Eleven free amino acids were identified in the grass weed loose silky-bent (Apera spica-venti) after exposure to four rates of the thiocarbamate herbicide prosulfocarb. A new HPTLC method was developed for quantification of amino acids in raw plant extracts. A pattern of significant changes in the content of glutamine, glycine, alanine, and proline due to exposure were detected. The content of all four amino acids increased at increasing exposure rates. Quantitative analysis 7 and 21 days after herbicide application showed that over time the amount of proline increased, the alanine and glycine content decreased, and that at high exposure rates the amount of glutamine increasing exposure rates. A significant correlation between the changes in the content of the four amino acids and the visual effects and the biomasses were found.