A new spectrophotometric method was developed for the simultaneous of ternary mixtures of nitrophenol isomers, without prior separation steps. This method is called the first derivative of the density ratio spectra. All factors affecting on the sensitivity were optimized and the linear dynamic range for determination of nitrophenol isomers was found. The method made use of a derivative of the double-divisor-ratio spectra of optical density. In this case, the linear determination ranges are 1.0-25.0 mg mL-1 for m-nitrophenol, 1.0-25.0 mg mL-1 for o-nitrophenol and 1.0-15.0 mg mL-1 for p-nitrophenol. The RMSEP for m-nitrophenol, o-nitrophenol and p-nitrophenol by proposed method was 0.4907, 0.4779 and 0.2068, respectively. The method developed in this paper was rapid, easy to apply, not expensive and it was suitable for analyzing to overlapping signals of compounds in their mixtures without any chemical pre-treatment and also, the proposed method was satisfactorily applied to the rapid simultaneous determination of m-nitrophenol, o-nitrophenol and p-nitrophenol in synthetic and water samples.