注射用绒促性素中纯度测定方法的研究

目的: 建立了分子排阻法测定注射用绒促性素中纯度的方法,并在2010年度国家药品质量评价性抽验工作中,按照所建方法对国产注射用绒促性素进行了测定和评价。 方法: 采用两根TSK G3000SWXL凝胶色谱柱(7.8 mm×300 mm,5 μm)串联,预柱为TSK SWXL(6.0 mm×40 mm,5 μm),以0.05 mol·L^-1磷酸二氢钠缓冲液(pH 7.0)-乙腈(80∶20)为流动相,流速0.6 mL·min^-1,检测波长214 nm,面积归一化法定量。 结果: 卵泡刺激素(FSH)和绒促性素(HCG)分离完全,各辅料均无干扰,绒促性素的效价比活和纯度呈正相关。目前法定检验标准中无此检查项,76批样品全部合格,没有发现问题;若按本法进行纯度检查,则样品的不合格率显著增加。 结论: 本法具有较高的选择性,结果稳定、准确可靠,通过测定绒促性素的纯度可达到监控制剂生产过程中所使用原料的效价比活的目的。在此次专项抽验中发现现行标准存在缺陷,仍有待进一步提高,可采用专属性较强的仪器分析法如分子排阻法检查该品种的纯度或进行鉴别。

Objective: To establish a Size Exclusion Chromatography method for the purity determination of Chorionic gonadotrophin for Injection.The method was applied for the domestic Chorionic Gonadotrophin for Injection of national drug evaluation in 2010. Methods: Two TSK G3000SWXL gel columns (7.8 mm×300 mm,5 μm) series connected,TSK SWXL (6.0 mm×40 mm,5 μm) as guard column.The mobile phase consisted of 0.05 mol·L^-1 NaH₂PO₄ (pH 7.0) - acetonitrile(80∶20) at the flow rate of 0.6 mL·min^-1 with the detection wavelength of 214 nm.The drug purity content was quantified by peak area normalization method. Results: FSH and HCG were separated completely,with no interference from excipients.The specific activity of HCG showed positive correlation with its purity.This test is not included in the official acceptance standard currently.In the 76 tested batches,many of them were found disqualified with this purity test while totally accordance with the official standard.Inclusion:The method showed high selectivity,reliability,and sensitivity.HCG purity test can be applied for specific activity control of materials in production.Further improvement are needed for the current standard.

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