕊异黄酮-7-O-β-D-吡喃葡萄糖苷、桂皮酸、芍药苷、芍药内酯苷、甘草苷和甘草酸质量浓度分别在0.2~10μg·mL^(-1)(r=0.9996)、0.5~25μg·mL^(-1)(r=0.9995)、2~100μg·mL^(-1)(r=0.9996)、0.5~25μg·mL^(-1)(r=0.9998)、0.5~25μg·mL^(-1)(r=0.9997)和0.5~25μg·mL^(-1)(r=0.9996)范围内与峰面积呈良好线性关系;平均回收率(n=9)分别为98.3%、98.4%、98.9%、98.0%、98.8%和99.2%。6批样品中上述6个成分的含量分别为0.114~0.128、0.118~0.144、4.19~4.52、1.03~1.12、0.277~0.345、0.679~0.798 mg·mL^(-1)。结论:本方法灵敏度高,稳定性好,可用于黄芪建中汤的质量控制。

Objective:To establish an ultra-fast liquid chromatography with tandem mass spectrometry(UFLC-MS/MS)method for simultaneous determination of six active components(calycosin-7-O-β-D-glucopyranoside,cinnamic acid,paeoniflorin,albiflorin,liquiritin and glycyrrhizic acid)in Huangqi Jianzhong decoction.Methods:Chromatographic separation was performed on a Shim-Pack XR-ODS column(75 mm×3.0 mm,2.2μm).The mobile phase consisted of 0.1%formic acid in water(A)and acetonitrile(B)at a flow rate of 0.4 mL·mL^(-1).The six analytes were determined with gradient elution(0-6 min,12%B→25%B;6-7 min,25%B→45%B;7-11 min,45%B→60%B;11-13 min,60%B→70%B),and the equilibration time was 5 min.The column temperature was maintained at 30℃,and the injection volume was 5μL.Detection was performed with electrospray ionization source in negative ionization mode.Quantification was performed using multiple reactions monitoring(MRM)mode.Results:The linear ranges of calycosin-7-O-β-D-glucopyranoside,cinnamic acid,paeoniflorin,albiflorin,liquiritin and glycyrrhizic acid were 0.2-10μg·mL^(-1)(r=0.9996),0.5-25μg·mL^(-1)(r=0.9995),2-100μg·mL^(-1)(r=0.9996),0.5-25μg·mL^(-1)(r=0.9998),0.5-25μg·mL^(-1)(r=0.9997)and 0.5-25μg·mL^(-1)(r=0.9996),respectively.The average recoveries(n=9)of the above-mentioned six components were 98.3%,98.4%,98.9%,98.0%,98.8%and 99.2%,respectively.The content ranges of six components in six batches were 0.114-0.128 mg·mL^(-1)for calycosin-7-O-β-D-glucopyranoside,0.118-0.144 mg·mL^(-1)for cinnamic acid,4.19-4.52 mg·mL^(-1)for paeoniflorin,1.03-1.12 mg·mL^(-1)for albiflorin,0.277-0.345 mg·mL^(-1)for liquiritin and 0.679-0.798 mg·mL^(-1)for glycyrrhizic acid.Conclusion:The established method is accurate,stable and reproducible,and can provide a research method for the quality control of Huangqi Jianzhong decoction.