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.

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.