目的: 建立沙格列汀原料药中差向异构体的定量分析方法。方法: 采用 Shim-pack VP-ODS 色谱柱（250 mm×4.6 mm,5 μm）,以 0.01 mol·L-1 磷酸二氢钾溶液（含 0.05 mol·L-1 高氯酸钠溶液）（取磷酸二氢 钾 1.3 g,一水合高氯酸钠 7.0 g,加水溶解并稀释至 1 000 mL）为流动相 A,乙腈 - 水 - 磷酸（90∶10∶0.1） 为流动相 B,梯度洗脱,流速为 1.0 mL·min-1,检测波长为 210 nm,进样量为 20 µL,柱温 35 ℃,采用加校正 因子的主成分外标法计算各差向异构体的含量,其中差向异构体 A、差向异构体 B 和差向异构体 C 的校正 因子分别为 1.07、1.07 和 1.09,均按 1.0 计。结果: 沙格列汀及差向异构体 A、差向异构体 B 和差向异构体 C 峰分离度良好,分离度均大于 1.5,差向异构体 A、差向异构体 B、差向异构体 C 分别在 0.245 4~2.944 5、 0.252 7~3.032 2、0.234 3~2.811 0 μg·mL-1 线 性 关 系 良 好（r 为 0.999 0~1.000）,回 收 率 分 别 为 96.8%、98.3%、97.4%（n=9）,进样精密度 RSD 均小于 2.0%,重复性 RSD 分别为 1.4%、1.6% 和 1.3%。检测下限分 别为 49.1、41.7、46.9 ng·mL-1。3 批样品中各差向异构体含量均小于检测下限。结论: 该方法简单,准确, 重复性好,可用于沙格列汀中差向异构体 A、差向异构体 B 和差向异构体 C 的测定。
Objective: To develop a method for the determination of three epimers in saxagliptin by high performance liquid chromatography. Methods: Chromatographic separation was carried out at 35 ℃,using Shim-pack VP-ODS（250 mm×4.6 mm,5 μm）column with mobile phases A（0.01 mol·L-1 potassium dihydrogen phosphate solution（containing 0.05 mol·L-1 sodium perchlorate））and B[acetonitrile-water-phosphoric acid （90∶10∶0.1）]. The flow rate was 1.0 mL·min-1,the injection volume was 20 μL,the detection wavelength was 210 nm. Principal component with correction factors as external standard was used to calculate the content of each epimer. The correction factors of epimer A,epimer B and epimer C were 1.07,1.07 and 1.09,respectively. Results: Saxagliptin,epimer A,epimer B and epimer C are well separated. The epimer A,epimer B and epimer C had good linear relationship in 0.245 4-2.944 5 μg·mL-1,0.252 7-3.032 2 μg·mL-1,0.234 3-2.811 0 μg·mL-1 （r was 0.999 0-1.000）,respectively. The recoveries of epimer A,epimer B and epimer C were 96.8%,98.3% and 97.4%（n=9）,respectively. The RSD of precisions were all less than 2.0%. The RSD of repeatability was 1.4%, 1.6% and 1.3%,respectively. The detection limit was 49.1,41.7 and 46.9 ng·mL-1,respectively. In the three batches of sample,the content of epimers did not exceed the detection limit（< LOD）. Conclusion: The method is simple,accurate and reproducible. It can be used for the determination of epimer A,epimer B and epimer C in saxagliptin.
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