Liquid chromatography-multistage tandem mass spectrometry for the quantification of dihydroxynonene mercapturic acid (DHN-MA), a urinary end-metabolite of 4-hydroxynonenal.
The mercapturic acid conjugate of 1,4-dihydroxynonene (DHN-MA) is a urinary metabolite of 4-hydroxynonenal (4-HNE), one of the main lipid peroxidation products occurring in vivo. To determine its level in urine, a combination of liquid chromatography with positive electrospray-multistage tandem mass spectrometry has been developed. A deuterated analog of the target compound (DHN-MA) with six deuterium atoms was synthesized and used as the internal standard. Three-stage tandem mass spectrometry was used, providing good selectivity for the detection of DHN-MA. The response of the system to DHN-MA was linear in the 5-100 ng range. Urine samples spiked with different levels of standard DHN-MA were used to evaluate the influence of matrix effects on the linearity. The repeatability of the method was also determined by using repeated 5 ng injections of DHN-MA, providing a RSD of 10%. The method was then applied to the determination of DHN-MA in rat urine samples; increased levels of urinary DHN-MA in urine from rats treated with BrCCl3 indicates that lipid peroxidation processes take place in such rats.
Rathahao
E
E
UMR 1089 Xénobiotiques, INRA/ENVT, BP 3, 31931 Toulouse Cedex 9, France.
Peiro
G
G
Martins
N
N
Alary
J
J
Guéraud
F
F
Debrauwer
L
L
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000652:urine*; D000447:Aldehydes / Q000378:metabolism*; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D003903:Deuterium; D016014:Linear Models; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity
C501116:1,4-dihydroxynonene mercapturic acid; D000447:Aldehydes; D003903:Deuterium; C027576:4-hydroxy-2-nonenal; D000111:Acetylcysteine