Identification of water‐soluble peptides for authenticity verification of commercial buffalo ricotta

Background, context or rationale Reliable methods to verify the authenticity of foods, such as buffalo ricotta, are essential for protecting brands and regulatory compliance. Prized for its sensory and nutritional qualities, buffalo ricotta is vulnerable to adulteration. Recent advances have developed precise, accessible technologies, broadening applications in the food industry and enhancing safety and quality. Aim (s) This study evaluated the authenticity of commercial buffalo ricotta adulterated with cow whey by investigating water‐soluble peptides using mid‐infrared spectroscopy, high‐performance liquid chromatography and electrophoresis, combined with chemometric analysis. Methods To evaluate the authenticity of ricotta, 11 experimental formulations were created by increasing the proportions of cow whey mixed with buffalo whey. These formulations were stored for up to 30 days. Water‐soluble peptides were analysed using mid‐infrared spectroscopy, high‐performance liquid chromatography and electrophoresis. The data obtained from mid‐infrared spectroscopy and high‐performance liquid chromatography were statistically processed using principal component analysis, analysis of covariance and multiple linear regression, with replication to ensure validity. Major Findings Principal component analysis of mid‐infrared spectra explained 97.46% of the variance, distinguishing pure and adulterated samples, while storage time significantly impacted spectral characteristics ( P < 0.05). High‐performance liquid chromatography identified 14 peptide peaks, with three recognized as specific markers for cow whey in adulterated samples. Principal component analysis explained 77.14% of the variance, distinguishing pure and adulterated ricotta. Multiple linear regression modelling of high‐performance liquid chromatography data predicted cow whey concentration with a correlation of R = 0.87. High‐performance liquid chromatography with chemometrics was effective for detecting buffalo ricotta adulteration. When applied to 14 commercial samples, the model revealed that nine contained adulteration ranging from 10% to 100% cow whey. Mid‐infrared spectroscopy and sodium dodecyl sulphate‐polyacrylamide gel electrophoresis were insufficient to identify adulteration. Scientific or Industrial Implications The study shows that the chromatographic method can detect cow whey in buffalo ricotta at low concentrations. This approach increases the reliability of detecting fraud in dairy products and strengthens quality control and regulation.