高效液相色谱法测定沙格列汀原料药中3种差向异构体的含量  

Determination of three epimers in saxagliptin by high performance liquid chromatography

作　　者：董兴华 吴欢 张灵芝 曹杰永 DONG Xing-hua;WU Huan;ZHANG Ling-zhi;CAO Jie-yong(Scientifc Research&Experiment Center,Anhui University of Chinese Medicine,Heifei 230038,China;Hefei Cosource Pharmaceuticals Inc.,Heifei 230088,China;Anhui Province Key Laboratory of Pharmaceutical Preparation Technology and Application,Heifei 230012,China)

机构地区：[1]安徽中医药大学科研实验中心,合肥230038 [2]合肥合源药业有限公司,合肥230088 [3]药物制剂技术与应用安徽省重点实验室,合肥230012

出　　处：《药物分析杂志》2021年第6期1072-1077,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：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,5μm),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1000 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20μL,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0.2454~2.9445、0.2527~3.0322、0.2343~2.8110μg·mL^(-1)线性关系良好(r为0.9990~1.000),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 Shimpack VP-ODS(250 mm×4.6 mm,5μm)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 20μL,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 0.2454-2.9445μg·mL^(-1),0.2527-3.0322μg·mL^(-1),0.2343-2.8110μg·mL^(-1)(r was 0.9990-1.000),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.

关 键 词：沙格列汀 差向异构体 原料药 高效液相色谱法 梯度洗脱 方法验证 

分 类 号：R917[医药卫生—药物分析学]