Quantitative determination of plasma C8-C26 total fatty acids for the biochemical diagnosis of nutritional and metabolic disorders
Quantitative determination of plasma C8-C26 total fatty acids for the biochemical diagnosis of nutritional and metabolic disorders
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DOI：
10.1006/mgme.2001.3170
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发表时间：
2001-05-01
影响因子：
3.8
通讯作者：
McConnell, JP
中科院分区：
文献类型：
--
作者：
Lagerstedt, SA;Hinrichs, DR;McConnell, JP
建立了血浆中C8-C26总脂肪酸的毛细管气相色谱-电子捕获负离子质谱（GC/MS）测定方法。在水解、己烷提取和用五氟苄基溴衍生化后，分两步分析脂肪酸酯：不分流进样和第二次分流进样（1：100），以定量更丰富的长链物质。通过选择离子监测与13种稳定同位素标记的内标物的比例定量14种饱和脂肪酸和25种不饱和脂肪酸。校准显示出一致的线性和重现性，批内（n = 17）和批间（n = 12）CV范围分别为2.5 - 13.2%和4.6 - 22.9%。回收率范围为76 - 106%。建立了四个年龄组（18岁）的参考范围，并与omega-3和omega-6多不饱和脂肪酸营养缺乏、线粒体脂肪酸氧化先天性缺陷和过氧化物酶体疾病患者的标本进行比较。回顾性评价了35例诊断为必需脂肪酸缺乏症的病例中亚油酸的浓度，这些病例先前通过气相色谱分析与火焰离子化检测（GC/FID）进行诊断，发现与GC/MS相比，GC/FID方法的特异性和灵敏度分别仅为55%和50%。（C）2001 Academic Press。
We have developed a capillary gas chromatography-electron-capture negative-ion mass spectrometry (GC/MS) method for the quantitative determination of C8-C26 total fatty acids in plasma. Following hydrolysis, hexane extraction, and derivatization with pentafluorobenzyl bromide, fatty acid esters are analyzed in two steps: a splitless injection and a second, split injection (1:100) far the quantitation of the more abundant long-chain species, Fourteen saturated and 25 unsaturated fatty acids are quantified by selected ion monitoring in ratio to 13 stable-isotope-labeled internal standards. Calibrations exhibit consistent linearity and reproducibility, Intraassay (n = 17) and interassay (n = 12) CVs ranged from 2.5 to 13.2% and from 4.6 to 22.9%, respectively. Recoveries ranged from 76 to 106%. Reference ranges were established for four age groups (18 years) and compared to specimens from patients with nutritional deficiency of omega -3 and omega -6 polyunsaturated fatty acids, inborn errors of mitochondrial fatty acid oxidation, and peroxisomal disorders. Retrospective evaluation of the concentration of linoleic acid in 35 cases with a diagnosis of essential fatty acid deficiency previously made by gas chromatographic analysis with flame ionization detection (GC/FID) found a specificity and sensitivity of only 55 and 50%, respectively, for the GC/FID method when compared to GC/MS. (C) 2001 Academic Press.