衍生化HPLC法测定低分子肝素钠中过氧化氢残留

Determination of Hydrogen Peroxide in Low Molecular Heparin Sodium by Derivatization HPLC

目的:建立测定低分子肝素钠中痕量过氧化氢残留的方法。方法:用4-氨基安替比林和苯酚溶液在过氧化物酶的作用下与过氧化氢的衍生化反应,以Thermo ODS-2 hypersil C18色谱柱(4.6 mm×250 mm,5μm)在流动相(甲醇-水=60∶40)的条件下,分离检测过氧化氢的浓度。结果:该方法专属性好,过氧化氢的最低检测限为0.0302μg·m L^(-1),定量下限为0.1007μg·m L^(-1),在0.1~2.0μg·m L^(-1)的浓度范围内线性关系良好;平均回收率为97.58%,RSD为4.3%。结论:建立的方法可对低分子肝素钠中痕量过氧化氢残留进行测定。

Objective： To establish a method for the determination of hydrogen peroxide in low molecular heparin sodium. Methods： A Thermo ODS-2 hypersil C18 column with the mobile phase mixture of methanol-water（60∶40） was used for the determination of hydrogen peroxide concentration by HPLC with a derivation reaction involving peroxidase, phenol and 4-aminoantipyrine（4-AAP）. Results： The method had a good specificity. The LOD and LOQ of hydrogen peroxide were 0.0302 μg·m L^-1 and 0.1007 μg·m L^-1, respectively. The calibration curve was linear in the concentration range 0.1-2.0 μg·m L^-1（r=0.993, n=5） and the average recovery was 97.58%（RSD=4.3%）. Conclusion： The method established was accurate and effective for qualitative determination of hydrogen peroxide in low molecular heparin sodium.