De Pascale Sabrina, Garro Giuseppina, Pellicano Silvia Ines, Scaloni Andrea, Carpino Stefania, Caira Simonetta, Addeo Francesco
Proteomics, Metabolomics & Mass Spectrometry Laboratory, Institute for the Animal Production System in the Mediterranean Environment, National Research Council, 80055 Portici, Italy.
Department of Agriculture, University of Naples "Federico II", 80055 Portici, Italy.
Foods. 2025 Mar 28;14(7):1193. doi: 10.3390/foods14071193.
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.
确保受保护的原产地名称（PDO）奶酪坎帕纳水牛奶酪（MdBC）的真实性，对于监管执法和消费者保护至关重要。本研究评估了一个多技术分析平台，该平台用于检测PDO水牛奶制品中因添加非水牛奶或非PDO水牛奶而导致的掺假情况。外围实验室使用凝胶电泳结合多克隆抗肽抗体进行初步筛查，能够检测出外来酪蛋白，包括那些源自PDO指定区域以外的酪蛋白。为了进行更精确的鉴定，基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）通过检测蛋白型肽来区分物种。在需要确认的情况下，中央州立实验室使用纳升液相色谱-电喷雾串联质谱（nano-LC-ESI-MS/MS）对PDO水牛奶MdBC奶酪中的外来乳蛋白进行高灵敏度检测。另一方面，对水牛奶蓝纹奶酪pH 4.6可溶部分的分析鉴定出2828种源自水牛的肽和几种牛特异性肽，证实了牛奶掺假。尽管胰蛋白酶水解后pH 4.6不溶部分的检测范围较低，但牛肽的存在仍足以验证掺假行为。这种结合了电泳和质谱技术的综合蛋白质组学方法显著提高了牛奶掺假检测能力，为应对日益复杂的欺诈行为提供了一个强大的工具。
