Collection and Storage of Human Plasma for Measurement of Oxylipins.
Collection and Storage of Human Plasma for Measurement of Oxylipins.
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DOI：
10.3390/metabo11030137
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发表时间：
2021-02-26
期刊：
影响因子：
4.1
通讯作者：
Norris JM
中科院分区：
文献类型：
--
作者：
Polinski KJ;Armstrong M;Manke J;Seifert J;Crume T;Yang F;Clare-Salzler M;Holers VM;Reisdorph N;Norris JM
来自ω-3和ω-6脂肪酸的氧脂素积极参与炎症和免疫过程，并在人类疾病中发挥重要作用。然而，随着对氧脂素的兴趣增加，关于哪些分子在血浆中可检测，收集样品的最佳方法以及分子在收集和储存期间是否稳定的问题仍然存在。因此，我们在现有研究的基础上，通过检查90种氧脂素（包括专门的促分解脂质介质（SPM））在人血浆（n = 5例受试者）中在样本采集、处理和-80 °C储存期间的稳定性。采用液相色谱-串联质谱法（LC/MS/MS）定量羟脂素。在乙二胺四乙酸（EDTA）或肝素中采集的血液样本，然后在处理前室温下储存长达2小时，与立即处理的样本（包括SPM脂氧素A4和消退素D1）相比，氧脂素浓度无显著差异。大多数分子，包括SPM，在储存长达1年后保持稳定。然而，为了支持以前的发现，在一小部分氧脂素中观察到了变化，包括12-HETE，TXB 2，14-HDHA和18-HEPE。总体而言，本研究表明，使用LC/MS/MS可以从储存的EDTA或肝素血浆样本中获得大多数氧脂素的准确测量值。
Oxylipins derived from omega-3 and -6 fatty acids are actively involved in inflammatory and immune processes and play important roles in human disease. However, as the interest in oxylipins increases, questions remain regarding which molecules are detectable in plasma, the best methods of collecting samples, and if molecules are stable during collection and storage. We thereby built upon existing studies by examining the stability of an expanded panel of 90 oxylipins, including specialized pro-resolving lipid mediators (SPMs), in human plasma (n = 5 subjects) during sample collection, processing, and storage at −80 °C. Oxylipins were quantified using liquid chromatography-tandem mass spectrometry (LC/MS/MS). Blood samples collected in ethylenediaminetetraacetic acid (EDTA) or heparin followed by up to 2 h at room temperature prior to processing showed no significant differences in oxylipin concentrations compared to immediately processed samples, including the SPMs lipoxin A4 and resolvin D1. The majority of molecules, including SPMs, remained stable following storage for up to 1 year. However, in support of previous findings, changes were seen in a small subset of oxylipins including 12-HETE, TXB2, 14-HDHA, and 18-HEPE. Overall, this study showed that accurate measurements of most oxylipins can be obtained from stored EDTA or heparin plasma samples using LC/MS/MS.
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10.4049/jimmunol.1601031
发表时间：
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期刊：
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影响因子：
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CLINICAL METABOLOMICS: METHODS AND PROTOCOLS
影响因子：
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发表时间：
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影响因子：
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DOI：
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发表时间：
2009-10-01
影响因子：
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