title: Quantification of whey proteins by reversed phase-HPLC and effectiveness of mid-infrared spectroscopy for their rapid prediction in sweet whey.
abstract: In the dairy industry, membrane filtration is used to reduce the amount of whey waste and, simultaneously, to recover whey proteins (WP). The composition of WP can strongly affect the filtration treatment of whey, and rapid determination of WP fractions would be of interest for dairy producers to monitor WP recovery. This study aimed to develop mid-infrared spectroscopy (MIRS) prediction models for the rapid quantification of protein in sweet whey, using a validated rapid reversed phase (RP)-HPLC as a reference method. Quantified WP included α-lactalbumin (α-LA), β-lactoglobulin (β-LG) A and B, bovine serum albumin, caseinomacropeptides, and proteose peptone. Validation of RP-HPLC was performed by calculating the relative standard deviation (RSD) in repeatability and reproducibility tests for WP retention time and peak areas. Samples of liquid whey (n=187) were analyzed by RP-HPLC and scanned through MIRS to collect spectral information (900 to 4,000 cm(-1)); statistical analysis was carried out through partial least squares regression and random cross-validation procedure. Retention times in RP-HPLC method were stable (RSD between 0.03 and 0.80%), whereas the RSD of peak area (from 0.25 to 8.48%) was affected by WP relative abundance. Higher coefficients of determination in validation for MIRS model were obtained for protein fractions present in whey in large amounts, such as β-LG (0.58), total identified WP (0.58), and α-LA (0.56). Results of this study suggest that MIRS is an easy method for rapid quantification of detail protein in sweet whey, even if better resolution was achieved with the method based on RP-HPLC. The prediction of WP in sweet whey by MIRS might be used for screening and for classifying sweet whey according to its total and individual WP contents.
authors.affiliation: Department of Agronomy, Food, Natural Resources, Animals, and Environment (DAFNAE), University of Padova, Viale dell'Università 16, 35020 Legnaro (PD), Italy.
authors.forename: Alba
authors.initials: A
authors.lastname: Sturaro
authors.affiliation: Department of Agronomy, Food, Natural Resources, Animals, and Environment (DAFNAE), University of Padova, Viale dell'Università 16, 35020 Legnaro (PD), Italy. Electronic address: massimo.demarchi@unipd.it.
authors.forename: Massimo
authors.initials: M
authors.lastname: De Marchi
authors.affiliation: Department of Agronomy, Food, Natural Resources, Animals, and Environment (DAFNAE), University of Padova, Viale dell'Università 16, 35020 Legnaro (PD), Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Masi
authors.affiliation: Department of Agronomy, Food, Natural Resources, Animals, and Environment (DAFNAE), University of Padova, Viale dell'Università 16, 35020 Legnaro (PD), Italy.
authors.forename: Martino
authors.initials: M
authors.lastname: Cassandro
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D015203:Reproducibility of Results; D013055:Spectrophotometry, Infrared / Q000379:methods*; D000067816:Whey Proteins / Q000032:analysis*
chemical_list: D000067816:Whey Proteins
keywords: mid-infrared spectroscopy; reversed phase (RP)-HPLC; whey protein quantification