高效液相色谱-电喷雾式检测器法同时测定恩格列净中D-葡萄糖-δ-内酯与D-葡萄糖-γ-内酯  

Simultaneous determination of D-glucose-δ-lactone and D-glucose-γ-lactone in empagliflozin by high performance liquid chromatography-electrospray detector(HPLC-CAD)

作　　者：王琴琴 王蓉蓉 杜鹏程 游凤 尤雪岩 毕森 WANG Qinqin;WANG Rongrong;DU Pengcheng;YOU Feng;YOU Xueyan;BI Sen(Nanjing Huawei Pharmaceutical Technology Group Co.,Ltd.,Nanjing 210046,China)

机构地区：[1]南京华威医药科技集团有限公司,南京210046

出　　处：《中国药品标准》2022年第5期546-550,共5页Drug Standards of China

摘　　要：目的:建立高效液相色谱-电喷雾式检测法(HPLC-CAD)测定恩格列净中两种可能存在的葡萄糖内酯含量的方法。方法:采用Waters Xbridge BEH Amide色谱柱(4.6 mm×150 mm,5μm),流动相A为甲酸盐缓冲液(20 mmol·L^(-1)甲酸铵溶液,用甲酸调pH为3.0),流动相B为乙腈,梯度洗脱,流速为0.8 mL·min^(-1);柱温为30℃;电喷雾检测器(蒸发温度35℃,滤光片3.6 s,采集频率2 Hz),进样量20μL。结果:在该色谱条件下,D-葡萄糖-δ-内酯与D-葡萄糖-γ-内酯峰的检测限分别为1.37μg·mL^(-1)和1.46μg·mL^(-1)(均相当于供试品浓度的0.01%),定量限分别为4.12μg·mL^(-1)和4.37μg·mL^(-1)(均相当于供试品浓度的0.04%);D-葡萄糖-δ-内酯与D-葡萄糖-γ-内酯之和的回收率为97.4%(RSD为6.6%<10.0%);3批恩格列净中均未检出D-葡萄糖-δ-内酯与D-葡萄糖-γ-内酯。结论:方法可用于恩格列净中葡萄糖内酯的质控。Objective:To establish a high performance liquid chromatography-electrospray detection(HPLC-CAD)method for the determination of two possible glucolactones in empagliflozin.Methods:The Waters XBridge BEH amide column(4.6 mm×150 mm,5μm)was used.The mobile phase A was formate buffer(20 mmol·L^(-1)ammonium formate solution,adjust to pH 3.0 with formic acid),the mobile phase B was acetonitrile,Gradient elution.The flow rate was 0.8 mL·min^(-1).The column temperature was 30℃.Electrospray detector(evaporation temperature 35℃,filter 3.6s,acquisition frequency of 2 Hz),and injection volume 20μL.Results:Under the chromatographic conditions,the detection limits of D-glucose-δ-lactone and D-glucose-γ-lactone peaks were 1.37μg·mL^(-1)and 1.46μg·mL^(-1)(both were equivalent to 0.01%of the concentration of the test substance),and the quatitative limits were 4.12μg·mL^(-1)and 4.37μg·mL^(-1)(both equivalent to 0.04%of the concentration of the sample solution).The recovery rate of the sum of lactones was 97.4%(RSD was 6.6%<10.0%).D-glucose-δ-lactone and D-glucose-γ-lactone were not detected in the three batches of empagliflozin.Conclusion:The method can be used for the quality control of glucolactone in empagliflozin.

关 键 词：高效液相色谱-电喷雾式检测器法 恩格列净 D-葡萄糖-δ-内酯 D-葡萄糖-γ-内酯 葡萄糖内酯 

分 类 号：R921.2[医药卫生—药学]