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目的 建立测定狗血清中格列美脲浓度的柱前衍生化反相高效液相色谱法。方法 血清样品用二氯甲烷萃取后挥干 ,以DNBF试剂衍生化 ,所得产物氮气挥干 ,用流动相 (含 1%冰醋酸 )溶解进样 ;色谱柱为HypersilBDSC18柱 ( 15 0mm×4 .6mm ,5 μm) ;流动相为乙腈 -水 ( 75∶30 ,V V) ,流速 0 .8mL·min- 1 ,检测波长 35 0nm。结果 血清中格列美脲浓度在 0 .0 2 8-0 .6 4 8μg·mL- 1 范围内线性关系良好 (R =0 .9998) ;高、中、低 3种浓度萃取回收率分别为 75 .9% - 83.2 % ;方法回收率在96 .5 % - 10 9.3%之间 ;高、中、低三种浓度日内精密度RSD分别为 1.5 %、2 3%、6 .3% ,日间精密度RSD分别为 2 9%、7.4 %、14 .8%。结论 本法简便易行、灵敏度高 ,衍生化反应速度适中 ,适用于格列美脲血药浓度测定及药代动力学研究
Objective To establish a pre-column derivatization reversed-phase high performance liquid chromatography (HPLC) for the determination of glimepiride in dog serum. Methods The serum samples were extracted with dichloromethane, evaporated, and derivatized with DNBF reagent. The resulting product was evaporated to dryness and dissolved in mobile phase (containing 1% glacial acetic acid). The column was Hypersil BDSC18 (150 mm × 4.6 mm , 5 μm). The mobile phase was acetonitrile-water (75:30, VV), the flow rate was 0.8 mL · min-1 and the detection wavelength was 35 0 nm. Results The concentration of glimepiride in serum ranged from 0.082 to 0.648 μg · mL-1 (R = 0.9998). The recoveries of high, medium and low concentrations were 75.9% -83.2%. The method recoveries ranged from 96.5% to 10 9.3%. The RSDs of daily high, medium and low concentrations were 1.5%, 23%, 6.3% The precision of RSD was 29%, 7.4%, 14.8% respectively. Conclusion This method is simple and easy to operate with high sensitivity and moderate derivatization reaction rate and is suitable for the determination of glimepiride and its pharmacokinetics