HPLC测定吡嗪酰胺的有关物质

Determination the related substances of Pyrazinamide by HPLC

目的采用HPLC测定吡嗪酰胺的有关物质。方法采用Cosmosil C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-5 mmol.L-1磷酸氢二钾溶液(磷酸调pH3)(10∶90),流速1.0 mL.min-1,检测波长268 nm。结果吡嗪-2-羧酸0.201～1.206μg.mL-1与峰面积的线性关系良好;2-氰基吡嗪0.202～1.214μg.mL-1与峰面积线性关系良好。加样回收率吡嗪-2-羧酸为99.92%,RSD=0.71%(n=9);2-氰基吡嗪为99.77%,RSD=0.96%(n=9)。结论所用方法准确、简便、快速,适用于吡嗪酰胺有关物质的检查。

OBJECTIVE To determine the related substances of pyrazinamide by HPLC.METHODS The C18（250 mm×4.6 mm,5 μm） was used as cosmosil column and the mobile phase was consisted of methanol-5 mmol·L-1 Dipotassium hydrogen phosphate solution（adjusted to pH3 by phosphate）10：90.Flow rate was 1.0 mL·min-1,and the detection wavelength set at 268 nm.RESULTS The calibration curve was linear（r=0.9999）within the range of 0.201-1.206 μg·mL-1 for 2-pyrazinecarboxylic acid,the calibration curve was linear（r=0.9999）within the range of 0.202-1.214 μg·mL-1 for pyrazinecarbonitrile and the average recovery of 2-pyrazinecarboxylic acid was 99.92%,RSD=0.71%（n=9）.The recovery of Pyrazinecarbonitrile was 99.77%,RSD=0.96%（n=9）.CONCLUSION The method is accurate,simple,rapid and suitable for anlaysis of related substances in pyrazinamide.