反相高效液相色谱法测定人血浆中格列吡嗪浓度

Determination of Glipizide in Human Plasma with RP-HPLC

目的 建立反相高效液相色谱法测定人血浆中格列吡嗪浓度的方法。方法 格列吡嗪血浆样品在酸性条件下以二氯甲烷 正己烷 (5 0∶5 0 )提取 ,以格列齐特为内标。色谱柱 :Nova pakC1 8柱 (4 6mm× 2 5 0mm ,4μm) ;流动相 :甲醇 0 0 3mol·L 1 磷酸二氢钾 (pH值 =3 0 ) (5 9∶41) ;流速 :0 8mL·min 1 ;检测波长 :2 2 8nm。结果 标准曲线线性范围 2 0～ 10 0 0 μg·L 1 ;回归方程 :Y =0 0 0 3 4X -0 5 12 ,r =0 9992 ,血浆中格列吡嗪最低检测限为 15 μg·L 1 。平均提取回收率为 (86 8± 5 7) % ,平均回收率为 (10 4 3± 4 3 ) % ,日内RSD≤ 2 9% ,日间RSD≤ 4 9%。结论 该方法具有良好的准确性、精密性和较高的灵敏度 ,适用于格列吡嗪的药动学研究和治疗监测。

Objective To set up an RP-HPLC method for th e determination of glipizide in human plasma. Method With gliclazide as the internal standard, glipiz ide was extracted from human plasma samples in an acidic milieu with a mixture o f dichloromethane -hexane(50∶50). A Nova-pak C 18 column(4.6 mm ×250 mm, 4 μm)served as the chrormatographic column. The mobile phase was meth anol - 0.03 mol·L -1 potassium dihydrogen phosphate (pH=3.0)(59 ∶41),and the flow rate , 0.8 mL·min -1 .The UV absorbance wavelength was 228 nm. Results The linear range of the standard curve was fro m 20 to 1 000μg·L -1. The regression equation: Y=0.003 4X-0.51 2, r=0.999 2. The assay limit for glipizide was 15 μg·L -1.The mean extraction recovery rate was (86.8±5.7)%. The mean analytical recovery ra te was (104.3±4.3)%. The intra-day and inter-day relative standard deviation s did not exceed 2.9% and 4.9%, respectively. Conclusion The method was shown to be stable, sensitiv e and accurate, and therefore suitable for the pharmacokinetic study of glipizid e.