Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud Detection

Milk is a nutritionally rich food and a frequent target of economically motivated adulteration, particularly through substitution with lower-cost milk types. Over the past decade, significant progress has been made in the authentication of milk using advanced proteomic and chemometric approaches, wi...

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Main Authors: Achilleas Karamoutsios, Pelagia Lekka, Chrysoula Chrysa Voidarou, Marilena Dasenaki, Nikolaos S. Thomaidis, Ioannis Skoufos, Athina Tzora
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Foods
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Online Access:https://www.mdpi.com/2304-8158/14/15/2588
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author Achilleas Karamoutsios
Pelagia Lekka
Chrysoula Chrysa Voidarou
Marilena Dasenaki
Nikolaos S. Thomaidis
Ioannis Skoufos
Athina Tzora
author_facet Achilleas Karamoutsios
Pelagia Lekka
Chrysoula Chrysa Voidarou
Marilena Dasenaki
Nikolaos S. Thomaidis
Ioannis Skoufos
Athina Tzora
author_sort Achilleas Karamoutsios
collection DOAJ
description Milk is a nutritionally rich food and a frequent target of economically motivated adulteration, particularly through substitution with lower-cost milk types. Over the past decade, significant progress has been made in the authentication of milk using advanced proteomic and chemometric approaches, with a focus on the discovery and application of protein and peptide biomarkers for species differentiation and fraud detection. Recent innovations in both top-down and bottom-up proteomics have markedly improved the sensitivity and specificity of detecting key molecular targets, including caseins and whey proteins. Peptide-based methods are especially valuable in processed dairy products due to their thermal stability and resilience to harsh treatment, although their species specificity may be limited when sequences are conserved across related species. Robust chemometric approaches are increasingly integrated with proteomic pipelines to handle high-dimensional datasets and enhance classification performance. Multivariate techniques, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), are frequently employed to extract discriminatory features and model adulteration scenarios. Despite these advances, key challenges persist, including the lack of standardized protocols, variability in sample preparation, and the need for broader validation across breeds, geographies, and production systems. Future progress will depend on the convergence of high-resolution proteomics with multi-omics integration, structured data fusion, and machine learning frameworks, enabling scalable, specific, and robust solutions for milk authentication in increasingly complex food systems.
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spelling doaj-art-acea8c8c505d4e59ad2e7a3f042f6f2e2025-08-20T03:36:35ZengMDPI AGFoods2304-81582025-07-011415258810.3390/foods14152588Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud DetectionAchilleas Karamoutsios0Pelagia Lekka1Chrysoula Chrysa Voidarou2Marilena Dasenaki3Nikolaos S. Thomaidis4Ioannis Skoufos5Athina Tzora6Laboratory of Animal Health, Hygiene and Food Quality, School of Agriculture, University of Ioannina, 47100 Arta, GreeceLaboratory of Food Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771 Athens, GreeceLaboratory of Animal Health, Hygiene and Food Quality, School of Agriculture, University of Ioannina, 47100 Arta, GreeceLaboratory of Food Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771 Athens, GreeceLaboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771 Athens, GreeceLaboratory of Animal Science, Nutrition and Biotechnology, School of Agriculture, University of Ioannina, 47100 Arta, GreeceLaboratory of Animal Health, Hygiene and Food Quality, School of Agriculture, University of Ioannina, 47100 Arta, GreeceMilk is a nutritionally rich food and a frequent target of economically motivated adulteration, particularly through substitution with lower-cost milk types. Over the past decade, significant progress has been made in the authentication of milk using advanced proteomic and chemometric approaches, with a focus on the discovery and application of protein and peptide biomarkers for species differentiation and fraud detection. Recent innovations in both top-down and bottom-up proteomics have markedly improved the sensitivity and specificity of detecting key molecular targets, including caseins and whey proteins. Peptide-based methods are especially valuable in processed dairy products due to their thermal stability and resilience to harsh treatment, although their species specificity may be limited when sequences are conserved across related species. Robust chemometric approaches are increasingly integrated with proteomic pipelines to handle high-dimensional datasets and enhance classification performance. Multivariate techniques, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), are frequently employed to extract discriminatory features and model adulteration scenarios. Despite these advances, key challenges persist, including the lack of standardized protocols, variability in sample preparation, and the need for broader validation across breeds, geographies, and production systems. Future progress will depend on the convergence of high-resolution proteomics with multi-omics integration, structured data fusion, and machine learning frameworks, enabling scalable, specific, and robust solutions for milk authentication in increasingly complex food systems.https://www.mdpi.com/2304-8158/14/15/2588milkspecies identificationproteomicspeptide biomarkerscaseinswhey proteins
spellingShingle Achilleas Karamoutsios
Pelagia Lekka
Chrysoula Chrysa Voidarou
Marilena Dasenaki
Nikolaos S. Thomaidis
Ioannis Skoufos
Athina Tzora
Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud Detection
Foods
milk
species identification
proteomics
peptide biomarkers
caseins
whey proteins
title Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud Detection
title_full Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud Detection
title_fullStr Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud Detection
title_full_unstemmed Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud Detection
title_short Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud Detection
title_sort assessing milk authenticity using protein and peptide biomarkers a decade of progress in species differentiation and fraud detection
topic milk
species identification
proteomics
peptide biomarkers
caseins
whey proteins
url https://www.mdpi.com/2304-8158/14/15/2588
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