Faster Protocol for Endogenous Fatty Acid Esters of Hydroxy Fatty Acid (FAHFA) Measurements.
Faster Protocol for Endogenous Fatty Acid Esters of Hydroxy Fatty Acid (FAHFA) Measurements.
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DOI：
10.1021/acs.analchem.8b00503
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发表时间：
2018-04-17
影响因子：
7.4
通讯作者：
Saghatelian A
中科院分区：
文献类型：
--
作者：
Kolar MJ;Nelson AT;Chang T;Ertunc ME;Christy MP;Ohlsson L;Härröd M;Kahn BB;Siegel D;Saghatelian A
羟基脂肪酸的脂肪酸酯（FAHFA）是最近发现的一类具有抗糖尿病和抗炎活性的内源性脂质。对这些脂质的兴趣是由于它们独特的生物活性以及观察到胰岛素抵抗的人具有较低的羟基硬脂酸棕榈酸酯（PAHSAs）水平，这表明FAHFA缺乏可能导致代谢疾病。对这一假设的严格检验将需要测量许多临床样本;然而，目前的分析工作流程太慢，无法快速分析样本。在这里，我们描述了一个显着更快的工作流程来测量FAHFA，优化这些脂质的分馏和色谱法的发展。我们可以在30分钟内测量FAHFA与这个新的协议与90分钟使用旧的协议具有可比的性能，在区域异构体检测和定量。我们还发现，通过这种优化，羟基硬脂酸的油酸酯（OAHSAs），另一个家庭的FAHFA，有一个更低的背景信号比PAHSAs，这使得他们更容易测量。我们更快的工作流程能够量化小鼠组织和人血浆中PAHSAs和OAHSAs的变化，突出了该方案在基础和临床应用中的潜力。
Fatty acid esters of hydroxy fatty acids (FAHFAs) are a recently discovered class of endogenous lipids with anti-diabetic and anti-inflammatory activities. Interest in these lipids is due to their unique biological activity and the observation that insulin resistant people have lower palmitic acid esters of hydroxystearic acid (PAHSAs) levels, suggesting that a FAHFA deficiency may contribute to metabolic disease. Rigorous testing of this hypothesis will require the measurement of many clinical samples; however, current analytical workflows are too slow to enable samples to be analyzed quickly. Here, we describe the development of a significantly faster workflow to measure FAHFAs that optimizes the fractionation and chromatography of these lipids. We can measure FAHFAs in 30 minutes with this new protocol versus 90 minutes using the older protocol with comparable performance in regioisomer detection and quantitation. We also discovered through this optimization that oleic acid esters of hydroxystearic acids (OAHSAs), another family of FAHFAs, have a much lower background signal than PAHSAs, which makes them easier to measure. Our faster workflow was able to quantify changes in PAHSAs and OAHSAs in mouse tissues and human plasma, highlighting the potential of this protocol for basic and clinical applications.
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