反相离子对HPLC法测定三磷酸胞苷二钠注射液的含量

Reversed phase ion-pair HPLC determination of cytidine disodium triphosphate in cytidine disodium triphosphate injection

目的:用反相离子对高效液相色谱法测定三磷酸胞苷二钠注射液中三磷酸胞苷二钠(CTP)的含量。方法:以 Luna ODSC_(18)为色谱柱(250 mm×4.60 mm,5～μm),含50 mmol·L^(-1)磷酸二氢钾的四丁基氢氧化铵(75 mmol·L^(-1))溶液(磷酸调 pH为4.0)-甲醇(75:25)为流动相,检测波长271 nm,进样量20μL。结果:CTP 浓度在40～200μg·mL^(-1)范围内,与峰面积呈良好的线性关系,回归方程为 Y=9.302×10~4X+1.000×10~6(r=0.9997);含量测定的重复性 RSD 为0.5%(n=6),回收率为99.96%,检出限为0.15 ng,定量限为0.5 ng。结论:方法灵敏度高,重现性好,专属性强,用反相离子对高效液相色谱法测定三磷酸胞苷二钠注射液中 CTP 的含量,国内未见报道。

Objective：To establish a reversed phase ion -pair HPLC method for determining the concentrations of sodium cytidine triphosphate（CTP） in cytidine disodium triphosphate injection. Method ： Luna ODS C18 （250 mm × 4.60 mm,5μm）column was applied ,using methanol -75 mmol · L^-1 tetrabutylammonium hydroxide solution（ contained 50 mmol · L^- 1 potassium dihydrogen phosphate, adjusted to pH 4.0 with phosphoric acid ） （25：75 ） as mobile phase. Injection volume was 20 μL and detection wavelength was 271 nm. Result： Peak area of CTP was in a good linear correlation with injection quantity within the range of 40 -200 μg · mL^-1. Regression equation was Y = 9. 302 × 10^4X ＋ 1. 000 × 10^6（r =0. 9997）. The recovery was 99.96% ,RSD was 0.5% （n =6）. The limit of detection was 0. 15 ng, and the limit of quantitation was 0. 5 ng. Conclusion：The method has high sensitivity and reproductivity. It can be used for quality control of cytidine disodium triphosphate injection. It has not been reported in China for determining the concentrations of CTP in cytidine disodium triphosphate injection with reversed phase ion - pair HPLC.