【摘要】 目的分析rHuEPO供试品及其对照品经胰酶酶解后分别在HPLC与UPLC上的肽图谱表现。方法rHuEPO供试品及对照品经透析、冻干、重新溶解和胰酶酶切后,以三氟醋酸/水,三氟醋酸/乙腈/水作为流动相梯度洗脱,分别以HPLC及UPLC连接反相色谱柱分析酶切后的肽段。结果rHuEPO供试品及其对照品经胰酶酶切后的肽段在HPLC上肽图谱表现一致,在UPLC上肽图谱也完全一致。UPLC分析rHuEPO肽图需要的进样量更少,且较传统HPLC的分析时间大为缩短,同时不损失分离度及灵敏度。结论UPLC与HPLC均能验证rHuEPO供试品及对照品肽图的一致性,相比于HPLC、UPLC分析肽图谱更方便快捷,更节省样品,更适合分析rHuEPO肽图。更多还原

【Abstract】 Objective To determine the tryptic peptide mapping of both test recombinant human erythropoietin(rhEPO) and standard rhEPO samples by HPLC and UPLC.Methods All of the rhEPO samples were dialysed,freeze-dried,redissolved,digested by trypsin,and then were subjected to peptide mapping by HPLC and UPLC.Each sample was chromatographed on a reversed-phase column using water and acetonitrile as eluents and trifluoroacetic acid as ion-pairing agent.Results The results of peptide mapping for test rhEPO samples were consistent with those of standard rhEPO samples by either HPLC or UPLC.Compared with the use of HPLC,the use of UPLC holds the advantages of shorter separation time and lower sample volume without compromising the separation degree and sensitivity.Conclusion Consistency of peptide mapping for test rhEPOs and standard rhEPOs can also be validated by UPLC as by conventional HPLC.The results from precision and comparative studies for rhEPOs by UPLC show that UPLC is a suitable technique for routine pharmaceutical analysis.It is a more advantageous,secure,sensitive and time-saving method suitable for separating peptide fragments of rhEPOs in comparison with traditional high performance liquid chromatography.更多还原