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A rapid, sensitive and selective HPLC separation with photodiode array detector was developed for the determination of the αketo acids from pharmaceutical tablet. α-keto acids (1) 3-methyl 2-oxobutyric acid (MKBA), (2) 3-methyl 2-oxovaleric acid (K3MVA), (3) 4-methyl-2-oxovaleric acid (K4MVA) and ( 4) Phenyl pyruvic acid (PPY) as derivatives with 4-dinitro 1,2phenylenediamine (NPD) eluted and separated from HPLC column Zorbax C-18 and the UV detection was carried out by photodiode array at 255 nm at pH 3. The isocratic elution was with methanol-water-acetonitrile (42:56:2V/V/V) with flow rate 0.9 ml/min. The standard curves were linear from 0.45-100 µg/ml and detection limits were within 0.1-0.266 µg/ml of each acid. The elution and separation of four keto acids was rapid within 10 min.The method was repeatable with relative standard deviation (RSD) within 0.1-2.9% for each of the α-keto acids. The method was applied for determination of α-keto acids from a pharmaceutical preparation. The result was further confirmed by standard addition technique.