Singh S.Thuanthong A.Mann B.Hooda A.Sharma R.Bajaj R.Agarwal A.Tripathi A.D.Rai H.K.Koirala P.Nirmal N.Mahidol University2026-03-122026-03-122026-01-01International Journal of Agriculture and Biosciences Vol.15 No.1 (2026) , 151-15823056622https://repository.li.mahidol.ac.th/handle/123456789/115641The study evaluated the influence of genetic variants and thermal processing on the release of β-casomorphins-7 (BCM-7) during simulated in-vitro digestion. Cross-bred Karan Fries bovine animals were segregated into A1A1 and A2A2 genotypes for the procurement of pure A1 and A2 milk. BCM-7-like peptides were detected only after gastrointestinal digestion from both genotypes, using reverse-phase high-performance liquid chromatography (RP-HPLC). The concentration of these peptides was 74-146ng/mL in digestive extracts of A2 variants, which was significantly lower than 900-1612 ng/mL in A1 variants. Pasteurization led to a reduction in peptide levels to 46-79ng/mL and 340-1132ng/mL in extracts from A2 and A1 variants, respectively. Upon sterilization, a marked decrease in peptide formation was observed, 84.79% in A1 and 84.56% in A2 milk. This indicated a strong negative correlation between heat severity and BCM-7-like peptide formation. Mass spectrometry analysis of selected RP-HPLC fractions confirmed the presence of BCM-7-like peptides in both variants, with a higher specificity toward histidine at position 67 compared to proline.Agricultural and Biological SciencesArticleSCOPUS10.47278/journal.ijab/2025.1442-s2.0-10503182490923063599