A simple and rapid procedure for the determination of quinalphos in human whole blood using liquid-liquid extraction and high-performance thin-layer chromatography was developed and validated. Seven different organic solvents were tested for optimum extraction of quinalphos from spiked blood samples. The effect of pH on the extraction yield of quinalphos was also examined. An average recovery of 93.61% was achieved from diethyl ether solvent at pH 3. Chromatographic separation was performed on silica gel 60F254 plates using mobile phase n-hexane-acetone in the ration 9:1 (v/v). Densitometric detection was carried out at 325 nm in absorbance mode. The interference of other organophosphorus pesticides of forensic relevance was not observed. The linear regression analysis in spiked whole blood samples resulted in linear calibration plot in the range 1 to 100 μg mL-1 with r2 = 0.9981. Sensitivity was represented by LLOQ at 1 μg mL-1. The within-day precision and between-day precision ranged from 0.18 to 1.04%, and 0.14 to 0.79% with an overall average recovery of 91.06% at three concentrations 1, 10, and 50 μg mL-1. No significant decrease in the concentration of quinalphos was observed for samples under different storage conditions. Finally, the developed procedure was applied to postmortem blood samples obtained in three fatal cases of poisoning by quinalphos.