We investigated the pH dependence of the fluorescence spectra of ADIFAB (FFA Sciences), a probe used for the quantification of free fatty acids (FFA). Data reports the change in the emission peak of ADIFAB and in the affinity for FFA as a function of pH. An algorithm based on spectral deconvolution allowed to correct ADIFAB fluorescence spectra for the spectroscopic effect caused by pH. Kd values were calculated at each pH based on a calibration with oleic acid. This method allows estimating FFA concentration by ADIFAB in media at different pH. The current data are related to the research article "Phospholipid components of the synthetic pulmonary surfactant CHF5633 probed by fluorescence spectroscopy" (Faggiano et al., 2018) .
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