柱前衍生化RP-HPLC测定人血清白蛋白中美西律对映体

 目的建立美西律对映体在人血清白蛋白中的测定方法。方法采用手性衍生化试剂2,3,4,6-四-O-乙酰基-β-D-吡喃葡萄糖异硫氰酸酯(GITC)对美西律进行柱前手性衍生化。衍生产物以乙腈-0.01mol·L^-1磷酸二氢钾(pH4.5)缓冲液(1∶1)为流动相,用C₁₈色谱柱分离,以R-艾司洛尔为内标,流速为1.0mL·min^-1,检测波长为250nm。结果实验表明,R-美西律与S-美西律的分离度良好。日内、日间RSD均小于15%。美西律对映体在0.1～40.0mg·L^-1内呈线性关系,r=0.9995。结论该方法结果准确,重现性好,灵敏度高,可用于美西律对映体的蛋白结合研究。

Determination of Mexiletine Enantiomers in Human Serum Albumin after Derivatization with GITC by RP-HPLC

OBJECTIVE To develope enantiomeric separation method for mexiletine enantiomers in human serum albumin.METHODS The mexiletine enantiomers were separated on a C18 column after chiral derivatization with 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl isothiocyanate (GITC).The mobile phase consisted of acetonitrile-0.01 mol·L^-1 potassium dihydrogen phosphate buffer (pH 4.5) (1∶1) at the flow rate of 1.0 mL·min^-1.R-esmolol was used as the internal standard.The UV detection wavelength was set at 250 nm.RESULTS The mexiletine enantiomers and internal standard were separated on base-line.The intra-day and inter-day variations were less than 15%.The assay linearity was determined over the range of 0.1～40.0 mg·L^-1,r=0.999 5. CONCLUSION The method established is accurate,precision,sensitive and specific.The validated method has been used for protein binding studies of mexiletine enantiomers.