To account for variations of the overall concentrations of samples, NMR spectra were processed using Topspin 3.6.1 and Amix 3.9.13 software (Bruker, Biospin, Italy) for simultaneous visual inspection and the successive bucketing process. NMR spectra were segmented in rectangular buckets of fixed 0.04 ppm width and integrated. For aqueous cell fractions, spectral regions between 5.00–4.50 ppm were discarded because of the residual peak of water signal, while for CCM NMR spectra, signals of Hepes and DMSO and its affected neighbouring regions between 3.87–3.77 and 3.20–2.55 ppm were discarded. For lipid extract, 1H-NMR spectra—regions between 7.75–6.75, 3.48–3.25, and 1.70–1.44 ppm—were excluded before analysis due to the residual peaks of solvents (chloroform, methanol, and residual water signals). For each of the resulting data sets, a matrix was obtained, made of the bucketed 1H-NMR spectra values (columns) measured for each sample (rows). The mean centred and Pareto scaling procedures were applied to the data before multivariate statistical analyses, to attenuate the effect of dominant variables and noise while amplifying weak signals to the largest possible32,59,60.

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