Li H, Lawson J A, Reilly M, Adiyaman M, Hwang S W, Rokach J, FitzGerald G A
Center for Experimental Therapeutics, University of Pennsylvania, Philadelphia, PA 19104, USA.
Proc Natl Acad Sci U S A. 1999 Nov 9;96(23):13381-6. doi: 10.1073/pnas.96.23.13381.
Isoprostanes (iPs) are free radical catalyzed prostaglandin isomers. Analysis of individual isomers of PGF(2alpha)-F(2)-iPs-in urine has reflected lipid peroxidation in humans. However, up to 64 F(2)-iPs may be formed, and it is unknown whether coordinate generation, disposition, and excretion of F(2)-iPs occurs in humans. To address this issue, we developed methods to measure individual members of the four structural classes of F(2)-iPs, using liquid chromatography/tandem mass spectrometry (LC/MS/MS), in which sample preparation is minimized. Authentic standards of F(2)-iPs of classes III, IV, V, and VI were used to identify class-specific ions for multiple reaction monitoring. Using iPF(2alpha)-VI as a model compound, we demonstrated the reproducibility of the assay in human urine. Urinary levels of all F(2)-iPs measured were elevated in patients with familial hypercholesterolemia. However, only three of eight F(2)-iPs were elevated in patients with congestive heart failure, compared with controls. Paired analyses by GC/MS and LC/MS/MS of iPF(2alpha)-VI in hypercholesterolemia and of 8, 12-iso-iPF(2alpha)-VI in congestive heart failure were highly correlated. This approach will permit high throughput analysis of multiple iPs in human disease.
异前列腺素（iPs）是自由基催化的前列腺素异构体。对尿液中前列腺素F2α - F2 - iPs的各个异构体进行分析，反映了人体内的脂质过氧化情况。然而，可能会形成多达64种F2 - iPs，目前尚不清楚F2 - iPs在人体内是否会协同生成、代谢和排泄。为了解决这个问题，我们开发了使用液相色谱/串联质谱法（LC/MS/MS）来测量F2 - iPs四类结构成员中各个成员的方法，该方法将样品制备过程减到最少。使用III、IV、V和VI类F2 - iPs的真实标准品来鉴定用于多反应监测的类别特异性离子。以iPF2α - VI作为模型化合物，我们证明了该检测方法在人尿液中的可重复性。在家族性高胆固醇血症患者中，所测量的所有F2 - iPs的尿液水平均升高。然而，与对照组相比，充血性心力衰竭患者中8种F2 - iPs中只有3种升高。通过气相色谱/质谱法（GC/MS）和液相色谱/串联质谱法（LC/MS/MS）对高胆固醇血症中的iPF2α - VI以及充血性心力衰竭中的8,12 - 异 - iPF2α - VI进行配对分析，结果高度相关。这种方法将允许对人类疾病中的多种iPs进行高通量分析。
