反向离子对色谱法测定格列吡嗪原料药中5-甲基-2-吡嗪羧酸的含量

Determination of 5-methyl-2-pyrazine carboxylic acid in glipizide by reverse ion pair chromatography

目的:建立格列吡嗪原料药中5-甲基-2-吡嗪羧酸的含量测定方法。方法:采用反向离子对色谱法。色谱柱为Agilent Eclipse XDB-C_(18),流动相:50 mmol·L^(-1)NaH_2PO_4溶液(含0. 05%四丁基氢氧化铵)-甲醇(梯度洗脱),流速为1. 0 m L·min^(-1),柱温为30℃,检测波长为276 nm,进样量为20μL。结果:5-甲基-2-吡嗪羧酸质量浓度线性范围为0. 038 54～1. 445 3μg·mL^(-1)(r=0. 999 9,n=5),检测限为0. 011 55μg·mL^(-1),定量限为0. 038 54μg·mL^(-1);平均回收率为100. 94%,RSD为1. 4%(n=9);系统精密度、重复性、中间精密度试验结果良好;对照品和供试品溶液8 h稳定; 3批原料药中5-甲基-2-吡嗪羧酸的量分别为0. 008 3%、0. 008 5%、0. 009 2%。结论:本方法操作简便、方法灵敏、结果准确可靠,可用于格列吡嗪原料药中微量杂质5-甲基-2-吡嗪羧酸的测定。

Objective: To establish a method for determination of 5-methyl-2-pyrazine carboxylic acid in glipizide. Methods: Reverse ion pair chromatography was used. The test was performed in Agilent Eclipse XDBC18 column under the gradient elution of 50 mmol· L^-1 NaH2PO4solution( containing 0. 05 % tetrabutylammonium hydroxide)-methanol. The flow rate was 1. 0 mL · min^-1,the column temperature was 30 ℃,the detection wavelength was as 276 nm,and the injection volume was 20 μL. Results: The linear range of 5-methyl-2-pyrazine carboxylic acid fell into 0. 038 54-1. 445 3 μg·mL^-1( r = 0. 999 9,n = 5),the limit of detection was 0. 011 55 μg·mL^-1,and the limit of quantitation was 0. 038 54 μg·mL^-1. The average recovery was 100. 94 % with RSD as 1. 4 %( n = 9). The system precision,repeatability and intermediate precision test results were good. The reference and test solutions were stable within 8 hours. The contents of 5-methyl-2-pyrazine carboxylic acid in 3 batches of API were 0. 008 3 %,0. 008 5 % and 0. 009 2 %. Conclusion: The established method is simple,sensitive,accurate and reliable. It can be used for determination of 5-methyl-2-pyrazine carboxylic acid in glipizide.