Methodology of Gliclazide Detection in Biological Samples

Aim: This research is dedicated to development of a methodology for the determination of gliclazide in biological samples for analytical diagnostics of lethal overdose caused by the drug. Materials and Methods: TLC method: chromatographic plates Merck silica gel 60 F254 10×10 cm in size, mobile phase - ethyl acetate, visualization reagent 1% vanillin solution or 5% solution of chloral hydrate. HPLC method: liquid chromatograph «Milichrom-A-02» with UV detecting (RF); Prontosil-120-5-C18-AQ reverse-phase column Ø2×75 mm in diameter, 5 μm grains (Bischoff Analysetechnik und Geräte GmbH, Germany); Gradient elution with a mixture of two eluents (eluent A – [0.2 M solution LiClO4 – 0.005 M solution of HClO4] and eluent B - acetonitrile); Wavelength 230 nm. Results: Methodology for the determination of gliclazide in biological samples is developed. It includes an effective method of gliclazide isolation from the objects of the study, specific and selective chromatographic detection methods and quantitative determination of toxicity in the extracts that had been received. It was established that while using TLC method gliclazide was assigned an Rf value of 0,48±0,01; detection limit is 3.0 μg. When applying HPLC method gliclazide holding time was 7.80 min (RSD = 0.13%), detection limit was 0.050 μg / ml, limit of the quantitative determination was 0.157 μg/ml, linearity of the procedure was in the range of 0.1-20.0 μg/ml. Under the developed conditions, gliclazide was determined at 17.48 ± 1.32 μg/ml (RSD = 6.06%). Conclusions: Results of the study indicate that methodology for the determination of gliclazide in biological samples that was developed can be used for analytical diagnostics of lethal overdose caused by the drug