UPLC-MS/MS法同时测定养胃汤中26个成分的含量

Simultaneous determination of twenty six components in Yangwei decoction by UPLC-MS/MS

目的:建立UPLC-MS/MS方法同时测定养胃汤中26个成分(厚朴酚、和厚朴酚、木兰花碱、木兰箭毒碱、木兰苷A、木兰苷B、橙皮苷、川陈皮素、橘皮素、甜橙黄酮、芸香柚皮苷、柚皮素、人参皂苷Rb_(1)、6-姜辣素、甘草苷、甘草酸、甘草素、异甘草素、异甘草苷、芹糖异甘草苷、毛蕊花糖苷、异毛蕊花糖苷、绿原酸、新绿原酸、咖啡酸、芦丁)的含量。方法:采用Shim-pack GIST C_(18)色谱柱(2.1 mm×100 mm,2.0μm),以乙腈-0.1%乙酸-1 mmol·L^(-1)乙酸铵水为流动相,进行梯度洗脱,柱温为30℃,流速为0.3 mL·min^(-1),进样量为1μL,分析时间为18 min。采用电喷雾电离源(ESI源),以多重反应监测(MRM)模式正负离子检测。结果:所测26个化学成分在测定浓度范围内线性关系良好,相关系数均大于0.9969;精密度、重复性和稳定性良好;平均加样回收率为96.5%~104.2%,RSD≤4.0%。6批样品中厚朴酚、和厚朴酚、木兰花碱、木兰箭毒碱、木兰苷A、木兰苷B、橙皮苷、川陈皮素、橘皮素、甜橙黄酮、芸香柚皮苷、柚皮素、人参皂苷Rb_(1)、6-姜辣素、甘草苷、甘草酸、甘草素、异甘草素、异甘草苷、芹糖异甘草苷、毛蕊花糖苷、异毛蕊花糖苷、绿原酸、新绿原酸、咖啡酸、芦丁的含量测定结果分别为2814.12~2951.41、2551.48~2675.66、288.12~312.36、439.88~494.60、452.00~500.74、329.76~358.68、5995.57~6283.13、70.16~73.81、22.95~24.21、5.56~6.39、997.32~1077.03、6.17~6.81、643.88~674.16、994.16~1063.20、1929.00~1984.85、2356.80~2518.96、247.68~256.64、38.02~43.18、228.84~241.4、227.17~234.87、129.61~150.39、28.97~31.90、622.85~643.23、526.05~542.73、70.09~78.64、152.41~166.3μg·g^(-1)。结论:本方法准确灵敏,稳定性和重复性好,可用于养胃汤的质量控制。

Objective:To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the simultaneous determination of twenty six components(magnolol,honokiol,magnolflorine,magnocurarine,magnoloside A,magnoloside B,hesperidin,nobiletin,tangeretin,sinensetin,narirutin,naringenin,ginsenoside Rb_(1),6-gingerol,liquiritin,glycyrrhizic acid,liquiritigenin,isoliquiritigenin,isoliquiritin,isoliquiritin apioside,verbascoside,isoacteoside,chlorogenic acid,neochlorogenic acid,caffeic acid and rutin)in Yangwei decoction.Methods:The separation was performed on a Shim-pack GIST C_(18) column(2.1 mm×100 mm,2.0μm)with a mobile phase consisting of acetonitrile and 0.1%acetic acid-1 mmol·L^(-1) ammonium acetate water solution with gradient elution at a flow rate of 0.3 mL·min^(-1).The column temperature was 30℃,the injection volume was 1μL and the analysis time was 18.0 min.The detection was carried out by electrospray ionization(ESI).Positive and negative electrospray ionization was performed in multiple reaction monitoring(MRM)mode.Results:All of the analytes showed good linearity(r≥0.9969)in the tested ranges.The precision,repeatability and stability of the method were good for the twenty six components.The average recoveries were in the range of 96.5%~104.2%with relative standard deviations RSD≤4.0%.The contents of magnolol,honokiol,magnolflorine,magnocurarine,magnoloside A,magnoloside B,hesperidin,nobiletin,tangeretin,sinensetin,narirutin,naringenin,ginsenoside Rb_(1),6-gingerol,liquiritin,glycyrrhizic acid,liquiritigenin,isoliquiritigenin,isoliquiritin,isoliquiritin apioside,verbascoside,isoacteoside,chlorogenic acid,neochlorogenic acid,caffeic acid and rutin in 6 batches of Yangwei decoction were 2814.12-2951.41,2551.48-2675.66,288.12-312.36,439.88-494.60,452.00-500.74,329.76-358.68,5995.57-6283.13,70.16-73.81,22.95-24.21,5.56-6.39,997.32-1077.03,6.17-6.81,643.88-674.16,994.16-1063.20,1929.00-1984.85,2356.80-2518.96,247.68-256.64,38.02-43.18,228.84-241.4,227.17-234.87,129.61-150.39,28.97-31.90,622.85-643.23,526.05-542.73,70.09-78.64 and 152.41-166.3μg·g^(-1),respectively.Conclusion:The established method is accurate,stable and reproducible,and can provide a research method for the quality control of Yangwei decoction.