柱前衍生化HPLC法测定异烟肼注射液中游离肼的含量

Determination of free hydrazine in isoniazid injection by HPLC with precolumn derivatization

目的:建立柱前衍生化高效液相色谱法测定异烟肼注射液中游离肼含量。方法:以苯甲醛为衍生化试剂,将游离肼衍生化为苯甲醛吖嗪,然后进行高效液相色谱测定。采用Ultimate XB-C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为0.1%乙二胺四乙酸二钠溶液-乙腈(30∶70),流速1.0 m L·min^(-1),检测波长为310 nm,柱温40℃。结果:游离肼质量浓度在2.5~100μg·m L^(-1)范围内与其衍生物的峰面积呈良好线性关系,在低、中、高3个添加水平范围内的平均回收率为96.9%~101.0%,RSD在0.2%~3.0%之间,游离肼的检出限为0.5μg·m L^(-1),定量限为2.5μg·m L^(-1)。3个批次样品的测定结果分别为0.014%、0.043%和0.095%。结论:本文方法为异烟肼注射液的质量控制提供了有效的检测方法。

Objective：To develop an HPLC method for determination of free hydrazine in isoniazid injection by precolumn derivatization.Methods：This method is based on precolumn derivatization with benzaldehyde to form benzaldehyde azine.Welch Ultimate XB-C18 column（250 mm×4.6 mm,5 μm） was adopted,the mobile phase consisted of 0.1% EDTA-acetonitrilr（30∶70） at a flow rate of 1.0 m L·min^-1,the detection wavelength was 310 nm,and the column temperature was 40 ℃.Results：The calibration curve of free hydrazine showed good linearity in the ranges of 2.5^（-1）00 μg·m L^-1.The average recoveries were 96.9%^（-1）01.0% with RSDs of 0.2%-3.0% at three spiked levels.The limit of detection was estimated to be 0.5 μg·m L^-1,and the limit of quatification was 2.5 μg·m L^-1.The contents of free hydrazine in three batches of samples were 0.014%,0.043% and 0.095%,respectively.Conclusion：Our study provides an effective method for the quality control of isoniazid injection.