FOODplus Research Centre, School of Agriculture, Food and Wine, the University of Adelaide, PMB 1, Glen Osmond, South Australia 5064, Australia.
FOODplus Research Centre, School of Agriculture, Food and Wine, the University of Adelaide, PMB 1, Glen Osmond, South Australia 5064, Australia.
Prostaglandins Leukot Essent Fatty Acids. 2017 Oct;125:1-7. doi: 10.1016/j.plefa.2017.08.010. Epub 2017 Aug 24.
Omega-3 and omega-6 polyunsaturated free fatty acids (PUFA-FFA) are precursors to potent downstream lipid mediators that are regulators of inflammation. We describe the development and validation of a novel and sensitive method for quantification of individual PUFA-FFA in a dried blood spot using liquid chromatography tandem mass spectrometry (LC-MS/MS). Lipids were extracted from dried blood spot and six individual PUFA-FFA were quantified by LC-MS/MS using stable isotope dilution analysis with deuterated internal standards. PUFA-FFA concentrations in blood samples from 30 subjects were measured using the new method and compared to the traditional approach of thin layer chromatography followed by gas chromatography with flame ionization detection (TLC-GC). Responses for each PUFA-FFA were linear throughout a range of concentrations expected in clinical samples. Intra-day and inter-day variations for all PUFA-FFA were ≤ 16%. The concentrations of all PUFA-FFA measured by LC-MS/MS were positively correlated with measures of the same PUFA-FFA obtained by a traditional TLC-GC method. This novel method for the quantification of PUFA-FFA extracted from dried blood is sensitive and precise, and accurately measures levels of biologically important PUFA-FFA in blood.
ω-3 和 ω-6 多不饱和游离脂肪酸（PUFA-FFA）是强效下游脂质介质的前体，这些介质是炎症的调节剂。我们描述了一种使用液相色谱串联质谱（LC-MS/MS）定量分析干血斑中单个 PUFA-FFA 的新型灵敏方法的开发和验证。从干血斑中提取脂质，并通过 LC-MS/MS 采用稳定同位素稀释分析和氘代内标定量 6 种单个 PUFA-FFA。使用新方法测量了 30 名受试者血液样本中的 PUFA-FFA 浓度，并与传统的薄层色谱（TLC）-气相色谱（GC）-火焰离子化检测（FID）方法进行了比较。在预期的临床样本浓度范围内，每种 PUFA-FFA 的响应均呈线性。所有 PUFA-FFA 的日内和日间变化均≤16%。通过 LC-MS/MS 测量的所有 PUFA-FFA 的浓度与通过传统 TLC-GC 方法获得的相同 PUFA-FFA 的测量值呈正相关。这种从干血中提取 PUFA-FFA 的新型定量方法灵敏、精确，能准确测量血液中生物重要的 PUFA-FFA 水平。
