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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-0.01mol·L-1磷酸二氢钾缓冲液(pH2.5)(50∶50)为流动相,在AgilentZorbaxC18(250mm×4.6mm,5μm)色谱柱上进行分离,流速为1.5mL·min-1,检测波长为225nm,柱温为30℃。结果R-米格列奈在0.3～3.0μg·mL-1内线性关系良好,检测限和最低定量下限分别0.1和0.3μg·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。
OBJECTIVE To establish a pre-column derivatization-RP-HPLC method for the determination of optical purity of S-mitiglinide calcium. Methods The chiral derivatization reagent S-Naphthylethylamine (S-NEA) was used in this study. The 1- (3- dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC · HCl) Hydroxybenzotriazole (HOBt) is a coupling agent for derivatization of sample S-mitiglinide. The derivative was separated on a Agilent Zorbax C18 (250 mm × 4.6 mm, 5 μm) column with a mobile phase of acetonitrile-0.01 mol·L-1 potassium dihydrogen phosphate buffer (pH 2.5) (50:50) at a flow rate of 1.5 mL · min-1, detection wavelength was 225nm, column temperature was 30 ℃. Results R-mitoxantrone showed a good linearity in the range of 0.3-3.0 μg · mL-1. The detection limits and the minimum limits of quantification were 0.1 and 0.3 μg · mL-1, respectively, with an average recovery of 102.4%. Intra-day and inter-day precision The RSDs were less than 5%. Conclusion The method has high sensitivity, stable derivatization product and good reproducibility. It can be used for the detection of S-mitiglinide calcium optical impurities.