Four sensitive, simple and specific methods were developed for the determination of desloratadine (DSL), a new antihistaminic drug in pharmaceutical preparations and biological fluids. Methods I and II are based on coupling DSL with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) in borate buffer of pH 7.6 where a yellow colored reaction product was obtained and measured spectrophotometrically at 485 nm (Method I). The same product could be measured spectrofluorometrically at 538 nm after excitation at 480 nm (Method II). Methods III and IV, on the other hand, involved derivatization of DSL with 2,4-dinitrofluorobenzene (DNFB) in borate buffer of pH 9.0 producing a yellow colored product that absorbs maximally at 375 nm (Method III). The same derivative was determined after separation adopting HPLC (Method IV). The separation was performed on a column packed with cyanopropyl bonded stationary phase equilibrated with a mobile phase composed of acetonitrile–water (60 : 40, v/v) at a flow rate of 1.0 ml min⁻¹ with UV detection at 375 nm. The calibration curves were linear over the concentration ranges of 0.5—6, 0.02—0.4, 1—10 and 1—30 μg ml⁻¹ for Methods I, II, III and IV, respectively. The lower detection limits (LOD) were 0.112, 0.004, 0.172 and 0.290 μg ml⁻¹, respectively, for the four methods. The limits of quantification (LOQ) were 0.340, 0.012, 0.522 and 0.890 μg ml⁻¹ for Methods I, II, III and IV, respectively. The proposed methods were applied to the determination of desloratadine in its tablets and the results were in agreement with those obtained using a reference method. Furthermore, the spectrofluorometric method (Method II) was extended to the in-vitro determination of the drug in spiked human plasma, with a mean percentage recovery (n=4) of 99.7±3.54. Interference arising from endogenous amino acids has been overcome using solid phase extraction. The proposed methods are highly specific for determination of DSL in the presence of the parent drug loratadine. A proposal for the reaction pathways is postulated.