VALIDATED HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) METHOD FOR ANALYSIS OF METHAMPHETAMINE IN HUMAN URINE USING LIQUID-LIQUID EXTRACTION
Abstract
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Objective: Methamphetamine (also known as pil kuda or syabu) is one of the illicit drugs, which are common among students and teenagers. In
Malaysia, methamphetamine was found to be the most commonly abused amphetamine-type stimulants compared with other types of amphetamines.
Therefore, fast and reliable detection of these drugs is important in toxicology and doping control. Many extraction methods such as solid phase (SPE)
and solid phase micro extraction (SPME) have been used for extraction of methamphetamine in the biological fluid. However, these methods are
expensive and may not be available in many laboratories. The objective of this study was to develop an effective liquid-liquid extraction method for
methamphetamine in human urine.
Methods: Optimization of the extraction method was done by varying the types of solvents used to extract the drug from the urine. Percentage
recovery of methamphetamine extraction was investigated using a variety of extracting solvents (ethyl acetate, dichloromethane, diethyl ether, and
chloroform) and solvents system (chloroform:isopropanol [9:1] and chloroform:ethyl acetate:ethanol [3:1:1]). The analysis of the extracted drug was
done by high performance liquid chromatography (HPLC) equipped with fluorescence detector. Optimization of chromatographic conditions was also
done by varying the pHs of mobile phase and the composition of the mobile phase.
Results: The extracting solvent system that consists of chloroform:ethyl acetate:ethanol (3:1:1) gave the highest recovery, which is 87%. The
optimized HPLC condition used for the analysis was a mixture of acetonitrile and deionized water at pH 2.4 in the ratio of 15:85 v/v with a flow rate
of 0.6 mL/minute.
Conclusion: This study shows that liquid-liquid extraction method is reliable and cheaper alternative to SPE and SPME.
Keywords: Methamphetamine, Liquid-liquid extraction, High performance liquid chromatography.
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