We present an experimental test of a new spectral approach that is aimed at quantifying the relative concentrations of two chromophores that are contained in a defect embedded in a turbid medium. The basic steps of our spectral approach are (a) perform a linear tandem scan of the source and detector across the defect; (b) measure the spectral dependence of the maximum change induced by the defect in the scanned intensity; (c) identify a set of appropriate pairs of wavelengths (, ) at which such maximum intensity changes are the same; and (d) measure the reduced scattering coefficient spectrum of the background medium. For each wavelength pair (, ), we obtain a measurement of the relative concentrations of the two chromophores, where the only required parameters are the extinction coefficients of the two chromophores and the ratio of the background scattering coefficients at and . In a mixture of two test chromophores (blue food coloring dye and black India ink) contained in a 0.78-cm diameter cylinder, our spectral approach yielded relative concentrations values that were within 6% of their actual values. Although our paired-wavelength spectral approach is not generally applicable to any pair of chromophores, it is suitable for oxyhemoglobin and deoxyhemoglobin and is thus appropriate for oximetry of localized lesions in biological tissues.