柱前衍生化高效液相色谱法测定预混剂中硫酸安普霉素

目的: 建立预混剂中硫酸安普霉素的柱前衍生化-高效液相色谱检测方法. 方法: 采用2,4-二硝基氟苯(DNFB)为衍生化试剂,以Thermo C₁₈(150 mm×4.6 mm,5 μm)色谱柱为分析柱,乙腈-水(含0.5%乙酸)为流动相,梯度洗脱,流速1.0 mL·min^-1,检测波长350 nm,柱温30 ℃.对3批次的预混剂中硫酸安普霉素含量进行了测定. 结果: 硫酸安普霉素浓度在0~100 μg·mL^-1范围内与其衍生物峰面积呈良好的线性关系(r=0.999 8);平均回收率(n=6)为98.70%,RSD=1.3%.3批次的预混剂中硫酸安普霉素含量的测定结果(n=3)分别为24.42、24.39和24.63 mg·g^-1,RSD分别为0.27%、0.49%和1.0%. 结论: 经方法学验证,本法能准确测定预混剂中硫酸安普霉素的含量.

Objective: To establish a pre-column derivatization-HPLC for determination of apramycin sulfate in premix. Methods: The separation was performed on a Thermo C₁₈ column(150 mm×4.6 mm,5 μm)with pre-column derivatization by DNFB and the mobile phase was acetonitrile-water(containing 0.5% acetic acid)using gradient elution program at the flow rate of 1.0 mL·min^-1.The detection wavelength was 350 nm and the column temperature was 30 ℃.And the contents of apramycin sulfate in three batches of premix were determined. Results: A linearity between derivative peak area and apramycin sulfate concentration was achieved in the range of 0-100.0 μg·mL^-1(r=0.999 8).The average recovery(n=6)was 98.70%,RSD=1.3%.The contents of apramycin sulfate in three batches(n=3)were 24.42 mg·g^-1,24.39 mg·g^-1 and 24.63 mg·g^-1, respectively,the corresponding RSDs were 0.27%,0.49% and 1.0%. Conclusion: It is proved by methodology validation that the method can accurately determine the content of apramycin sulfate in the premix.

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