UPLC-MS/MS法同时测定乳核内消液中柴胡皂苷a、柴胡皂苷d、柴胡皂苷b_(1)、柴胡皂苷b_(2)和柴胡皂苷k的含量

Simultaneous determination of saikosaponin a,saikosaponin d,saikosaponin b_(1),saikosaponin b_(2) and saikosaponin k in Ruhe Neixiao liquids by UPLC-MS/MS

目的:采用超高效液相色谱-串联质谱(UPLC-MS/MS)法同时测定乳核内消液中柴胡皂苷a、柴胡皂苷d、柴胡皂苷b_(1)、柴胡皂苷b_(2)、柴胡皂苷k的含量。方法:采用YMC-Triart C18(150 mm×2.1 mm,3μm)色谱柱,流动相为乙腈-0.1%甲酸水溶液,梯度洗脱,流速0.3 mL·min^(-1),柱温35℃;采用ESI-离子源,多反应监测(SRM)模式检测。结果:柴胡皂苷a、柴胡皂苷d、柴胡皂苷b_(1)、柴胡皂苷b_(2)、柴胡皂苷k线性范围分别为0.106~10.622μg·mL^(-1)(r=0.9996)、0.101~10.097μg·mL^(-1)(r=0.9998)、0.098~9.840μg·mL^(-1)(r=0.9999)、0.343~34.339μg·mL^(-1)(r=0.9980)、0.126~12.640μg·mL^(-1)(r=0.9983),平均回收率(n=6)分别为98.6%(RSD=1.4%)、99.9%(RSD=1.2%)、101.1%(RSD=1.8%)、101.6%(RSD=1.8%)、98.6%(RSD=1.9%)。10个厂家25批次乳核内消液样品中柴胡皂苷a、柴胡皂苷d、柴胡皂苷b_(1)、柴胡皂苷b_(2)、柴胡皂苷k含量范围分别为1.59~94.62、0~0.52、21.29~286.19、14.78~156.64、0~66.01μg·mL^(-1)。结论:该方法可同时测定乳核内消液中柴胡5个成分的含量,方法灵敏、准确,重现性好,可为乳核内消液的质量控制和综合评价提供方法依据。

Objective:To establish a method for simultaneous determination of saikosaponin a,saikosaponin d,saikosaponin b_(1),saikosaponin b_(2) and saikosaponin k in Ruhe Neixiao liquids by UPLC-MS/MS.Methods:The samples were separated on a YMC-Triart C_(18)column(150 mm×2.1 mm,3μm)by gradient elution at a flow rate of 0.3 mL·min^(-1)using acetonitrile and 0.1% aqueous formic acid as the mobile phase.The column temperature was maintained at 35℃.Multiple reaction monitoring(SRM)scanning was employed for the quantification with switching electrospray ion(ESI)source polarity in negative mode.Results:The linear ranges of saikosaponin a,saikosaponin d,saikosaponin b_(1),saikosaponin b_(2) and saikosaponin k were 0.106-10.622μg·mL^(-1)(r=0.9996),0.101-10.097μg·mL^(-1)(r=0.9998),0.098-9.840μg·mL^(-1)(r=0.9999),0.343-34.339μg·mL^(-1)(r=0.9980)and 0.126-12.640μg·mL^(-1)(r=0.9983),respectively.The average recoveries(n=6)were 98.6%(RSD=1.4%),99.9%(RSD=1.2%),101.1%(RSD=1.8%),101.6%(RSD=1.8%)and 98.6%(RSD=1.9%),respectively.The content ranges of saikosaponin a,saikosaponin d,saikosaponin b_(1),saikosaponin b_(2) and saikosaponin k in Ruhe Neixiao liquids were 1.59-94.62μg·mL^(-1),0-0.52μg·mL^(-1),21.29-286.19μg·mL^(-1),14.78-156.64μg·mL^(-1)and 0-66.01μg·mL^(-1),respectively.Conclusion:The method can be used for simultaneous determination of saikosaponin a,saikosaponin d,saikosaponin b_(1),saikosaponin b_(2) and saikosaponin k in 25 samples from 10 manufacturers.This method is sensitive,accurate and reproducible,and provides a scientific reference for quality control and comprehensive evaluation of Ruhe Neixiao liquids.