丹红注射液大分子物质筛查方法研究

目的：采用高效分子排阻色谱法（HPSEC法）对丹红注射液中大分子物质进行筛查。方法：采用HPSEC-RID技术，色谱柱为Shodex OHPak SB-803HQ凝胶色谱柱（6.8 mm×300 mm，10 μm），以0.71%硫酸钠水溶液（内含0.02%叠氮钠）为流动相，系列分子量右旋糖酐标准品为对照，对丹红注射液及其中间体中多糖类大分子物质进行筛查；再采用HPSEC-UV技术，色谱柱为TSK-GEL G2000 SWXL凝胶柱（7.8 mm×300 mm，10 μm），以乙腈-水-三氟乙酸（45：55：0.05）为流动相，核糖核酸酶A、人胰岛素、胸腺肽1为对照，在214 nm波长处对丹红注射液及其中间体中蛋白质类大分子物质进行筛查。结果：丹红注射液未检出大分子物质，中间体检出相对分子质量在4 500～9 000的大分子糖类物，说明经过生产过程的逐步除杂，大分子物质在丹红注射液成品中已完全去除。结论：该方法对丹红注射液质量控制具有重要意义，对其他注射液中大分子杂质的检查亦有借鉴作用。

Objective: To establish methods for screening of macromolecules in Danhong injections by high performance size exclusion chromatography(HPSEC).Methods: Using HPSEC-RID technology,the Shodex OHPak SB-803HQ gel chromatographic column(6.8 mm×300 mm,10 μm) was used as chromatographic column,the mobile phase was 0.7% Na₂SO₄(containing 0.02% NaN₃),with a series of dextran of different molecular weight as control to screen the polysaccharide macromolecular material in Danhong injection and its intermediates;then with HPSEC-UV technology,the TSK-GEL G2000 SWXL gel chromatographic column(7.8 mm×300mm,10 μm) was used as chromatographic column,the mobile phase was acetonitrile-water-trifluoroacetic acid(45:55:0.05),ribonuclease A,human insulin,and thymosin alpha 1 were used as control,to screen the protein macromolecules in Danhong injection and its intermediates at 214 nm.Results: Macromolecules were not detected in Danhong injection;4 500-9 000 polysaccharide macromolecules were detected in intermediates.The result indicated that macromolecules were completely removed in Danhong injection after the removal of impurities in production process.Conclusion: The method is of great significance to Danhong injection quality control,and can be used as reference for impurity limit test of other injections.