LC-MS/MS法同时测定中药注射剂中3种成分的含量

Simultaneous determination of the contents of 3 components in traditional Chinese medicine injections by LC-MS/MS

目的建立高效液相色谱-质谱联用(LC-MS/MS)法测定注射用益气复脉和生脉注射液中5-羟甲基糠醛(5-HMF)、人参皂苷Rd与人参皂苷Rg 3的含量。方法采用SHIMADZU VP-ODS色谱柱(150 mm×2.0 mm,5μm)。流动相:1 mL·L^-1甲酸(A)-乙腈(B),梯度洗脱,梯度程序:0~6 min,5%B;6~20 min,5%B~80%B;20~25 min,80%B。柱温:35℃;流速:0.2 mL·min^-1;进样体积:5μL。质谱条件:ESI源;脱溶剂管线温度:250℃,加热块温度:400℃;雾化气流速:3 L·min^-1;干燥气流速:15 L·min^-1;正离子模式检测电压:+4.5 kV;负离子模式检测电压-3.5 kV。结果3种成分质量浓度分别在53.76~2204.00(r 1=0.9972),61.60~2464.00(r 2=0.9983),51.05~2042.00(r 3=0.9998)ng·mL^-1范围内与峰面积呈良好的线性关系;平均回收率分别为101.41%,101.44%,99.11%,RSD值分别为4.93%,3.65%,3.79%。结论建立的LC-MS/MS法专属性强、重复性好、稳定性好、精密度高,回收率满足实验要求,可用于中药注射剂中3种成分的含量测定。

Objective To establish an LC-MS/MS quantitative method for determining 5-hydroxymethylfurfural(5-HMF),ginsenoside Rd and ginsenoside Rg 3 in Yiqi Fumai Lyophilized Injections and Shengmai Injections.Methods SHIMADZU VP-ODS(150 mm×2.0 mm,5μm)was adopted as analytical column.The mobile phase was 1 mL·L^-1 formic acid(A)and acetonitrile(B),gradient program:0-6 min,5%B;6-20 min,5%B-80%B;20-25 min,80%B.The column temperature was 35℃,the flow rate was 0.2 mL·min^-1,and the volume of injection was 5μL.MS conditions:ESI ion source was used;dissolvent line temperature was 250℃,the temperature of heating block was 400℃,nebulizing gas flow rate was 3 L·min^-1 and drying gas flow rate was 15 L·min^-1.Positive ion mode detection voltage:+4.5 kV,negative ion mode test voltage:-3.5 kV.Results A good linear relationship was observed between the concentration and the peak area of 3 components in the ranges of 53.76-2204.00(r 1=0.9972),61.60-2464.00(r 2=0.9983)and 51.05-2042.00 ng·mL^-1(r 3=0.9998),respectively.The average recovery rates were 101.41%,101.44%and 99.11%,respectively.The RSDs were 4.93%,3.65%and 3.79%,respectively.Conclusion The established LC-MS/MS method showed good specificity,repeatability,stability,precision and recovery rate,and can accurately determine the contents of 3 components in traditional Chinese medicine injections.