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Automated high-throughput fatty acid analysis of umbilical cord serum and application to an epidemiological study
Yu Hong Lin1, Norman Salem, Ellen M Wells
1Section of Nutritional Neuroscience, Laboratory of Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism, NIH, 5625 Fishers Lane, Room 3N-07, MSC 9410, Bethesda, MD 20892-9410, USA. yulin@mail.nih.gov
Abstract:
Large population studies show that polyunsaturated fatty acids are important for human health, but determining relationships between the health benefits and the fatty acid content has been hampered by the unavailability of labor-effective high-throughput technologies. An automated high throughput fatty acid analysis was developed from a previous procedure based on direct transesterification including the automation of chemical procedures, data acquisition and automatic data processing. The method was validated and applied to umbilical cord serum samples in an epidemiological study. The method was linear in the range of 1-600 microg/mL serum with r² ≥ 0.99. The within-run CV was <5.4% for 23 fatty acids and a range of recoveries over three concentrations were 76-119% in a low-lipid matrix with the exception of 14:0. The fatty acid concentration as measured by the robotic method for human plasma was in good agreement with the Lepage & Roy method. The fatty acid profile in umbilical cord serum from American subjects (n = 287) showed an average of 38.0, 24.9, 32.0 and 4.6% of total fatty acids for saturates, monounsaturates, n-6 and n-3 polyunsaturates, respectively. This is the first report of a complete, validated, cost-effective, automated, high throughput fatty acid measurement method along with application to a population-based study. Automated fatty acid analysis coupled with automated data processing greatly facilitates the high throughput, 72 samples transesterified in 6 h, required for large population-based studies.
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