HPLC-MS/MS法测定桑菊感冒片中5种成分的含量

Determinations of Five Components in Sangju Ganmao Tablets by HPLC-MS/MS

目的:建立高效液相色谱-串联质谱法同时测定桑菊感冒片中芦丁、绿原酸、连翘苷、桔梗皂苷D和苦杏仁苷的含量。方法:采用Waters Atlantis C_(18)色谱柱(2.1 mm×150 mm,5μm),以乙腈(A)-0.1%甲酸(B)为流动相,梯度洗脱(0~3 min,20%A;3~15 min,20%A→60%A;15~23 min,60%A;23~24 min,60%A→20%A;24~30 min,20%A),流速0.2 m L·min^(-1),柱温35℃,采用电喷雾离子源(ESI),以多反应监测(MRM)方式进行正离子扫描。结果:芦丁、绿原酸、连翘苷和桔梗苷D分别在浓度0.01~2μg·m L^(-1)的线性范围内线性关系良好(r>0.999),苦杏仁苷在浓度0.25~50μg·m L^(-1)的线性范围内线性关系良好(r>0.999),平均回收率(n=6)分别为98.89%、99.80%、100.61%、99.35%、99.51%,RSD%分别为0.84%、0.90%、1.19%、1.28%、0.86%。测定样品3批,5个成分的测定结果分别为:批号S154601:0.2605、0.2721、0.1848、0.1693、4.8203 mg·g^(-1);批号S154602:0.2440、0.2573、0.1709、0.1712、4.7639 mg·g^(-1);批号S154603:0.2679、0.2788、0.1900、0.1778、4.8725 mg·g^(-1)。结论:所建立的方法准确、可靠、重复性好,灵敏度高,为桑菊感冒片的全面质量控制提供科学依据。

Objective：To establish the method for determinations of rutin, chlorogenic acid, forsythin, platycodin D and amygdalin in Sangju Ganmao Tablets by HPLC-MS/MS. Methods：The determination was carried out with Waters Atlantis C18 column （2.1×150mm,5μm）.The mobile phase consisted of acetonitrile（A）-0.1% formic acid solution（B） with gradient elution（0～3min,20%A;3～15min,20%A→60%A;15～23min,60%A;23～24min,60%A→20%A;24～30min,20%A） at a flow rate of 0.2 mL.min-1 at 35℃.The analytes were detected by tandem mass spectrometry with the electrospray ionization（ESI） source combined with multiple reaction monitoring（MRM） mode. Results：Rutin, chlorogenic acid, forsythin and platycodin D showed a good linearity in the range of 0.01～2μg.mL-1（r〉0.999）.The amygdalin showed a good linearity in the range of 0.25～50μg.mL-1（r〉0.999）. The average recoveries （n=6） were 98.86%,99.80%,100.61%,99.35%,99.60% with the RSD% were 0.84%、0.90%,1.19%,1.28%,0.86%.Conclusion：This method is reliable, accurate, which offers advantages of highy sensitivity and precision. It can be used for the overall assessment of the quality of Sangju Ganmao Tablets.