Abstract
BackgroundOpioids abuse and related deaths are increasing in the world. Therefore, the design of new analytical methods for detection of opioids in biological samples is necessary for clinical and forensic settings.MethodsIn this study, dispersive liquid-liquid microextraction (DLLME) combined with high-performance liquid chromatography with photo diode array detector (HPLC-PDA), as a new and sensitive method were examined for the extraction and determination of morphine, codeine and methadone in postmortem urine samples. Effective factors on DLLME were optimized. The extracts were analyzed by HPLC-PDA using a Eurospher® C18 column (250 mm × 4.6 mm, particle size: 5 μm).ResultsThe volumes of chloroform as the extraction solvent and acetone as the dispersive solvent were selected 300 μl and 500 μl, respectively. The optimum pH 9.8 and extraction time was 0.5 min were selected. Under optimum condition, the enrichment factor and the recovery of morphine, codeine, and methadone spiked into postmortem urine samples were in the range of 175–215.8 and 87.5–107.9%, respectively. Calibration curves for each analyte are linear in the range of 0.5–100 μg ml− 1. Limit of detection (LOD) for the analytes was in the range of 10–25 μg l− 1. Finally, the proposed method was successfully applied to 50 postmortem urine samples for determination of the opioids.ConclusionsThe proposed method is an easy, fast, low cost and efficient for the extraction and determination of opioids in postmortem urine samples and should be considered as analytical method for determination of opioids in forensic and clinical toxicology labs.
Highlights
Opioids abuse and related deaths are increasing in the world
Analysis of the opioids in biological samples has been considered as an important issue in the forensic and clinical toxicology (Gergov et al 2009; Shamsipur and Fattahi 2011)
In this study, we optimized a dispersive liquid-liquid microextraction (DLLME)-HPLCPAD for the extraction and determination of morphine, codeine, and methadone in postmortem urine samples
Summary
Chemicals Standard morphine, codeine and methadone were obtained from Darou Pakhsh Pharmaceutical Co. (Tehran, Iran). Optimization of DLLME Affecting factors the DLLME procedure including the type and volume of extraction solvent, type and volume of disperser solvent, pH and extraction time were optimized in this study Optimization of these factors was done by using postmortem blank urine samples spiked with morphine, codeine, and methadone. Accuracy, enrichment factor, recovery, and relative recovery Inter-day and intra-day precisions method, and the enrichment factors and recovery for morphine, codeine and methadone were studied by extracting the spiked blank postmortem urine samples with 2, 10 and 30 μg ml− 1 concentrations. The relative recovery was studied by extracting the spiked postmortem urine samples (with suspected drug abuse) with two concentrations of morphine, codeine, and methadone (5 and 20 μg ml− 1).
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