Kulig Clark C, Beresford Thomas P, Everson Gregory T
Division of Gastroenterology and Hepatology, University of Colorado Health Sciences Center, Denver, Colorado.
J Lab Clin Med. 2006 Mar;147(3):133-8. doi: 10.1016/j.lab.2005.11.006.
Fatty acid ethyl esters (FAEEs) are useful markers of ongoing alcohol use and may be associated with alcohol-induced damage to the liver and pancreas. In this article, we describe a novel method for rapid determination of the three major FAEEs found in human plasma.
Internal standard, ethyl heptadecanoate, was added to plasma samples, and FAEEs were isolated by acetone precipitation, hexane lipid extraction, and amino-propyl silica solid phase extraction. FAEEs were quantitated by gas chromatography-mass spectrometry (GC-MS) using a nonpolar dimethylpolysiloxane column. The accuracy, precision, specificity, and sensitivity of the assay were defined from plasma samples from recently drinking and abstinent persons, with and without the addition of FAEEs.
Individual FAEE peaks demonstrated excellent resolution. Instrument time was reduced by more than 60%. The lower limit of detection was 5 to 10 nM, and the lower limit of quantitation for each FAEE was 60 nM (for 22 samples with known concentration 60 nM, x +/-SD: 61 +/- 5.7, 57 +/- 5.7, and 57 +/- 5.9 nM, for ethyl palmitate, ethyl oleate, and ethyl stearate, respectively). Instrument precision (coefficient of variance, CV) for these three FAEEs was 0.3%, 0.4%, and 0.7%, respectively. Intra-assay precision (CV) for total FAEEs was less than 7%. FAEEs were absent in 49 samples from abstinent persons. FAEEs were detected in all 76 samples with associated positive blood alcohol levels.
Our method of FAEE analysis is rapid and potentially useful in research and clinical studies. FAEE determination using this method is precise, accurate, sensitive, and specific and deserves broader application.
脂肪酸乙酯（FAEEs）是当前酒精使用的有用标志物，可能与酒精引起的肝脏和胰腺损伤有关。在本文中，我们描述了一种快速测定人血浆中三种主要FAEEs的新方法。
将内标十七烷酸乙酯添加到血浆样本中，通过丙酮沉淀、己烷脂质萃取和氨丙基硅胶固相萃取分离FAEEs。使用非极性二甲基聚硅氧烷柱通过气相色谱-质谱联用（GC-MS）对FAEEs进行定量。通过来自近期饮酒者和戒酒者的血浆样本（添加和未添加FAEEs）确定该检测方法的准确性、精密度、特异性和灵敏度。
各个FAEE峰显示出极佳的分辨率。仪器分析时间减少了60%以上。检测下限为5至10 nM，每种FAEE的定量下限为60 nM（对于22个已知浓度为60 nM的样本，x±标准差：棕榈酸乙酯、油酸乙酯和硬脂酸乙酯分别为61±5.7、57±5.7和57±5.9 nM）。这三种FAEEs的仪器精密度（变异系数，CV）分别为0.3%、0.4%和0.7%。总FAEEs的批内精密度（CV）小于7%。49份戒酒者的样本中未检测到FAEEs。在所有76份血酒精水平呈阳性的样本中均检测到了FAEEs。
我们的FAEE分析方法快速，在研究和临床研究中可能有用。使用该方法测定FAEEs精确、准确、灵敏且特异，值得更广泛应用。
