A simple, selective and sensitive spectrophotometric method is described for the determination of sodium [2-[2, 6-dichlorophenyl [(4-amino n-acetyl) phenyl acetate]-amino] phenyl acetate] (DPAAPA) in rat blood sample. The assay was based on protein precipitation with mixture of methanol and ethyl acetate (1:2) then alkaline oxidation reaction with potassium ferricyanide. DPAAPA reacts with potassium ferricyanide in alkaline medium to give a yellow color after 5 min at 30 ˚C temperature having maximum absorbance at 451 nm. The reaction obeys Beer’s Law from 2-12 µg mL^‑1 for DPAAPA with molar absorptivity 4.2837×10⁴ L mol^-1 cm^-1. The recoveries of DPAAP from spiked blood samples were 99.20-101.20 µg mL^‑1 with relative standard derivation was less than 2.0%. The analytical recovery, sensitivity and accuracy of this assay method were adequate for characterization of the pharmacokinetics of oral administration of DPAAPA to Wistar albino rats and the assay has been successfully applied to provide pharmacokinetic data.