APPLICATION OF DISPERSIVE LIQUID LIQUID MICROEXTRACTION FOR THE QUANTITATIVE ANALYSIS OF TRAMADOL AND METHADONE IN VITREOUS HUMOR SPECIMEN USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Background and Aim : Background: Non-medical use and abuse of opioids is becoming a major threat to public health.Many toxicologists attempt to setup innovate and efficient methods for the sample clean-up prior to toxicological analysis. Dispersive liquid-liquid microextraction (DLLME) is one of the efficient methods for the preparation of biological samples. Alternative matrices such as vitreous humor are good substitutes for conventional samples due to resistance to autolysis and contamination. Moreover alternative samples may provide additional information about the route of drug administration and timing of exposure in cases of limited, unavailable routine samples in burnt cadavers, cases with multiple trauma and also putrefied bodies. The aim of the present study was to develop a reliable DLLME method for the extraction of methadone and tramadol from vitreous humor samples.Methods : Methods: Ten mL of distilled water was added to 2 mL of vitreous humor sample. The pH of the experiment medium was adjusted to pH=10 using sodium hydroxide. Chloroform and methanol mixture was pushed to the prepared sample by force. After centrifugation, the chloroform phase was gathered and evaporated to dryness. The residue was dissolved in methanol and injected to HPLC.Results : Result: Results of the present study showed that DLLME method is a valid and sensitive sample preparation method. LOD and LOQ were 3 and 8 ng/mL for methadone respectively. Whereas LOD and LOQ were 6 and 16 ng/mL for tramadol.Conclusion : Conclusion: Sample clean-up and extraction methods are among the crucial steps in forensic toxicology analysis.

کلیدواژه ها:

Vitreous Humor ، DLLME ، Methadone ، Tramadol ، Forensic toxicology