Trans‐fatty acids (TFA) intake has been associated with risk factors for cardiovascular disease. However, little is known about the TFA levels in humans or about how these levels relate to other fatty acids (FA).In order to assess TFA levels in humans we have developed and validated an isotope dilution‐gas chromatography‐negative chemical ionization‐mass spectrometry method for the quantitation of 28 FA in human plasma, including 4 major TFA. We analyzed 73 convenience samples from adults. All samples contained TFA, with average levels ± standard deviation of 2.66 ± 1.29 μM for palmitelaidic acid 12.70 ± 8.31 μM for elaidic acid, 16.59 ± 11.10 μM for vaccenic acid, and 1.15 ± 0.52 μM for linoelaidic acid. The sum of all FA was 9647 ± 2913 μM. The sum of the 4 TFA accounted for 0.34 ± 0.20 % of total FA. FA values expressed as percent of total FA showed negative association between monounsaturated FA and polyunsaturated fatty acids (PUFAs), ω‐6 PUFAs, and ω‐3 PUFAs. Negative trends were also seen between certain TFA and the summed PUFAs. We have developed an analytical method that enables the assessment of major TFA and other FA in 100 μL of plasma, which allows the investigation of FA concentrations as well as differences in the composition of these FA in plasma. This method is being used to analyze samples from the National Health and Nutrition Examination Survey (NHANES).
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