高效液相色谱测定依达拉奉中苯肼残留方法的优化

Optimization of HPLC determination of phenylhydrazine residues in edaravone

目的优化HPLC测定依达拉奉中苯肼残留的方法。方法采用HPLC法,以Inertsil C18（150mm×4.6 mm,5μm）为色谱柱,以0.05 mol·L-1磷酸二氢铵溶液（用磷酸调pH至3.5）-甲醇（65：35）为流动相,流速为1.0 mL·min-1,柱温30℃,检测波长为226 nm,进样量为20μL,以0.05 mol·L-1磷酸二氢铵溶液-甲醇（65：35）（用磷酸调pH至2.0）作为溶剂。结果苯肼在0.17~0.68μg·mL-1与峰面积线性关系良好（r=0.999 9,n=6）,平均回收率〉95%（n=3）,苯肼溶液在12 h内稳定,最低检测限为0.34 ng。结论本测定方法可靠、简便、结果准确、稳定性好,适于该制剂中苯肼残留的测定。

Objective To establish an HPLC method to determine phenylhydrazine residues in edaravone. Methods The ingredient was separated on an Inertsil C18（150 mm×4.6 mm, 5 μm） column. The mobile phase was 0.05 mol·L- 1 ammonium acetate（adjusted pH to 3.5 by phosphoric acid）-methanol（65 ：35）. The fl ow rate was 1.0 mL·min- 1. The column temperature was maintained at 30 ℃, the detection wavelength was 226 nm, the injection volume was 20 μL. Ammonium acetate（0.05 mol·L- 1）-methanol（65 ：35）（adjusted pH to 2.0 by phosphoric acid） was used as the solvent. Results The linearity of phenylhydrazine was at 0.17- 0.68 μg·mL- 1（r = 0.999 9, n= 6）, the average recovery was more than 95%（n = 3）, the test solution was stable in 12 h, and the detection limit was 0.34 ng. Conclusion This method is simple, reliable, accurate, stable, and is suitable for the determination of phenylhydrazine residues in edaravone.