HPLC-MS/MS测定当归六黄汤中4种不同成分的含量

Determination of four different components in Danggui Liuhuang decoction by HPLC-MS/MS

目的建立HPLC-MS/MS的分析方法同时测定当归六黄汤中4种主要成分黄柏碱、巴马汀、毛蕊异黄酮、阿魏酸的含量,为当归六黄汤的质量控制提供方法学参考和借鉴。方法基于HPLC-MS/MS的分析方法,质谱检测采用ESI源MRM的正离子数据采集模式。色谱柱为Agilent Extend-C_(18)(5μm,4.6 mm×250 mm),甲醇和含0.5%的甲酸水溶液进行梯度洗脱。结果黄柏碱的线性范围为2~200 nmol/ml,巴马汀、毛蕊异黄酮、阿魏酸的线性范围均为20~2000 nmol/ml,当归六黄汤7批样品中4种成分的含量相对稳定,其中,阿魏酸主要存在于黄柏和黄连中;黄柏碱只在黄柏中测到;毛蕊异黄酮在黄芩和黄芪中含量较高;巴马汀在黄柏和黄芪中都能检测到。结论该方法灵敏度高,专属性好,样品的稳定性良好,能够满足中药复方定量分析的要求,可用于当归六黄汤的质量控制和评价研究。

Objective To establish the method of simultaneous determination of four main components of Danggui Liuhuang Decoction,including phellodendrine,palmatine,calycosin,and ferulic acid and provide reference for the quality control of Danggui Liuhuang Decoction.Methods Based on the HPLC-MS/MS analysis method,the positive ion data acquisition mode were adopted for the mass spectrometry detection and the four main components were quantified with multiple reaction monitoring mode(MRM)by ESI source.The chromatographic column was Agilent Extend-C_(18)(5μm,4.6 mm×250 mm),and gradient elution was performed with methanol and 0.5%formic acid in water.Results The linear range of phellodendrine was from 2-200 nmol/ml,and the linear range of palmatine,calycosin and ferulic acid was from 20-2000 nmol/ml.The contents of the four components in the seven batches of Danggui Liuhuang Decoction were relatively stable,among which ferulic acid was mainly found in Phellodendrine and Coptidis;Phellodendrine was only detected in cortex phellodendri;the content of calycosin in Scutellaria baicalensis and Astragalus was higher;palmatine was detected in both Phellodendron and Astragalus.Conclusion The method had high sensitivity,good specificity and sample stability,which could meet the requirements of quantitative analysis of Traditional Chinese Medicine compounds,and could provide reference for further pharmacokinetics study on the content changes of traditional Chinese medicine compounds in biological samples.