肝素中残留核苷酸检测方法国际协作研究

Round robin study on methods for detection of residual nucleotides in heparin

目的:美国药典会肝素原料个论中拟增加核苷酸杂质的检查方法,全球18家实验室参加了该方法的协作研究以确定检测方法和限度可行性。方法:征集国内2家企业共10批肝素钠样品,及USP提供的测试样品。使用Phenomenex Synergi FUSION-RP C18色谱柱(250 mm×4.6 mm,5μm),样品中的核酸杂质经酶解后,用0.02 mol·L-1醋酸铵和乙腈梯度洗脱分离,流速为1 mL·min-1,检测波长为260 nm。结果:系统适用性及测试样品结果均符合USP的要求,本实验室提交的数据全部被采纳,国内10批样品结果均符合USP拟定限度(<0.1%)。结论:该方法基本可行,较现有方法专属性高。

Objective： To evaluate the proposed methods for detection of nucleotidic impurities that would be added into the monograph of heparin raw materials in USP as part of a round robin study involving 18 laboratories worldwide to determine the proposed detection methods and limited feasibility. Methods： Ten different batches were collected from two heparin API manufactures representing the domestic market,and the proficiency sample given by USP was prepared.The Phenomenex Synergi FUSION-RP C18 column（250 mm×4.6 mm,5 μm）was used to separate the samples which were processed by enzymatic hydrolysis.The mobile phase consisted of 0.02 mol·L-1 ammonium acetate（A）-100%acetonitrile（B）with gradient elution,at the flow rate of 1 mL·min-1 and the detection wavelength was 260 nm. Results： The system suitability and results of proficiency samples all met the USP requirements,the data submitted by our laboratory were all accepted,and results of all the 10 batches of the domestic samples were in accordance with the proposed acceptance limit（〈0.1%）by USP. Conclusion： The proposed method is generally feasible with a higher specificity than the current method.