Proteomic Approach Based on MALDI-TOF MS To Detect Powdered Milk in Fresh Cow’s Milk

• Published in: Journal of agricultural and food chemistry 2013-02, Vol.61 (8), p.1609-1617
• Main Authors: Calvano, Cosima Damiana, Monopoli, Antonio, Loizzo, Pasqua, Faccia, Michele, Zambonin, Carlo
• Format: Article
• Language: English
• Subjects: adulterated products; Animal productions; Animals; Biological and medical sciences; Cattle; cheese milk; chemical composition; deamidation; digestion; dried milk; Food Contamination - analysis; Food industries; Fundamental and applied biological sciences. Psychology; gels; glycation; heat treatment; mass spectrometry; milk; Milk - chemistry; Milk and cheese industries. Ice creams; nutritive value; oxidation; peptides; Powders - chemistry; proteins; proteomics; Proteomics - methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; temperature; Terrestrial animal productions; two-dimensional gel electrophoresis; Vertebrates; whey
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf302999s

Milk and cheese are expensive foodstuffs, and their consumption is spread among the population because of their high nutritional value; for this reason they are often subjected to adulterations. Among the common illegal practices, the addition of powdered derivatives seems very difficult to detect because the adulterant materials have almost the same chemical composition of liquid milk. However, the high temperatures (180–200 °C) used for milk powder production could imply the occurrence of some protein modifications (e.g., glycation, lactosylation, oxidation, deamidation, dehydration). The modified proteins or peptides could then be used as markers for the presence of powdered milk. In this work, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was employed to analyze tryptic digests relevant to samples of raw liquid (without heat treatment), commercial liquid, and powdered cow's milk. Samples were subjected to two-dimensional gel electrophoresis (2-DE); differences among liquid and powder milk were detected at this stage and eventually confirmed by MALDI analysis of the in gel digested proteins. Some diagnostic peptides of powdered milk, attributed to modified whey proteins and/or caseins, were identified. Then, a faster procedure was optimized, consisting of the separation of caseins from milk whey and the subsequent in-solution digestion of the two fractions, with the advantage of obtaining almost the same information in a limited amount of time. Finally, analyses were carried out with the fast procedure on liquid milk samples adulterated with powdered milk at different percentages, and diagnostic peptides were detected down to 1% of adulteration level.