NONCYCLOOXYGENASE OXIDATIVE FORMATION OF A SERIES OF NOVEL PROSTAGLANDINS - ANALYTICAL RAMIFICATIONS FOR MEASUREMENT OF EICOSANOIDS
NONCYCLOOXYGENASE OXIDATIVE FORMATION OF A SERIES OF NOVEL PROSTAGLANDINS - ANALYTICAL RAMIFICATIONS FOR MEASUREMENT OF EICOSANOIDS
复制标题
DOI：
10.1016/0003-2697(90)90002-q
复制
发表时间：
1990-01-01
影响因子：
2.9
通讯作者：
ROBERTS, LJ
中科院分区：
文献类型：
--
作者：
MORROW, JD;HARRIS, TM;ROBERTS, LJ
通过负离子化学电离GC/MS对正常志愿者新鲜血浆进行分析，发现似乎存在多种PGF2化合物，其含量范围约为5至40 pg/ml。有趣的是，在-20℃的环境下储存血浆数月后，发现这些化合物的含量显著增加到约1000-4000 pg/ml，比新鲜血浆中检测到的含量高出约50倍。旨在理解这一观察结果的进一步研究表明，血浆脂质的碱性水解也产生了在储存血浆中检测到的数量范围内的这些化合物。采用许多方法，如氘化衍生物，氢化，免疫反应与抗9. α,11. β。通过电子电离质谱分析，获得了令人信服的证据，证明这些化合物在储存和碱处理的血浆中实际上是PGF2化合物。发现这些化合物的形成是通过非酶氧化过程发生的，其中抗氧化剂丁基羟基甲苯和还原剂三苯基膦明显抑制了它们的形成。有证据表明，这些化合物的氧化形成涉及内过氧化物中间体的形成，这些中间体被天然存在的生物物质直接还原为PGF2化合物。这些化合物在生物流体中很容易形成。这一发现不仅对酶衍生的PGF2化合物的分析具有重要意义，而且对可以通过相同的非酶过程形成的其他二十烷类化合物的分析也具有重要意义。这些分析问题适用于免疫分析方法和物理分析方法，如气相色谱法/质谱法。
Analysis of fresh plasma from normal volunteers by negative ion chemical ionization GC/MS reveals what appear to be multiple PGF2 compounds with levels ranging from approximately 5 to 40 pg/ml. Interestingly, storage of plasma at -20.degree.C for several months was found to markedly increase the levels of these compounds to about 1000-4000 pg/ml, approximately 50-fold higher than levels detected in fresh plasma. Further studies aimed at understanding this observation revealed that alkaline hydrolysis of plasma lipids also yielded quantities of these compounds in the range that were detected in stored plasma. Employing a number of approaches such as deuteriated derivatives, hydrogenation, immunoreactivity with an anti-9.alpha.,11.beta.-PGF2 antibody, and electron ionization mass spectral analysis, convincing evidence was obtained that these compounds in both stored and base-treated plasma were in fact PGF2 compounds. Formation of these compounds was found to occur by a nonenzymatic oxidative process in that the antioxidant, butylated hydroxytoluene, and the reducing agent, triphenylphosphine, markedly suppressed their formation. Evidence is presented to support a proposed mechanism that oxidative formation of these compounds involves the formation of endoperoxide intermediates which are directly reduced by naturally occurring biological substances to PGF2 compounds. Formation of these compounds occurs very readily in biological fluids. This finding has important ramifications not only for analysis of enzymatically derived PGF2 compounds but also for other eicosanoids which can be formed by this same nonenzymatic process. These analytical concerns apply to both immunoassay methods and physical methods of analysis such as gas chromatography/mass spectrometry.