Abstract:

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 serun using dichlromethane followed by derivatization with DNBF for 20 min at 100 °C. The solvent was then evaporated at 60 °C 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 350nm. Results Extraction recovery ranged from 75.9% to83.2%, 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. reaction of derivatization was reproducible.

Key words: Glimepiride, Glimepiride, pre column derivatization, pre column derivatization, DNBF, DNBF, serum concentration, serum concentration