Deconvolution of complex steady-state fluorescence spectra is a key subject in analytical fluorescence spectroscopy. The shape of the spectra is generated by the presence in the analyzed solution of a mixture of several fluorophores or by a single fluorophore found in different excited states. The spectra shape of most of the fluorophores is asymmetric, even in a homogeneous solution, where only one excited state is presumed to be present. Due to this, fluorescence spectra can be analyzed much better by a log-normal (LN) distribution than by a Gaussian one. Laurdan is a membrane fluorescent probe who has the advantage of detecting changes in bilayer phase properties. Laurdan typical red-shift (~ 50 nm) is observed during the phospholipid phase transition, and is originating from the probe sensitivity to its environment polarity. In this study, we propose a comparison between Gaussian and LN deconvolution of fluorescence spectra of Laurdan, inserted in large unilamellar vesicles, prepared from lipids with different hydrocarbon tails. We used a new parameter, namely, the difference of relative areas of the elementary peaks (∆Sr) to assess lipid membrane fluidity. We found that the results give a better characterization of the hydration level of the environment surrounding Laurdan.