A simple and sensitive spectrophotometric method has been developed for the determination of palladium(II) using newly synthesized reagent 3,5-Dimethoxy-4-hydroxy benzaldehydeisonicotinoylhydrazone (DMHBIH) reagent in a micellar medium of Triton X-100 (neutral surfactant). Palladium(II) forms a bright yellow coloured water soluble complex with the reagent in acidic buffer medium pH 5.5. Beer’s law obeyed in the range 0.1064 to 2.1284 μg/mL of Pd(II) at λmax 382 nm. The molar absorptivity and Sandell’s sensitivity of coloured species are 2.44 x 104 L.mol-1.cm-1 and 0.0044 μg/cm2 respectively. Palladium(II) forms (M: L) 1:1 complex and stability constant of the complex is 2.2 x 107. The developed derivative spectrophotometric method was employed for the determination of palladium(II). This method has been satisfactorily applied for the determination of Pd(II) in alloy samples, hydrogenation catalyst samples and real water samples.