Liang Yuanling, Wei Ping, Duke Russell W, Reaven Peter D, Harman S Mitchell, Cutler Richard G, Heward Christopher B
Kronos Science Laboratory, Phoenix, AZ, USA.
Free Radic Biol Med. 2003 Feb 15;34(4):409-18. doi: 10.1016/s0891-5849(02)01018-3.
Quantification of 8-iso-prostaglandin F(2alpha) (8-iso-PGF(2alpha)) has been suggested to be a reliable indicator of lipid peroxidation that may be related to in vivo free radical generation, oxidative damage, and antioxidant deficiency. We have developed a LC-MS/MS method to quantify 8-iso- PGF(2alpha) and its dinor metabolite, 2,3-dinor-8-iso-prostaglandin F(2alpha) (2,3-dinor-8-iso-PGF(2alpha)), in human urine samples. After an initial purification step using an automated C18 solid phase extraction procedure, the urine sample was injected directly into a liquid chromatography (LC) system and detected with tandem mass spectrometry. The detection limit of the assay was 9 pg for 8-iso-PGF(2alpha) and 3 pg for 2,3-dinor-8-iso-PGF(2alpha) with both inter- and intraday variations of less than 12%. The inaccuracies were less than 3% for both analytes at three different levels. The urinary excretion rate of 2,3-dinor-8-iso-PGF(2alpha) was higher than that of 8-iso-PGF(2alpha), and changed in proportion to the parent compound (R = 0.70, n = 60). Values obtained with this method showed good linear correlation to duplicate 8-iso-PGF(2alpha) measurements performed with GCMS (R = 0.97, n = 15). The mean excretion rates of 8-iso-PGF(2alpha) and 2,3-dinor-8-iso-PGF(2alpha) were significantly higher in smokers than in nonsmokers (0.53 +/- 0.37 vs. 0.25 +/- 0.15 microg/g creatinine, p = 0.002 for 8-iso-PGF(2alpha) and 8.9 +/- 3.8 vs. 4.6 +/- 2.6 microg/g creatinine, p = 0.003 for 2,3-dinor-8-iso-PGF(2alpha), respectively). The excellent accuracy, reproducibility, and high throughput of this method should permit it to be used in large clinical studies and standard clinical laboratories.
8-异前列腺素F(2α)（8-iso-PGF(2α)）的定量分析被认为是脂质过氧化的可靠指标，这可能与体内自由基生成、氧化损伤和抗氧化剂缺乏有关。我们开发了一种液相色谱-串联质谱法（LC-MS/MS），用于定量分析人尿液样本中的8-异前列腺素F(2α)及其双降代谢物2,3-双降-8-异前列腺素F(2α)（2,3-dinor-8-iso-PGF(2α)）。在使用自动C18固相萃取程序进行初步纯化步骤后，将尿液样本直接注入液相色谱（LC）系统，并通过串联质谱进行检测。该检测方法对8-异前列腺素F(2α)的检测限为9皮克，对2,3-双降-8-异前列腺素F(2α)的检测限为3皮克，日内和日间变异均小于12%。在三个不同水平下，两种分析物的误差均小于3%。2,3-双降-8-异前列腺素F(2α)的尿排泄率高于8-异前列腺素F(2α)，且与母体化合物成比例变化（R = 0.70，n = 60）。用该方法获得的值与用气相色谱-质谱法（GCMS）进行的重复8-异前列腺素F(2α)测量结果显示出良好的线性相关性（R = 0.97，n = 15）。吸烟者中8-异前列腺素F(2α)和2,3-双降-8-异前列腺素F(2α)的平均排泄率显著高于非吸烟者（8-异前列腺素F(2α)：0.53±0.37 vs. 0.25±0.15微克/克肌酐，p = 0.002；2,3-双降-8-异前列腺素F(2α)：8.9±3.8 vs. 4.6±2.6微克/克肌酐，p = 0.003）。该方法具有出色的准确性、重现性和高通量，应使其能够用于大型临床研究和标准临床实验室。
