高效液相色谱法测定腺苷钴胺片中的羟钴胺素含量

Determination of cyanokit in cobamamide tablets by HPLC

目的建立高效液相色谱法测定腺苷钴胺片中的羟钴胺素含量。方法采用DIKMA C8色谱柱（150mm×4.6 mm,5μm）,流动相为柠檬酸-磷酸氢二钠缓冲液（pH=4.0）-甲醇（80：20）,柱温为35℃,流速为1.0 mL·min-1,检测波长为351 nm。结果羟钴胺素在1.0~20.0μg·mL-1与峰面积呈良好线性关系（r=0.999 7）,低、中、高浓度平均回收率分别为99.0%、100.2%、100.0%,RSD分别为0.17%、0.21%、0.23%（n=3）,检出限为4.0 ng,日内、日间精密度RSD分别为0.58%和0.69%（n=6）。结论该方法灵敏、准确,专属性强,可作为腺苷钴胺片中的羟钴胺素含量测定。

Objective To determine cyanokit in cobamamide tablets by HPLC.Methods The chromatography separation was performed on a DIKMA C8 column（150 mm×4.6 mm,5 μm） at 35 ℃,The flow rate was at 1.0 mL·min-1.The mobile phase was lemon acid-sodium hydrogen phosphate buffer solution（pH = 4.0）-methanol = 80 ：20.The detection wavelength was 351 nm.Results Cyanokit showed a good linearity at 1.0-20.0 μg · mL-1.The average recovery of low,medium,high concentration of cyanokit was 99.0%,100.2% and 100.0%,RSD was 0.17%,0.21% and 0.23%,respectively.The limit for determination was 4.0 ng and the RSD of intra-day and inter-day was 0.58% and 0.69%（n=6）,respectively.Conclusion The method is simple and accurate for the determination of cyanokit in cobamamide tablets.