超临界流体色谱法同时测定格列美脲片中手性和非手性杂质含量

Separation and determination of chiral and achiral impurities in glimepiride tablets by supercritical fluid chromatography

建立了一种超临界流体色谱法同时分离和测定格列美脲片中手性和非手性杂质的含量。本方法选用Waters TrefoilTMCEL1色谱柱(150 mm×3.0 mm,2.5μm);流动相A (CO_(2))-流动相B (甲醇-异丙醇,1∶1),梯度洗脱流速为1 mL·min^(-1);检测器波长为228 nm;柱温:30℃;背压:13.8 MPa;进样量:5μL。系统适用性溶液中出峰顺序依次为杂质Ⅳ、顺式异构体(杂质Ⅴ)、格列美脲、杂质Ⅲ、杂质Ⅰ和杂质Ⅱ,6个成分在6 min内完全分离,分离度依次为2.9、1.6、3.0、2.0、6.4。杂质Ⅰ~Ⅴ的检出限分别为0.17、0.10、0.06、0.15、0.10μg·mL^(-1);线性范围均在0.48~51.30μg·mL^(-1),平均加标回收率分别为99.9%、98.9%、102.1%、100.1%、96.3%(n=9)。11批次样品有关物质和含量测定结果与中国药典HPLC方法的测定结果基本一致。相较于中国药典两种HPLC方法,建立的超临界流体色谱方法能一次同时分离格列美脲及其5个杂质,具有前处理简单、极大幅度减少有机溶剂使用量、对环境友好、准确度高、重现性好等优点。本方法可同时测定格列美脲中手性和非手性杂质含量。

Separation and determination of chiral and achiral impurities in glimepiride tablets by supercritical fluid chromatography.Chiral and achiral impurities were separated on a ACQUITY UPC2 TrefoilTM CEL1 column(150 mm×3.0 mm,2.5μm)maintained at 30℃with the mobile phase containing a mixture of CO_(2) and methanol−isopropanol(1∶1)at 1 mL·min^(-1),and the detection wavelength was set at 228 nm.The back pressure was set at 13.8 MPa.The injection volume was 5μL.In the chromatogram of the system suitability solution,the peaks elute in the following order:impurityⅣ,impurityⅤ,glimepiride,impurityⅢ,impurityⅠand impurityⅡ.The six substances were separated successfully in 6 min using the proposed method with a resolution factor of 2.9,1.6,3.0,2.0,6.4.The impurityⅠ−Ⅴdetection limit(S/N=3)was 0.17,0.10,0.06,0.15,0.10μg·mL^(-1),respectively.Good linear relationship was established between the peak response and the concentration in the range of 0.48−51.30μg·mL^(-1)for all impurities.The spiked recovery of impurityⅠ−Ⅴwas found to be acceptable for 99.9%,98.9%,102.1%,100.1%,96.3%(n=9),respectively.The related substance and assay results of 11 sample batches are consistent with the results obtained using the HPLC method in the Chinese Pharmacopoeia.Compared to the two HPLC methods in the Chinese Pharmacopoeia,the established supercritical fluid chromatography method can simultaneously separate glimepiride and its 5 impurities in a single run,and it has the following advantages:simplified sample preparation,greatly reducing the volumes of organic solvents,environmentally friendly,high accuracy and good reproducibility.It can be employed for the quality control of the chiral and achiral impurities in glimepiride tablets.