ABSTRACTa simple, rapid, and with high specificity method has been proposed for the determination of colchicine from biological samples as well from tablet. The method based on the reaction of the colchicine with quaternary ammonium salt in aqueous solution. The analytical parameters such as type of organic solvent (extraction solvent), volume of aqueous solvent, type of salt (ion-pairing agent), concentration of salt and effect pH have been investigated. The calibration graph was linear in the range of 0.3-6 µg mL -1 with the correlation coefficient of 0.999 under the optimum conditions. Good repeatability (R.S.D, %) 2.45, 1.33 and 2.36 for urine and 2.20, 1.97 and 2.79 for plasma in three spiked level 5, 2 and 0.5 µg mL -1 respectively and 2.26 for tablet was obtained.