HPLC测定吡拉西坦及其注射液有关物质的研究

Improvement of determination of related substances in piracetam and its injection by HPLC

目的 改进并建立高效液相色谱法测定吡拉西坦及其注射液有关物质。方法 以Thermo Syncronis C18（4.6 mm×250mm,5μm）为色谱柱;以磷酸盐缓冲液（取1.0 g/L的磷酸氢二钾溶液,用磷酸调节pH值至6.0）：乙腈（95∶5）为初始比例,梯度洗脱;检测波长205 nm;流速1.0 mL/min;分别测定5种已知杂质的校正因子,采用加校正因子的自身成分对照法对有关物质进行定量测定。结果 主成分与各有关物质分离度良好;已知杂质A～E分别在各自范围内线性关系良好;平均回收率分别为99.46%、98.21%、97.22%、100.23%、97.58%（n=9）,其RSD均小于2.0%;杂质A^E的校正因子分别为1.1、1.4、1.5、0.8、1.4。结论 本方法可用于吡拉西坦原料药及其注射剂中有关物质的测定,采用加校正因子的主成分自身对照法可以准确地测定其有关物质的含量。

Objective To improve and establish an RP-HPLC method for the determination of related substances of piracetam and its injection. Methods Using thermo syncronis C18（4.6 mm ＆#215;250 mm,5μm）chromatographic column, mobile phase was acetonitrile-phosphate buffer （adjust pH of 1.0 g/L potassium hydrogen phosphate solution to 6.0 with phosphoric acid）, gradient elution with initial mobile phase 5% acetonitrile.The flow rate was 1 mL/min, detected at 205 nm.Five calibration factors of the known impurities were separately measured.the related substances were determined using main component self-compare with calibration factor.Results Piracetam and their related substances were well separated.The calibration curves of five known impurity A^E were linear within the concentration range measured.The average recoveries were 99.46%,98.21%,97.22%,100.23% and 97.58%, respectively（n=9）, RSDs of the average recoveries were lower than 2.0%.The calibration factors of five known impurity A ~E were 1.1, 1.4,1.5,0.8, 1.4, respectively.Conclusion The method can be used to determine the related substances in piracetam and its injection.The related substances can be determined accurately using main component self-compare with calibration factor.