고속액체크로마토그라피법을 이용한 사람 혈장 중 알리벤돌(Alibendol)의 정량 및 검증

Determiniation and Validation of Alibendol using High Pressure Liquid Chromatography in Human plasma

The aim of this study was to develop and validate for determination of alibendol in human plasma by HPLC method. After precipitation of 500 μl plasma samples by 50% methanol 50 μl and 60% perchloric acid 30 μl and the supernatant 50 μl was injected into HPLC. The assay was performed isocratically using 10 mM potassium phosphate (pH 3.0) and acetonitrile (80 : 20, v/v) as mobile phase. The C18 column (particle size 3.5 μm, 4.6×50 mm, Zorbax Eclipse) was used as a solid phase. The mobile phase was delivered at a flow-rate of 1.7 ml/min, detection was by ultraviolet absorption at 232nm and concentrations were calculated on the basis of peak areas. In these conditions, alibendol can be separated from ethylparaben, the internal standard, and endogenous substances. The retention times of alibendol and ethylparaben were just about 2.6 and 3.5 minutes, respectively. This rapid HPLC method was validated by examining the precision and accuracy for inter- and intra-day analysis. The standard curve was linear (R2=1.0000) over the concentration range of 0.05~20 μg/ml. The inter-day relative standard deviation (R.S.D.) and accuracy were 0.2~12.2% and 94.4~101.2% (82.7% at the lower limit of quatitation). The intra-day R.S.D. and accuracy were 0.1~11.8% and 98.8~102.5%, respectively. The method was successfully applied to the determination of alibendol in plasma for a pharmacokinetic study.