Fatty Acid- and Amino Acid-Specific Isotope Analysis for Accurate Authentication and Traceability in Organic Milk

The present study describes compound-specific δ¹³C and δ¹⁵N analyses of fatty acids and amino acids for improving the accurate authentication of organic milk (OM) against conventional milk (CM) collected in Korea. Most δ¹³C_fatty-acid and δ¹³C_amino-acid values were lower in OM than in CM ( P < 0.05); however, most δ¹⁵N_amino-acid values displayed weak discriminative power for OM authentication. Higher isotopic fractionation was observed in δ¹³C_fatty-acid than in δ¹³C_amino-acid and δ¹⁵N_amino-acid, with fractionation trends differing with individual amino acids. In particular, δ¹³C_{linoleic-acid} of -33.5‰ and δ¹³C_{myristic-acid} of -28‰ were determined to be promising year-round threshold values for Korean OM authentication. The δ¹³C_bulk was highly correlated with δ¹³C_Ala ( r = 0.92) and δ¹³C_{oleic-acid, trans} ( r = 0.77), and strong positive correlations were observed between δ¹³C_Val and δ¹³C_Ile ( r = 0.98) and between δ¹⁵N_Thr and δ¹⁵N_Ser ( r = 0.90). Chemometric modeling for OM authentication produced a high quality model ( R² X = 0.547, R² Y = 0.865, and Q² = 0.689) with reliable chemical markers, notably δ¹³C_{myristic-acid}, δ¹³C_{linoleic-acid}, and δ¹³C_stearic-acid. Furthermore, the models developed for seasonal separation in OM ( Q² = 0.954) and CM ( Q² = 0.791) were of good quality. Our findings, based on compound-specific isotope data, improve the reliability of OM authentication in cases where bulk stable isotope ratio analysis alone is insufficient. They also provide valuable insight into the control of fraudulent OM labeling in Korea, with potential application in other countries.