Novel method for metalloproteins determination in human breast milk by size exclusion chromatography coupled to inductively coupled plasma mass spectrometry

ACOSTA, MARIANO; TORRES, SABIER; MARIÑO-REPIZO, LEONARDO; MARTINEZ, LUIS D.; GIL, RAÚL A.

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS

ELSEVIER SCIENCE BV

Año: 2018 vol. 158 p. 209 - 213

0731-7085

tLevels of essential metals in human breast milk (HBM) have been determined by different analyticaltechniques, but there is few woks about human whey milk fractions. However, the current trend lies inmetalloproteomic and identification of different metalloproteins. In this sense, native separative tech-niques (N-PAGE and SEC) coupled to ICP-MS provide us with valuable information. Besides it is necessarythe development of new methodologies in order to determine with accuracy and precision the profileof such metals and metalloproteins in the different whey protein fractions of HBM. Thus, the aim of thiswork was to develop a new method for metals and metalloproteins determination by SEC-ICP-MS in wheyprotein fractions of HBM. Human whey fractions were obtained of HBM samples by ultracentrifugation.Then, protein fractions of whey milk were separated by SEC coupled to ICP-MS for metalloproteins andMn, Co, Cu and Se quantification. Besides, protein profile of whey milk was determined by N-PAGE andcomputer assisted image analysis. SEC-ICP-MS results indicated that first and second protein fractionsshowed detectable levels of the Mn, Co, Cu, and Se. Protein profile determined by N-PAGE and imageanalysis showed that molecular weight of protein fractions ranged between 68,878?1,228,277 kDa. Inthis work, metalloproteins were analyzed by SEC coupled to ICP-MS, with adequate sensitivity and accu-racy. Our study has shown the presence of Mn, Co, Cu and Se bound to two protein fractions in whey milkof HBM. Metals levels analyzed were within the ranges reported in the literature.