反相高效液相色谱法测定氯高铁血红素的有关物质

RP-HPLC determination of related substances of hemin

目的:建立反相高效液相色谱法测定氯高铁血红素原料药的有关物质。方法:采用ODS色谱柱(150 mm×4.6 mm,5μm);以甲醇-乙腈-0.05 mol.L-1醋酸铵缓冲液(冰醋酸调pH 3.0)-吡啶(31∶31∶38∶0.4)为流动相;检测波长为399 nm;流速1.0 mL.min-1;柱温30℃。结果:氯高铁血红素浓度在0.0205～0.1025μg.mL-1范围内,浓度与主要杂质峰面积呈良好的线性关系(r=0.9999),重复性试验中单个杂质含量的RSD为1.8%(n=6)。结论:该方法简便、快速、准确且重复性好,可作为氯高铁血红素原料药的有关物质控制方法。

Objective： To establish a RP - HPLC method for the related substances of hemin. Methods： An ODS col- umn （ 150 mm ×4. 6 mm, 5 txm） was used with a mobile phase of methanol - acetonitrile -0. 05% ammonium acetate buffer solution （adjusted to pH3.0 with glacial acetic acid） -pyridine （31：31：38： 0. 4） ;the detection wavelength was set at 399 nm; the flow rate was 1.0 mL · min^ - 1 and the column temperature was maintained at 30 ℃. Results： A good linearity of the concentration and main impurity peak area was achieved in the concentration range of 0. 0205 - 0. 1025 μg · mL^-1 （ r = 0. 9999 ）, RSD of repeatability for single impurity was 1.8% （ n = 6 ）. Conclusion： The method is simple, rapid, precise and reproducible. It can be used for quality control of hemin.