注射用腺苷钴胺有关物质测定方法改进

Improved method for determination of related substances in cobamamide for injection

目的建立高效液相色谱法,用外标法加主成分自身对照法测定注射用腺苷钴胺有关物质的含量。方法采用C18色谱柱(250mm×4.6 mm,5μm),以0.05mol·L-1磷酸二氢钾-乙腈(85∶15)(用磷酸调节pH至3.2)为流动相,流速为1.0mL·min-1,柱温为30℃,检测波长为260nm;进样量为10μL;测定有关物质的含量。结果腺苷钴胺及氰钴胺与相邻峰分离度良好,线性范围分别为100～0.2μg·mL-1(R=0.9999)和10～0.1μg·mL-1(R=1.0000),精密度、准确度等均符合分析样品的要求。结论本方法操作简便快速,重复性好,专属性强,可用于该制剂中有关物质的含量控制。

OBJECTIVE To establish a high performance liquid chromatographic method, standard and control method for the determination of the main component of injection related substances content with cobamamide. METHODS C18 chromatographic column （250mm×4.6 mm,5μm）,with 0. 05mol·L^-1 potassium dihydrogen phosphate （adjusted to pH 3.2 with phosphoric acid） and acetonitrile（85：15 ） as mobile phase, the flow rate was 1.0mL · min^-1, the column temperature was 30℃, the detection wavelength was 260nm; 10μL sample;determination of content of related substances. RESULTS Cobamamide and cyanocobalamin and adjacent peak separation is good,the linear range was 100 ~0. 2μg · mL^-1 （ R =0. 9999） and 10 ~0.1μg · mL^-1 （ R = 1. 0000） ,precision, accuracy can meet the requirements of sample. CONCLUSION This method was simple,rapid,repeatable,reproducible, and can be used for the control of related substances in the preparation.