Integrated Gel Electrophoresis and Mass Spectrometry Approach for Detecting and Quantifying Extraneous Milk in Protected Designation of Origin Buffalo Mozzarella Cheese
Ensuring the authenticity of Mozzarella di Bufala Campana (MdBC), a Protected Designation of Origin (PDO) cheese, is essential for regulatory enforcement and consumer protection. This study evaluates a multi-technology analytical platform developed to detect adulteration due to the addition of non-b...
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MDPI AG
2025-03-01
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| author | Sabrina De Pascale Giuseppina Garro Silvia Ines Pellicano Andrea Scaloni Stefania Carpino Simonetta Caira Francesco Addeo |
| author_facet | Sabrina De Pascale Giuseppina Garro Silvia Ines Pellicano Andrea Scaloni Stefania Carpino Simonetta Caira Francesco Addeo |
| author_sort | Sabrina De Pascale |
| collection | DOAJ |
| description | Ensuring the authenticity of Mozzarella di Bufala Campana (MdBC), a Protected Designation of Origin (PDO) cheese, is essential for regulatory enforcement and consumer protection. This study evaluates a multi-technology analytical platform developed to detect adulteration due to the addition of non-buffalo milk or non-PDO buffalo milk in PDO dairy buffalo products. Peripheral laboratories use gel electrophoresis combined with polyclonal antipeptide antibodies for initial screening, enabling the detection of foreign caseins, including those originating outside the PDO-designated regions. For more precise identification, Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) differentiates species by detecting proteotypic peptides. In cases requiring confirmation, nano-liquid chromatography coupled to electrospray tandem mass spectrometry (nano-LC-ESI-MS/MS) is used in central state laboratories for the highly sensitive detection of extraneous milk proteins in PDO buffalo MdBC cheese. On the other hand, analysis of the pH 4.6 soluble fraction from buffalo blue cheese identified 2828 buffalo-derived peptides and several bovine specific peptides, confirming milk adulteration. Despite a lower detection extent in the pH 4.6 insoluble fraction following tryptic hydrolysis, the presence of bovine peptides was still sufficient to verify fraud. This integrated proteomic approach, which combines electrophoresis and mass spectrometry technologies, significantly improves milk adulteration detection, providing a robust tool to face increasingly sophisticated fraudulent practices. |
| format | Article |
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| institution | DOAJ |
| issn | 2304-8158 |
| language | English |
| publishDate | 2025-03-01 |
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| series | Foods |
| spelling | doaj-art-df1aae871d87415cb5bfff10a99138922025-08-20T03:08:55ZengMDPI AGFoods2304-81582025-03-01147119310.3390/foods14071193Integrated Gel Electrophoresis and Mass Spectrometry Approach for Detecting and Quantifying Extraneous Milk in Protected Designation of Origin Buffalo Mozzarella CheeseSabrina De Pascale0Giuseppina Garro1Silvia Ines Pellicano2Andrea Scaloni3Stefania Carpino4Simonetta Caira5Francesco Addeo6Proteomics, Metabolomics & Mass Spectrometry Laboratory, Institute for the Animal Production System in the Mediterranean Environment, National Research Council, 80055 Portici, ItalyDepartment of Agriculture, University of Naples “Federico II”, 80055 Portici, ItalyCentral Inspectorate for Fraud Repression and Quality Protection of the Agrifood Products and Food, Ministry of Agricultural, Food and Forestry Policies, 06128 Perugia, ItalyProteomics, Metabolomics & Mass Spectrometry Laboratory, Institute for the Animal Production System in the Mediterranean Environment, National Research Council, 80055 Portici, ItalyCentral Inspectorate for Fraud Repression and Quality Protection of the Agrifood Products and Food, Ministry of Agricultural, Food and Forestry Policies, 00187 Roma, ItalyProteomics, Metabolomics & Mass Spectrometry Laboratory, Institute for the Animal Production System in the Mediterranean Environment, National Research Council, 80055 Portici, ItalyDepartment of Agriculture, University of Naples “Federico II”, 80055 Portici, ItalyEnsuring the authenticity of Mozzarella di Bufala Campana (MdBC), a Protected Designation of Origin (PDO) cheese, is essential for regulatory enforcement and consumer protection. This study evaluates a multi-technology analytical platform developed to detect adulteration due to the addition of non-buffalo milk or non-PDO buffalo milk in PDO dairy buffalo products. Peripheral laboratories use gel electrophoresis combined with polyclonal antipeptide antibodies for initial screening, enabling the detection of foreign caseins, including those originating outside the PDO-designated regions. For more precise identification, Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) differentiates species by detecting proteotypic peptides. In cases requiring confirmation, nano-liquid chromatography coupled to electrospray tandem mass spectrometry (nano-LC-ESI-MS/MS) is used in central state laboratories for the highly sensitive detection of extraneous milk proteins in PDO buffalo MdBC cheese. On the other hand, analysis of the pH 4.6 soluble fraction from buffalo blue cheese identified 2828 buffalo-derived peptides and several bovine specific peptides, confirming milk adulteration. Despite a lower detection extent in the pH 4.6 insoluble fraction following tryptic hydrolysis, the presence of bovine peptides was still sufficient to verify fraud. This integrated proteomic approach, which combines electrophoresis and mass spectrometry technologies, significantly improves milk adulteration detection, providing a robust tool to face increasingly sophisticated fraudulent practices.https://www.mdpi.com/2304-8158/14/7/1193buffalo milk adulterationgel electrophoresisMALDI-TOF-MSnano-LC-ESI-MS/MSpeptidomicsproteomics |
| spellingShingle | Sabrina De Pascale Giuseppina Garro Silvia Ines Pellicano Andrea Scaloni Stefania Carpino Simonetta Caira Francesco Addeo Integrated Gel Electrophoresis and Mass Spectrometry Approach for Detecting and Quantifying Extraneous Milk in Protected Designation of Origin Buffalo Mozzarella Cheese Foods buffalo milk adulteration gel electrophoresis MALDI-TOF-MS nano-LC-ESI-MS/MS peptidomics proteomics |
| title | Integrated Gel Electrophoresis and Mass Spectrometry Approach for Detecting and Quantifying Extraneous Milk in Protected Designation of Origin Buffalo Mozzarella Cheese |
| title_full | Integrated Gel Electrophoresis and Mass Spectrometry Approach for Detecting and Quantifying Extraneous Milk in Protected Designation of Origin Buffalo Mozzarella Cheese |
| title_fullStr | Integrated Gel Electrophoresis and Mass Spectrometry Approach for Detecting and Quantifying Extraneous Milk in Protected Designation of Origin Buffalo Mozzarella Cheese |
| title_full_unstemmed | Integrated Gel Electrophoresis and Mass Spectrometry Approach for Detecting and Quantifying Extraneous Milk in Protected Designation of Origin Buffalo Mozzarella Cheese |
| title_short | Integrated Gel Electrophoresis and Mass Spectrometry Approach for Detecting and Quantifying Extraneous Milk in Protected Designation of Origin Buffalo Mozzarella Cheese |
| title_sort | integrated gel electrophoresis and mass spectrometry approach for detecting and quantifying extraneous milk in protected designation of origin buffalo mozzarella cheese |
| topic | buffalo milk adulteration gel electrophoresis MALDI-TOF-MS nano-LC-ESI-MS/MS peptidomics proteomics |
| url | https://www.mdpi.com/2304-8158/14/7/1193 |
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