Comprehensive quantitation of multi-signature peptides originating from casein for the discrimination of milk from eight different animal species using LC-HRMS with stable isotope labeled peptides.
Abstract: Milk species adulteration has become an altering issue worldwide. In this study, a robust quantification method based on LC-HRMS for the simultaneous detection and differentiation of milk type from eight different animal species (namely: cow, water buffalo, wild yak, goat, sheep, donkey, horse, and camel) was established by detecting nine signature peptides originating from casein. The developed method was in-house validated in terms of sensitivity, accuracy, and precision. As a result, limits of quantification (LOQ) were ranging from 5 to 30 µg/L, recoveries ranged from 95.2% to 104.5%, and intra-day and inter-day variability were lower than 11.4% and 12.6%, respectively, for all the targeted peptides. Furthermore, this method was successfully applied to 46 commercial minor species' milk, in which 15 samples were false labeling. The obtained results indicate the necessity to monitor milk species adulteration in order to protect consumers from consuming misleading labeled minor species animal's milk.
Copyright © 2022 Elsevier Ltd. All rights reserved.
Publication Date: 2022-05-02 PubMed ID: 35567972DOI: 10.1016/j.foodchem.2022.133126Google Scholar: Lookup
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Summary
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This research study presents a method for detecting and differentiating milk types from eight different animal species. Focusing on nine signature peptides that originate from casein, this method was applied to 46 commercial minor species milk samples, finding that 15 were falsely labelled.
Study Objective and Methodology
- The main objective of this study was to establish a method for differentiating and simultaneously detecting milk types from eight different animals due to increasing concerns of milk species adulteration.
- The animals used to source the milk for testing in this study include cow, water buffalo, wild yak, goat, sheep, donkey, horse, and camel.
- The method is based on the detection of nine signature peptides originating from casein, a protein found in milk.
- The researchers used Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS), a sensitive technique for the separation and identification of compounds, in conjunction with stable isotope labeled peptides.
Findings and Validation
- The method was validated internally for sensitivity, accuracy, and precision.
- Findings showed that the limits of quantification (LOQ) – the lowest concentration level that can be determined with acceptable precision and accuracy – were between 5 to 30 µg/L.
- The recovery rates, indicating the ability to extract the peptide back after a known amount was added, were between 95.2% and 104.5%.
- Both intra-day (day-to-day) and inter-day (between different days) variations were less than 11.4% and 12.6% respectively. This indicates good repeatability and reproducibility of the method.
Application to Commercial Milk
- The developed methodology was applied to 46 commercial samples of minor species’ milk.
- Interestingly, it was found that 15 out of these 46 samples were falsely labelled, highlighting an issue with misrepresentation or adulteration in commercially available milk products.
- The study therefore further underscores the importance of monitoring adulteration in milk from various animal species to protect consumers from mislabelled products.
Cite This Article
APA
Zhang H, Abdallah MF, Zhang J, Yu Y, Zhao Q, Tang C, Qin Y, Zhang J.
(2022).
Comprehensive quantitation of multi-signature peptides originating from casein for the discrimination of milk from eight different animal species using LC-HRMS with stable isotope labeled peptides.
Food Chem, 390, 133126.
https://doi.org/10.1016/j.foodchem.2022.133126 Publication
Researcher Affiliations
- State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; Scientific Observing and Experiment Station of Animal Genetic Resources and Nutrition in North China of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
- Department of Food Technology, Safety and Health, Ghent University, Coupure Links 653, 9000 Ghent, Belgium; Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Assiut University, Assiut 71515, Egypt.
- State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; Scientific Observing and Experiment Station of Animal Genetic Resources and Nutrition in North China of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
- State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; Scientific Observing and Experiment Station of Animal Genetic Resources and Nutrition in North China of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
- State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; Scientific Observing and Experiment Station of Animal Genetic Resources and Nutrition in North China of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
- State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; Scientific Observing and Experiment Station of Animal Genetic Resources and Nutrition in North China of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
- State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; Scientific Observing and Experiment Station of Animal Genetic Resources and Nutrition in North China of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
- State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; Scientific Observing and Experiment Station of Animal Genetic Resources and Nutrition in North China of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: zhangjunmin@caas.cn.
MeSH Terms
- Allergens
- Animals
- Buffaloes
- Camelus
- Caseins / chemistry
- Cattle
- Equidae
- Female
- Goats
- Horses
- Isotopes
- Milk / chemistry
- Peptides / chemistry
- Sheep
Citations
This article has been cited 5 times.- Xu DK, Yang Y, Huang HL, Sun XJ, Hu WY, Liu XM, Yang J. [Quantitation of cow's milk allergen in hypoallergenic infant formulas using ultra-high performance liquid chromatography- tandem mass spectrometry]. Se Pu 2025 Nov;43(11):1244-1251.
- Liu D, Xu Y, Zhang X, Yin F, Cao J, Liu Z, Zhou D, Feng A, Li C. Organic Fusion of Molecular Simulation and Wet-Lab Validation: A Promising High-Throughput Strategy for Screening Bioactive Food Peptides. Foods 2025 Aug 20;14(16).
- Karamoutsios A, Lekka P, Voidarou CC, Dasenaki M, Thomaidis NS, Skoufos I, Tzora A. Assessing Milk Authenticity Using Protein and Peptide Biomarkers: A Decade of Progress in Species Differentiation and Fraud Detection. Foods 2025 Jul 23;14(15).
- Magdas DA, Hategan AR, David M, Berghian-Grosan C. The Journey of Artificial Intelligence in Food Authentication: From Label Attribute to Fraud Detection. Foods 2025 May 19;14(10).
- Ma X, Xia H, Pan Y, Huang Y, Xu T, Guan F. Double-Tube Multiplex TaqMan Real-Time PCR for the Detection of Eight Animal-Derived Dairy Ingredients. J Agric Food Chem 2024 May 22;72(20):11640-11651.
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