A simple UV spectrophotometric differential derivatization method was performed for the simultaneous quantification of three aromatic amino acids of tryptophan, the polar tyrosine and phenylalanine TRP, TYR and PHE respectively. The avoidance of the time and reagents consuming steps of sample preparation or analyze separation from its bulk of interferences made the approach environmentally benign, sustainable and green. The linear calibration curves of differential second derivative were built at the optimum wavelength for each analyze (218.9, 236.1 and 222.5 nm) for PHE, TRP and TYR respectively. Quantification for each analyze was in the concentration range of (1.0– 45, 0.1–20.0 and 1.0– 50.0 μg/ml) at replicates of (n=3) with a reasonable linearity R2 value of (0.9983, 0.9970 and 0.9990) for PHE, TRP and TYR, respectively. The good repeatability of the approach was expressed by the low values of relative standard deviations which were less than 1.03%. Recovery study was implemented to confirm the accuracy of the method which was (97.35-99.65, 99.90-96.10 and 98.30-99.03) for PHE, TRP and TYR, respectively
