Lin Yu Hong, Salem Norman, Wells Ellen M, Zhou Weiyin, Loewke James D, Brown James A, Lands William E M, Goldman Lynn R, Hibbeln Joseph R
Section 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.
Lipids. 2012 May;47(5):527-39. doi: 10.1007/s11745-012-3661-6. Epub 2012 Mar 21.
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 μg/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.
大规模人群研究表明,多不饱和脂肪酸对人类健康很重要,但由于缺乏高效的高通量技术,确定健康益处与脂肪酸含量之间的关系受到了阻碍。基于直接酯交换反应的先前方法开发了一种自动化高通量脂肪酸分析方法,包括化学程序自动化、数据采集和自动数据处理。该方法经过验证并应用于一项流行病学研究中的脐带血清样本。该方法在血清浓度为1 - 600μg/mL范围内呈线性,r²≥0.99。23种脂肪酸的批内变异系数<5.4%,在低脂质基质中,除14:0外,三种浓度的回收率范围为76 - 119%。机器人法测定的人血浆脂肪酸浓度与勒帕热和罗伊法结果高度一致。美国受试者(n = 287)脐带血清中的脂肪酸谱显示,饱和脂肪酸、单不饱和脂肪酸、n - 6和n - 3多不饱和脂肪酸分别占总脂肪酸的38.0%、24.9%、32.0%和4.6%。这是首次报道一种完整、经过验证、经济高效、自动化的高通量脂肪酸测量方法及其在基于人群的研究中的应用。自动化脂肪酸分析与自动数据处理相结合,极大地促进了大规模基于人群研究所需的高通量分析,6小时内可对72个样本进行酯交换反应。