目的 建立一种超临界流体色谱方法分离降糖药物格列美脲及其顺式异构体杂质。方法 采用ACQUITY UPC2 Trefoil CEL1色谱柱(3.0 mm×150 mm,2.5 μm),以超临界二氧化碳-甲醇(82∶18,V/V)为流动相,背压为13.8 MPa,进样量为4 μL,柱温为40 ℃,流速为1.5 mL·min-1,于228 nm波长处分离格列美脲及其顺式异构体。结果 格列美脲与其顺式异构体在5 min内分别被洗脱,分离度达1.6,顺式异构体杂质在1.5~15 μg·mL-1内线性关系良好,相关系数为0.999 9(n=6)。顺式异构体杂质的检测限与定量限分别为2 ng(S/N=3)和6 ng(S/N=10);格列美脲原料及片剂中顺式异构体杂质的平均加样回收率分别为99.4%和100.0% (n=9)。结论 与常规液相色谱方法相比,采用超临界流体色谱分离格列美脲及其顺式异构体杂质,可显著提高分离效率,缩短分离时间,大量减少烷烃类有机溶剂的使用量,重现性好,可有效用于格列美脲原料及制剂中顺式异构体的质量控制。
Abstract
OBJECTIVE To develop a supercritical fluid chromatography method for the separation of glimepiride and its cis-isomer. METHODS Glimepiride and its cis-isomer were separated on an ACQUITY UPC2 Trefoil CEL1 column (3.0 mm×150 mm,2.5 μm) maintained at 40 ℃ with mobile phase containing a mixture of CO2 and methanol (82∶18, V/V) at 1.5 mL·min-1. The detection wavelength was set at 228 nm,and the back pressure was set at 13.8 MPa. RESULTS The cis-isomer impurity and glimepiride were successfully separated in 5 min with a resolution factor of 1.6. Good linear relationship was established by plotting the peak area versus concentration in the range of 1.5-15 μg·mL-1 for cis-isomer impurity (r2=0.999 9, n=6). The quantitative limit (S/N=10) was 6 ng, and the detection limit (S/N=3) was 2 ng. The spiked recovery of cis-isomer impurity in glimepiride crude drug and tablets were 99.4% and 100.0%, respectively (n=9). CONCLUSION The proposed method shows high efficiency, accuracy,repeatability and stability. It could be employed for the quality control and stability research of the cis-isomer impurity in glimepiride crude drug and its tablet preparation.
关键词
格列美脲 /
顺式异构体 /
超临界流体色谱法 /
异构体分离
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Key words
glimepiride /
cis-isomer /
supercritical fluid chromatography /
isomer separation
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中图分类号：
R917
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参考文献
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[6] SONG Y, WANG D F, NIU L P, et al. Direct separation and quantitative determination of transglimepiride by high performance liquid chromatography using an amide type chiral stationary phase [J]. Chin J Anal Chem(分析化学), 2004, 32(4):435-438.
[7] ZHANG Q S, FANG K Z, LIU A L. Determination of the cis-isomer in Glimpiride Pills by RP-HPLC [J]. China Pharm (中国药师), 2012,15(8):1148-1150.
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脚注
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基金
国家科技重大专项资助(2017ZX09101001)
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