Divergence in urinary 8-iso-PGF2x (iPF2x-III 15-F2t-IsoP) levels from gas chromatography tandem mass spectrometry quantification after thin-layer chromatography and immunoaffinity column chromatography reveals heterogeneity of 8-iso-PGF2x -: Possible methodological, mechanistic and clinical implications

被引:89
作者
Tsikas, D
Schwedhelm, E
Suchy, MT
Niemann, J
Gutzki, FM
Erpenbeck, VJ
Hohfeld, JM
Surdacki, A
Frölich, JC
机构
[1] Hannover Med Sch, Inst Clin Pharmacol, D-30625 Hannover, Germany
[2] Hannover Med Sch, Dept Resp Med, D-30623 Hannover, Germany
[3] Fraunhofer Inst Toxicol & Expt Med, Dept Immunol Allergol & Clin Inhalat, D-30623 Hannover, Germany
[4] Jagiellonian Univ, Inst Cardiol, Krakow, Poland
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2003年 / 794卷 / 02期
关键词
prostaglandins;
D O I
10.1016/S1570-0232(03)00457-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Free radical-catalysed oxidation of arachidonic acid esterified to lipids leads to the formation of the F-2-isoprostane family which may theoretically comprise up to 64 isomers. We have previously shown that the combination of TLC and GC-tandem MS (refer-red to as method A) allows for the accurate and highly specific quantification of 8-iso-PGF(2alpha) (iPF(2alpha)-III, 15-F-2t-IsoP) in human urine. Immunoaffinity column chromatography (IAC) with immobilized antibodies raised against 8-iso-PGF(2alpha) (i.e. 15(S)-8-iso-PGF(2alpha)) has been shown by others to be highly selective and specific for this 8-iso-PGF(2alpha) isomer when quantified by GC-MS. In the present study we established LAC for urinary 8-iso-PGF(2alpha) for subsequent quantification by GC-tandem MS (referred to as method B). This method was fully validated and found to be highly accurate and precise for urinary 15(S)-8-iso-PGF(2alpha). 8-iso-PGF(2alpha) was measured in urine of 10 young healthy humans by both methods. 8-iso-PGF(2alpha) was determined to be 291+/-102 pg/mg creatinine by method A and 141+/-41 pg/mg creatinine by method B. Analysis of the combined through and wash phases of the IAC step, i.e. of the unretained compounds, by method A showed the presence of non-immunoreactive 8-iso-PGF(2alpha) at 128+/-55 pg/mg creatinine. This finding suggests that urinary 8-iso-PGF(2alpha) is heterogenous, with 15(S)-8-iso-PGF(2alpha) contributing by similar to50%. PGF(2alpha) and other 8-iso-PGF(2alpha) isomers including 15(R)-8-iso-PGF(2)alpha are not IAC-immunoreactive and are chromatographically separated from 15(S)-8-iso-PGF(2alpha). We assume that ent-15(S)-8-iso-PGF(2alpha) is also contributing by similar to50% to urinary 8-iso-PGF(2alpha). This finding may have methodological, mechanistic and clinical implications. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:237 / 255
页数:19
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