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Size exclusion and reversed-phase high-performance liquid chromatography/UV for routine control of thermal processing of cows' and donkey milk major proteins

Published online by Cambridge University Press:  16 March 2012

Carina Pinho
Affiliation:
REQUIMTE, Laboratório de Bromatologia e Hidrologia, Departamento de Ciências Químicas, Faculdade de Farmácia da Universidade do Porto, Portugal
Zita E. Martins
Affiliation:
REQUIMTE, Laboratório de Bromatologia e Hidrologia, Departamento de Ciências Químicas, Faculdade de Farmácia da Universidade do Porto, Portugal
Catarina Petisca
Affiliation:
REQUIMTE, Laboratório de Bromatologia e Hidrologia, Departamento de Ciências Químicas, Faculdade de Farmácia da Universidade do Porto, Portugal
Agata M. Figurska
Affiliation:
Medical University, 1 Chodźki St., 20-093 Lublin, Poland
Olívia Pinho
Affiliation:
REQUIMTE, Laboratório de Bromatologia e Hidrologia, Departamento de Ciências Químicas, Faculdade de Farmácia da Universidade do Porto, Portugal Faculdade de Ciências da Nutrição e Alimentação da Universidade do Porto, Portugal
Isabel M.P.L.V.O. Ferreira*
Affiliation:
REQUIMTE, Laboratório de Bromatologia e Hidrologia, Departamento de Ciências Químicas, Faculdade de Farmácia da Universidade do Porto, Portugal
*
*For correspondence; e-mail:[email protected]

Abstract

Cows' and donkey milks (raw and thermally processed) and respective whey were analysed for quantification of major proteins. Two different chromatographic approaches, size exclusion (SE-HPLC) and reversed-phase high performance liquid chromatography (RP-HPLC) both coupled to UV detection were used. Usefulness of these methods for routine control of the effect of thermal processing was evaluated. The external standard method was used to calibrate the SE-HPLC and RP-HPLC systems. Concerning quantification of β-lactoglobulin (β-lg), α-lactalbumin (α-la), lysozyme (lys), and total casein (cn), no significant differences between results obtained by SE-HPLC and by RP-HPLC (t-test, P>0·05) were observed for raw milks and whey. Heating of cows' milk promoted aggregation of denatured proteins as observed by SE-HPLC, whereas α-la and β-lg from donkey milk were stable to thermal processing at 100°C (5 min). Lys was quantified in donkey raw milk and whey however, in thermally processed donkey milk lys was denatured and could not be quantified by HPLC.

Type
Research Article
Copyright
Copyright © Proprietors of Journal of Dairy Research 2012

