Rapid preparation of human urine and plasma samples for analysis of F2-isoprostanes by gas chromatography-mass spectrometry

Biochemical and Biophysical Research Communications
Chung-Yung J LeeB Halliwell

Abstract

Reliable MS-based methods have been developed for the measurement of free and esterified F2-isoprostanes. However, prior to sample analysis several steps of purification, including solid-phase extraction followed by TLC or HPLC, are usually required, making it tedious to analyze large sample numbers, e.g., for population studies. We report a quick sample purification method using anion exchange solid phase extraction (SPE), which is highly selective for acidic compounds. Urine and hydrolyzed plasma of healthy individuals were acidified before SPE extraction, washed with 4 different solvent mixtures and finally eluted with ethyl acetate. The eluted samples were first derivatized with pentafluorobenzyl bromide followed by a second derivatization with bis-(trimethylsilyl)trifluoroacetamide. F2-isoprostanes were analyzed by GC-MS-NCI. The method was highly sensitive; the limit of detection at 5:1 signal-to-noise ratio was 0.037 ng/ml and 0.007 ng/mg creatinine for plasma and urine, respectively. Anion exchange SPE extraction for F2-isoprostane showed recovery of 55-65% and high linearity for concentration 0-1.0 ng/ml for urine (CV=4.08%, r2=0.990) and 0-0.5 ng/ml for plasma (CV=4.07%, r2=0.998). Fasting for 6h significantly increas...Continue Reading

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Citations

Oct 22, 2011·Analytical and Bioanalytical Chemistry·Magdalena LigorBogusław Buszewski
Dec 17, 2009·Antioxidants & Redox Signaling·Barry Halliwell, Chung Yung J Lee
Apr 29, 2008·American Journal of Respiratory Cell and Molecular Biology·Luke J Janssen
May 13, 2014·Analytical and Bioanalytical Chemistry·Julia KuligowskiMáximo Vento
Jan 30, 2008·Free Radical Biology & Medicine·Chung-Yung J LeeBarry Halliwell
Jan 30, 2008·Free Radical Biology & Medicine·Siu-Wai ChoiBernadette M Hannigan
Dec 28, 2010·Nutrition, Metabolism, and Cardiovascular Diseases : NMCD·C M HughesP P McKeown
May 12, 2009·Free Radical Biology & Medicine·Raymond C S SeetBarry Halliwell
Dec 30, 2008·Free Radical Biology & Medicine·Barry Halliwell
Nov 27, 2007·Free Radical Biology & Medicine·Chutima MatayatsukBarry Halliwell
Apr 25, 2006·Biochemical and Biophysical Research Communications·Chung-Yung J LeeBarry Halliwell
Dec 12, 2012·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Ming Long Sirius ChungChung-Yung Jetty Lee
Dec 9, 2014·Biochimica Et Biophysica Acta·Giuseppe AstaritaAnna Nicolaou
Jul 4, 2016·Clinica Chimica Acta; International Journal of Clinical Chemistry·Yi XiaoMaurice R O'Gorman
Apr 26, 2007·Journal of Lipid Research·Weiying YanMichael W Ogden
Apr 28, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ivan LiakhAdriana Mika
Sep 30, 2008·Free Radical Research·Delphine DelampleChristian Prefaut
Sep 15, 2010·Free Radical Research·Corinne M SpickettNeven Zarkovic
Sep 17, 2008·Biochemical Society Transactions·Jaffar Nourooz-Zadeh
Jan 19, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ivan LiakhAdriana Mika
Sep 1, 2013·Comprehensive Reviews in Food Science and Food Safety·Alaa El-Din A BekhitEric N Ponnampalam
Sep 29, 2012·Annals of Clinical Biochemistry·Zhi-Jiang Zhang

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