LC-ESI-MS/MS法测定清开灵注射液中5种有效成分的含量

Determination of 5 active components in Qingkailing injection by LC-ESI-MS/MS

目的建立同时测定清开灵注射液中猪去氧胆酸、胆酸、栀子苷、黄芩苷、绿原酸含量的LC-ESI-MS/MS法。方法采用ACE UltraCore 2.5 Super C_(18)柱(100 mm×2.1 mm,2.5μm),流动相:甲醇-含0.1%甲酸的5 mmol·L^(-1)甲酸铵溶液,梯度洗脱,流速:0.4 mL·min^(-1),进样量:2μL,柱温:40℃。采用电喷雾离子源,正负离子切换模式,多反应监测模式进行扫描定量分析,结果猪去氧胆酸、胆酸、栀子苷、黄芩苷、绿原酸分别在7.44~44.64μg·mL^(-1),7.22~43.32μg·mL^(-1),1.88~11.28μg·mL^(-1),11.26~67.56μg·mL^(-1),1.10~6.60μg·mL^(-1)与测定值呈现良好的线性关系(r≥0.9994),平均加样回收率在98.5%~99.8%,RSD在0.83%~2.0%(n=6)。结论该方法简便准确,灵敏度高,专属性高,干扰小,重现性好,可用于清开灵注射液中5种主要成分的含量测定。

Objective To determine the content of hyodeoxycholic acid,cholic acid,jasminoidin,baicalin,and chlorogenic acid in Qingkailing injection by LC-ESI-MS/MS.Methods ACE UltraCore 2.5 Super C_(18)(100 mm×2.1 mm,2.5μm)column was used with methanol-5 mmol·L^(-1) ammonium formate(containing 0.1%formic acid)as the mobile phase,and gradient elution at 0.4 mL·min^(-1).The injection volume was 2μL and the column temperature was 40℃.MS detection was performed in multi-reaction monitoring mode with positive/negative ion switching electrospray ionization.Results Hyodeoxycholic acid,cholic acid,jasminoidin,baicalin,and chlorogenic acid in Qingkailing injection had good linearity with the peak area at 7.44-44.64μg·mL^(-1),7.22-43.32μg·mL^(-1),1.88-11.28μg·mL^(-1),11.26-67.56μg·mL^(-1),and 1.10-6.60μg·mL^(-1),respectively(r≥0.9994).The average recovery was 98.5%-99.8%,and RSD was 0.83%-2.0%(n=6).Conclusion The method is simple,accurate,sensitive,specific,and reproducible with little interference,which can be used for the quantitative determination of 5 components in Qingkailing injection.