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This study reports a simple and rapid high-performance liquid chromatographic (HPLC) method for the determination of the insecticide diazinon ( O , O -diethyl- O [2-isopropyl-6- methylpyridimidinyl] phosphorothioate), its metabolites diazoxon ( O , O -diethyl- O -2-isopropyl-6-methylpyridimidinyl phosphate) and 2-isopropyl-6-methyl-4-pyrimidinol, the insecticide chlorpyrifos ( O , O -diethyl- O [3,5,6-trichloro-2-pyridinyl] phosphorothioate) and its metabolites chlorpyrifos-oxon ( O , O -diethyl- O [3,5,6-trichloro-2-pyridinyl] phosphate), and TCP (3,5,6-trichloro-2-pyridinol) in rat plasma and urine samples. The method is based on using C 18  Sep-Pak cartridges for solid-phase extraction and HPLC with a reversed-phase C 18  column and programmed UV detection ranging between 254 and 280 nm. The compounds are separated using a gradient of 1% to 80% acetonitrile in water (pH 3.0) at a flow rate ranging between 1 and 1.5 mL/min in a period of 16 min. The limits of detection ranged between 50 and 150 ng/mL, and the limits of quantitation were 100 to 200 ng/mL. The average percentage recovery of five spiked plasma samples were 86.3 ± 8.6, 77.4 ± 7.0, 82.1 ± 8.2, 81.8 ± 8.7, 73.1 ± 7.4, and 80.3 ± 8.0 and from urine were 81.8 ± 7.6, 76.6 ± 7.1, 81.5 ± 7.9, 81.8 ± 7.1, 73.7 ± 8.6, and 80.7 ± 7.7 for diazinon, diazoxon, 2-isopropyl-6-methyl-4-pyrimidinol, chlorpyrifos, chlorpyrifos-oxon, and TCP, respectively. The relationship between the peak area and concentration was linear over a range of 200 to 2000 ng/mL. This method was applied in order to analyze these chemicals and metabolites following dermal administration in rats.

Determination of Diazinon, Chlorpyrifos, and Their Metabolites in Rat Plasma and Urine by High-Performance Liquid Chromatography