A simple and sensitive spectrophotometric method for simultaneous determination of nitrite and nitrate was introduced. The method is based on the reaction of nitrite and nitrate with 2,4-xylenol as reagent in the concentrated sulfuric acid. Calibration curves were linear in the range of 0.2-20.0 and 0.5-7.0 mg L -1 for nitrite and nitrate, respectively. Due to the severe spectral overlapping of the reaction products of two anions with 2,4-xylenol, partial least squares (PLS) was used for simultaneous determination of nitrite and nitrate in the mixtures. For calibration and validation, 25 binary mixtures of nitrite and nitrate (17 calibration and eight external test samples) were designed by simplex lattice design with lattice degree of one. PLS models with Q 2 values of 0.988 and 0.976 were constructed for nitrite and nitrate, respectively. The validated models were used to analyze real water samples. With the recommended method, simultaneous determination of nitrite and nitrate in water samples is possible with satisfactory results.