Urinalysis of 4-hydroxynonenal, a marker of oxidative stress, using stir bar sorptive extraction-thermal desorption-gas chromatography/mass spectrometry.
A simple and fast method for the measurement of 4-hydroxynonenal (4HNE), a highly toxic end-product of lipid peroxidation, in urine samples is described. The method combines stir bar sorptive extraction (SBSE) with two derivatization steps, followed by thermal desorption and GC/MS. 4HNE is derivatized in situ with O-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine and the oxime is extracted from the aqueous phase with SBSE. The 4HNE-oxime is further acylated by headspace derivatization prior to thermal desorption. Derivatization reactions and extraction were optimized in terms of reagent quantities, temperature and time. The method is linear over a concentration range of 0.5-5 ng mL(-1) with a correlation coefficient of 0.997. The limit of detection and limit of quantitation are 22 and 75 pg mL(-1) urine, respectively. The high sensitivity of the method allows the measurement of physiological concentrations of 4HNE in urine samples.
Stopforth
Adriana
A
Department of Chemistry, University of Stellenbosch, Private Bag X1, Matieland 7602, South Africa.
Burger
Ben V
BV
Crouch
Andrew M
AM
Sandra
Pat
P
D000447:Aldehydes / Q000652:urine*; D015415:Biomarkers / Q000652:urine*; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D008297:Male; D018384:Oxidative Stress
D000447:Aldehydes; D015415:Biomarkers; C027576:4-hydroxy-2-nonenal