Due to the diversity and limitation of determination methods, published data on the fatty acid (FA) compositions of different milk samples have contributed to inaccurate comparisons. In this study, we developed a high-throughput gas chromatography-mass spectrometry method to determinate milk FA, and the proposed method had satisfactory linearity, sensitivity, accuracy, and precision. We also analyzed the FA compositions of 237 milk samples from Holstein cows, Jersey cows, buffalos, yaks, humans, goats, donkeys, and camels. Holstein, Jersey, goat, and buffalo milks contained high content of even-chain saturated FA, whereas goat milk had higher content of medium-and short-chain FA (MSCFA). Yak and camel milk are potential functional foods due to their high levels of odd-and branchedchain FA and low ratios of n-6 to n-3 polyunsaturated FA (PUFA). Human milk contained lower levels of saturated FA, MSCFA, and conjugated linoleic acid, and higher levels of monounsaturated FA and PUFA. As a special nonruminant milk, donkey milk contained low levels of monounsaturated FA and high levels of PUFA and MSCFA. Based on the FA profiles of 8 types of milk, nonruminant milk was distinct from ruminant milk, whereas camel and yak milk were different from other ruminant milks and considered as potential functional foods for balanced human diet.

The determination of C18 fatty acids (FAs) is a key and difficult aspect in FA profiling, and a qualified method with good chromatographic separation and high sensitivity, as well as easy methylation, is required. A GC-MS method was established to simultaneously determine C18 FAs in milk. To simplify the methylation protocol for milk samples, besides a base-catalyzation methylation (50 °C for 20 min), the necessity of an additional acid-catalyzation was also studied using different temperatures (60 °C, 70 °C, 80 °C, and 90 °C) and durations (90 min and 150 min). The results showed that the chromatographic resolution was improved, although three co-eluted peaks existed. The base-catalyzation was sufficient, and an additional acid-catalyzation was not necessary. The proposed method was validated with good sensitivity, linearity, accuracy, and precision, and then applied in determining C18 FAs in 20 raw milk and 30 commercial milk samples. UHT milk presented a different profile of C18 FAs from raw milk and PAS milk samples, which indicated that excessive heating could change the profile. Overall, the proposed method is a high-throughput and competent approach for the determination of C18 FAs in milk, and which presents an improvement in chromatographic resolution and sensitivity, as well as a simplification of methylation.