Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, Córdoba, Spain; Maimónides Institute of Biomedical Research (IMIBIC), Reina Sofía University Hospital, University of Córdoba, Córdoba, Spain; University of Córdoba Agroalimentary Excellence Campus, ceiA3, Córdoba, Spain; CIBER Fragilidad y Envejecimiento Saludable (CIBERfes), Instituto de Salud Carlos III, Spain.
Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, Córdoba, Spain; Maimónides Institute of Biomedical Research (IMIBIC), Reina Sofía University Hospital, University of Córdoba, Córdoba, Spain; University of Córdoba Agroalimentary Excellence Campus, ceiA3, Córdoba, Spain; CIBER Fragilidad y Envejecimiento Saludable (CIBERfes), Instituto de Salud Carlos III, Spain.
Talanta. 2019 Feb 1;193:29-36. doi: 10.1016/j.talanta.2018.09.088. Epub 2018 Sep 25.
The recent growing interest in primary fatty acid amides (PFAMs) is due to the broad range of physiological effects they exhibit as bioindicator of pathological states. These bioactive lipids are usually in biological samples at the nanomolar level, making their detection and identification a challenging task. A method for quantitative analysis of seven main PFAMs (lauramide, myristamide, linoleamide, palmitamide, oleamide, stearamide and behenamide) in four human biofluids -namely, urine, plasma, saliva and sweat- is here reported. Two sample preparation procedures were compared to test their efficiency in each biofluid: solid-phase extraction (SPE) and protein precipitation. The latter was the best for plasma and urine, while the analysis of saliva and sweat required an SPE step for subsequent suited determination of PFAMs. Detection of the seven metabolites was performed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in multiple reaction monitoring (MRM) mode. Quantitative analysis was supported on the use of stable isotopically labeled internal standards (SIL-ISs) in the calibration method, which required the synthesis of each IS from the precursor deuterated fatty acids. Detection limits for the target analytes were within 0.3-3 ng mL. The method was applied to a small cohort of male and female volunteers (n = 6) to estimate the relative concentration profiles in the different biofluids. The analytical features of the method supported its applicability in clinical studies aimed at elucidating the role of PFAMs metabolism.
最近，人们对初级脂肪酸酰胺 (PFAMs) 的兴趣日益浓厚，这是因为它们表现出广泛的生理效应，可作为病理状态的生物标志物。这些生物活性脂质通常以纳摩尔级别的浓度存在于生物样本中，这使得它们的检测和鉴定成为一项具有挑战性的任务。本文报道了一种在四种人体生物流体（即尿液、血浆、唾液和汗液）中定量分析七种主要 PFAMs（月桂酰胺、肉豆蔻酰胺、亚油酸酰胺、棕榈酰胺、油酸酰胺、硬脂酰胺和山嵛酰胺）的方法。比较了两种样品前处理方法，以测试它们在每种生物流体中的效率：固相萃取 (SPE) 和蛋白质沉淀。后一种方法最适合于血浆和尿液，而唾液和汗液的分析则需要 SPE 步骤，以随后适合地测定 PFAMs。通过液相色谱-串联质谱（LC-MS/MS）在多重反应监测（MRM）模式下检测七种代谢物。定量分析支持在校准方法中使用稳定同位素标记的内标物（SIL-ISs），这需要从前体氘化脂肪酸合成每种 IS。目标分析物的检测限在 0.3-3ng/mL 范围内。该方法应用于一小批男性和女性志愿者（n=6），以估计不同生物流体中的相对浓度分布。该方法的分析特点支持其在旨在阐明 PFAMs 代谢作用的临床研究中的适用性。