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References

Andrews, AT, Taylor, MD & Owen, AJ 1985 Rapid analysis of bovine milk proteins by fast protein liquid chromatography. Journal of Chromatography 348 177185CrossRefGoogle ScholarPubMed
Bonfatti, V, Grigoletto, L, Cecchinato, A, Gallo, L & Carnier, P 2008 Validation of a new reversed-phase high-performance liquid chromatography method for separation and quantification of bovine milk protein genetic variants. Journal of Chromatography A 1195 101106CrossRefGoogle ScholarPubMed
Bradford, MM 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye-binding. Analytical Biochemistry 72 248254CrossRefGoogle ScholarPubMed
Carretero, C, Trujillo, AJ, Mor-Mur, M, Pla, R & Guamis, B 1994 Electrophoretic study of casein breakdown during ripening of goat's milk cheese. Journal of Agricultural and Food Chemistry 42 15461550CrossRefGoogle Scholar
Carroccio, A, Cavataio, F, Montalto, G, D'Amico, D, Alabrese, L & Iacono, G 2000 Intolerance to hydrolysed cows' milk proteins in infants: clinical characteristics and dietary treatment. Clinical and Experimental Allergy 30 15971603CrossRefGoogle ScholarPubMed
Clément, P, Agboola, SO & Bencini, R 2006 A study of polymorphism in milk proteins from local and imported dairy sheep in Australia by capillary electrophoresis. LWT 39 6369CrossRefGoogle Scholar
Criscione, A, Consolo, V, Bordonaro, S, Guastella, AM, Saletti, R, Zuccaro, A, D'Urso, G & Marletta, D 2009 Donkey's milk protein fraction investigated by electrophoretic methods and mass spectrometric analysis. International Dairy Journal 19 190197CrossRefGoogle Scholar
Cunsolo, V, Saletti, R, Muccilli, V & Foti, S 2007 Characterization of the protein profile of donkey's milk whey fraction. Journal of Mass Spectrometry 42 11621174CrossRefGoogle ScholarPubMed
Curda, L, Belhácova, L, Uhrova, M, Stetina, J & Fukal, L 1997 Assessment of heat-induced denaturation of whey proteins. Journal of Chromatography A 772 231234CrossRefGoogle Scholar
D'Alessandro, AG, Martemucci, G, Jirillo, E & Leo, VD 2011 Major whey proteins in donkey's milk: effect of season and lactation stage. Implications for potential dietary interventions in human diseases. Immunopharmacology and Immunotoxicology 33(2) 259265CrossRefGoogle ScholarPubMed
De Block, J, Merchiers, M, Mortier, L, Braekman, A & Ooghe, W 2003 Monitoring nutritional quality of milk powders: capillary electrophoresis of the whey protein fraction compared with other methods. International Dairy Journal 13 8794CrossRefGoogle Scholar
Ferreira, IMPLVO 2005 Reversed-Phase-HPLC Methods for Separation of Milk Proteins: Application on Quality Control and Detection of Species Adulteration in Dairy Products in Encyclopedia of Chromatography DOI: 10.1081/E-ECHR-120041312, Taylor & FrancisCrossRefGoogle Scholar
Ferreira, IMPLVO 2007 Chromatographic separation and quantification of major human milk proteins. Journal of Liquid Chromatography and Related Technologies 30(4) 499507CrossRefGoogle Scholar
Ferreira, IMPLVO & Caçote, H 2003 Detection and quantification of bovine, ovine and caprine milk percentages in protected denomination of origin cheeses by reversed-phase high-performance liquid chromatography of beta-lactoglobulins. Journal of Chromatography A 1015 111118CrossRefGoogle ScholarPubMed
Ferreira, IMPLVO, Mendes, E & Ferreira, M 2001 HPLC/UV analysis of Proteins in Dairy Products using hydrophobic interaction Chromatographic column. Analytical Sciences 17 499501CrossRefGoogle ScholarPubMed
Giambra, IJ, Jäger, S & Erhardt, G 2010 Isoelectric focusing reveals additional casein variants in German sheep breeds. Small Ruminant Research 90 1117CrossRefGoogle Scholar
Goheen, SC & Gibbins, BM 2000 Protein losses in ion-exchange and hydrophobic interaction high-performance liquid chromatography. Journal of Chromatography A 890 7380CrossRefGoogle ScholarPubMed
Grupta, BB 1983 Determination of native and denatured milk proteins by high-performance size exclusion chromatography. Journal of Chromatography 282 463475CrossRefGoogle Scholar
Guo, HY, Pang, K, Zhang, XY, Zhao, L, Chen, SW, Dong, ML & Ren, FZ 2007 Composition, physiochemical properties, nitrogen fraction distribution, and amino acid profile of donkey milk. Journal of Dairy Science 90 16351643CrossRefGoogle ScholarPubMed
Haza, AI, Morales, P, Martin, R, Garcia, T, Anguita, G, Gonzalez, I & Sanz, B 1996 Development of monoclonal antibodies against Caprine αS2-Casein and their potential for detecting the substitution of ovine milk by caprine milk by an indirect ELISA. Journal of Agricultural and Food Chemistry 44 17561761CrossRefGoogle Scholar
Iacono, G, Carroccio, A, Cavataio, F, Montalto, G, Soresi, M, Balsamo, V 1992 Use of ass's milk in multiple food allergy. Journal of Pediatric Gastroenterology and Nutrition 14 177181Google Scholar
Miralles, B, Krause, I, Ramos, M & Amigo, L 2006 Comparison of capillary electrophoresis and isoelectric focusing for analysis of casein/caseinate addition in processed cheeses. International Dairy Journal 16 14481453CrossRefGoogle Scholar
Park, YW & Jin, YK 1998 Proteolytic patterns of Caciotta and Monterey Jack hard goat milk cheeses as evaluated by SDS–PAGE and densitometric analyses. Small Ruminant Research 28 263272CrossRefGoogle Scholar
Polidori, P, Beghelli, D, Mariani, P & Vincenzetti, S 2009 Donkey milk production: state of the art. Italian Journal of Animal Science 8(2) 677683CrossRefGoogle Scholar
Polidori, P & Vincenzetti, S 2010 Differences of protein fractions among fresh, frozen and powdered donkey milk. Recent Patents on Food, Nutrition and Agriculture 2 5660Google ScholarPubMed
Salimei, E, Fantuz, F, Coppola, R, Chiofalo, B, Polidori, P & Varisco, G 2004 Composition and characteristics of ass's milk. Animal Research 53 6778CrossRefGoogle Scholar
Santos, LHLMLM & Ferreira, IMPLVO 2007 Quantification of a-lactalbumin in human milk: Method validation and application. Analytical Biochemistry 362 293295CrossRefGoogle Scholar
Uniacke-Lowe, T, Huppertz, T & Fox, PF 2010 Equine milk proteins: chemistry, structure and nutritional significant. International Dairy Journal 20 609629CrossRefGoogle Scholar
Veloso, ACA, Teixeira, N, Peres, AM, Mendonça, A & Ferreira, IMPLVO 2004 Evaluation of cheese authenticity and proteolysis by HPLC and urea–polyacrylamide gel electrophoresis. Food Chemistry 87 289295CrossRefGoogle Scholar
Vincenzetti, S, Polidori, P, Mariani, P, Cammertoni, N, Fantuz, F & Vita, A 2008 Donkey's milk protein fractions characterization. Food Chemistry 106 640649CrossRefGoogle Scholar
Walstra, P 1990 On the stability of casein micelles. Journal of Dairy Science 73 19651979CrossRefGoogle Scholar