高效液相色谱-串联质谱法测定蜂王浆中红霉素及其代谢物

Determination of erythromycin A and its metabolites in royal jelly by high performance liquid chromatography-tandem mass spectrometry

目的建立蜂王浆中红霉素A、红霉素A烯醇醚、脱水红霉素A和N-脱甲基红霉素A的高效液相色谱-串联质谱检测方法。方法以pH 6.0磷酸盐缓冲液为提取剂,经HLB小柱净化富集后,采用高效液相色谱-串联质谱仪分析。采用Agilent Polaris C18色谱柱(100 mm×2.0 mm,5μm),以甲醇和0.1%甲酸为流动相,质谱模式为电喷雾正离子监测。结果该方法前处理简单,线性范围为2.0~100μg/L,相关系数在0.99以上,四种化合物的检出限和定量限分别为0.5μg/kg和5.0μg/kg。方法采用内标法定量,回收率范围为70.0~118.1%,相对标准偏差小于10%。结论方法回收率稳定且重现性较好,能够满足日常检测工作的需要。

Objective To established a method for determination of erythromycin A, erythromycin A enol ether, anhydroerythromycin A and N-demethyl erythromycin A by high performance liquid chromatogra-phy-tandem mass spectrometry.Methods The sample was extracted with pH 6.0 phosphate buffer and cleaned-up by HLB column and then analyzed by LC-MS with electrospray positive ionization. Separations were performed on an Agilent Polaris C18 column (100 mm×2.0 mm, 5μm). The mobile phase was methanol- 0.1% formic acid.Results Under the optimal conditions, the calibration curves were linear in the ranges of 2.0~100.0μg/L with the correlation coefficients more than 0.99 while the limit of detection and limit of quantitation were 0.5 and 5.0μg/kg, respectively. The internal standard was used for quantitative analysis. The recoveries ranged from 70.0 % to 118.1 % with RSDs less than 10%.Conclusion The method with higher recoveries and better precision could meet the needs of practical analysis.