Caracterização e identificação de adulterações em Whey Protein por espectroscopia de fluorescência estacionária e resolvida no tempo

Abstract

The objective of this work was to evaluate the characterization and efficacy of different fluorescence spectroscopy techniques in the detection of adulterations in formulations of Whey Protein Concentrate (WPC) powder, from its mixture with substances of different origins. Two different batches of WPC from the same supplier were studied. Adulterations were performed from the individual addition of Caffeine (Treatment 1), Creatine (Treatment 2) and Lactose (Treatment 3) at 30% (w / w) in WPC and subjected to stationary fluorescence spectroscopy and time-resolved fluorescence, using the excitation wavelengths of 275 and 335 nm. When excited at 275 nm, the samples showed an emission peak at approximately 335 nm, with an emission band around 380 nm, characteristic only in the sample containing caffeine, while lactose and creatine did not induce alterations in the WPC spectrum. When excited at 335 nm, the samples showed peak emission at 425 and 470 nm, without differentiation by simple observation of the spectrum. Euclidean distance analysis showed that, when excited at 275 nm, the complete spectra of treatments 2 and 3 were similar to Control 1 while Treatment 1 was different. Regarding the excitation at 335 nm, all spectra were similar. Principal Component Analysis (PCA) confirmed the differentiation of Treatment 1 at 275 nm but it was inconclusive with 335 nm excitation, however, it determined points of interest for spectra derivative studies. With the derivatives, it was possible to differentiate between Treatments 2 and 3 in the two wavelengths, focusing on specific wavelengths that can be decisive in the differentiation of adulterations. In relation to time resolved fluorescence spectroscopy, Treatment 1 demonstrated values of the mean emission lifetime higher over Treatments 2 and 3 at the two excitation wavelengths employed. The caffeine adulteration was performed in the Control 2 sample and a similar result was observed when compared to Control 1. In general, the application of stationary and time resolved fluorescence spectroscopy techniques allowed the characterization of the samples used in the study. In addition, it made possible the observation of differences between the control and adulterated samples, especially the one with caffeine added and excited at the wavelength 275 nm, with the help of mathematical tools. The results obtained here corroborate the fact that the techniques employed may be important in the detection of fraud in dairy products.