柱前衍生化高效液相色谱法分析氨基酸原料药中的4种还原糖类杂质

Analysis of four reducing sugar impurities in amino acid bulk drug by high performance liquid chromatography with precolumn derivatization

建立了一种利用柱前衍生高效液相色谱测定氨基酸原料药中4种还原糖类杂质的分析方法。氨基酸样品中的糖经阳离子交换树脂分离富集后用1-苯基-3-甲基-5-吡唑啉酮进行衍生,采用Waters Atlantis C18色谱柱(4. 6mm×250mm,5μm)进行分离,以0. 05 mol/L磷酸盐缓冲液-乙腈为流动相进行梯度洗脱。结果表明,麦芽糖、乳糖、葡萄糖、半乳糖在线性范围内线性关系良好,相关系数(R2)均大于0. 999,检出限为1. 1~2. 2 mg/kg,定量限为2. 9~6. 6 mg/kg,加标回收率在83. 0%~102. 7%之间。该方法可用于氨基酸原料药的质量控制。

The method was developed for the determination of four reducing sugar impurities in amino acid bulk drug by high performance liquid chromatography with precolumn derivatization. The saccharides in the amino acid were separated and enriched with cation exchange resin,and then derivatized with 1-phenyl-3-methyl-5-pyrazolinone. A Waters Atlantis C18 column( 4. 6 mm × 250 mm,5 μm) was used for gradient elution with0. 05 mol/L phosphate buffer-acetonitrile as mobile phase. The results showed good linear relationship in the linear range( R2> 0. 999) for maltose,lactose,glucose and galactose. The limits of detection were in the range of1. 1-2. 2 mg/kg and the limits of quantitation were in the range of 2. 9-6. 6 mg/kg. The recoveries ranged from 83. 0% to 102. 7%. The method can be used for the quality control of amino acid bulk drug.