A sensitive extraction method for the concentration of the triazines simazine, atrazine, and ametryn was developed and applied to a biological sample (urine). After protein precipitation with acetonitrile, a further purification using solid-phase extraction (SPE) was carried out. The samples were then analysed by high performance liquid chromatography (HPLC) using a C-18 column and an acetonitrile-water (40: 60, v/v) mobile phase. The pH of the mobile phase was adjusted to 9.0 with NH4OH, the flow-rate was 0.5 mL/min and UV detection was at 220 nm. Recovery values were satisfactory and the method developed can be used for monitoring urine samples of workers exposed to simazine, atrazine, and ametryn, in forensic, veterinary, and environmental toxicologies.