摘要
Aim A simple, sensitive and rapid RP HPLC method with pre column derivatization has been developed for the determination of sulphonylurea glimepiride in dog serum. Methods The sulphonylurea glimepiride was extracted from the dog serum using dichlromethane followed by derivatization with DNBF for 20 min at 100℃. The solvent was then evaporated at 60℃ under nitrogen, and the residue was taken up in 100 μL of mobile phase consisting of acetonitrile water (75∶30, v/v). The separation was performed on a Hypersil BDS C18 column with a flow rate of 0 8 mL·min -1 , and the ultraviolet detector wavelength was set at 350 nm. Results Extraction recovery ranged from 75.9% to 83.2%, and methodological recovery was between 96.5% and 109.3%. Within day RSD ranged from 1.5% to 6.3%, and inter day RSD was between 2 9% and 14.8%. The method showed good linearity (R=0.9998). Conclusion The method was simple, convenient and sensitive. The reaction of derivatization was reproducible.
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 。
