Detection of Two Antibiotic Residues in Human Erythropoietin Injection and Human Erythropoietin for Injection by UPLC-MS/MS
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,2.7 μm),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 5 μL.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.
Human Erythropoietin InjectionHuman Erythropoietin for InjectionUPLC-MS/MSampicillinbleomycinresiduecontent determination