超高效液相色谱串联质谱法检测人促红素注射剂中2种抗生素残留

Detection of Two Antibiotic Residues in Human Erythropoietin Injection and Human Erythropoietin for Injection by UPLC-MS/MS

目的建立检测人促红素注射剂中氨苄西林和博来霉素残留的超高效液相色谱串联质谱(UPLC-MS/MS)法。方法色谱柱为Waters Cortecs T3柱(100 mm×2.1 mm,2.7μm);流动相为0.1%甲酸水溶液-[乙腈-甲醇(95∶5,V/V)],梯度洗脱;流速为0.2 mL/min;柱温为40℃;进样量为5μL。采用电喷雾离子源正离子模式和多反应监测扫描模式。结果氨苄西林和博来霉素质量浓度分别在19.38~484.5 ng/mL(r=0.9979,n=5)和21.26~212.6 ng/mL(r=0.9988,n=5)范围内与峰面积线性关系良好,检测限分别为0.00969,10.63 ng/mL,定量限分别为0.0969,21.26 ng/mL;精密度、稳定性试验结果的RSD均小于4.0%;平均加样回收率分别为102.11%和101.77%,RSD为分别为7.46%和13.35%;基质效应均不明显(分别为95.03%和84.51%)。9家国内生产企业和1家国外生产企业的39批人促红素注射剂均未检出氨苄西林和博来霉素。结论所建立的方法操作简单、检测灵敏,结果准确,可用于人促红素注射剂中氨苄西林和博来霉素残留的检测。

Objective To establish an ultra-high-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determining the ampicillin and bleomycin residues in Human Erythropoietin Injection and Human Erythropoietin for Injection.Methods The chromatographic column was the Waters Cortecs T3 column(100 mm×2.1 mm,2.7μm),the mobile phase was 0.1%formic acid aqueous solution-[acetonitrile-methanol(95∶5,V/V)]with gradient elution,the flow rate was 0.2 mL/min,the column temperature was 40℃,and the injection volume was 5μL.The electron spray ionization with positive ion mode and multiple reaction monitoring mode was adopted.Results The linear ranges of ampicillin and bleomycin were 19.38-484.5 ng/mL(r=0.9979,n=5)and 21.26-212.6 ng/mL(r=0.9988,n=5)respectively.The limits of detection of ampicillin and bleomycin were 0.00969,10.63 ng/mL,respectively,and their limits of quantification were 0.0969,21.26 ng/mL,respectively.The RSDs of precision and stability tests were lower than 4.0%.The average recovery rates of the above two antibiotics were 102.11%and 101.77%,with RSDs of 7.46%and 13.35%respectively.The matrix effects of ampicillin and bleomycin were not obvious,with 95.03%and 84.51%,respectively.No ampicillin or bleomycin was detected in 39 batches of Human Erythropoietin Injection and Human Erythropoietin for Injection from nine domestic manufacturing enterprises and one foreign manufacturing enterprise.Conclusion The established method is simple,sensitive and accurate,which can be used to detect ampicillin and bleomycin residues in Human Erythropoietin Injection and Human Erythropoietin for Injection.