柱前衍生化HPLC法测定异烟肼注射液中游离肼的含量

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-C₁₈色谱柱（250 mm×4.6 mm，5 μm），流动相为0.1%乙二胺四乙酸二钠溶液-乙腈（30：70），流速1.0 mL·min^-1，检测波长为310 nm，柱温40℃。结果：游离肼质量浓度在2.5~100 μg·mL^-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0.5 μg·mL^-1，定量限为2.5 μg·mL^-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 mL·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 2.5-100 μg·mL^-1. The average recoveries were 96.9%-101.0% with RSDs of 0.2%-3.0% at three spiked levels. The limit of detection was estimated to be 0.5 μg·mL^-1, and the limit of quatification was 2.5 μg·mL^-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.

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