Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.bbrc.2004.06.015
Title: Rapid preparation of human urine and plasma samples for analysis of F2-isoprostanes by gas chromatography-mass spectrometry
Authors: Lee, C.-Y.J.
Jenner, A.M. 
Halliwell, B. 
Keywords: 8-Iso-PGF2α
F2-isoprostane
GC-MS-NCI
iPF2α
Oxidative stress
SPE
Issue Date: 2004
Source: Lee, C.-Y.J., Jenner, A.M., Halliwell, B. (2004). Rapid preparation of human urine and plasma samples for analysis of F2-isoprostanes by gas chromatography-mass spectrometry. Biochemical and Biophysical Research Communications 320 (3) : 696-702. ScholarBank@NUS Repository. https://doi.org/10.1016/j.bbrc.2004.06.015
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. F 2-isoprostanes were analyzed by GC-MS-NCI. The method was highly sensitive; the limit of detection at 5:1 signal-to-noise ratio was 0.037ng/ml and 0.007ng/mg creatinine for plasma and urine, respectively. Anion exchange SPE extraction for F2-isoprostane showed recovery of 55-65% and high linearity for concentration 0-1.0ng/ml for urine (CV=4.08%, r2=0.990) and 0-0.5ng/ml for plasma (CV=4.07%, r2=0.998). Fasting for 6h significantly increased plasma F2-isoprostanes levels, which has implications for the design of intervention studies using this biomarker. © 2004 Elsevier Inc. All rights reserved.
Source Title: Biochemical and Biophysical Research Communications
URI: http://scholarbank.nus.edu.sg/handle/10635/38161
ISSN: 0006291X
DOI: 10.1016/j.bbrc.2004.06.015
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

56
checked on Dec 11, 2017

WEB OF SCIENCETM
Citations

55
checked on Dec 11, 2017

Page view(s)

47
checked on Dec 9, 2017

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.