柱前衍生化RP-HPLC测定米格列奈钙光学纯度

Enantiospecific Analysis of Mitiglinide Calcium by RP-HPLC with Precolumn Derivatization

关键词:  米格列奈钙  光学纯度  高效液相色谱法  柱前衍生化

Key words:  mitiglinide calcium  optical purity  HPLC  precolumn derivatization

摘要:

目的建立柱前衍生化-反相高效液相色谱法测定S-米格列奈钙的光学纯度。方法采用手性衍生化试剂S-萘乙胺(S-NEA)，以1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC·HCl)和1-羟基苯并三唑(HOBt)为偶联剂对样品S-米格列奈钙进行衍生化。衍生产物以乙腈- 0.01 mol·L-1磷酸二氢钾缓冲液(pH 2.5)(50∶50)为流动相，在Agilent Zorbax C18(250 mm × 4.6 mm，5 μm)色谱柱上进行分离，流速为1.5 mL·min-1，检测波长为225 nm，柱温为30 ℃。结果R-米格列奈在0.3～3.0 μg·mL-1内线性关系良好，检测限和最低定量下限分别0.1和0.3 μg·mL-1，平均回收率为102.4%，日内日间精密度考察中RSD均小于5%。结论该方法灵敏度高，衍生化产物稳定，重复性好，可用于S-米格列奈钙光学杂质的检测。

Abstract:

OBJECTIVE To establish a chiral precolumn derivatization method for optical purity test of mitiglinide calcium.METHODS The enantiomers of mitiglinide calcium were derivatized with a precolumn chiral derivatization reagent S-(-)-1-(1-naphthyl) ethylamine (S-NEA). The diastereoisomers produced were separated on an Agilent Zorbax C18 (250 mm × 4.6 mm，5 μm) column with a mobile phase composed of acetonitrile-0.01 mol·L-1 phosphate buffer (pH 2.5) (50∶50). The flow rate was set at 1.5 mL·min-1 and the detection wavelength was set at 225 nm. RESTULTS The method was linear in the range of 0.30-3.0 μg·mL-1 for the R-mitiglinide calcium. The lower limit of detection for R-mitiglinide calcium was 0.1 μg·mL-1 and the lower limit of quantification was 0.3 μg·mL-1. The inter- and intra- day precision (RSD) was below 5.0% and the average recovery was 102.4%. CONCLUSION The derivatives are stable in the mobile phase. The method is sensitive and suitable for the test of optical purity of mitiglinide calcium.