Penetapan Kadar Glibenklamid dan Metabolitnya 4-Trans-Hidroksiglibenklamid Secara Spektrofotometri Ultraviolet Mean Centering Of Ratio Spectra (Mcr) dalam Plasma Kelinci (Oryctolagus cuniculus)

Abstract

Glibenclamide is a second generation sulfonylurea derivative that is used as one of the oral antidiabetic drugs (ADO). is 4-trans-hydroxyglibenclamide is the main metabolite of glibenclamide which also has a hypoglycemic effect. Drugs and their metabolites have a high possibility of causing hypoglycemia, so it is important to monitor drug levels in the blood or Therapeutic Drug Monitoring (TDM). Drugs and their metabolites have similar chemical structures so they are in an overlapping state. The aim of this study was to develop a spectrophotometric method using mean centering ratio spectra (MCR) to determine the levels of glibenclamide and 4-trans-hydroxyglibenclamide in rabbit plasma simultaneously, without any separation. This study used six rabbits with an average body weight of ± 2 kg and blood samples were drawn every hour for four hours after drug administration. Determination with this method is carried out in stages, namely making each absorption spectrum, making a ratio absorption spectrum, and making an MCR absorption spectrum. Then the validity was tested based on the validation parameters, namely linearity, LOD, LOQ, accuracy and precision. Then, this method was applied to determine the levels of glibenclamide and 4-trans hydroxyglibenclamide in rabbit plasma. The results showed that the MCR application for assay was carried out at 301 nm and 336 nm for glibenclamide and 4-trans-hydroxyglibenclamide, with the highest concentrations of glibenclamide being 1,528 g/ml and 4-trans hydroxyglibenclamide 2,066 g/ml. The obtained validation parameters are eligible. The conclusion is that the MCR method meets the validation requirements, there has not been a breakdown reaction of glibenclamide into its metabolite 4- trans hydroxyglibenclamide at 1 hour after drug administration, and the bioanalytic MCR method can be used to determine the levels of glibenclamide and its metabolite 4-trans hydroxyglibenclamide in rabbit plasma.