RESUMEN
Urea is added as an adulterant to give milk whiteness and increase its consistency for improving the solid not fat percentage, but the excessive amount of urea in milk causes overburden and kidney damages. Here, an innovative sensitive methodology based on near-infrared spectroscopy coupled with multivariate analysis has been proposed for the robust detection and quantification of urea adulteration in fresh milk samples. In this study, 162 fresh milk samples were used, those consisting 20 nonadulterated samples (without urea) and 142 with urea adulterant. Eight different percentage levels of urea adulterant, that is, 0.10%, 0.30%, 0.50%, 0.70%, 0.90%, 1.10%, 1.30%, and 1.70%, were prepared, each of them prepared in triplicates. A Frontier NIR spectrophotometer (BSEN60825-1:2007) by Perkin Elmer was used for scanning the absorption of each sample in the wavenumber range of 10,000-4,000 cm-1, using 0.2 mm path length CaF2 sealed cell at resolution of 2 cm-1. Principal components analysis (PCA), partial least-squares discriminant analysis (PLS-DA), and partial least-squares regressions (PLSR) methods were applied for the multivariate analysis of the NIR spectral data collected. PCA was used to reduce the dimensionality of the spectral data and to explore the similarities and differences among the fresh milk samples and the adulterated ones. PLS-DA also showed the discrimination between the nonadulterated and adulterated milk samples. The R-square and root mean square error (RMSE) values obtained for the PLS-DA model were 0.9680 and 0.08%, respectively. Furthermore, PLSR model was also built using the training set of NIR spectral data to make a regression model. For this PLSR model, leave-one-out cross-validation procedure was used as an internal cross-validation criteria and the R-square and the root mean square error (RMSE) values for the PLSR model were found as 0.9800 and 0.56%, respectively. The PLSR model was then externally validated using a test set. The root means square error of prediction (RMSEP) obtained was 0.48%. The present proposed study was intended to contribute toward the development of a robust, sensitive, and reproducible method to detect and determine the urea adulterant concentration in fresh milk samples.
RESUMEN
Cows' butterfat may be adulterated with animal fat materials like tallow which causes increased serum cholesterol and triglycerides levels upon consumption. There is no reliable technique to detect and quantify tallow adulteration in butter samples in a feasible way. In this study a highly sensitive near-infrared (NIR) spectroscopy combined with chemometric methods was developed to detect as well as quantify the level of tallow adulterant in clarified butter samples. For this investigation the pure clarified butter samples were intentionally adulterated with tallow at the following percentage levels: 1%, 3%, 5%, 7%, 9%, 11%, 13%, 15%, 17% and 20% (wt/wt). Altogether 99 clarified butter samples were used including nine pure samples (un-adulterated clarified butter) and 90 clarified butter samples adulterated with tallow. Each sample was analysed by using NIR spectroscopy in the reflection mode in the range 10,000-4000 cm-1, at 2 cm-1 resolution and using the transflectance sample accessory which provided a total path length of 0.5 mm. Chemometric models including principal components analysis (PCA), partial least-squares discriminant analysis (PLSDA), and partial least-squares regressions (PLSR) were applied for statistical treatment of the obtained NIR spectral data. The PLSDA model was employed to differentiate pure butter samples from those adulterated with tallow. The employed model was then externally cross-validated by using a test set which included 30% of the total butter samples. The excellent performance of the model was proved by the low RMSEP value of 1.537% and the high correlation factor of 0.95. This newly developed method is robust, non-destructive, highly sensitive, and economical with very minor sample preparation and good ability to quantify less than 1.5% of tallow adulteration in clarified butter samples.
