二丁酰环磷腺苷钠有关物质检测方法优化及评价

Optimization and evaluation of detection method for related substances in bucladesine sodium

目的:建立并优化二丁酰环磷腺苷钠有关物质检测方法。方法:在日本药局方基础上,对各实验条件进行优化,得到最优检测条件,色谱柱为CAPCELL PAK C 18,流动相为pH 6.0磷酸盐缓冲液-甲醇,梯度洗脱,检测波长为258 nm,并进行了方法学验证。结果:系统适用性溶液中,各破坏溶液中主峰与相邻峰之间的分离度均大于1.5;溶液在48 h内稳定;各杂质含量在其限度浓度的50%~150%范围内与峰面积均呈现良好的线性关系,R 2>0.9994;加标实验中,N6-丁酰3′,5-环磷腺苷钠平均回收率为99.4%~106.6%;2′-O-丁酰3′,5-环磷腺苷钠平均回收率为88.8%~103.6%,环磷腺苷钠平均回收率96.9%~102.5%,均符合要求,对3批样品进行检测,均检出上述杂质。结论:该方法灵敏度高,分离效果、准确性、耐用性好,可用于二丁酰环磷腺苷钠有关物质检测,为其质量控制提供依据。

Objective:To establish and optimize a detection method of dibutyryl cyclic adenosine phosphate related substances.Methods:On the basis of the JP,the experimental conditions were optimized to obtain the optimal detection conditions.The column was CAPCEILPAK C 18;mobile phase was pH 6.0 phosphate buffer solution-methanol with gradient elution;detection wavelength was 258 mm.The method validation was carried out.Results:In the system applicability solution,the separation degree of the main peak and adjacent peaks in each destruction solution was greater than 1.5;the solution was stable within 48 h.The content of each impurity showed a good linear relationship with the peak area in the range of 50%-150%of the limit concentration,R 2>0.9994.In the spike experiment,the average recovery rate of N6-butyryl 3′,5-cyclophosphatin sodium was 99.4%-106.6%.The average recoveries of 2′-0 butyryl 3,5-cycloadenosine monophosphate sodium were 88.8%-103.6%,and the average recoveries of cyclophosphaminate sodium were 96.9%-102.5%,which met the requirements.Three batches of samples were tested,and the above impurities were detected.Conclusion:This method has high sensitivity,good separation effect,accuracy and durability,and can be used for the detection of bucladesine sodium,providing a basis for its quality control.