反相高效液相色谱法分离测定来氟米特3-甲基异构体及有关物质

Determination of 3-methyl-isomer and related substances in leflunomide by RP-HPLC

目的 建立高效液相色谱法分离测定来氟米特 3 甲基异构体及有关物质的方法。方法采用HypersilODS2 (4 6mm×2 5 0mm ,5 μm)色谱柱 ;流动相 :乙腈 甲醇 0 0 1mol·L-1磷酸二氢钾溶液 (2 5∶2 5∶5 0 ) ;流速 :1 0mL·min-1;检测波长为 2 6 1nm ;进样量 2 0 μL。 结果来氟米特中的 3 甲基异构体能很好地被分离。 3 甲基异构体在 0 2 6 4～ 1 32 μg·mL-1浓度内 ,峰面积与浓度线性关系良好 ,相关系数r=0 9995 ;平均回收率为 99 3% (n =9) ,RSD为 0 9%。来氟米特与有关物质能完全分离。结论本方法简便、准确、专属性强 ,可用于来氟米特 3

OBJECTIVE: To develop a RP-HPLC method for the determination of 3-methyl-isomer and its related substances in leflunomide. METHODS: The chromatographic analysis was performed on Hypersil ODS2 column (4.6 mm × 250 mm, 5 μ). The mobile phase consisted of acetonitrile-methanol- 0.01 mol&middotL-1 phosphate buffer(25:25:50). The flow rate was 1.0 mL&middotmin-1. The detection wavelength was at 261 nm, with injection volume 20 μL. RESULTS: The 3-methyl-isomer was separated from leflunomide. A good linearity was obtained over the range of 0.264∼1.32 μg&middotmL-1(r = 0.999 5). The average recovery of 3-methyl-isomer was 99.3% (n = 9) with RSD 0.9%. The related substances was separated from leflunomide. CONCLUSION: This method was simple, accurate, specific and will be used for the quality control of leflunomide.