This article reports a targeted metabolomic method for total plasma fatty acids (FAs) of clinical or nutritional relevance. Thirty-six saturated, unsaturated, or branched-chain FAs with a chain length of C8-C28 were quantified using reversed-phase liquid chromatography-tandem mass spectrometry. FAs in plasma (10 μL) were acid-hydrolyzed, extracted, and derivatized with DAABD-AE (4-[2-(N,N-Dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole) at 60 °C for 1 h. Derivatization resulted in a staggering nine orders of magnitude higher sensitivity compared to underivatized analytes. FAs were measured by multiple-reaction monitoring using stable isotope internal standards. With physiological and pathological analyte levels in mind, linearity was established using spiked plasma. Intra-day (n = 15) and inter-day (n = 20) imprecisions expressed as variation coefficient were ≤10.2% with recovery ranging between 94.5–106.4%. Limits of detection and limit of quantitation ranged between 4.2–14.0 and 15.1–51.3 pmol per injection, respectively. Age-stratified reference intervals were established in four categories: <1 month, 1–12 month, 1–18 year, and >18 year. This method was assessed using samples from patients with disorders affecting FAs metabolism. For the first time, C28:0 and C28:0/C22:0 ratio were evaluated as novel disease biomarkers. This method can potentially be utilized in diagnosing patients with inborn errors of metabolism, chronic disease risk estimation, or nutritional applications.

中文翻译：

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液相色谱-串联质谱分析人血浆中总脂肪酸的目标代谢组学

本文报道了一种具有临床或营养意义的血浆总脂肪酸（FA）靶向代谢组学方法。使用反相液相色谱-串联质谱法对链长为C8-C28的36个饱和，不饱和或支链FA进行定量。血浆中的FAs（10μL）被酸水解，萃取并用DAABD-AE（4- [2-（N，N-二甲基氨基）乙基氨基磺酰基] -7-（2-氨基乙基氨基）-2,1,3-苯并恶二唑）在60℃下1小时。与未衍生化的分析物相比，衍生化导致的灵敏度高出惊人的九个数量级。通过使用稳定同位素内标进行多反应监测来测量FA。考虑到生理和病理分析物的水平，使用加标血浆建立了线性关系。日内（n = 15）和日间（n = 20）的不精确度以变异系数表示，≤10.2％，回收率在94.5-106.4％之间。每次进样的检测限和定量限分别在4.2-14.0和15.1-51.3 pmol之间。按年龄分层的参考间隔分为四个类别：<1个月，1-12个月，1-18岁和> 18岁。使用来自影响FA代谢异常的患者的样品评估了该方法。首次将C28：0和C28：0 / C22：0的比率评估为新型疾病生物标记。该方法可潜在地用于诊断患有先天性代谢错误，慢性疾病风险估计或营养应用的患者。