HPLC-ELSD法测定氨基酸原料药中5种糖类杂质

Determination of five carbohydrate impurities in amino acid bulk drug by HPLC-ELSD

建立一种高效液相色谱-蒸发光散射检测(HPLC-ELSD)测定氨基酸原料药中5种糖类杂质的分析方法。利用离子交换树脂分离富集氨基酸样品中的残留糖类杂质,并采用Lichropher NH2色谱柱(4.6 mm×250 mm,5μm)进行分离,以乙腈-水为流动相进行梯度洗脱,漂移管温度为40℃;增益值为8;载气为氮气(压力为350 kPa)。经验证,果糖、葡萄糖、蔗糖、麦芽糖、乳糖检测限为20.8~75.0 mg/kg,定量限为96.2~238.8 mg/kg,5种糖在线性范围内线性关系良好,相关系数均大于0.999,加样回收率在84.9%~107.8%之间。该方法操作简便、灵敏度高、精密度好、准确可靠,可用于氨基酸原料药的残糖杂质分析。

An analytical method was developed for the determination of five carbohydrate impurities in amino acid drug substances by high performance liquid chromatography-evaporative light scattering detection(HPLCELSD).Sugar impurities in the amino acid sample were separated and enriched by cation exchange resin.A Lichropher NH2 column(4.6 mm×250 mm,5μm)was used for chromatographic separation,and a gradient elution was performed using acetonitrile-water as mobile phase.The drift tube temperature was 40 oC,the gain value was 8,and nitrogen(350 kPa)was auxiliary gas.Method validation results showed that the limits of detection for fructose,glucose,sucrose,maltose and lactose were in the range of 20.8-75.0 mg/kg and that the limits of quantitation were in the range of 96.2-238.8 mg/kg.Good linear relationship(r≥0.999)were in the linear range for the five sugars,and the recoveries ranged from 84.9%-107.8%.With easy operation,high sensitivity,good precision and reliable accuracy,the method can be used for analysis of residual sugar impurities in amino acid drug bulk drug.