Determination of F2-Isoprostanes in Urine by Online Solid Phase Extraction Coupled to Liquid Chromatography with Tandem Mass Spectrometry

F2-isoprostanes are a unique class of prostaglandin-like compounds formed in vivo, which have been established as biomarkers of oxidative stress. Accurate analysis has been challenging due to lack of specificity for the isoforms of isoprostanes and lengthy sample preparation procedures to enable tra...

Full description

Saved in:
Bibliographic Details
Published in:Journal of agricultural and food chemistry 2010-06, Vol.58 (11), p.6614-6620
Main Authors: Langhorst, Marsha L, Hastings, Michael J, Yokoyama, Wallace H, Hung, Shao-Ching, Cellar, Nicholas, Kuppannan, Krishna, Young, Scott A
Format: Article
Language:English
Subjects:
accuracy
Analytical Methods
animal models
Animals
Biological and medical sciences
biomarkers
Chromatography, Liquid - methods
Cricetinae
detection
diet
extraction
F2-Isoprostanes - urine
Female
Food industries
Fundamental and applied biological sciences. Psychology
hamsters
high fat diet
human nutrition
liquid chromatography
low fat diet
Male
mass spectrometry
Mesocricetus
oxidative stress
prostaglandins
quantitative analysis
Rats
Solid Phase Extraction - methods
Tandem Mass Spectrometry - methods
urine
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:F2-isoprostanes are a unique class of prostaglandin-like compounds formed in vivo, which have been established as biomarkers of oxidative stress. Accurate analysis has been challenging due to lack of specificity for the isoforms of isoprostanes and lengthy sample preparation procedures to enable trace quantitative analysis. A quantitative analytical method was developed for the determination of F2-isoprostanes in rat and hamster urine by online solid phase extraction (SPE) coupled with liquid chromatography and tandem mass spectrometry (LC-MS/MS). The online SPE LC-MS/MS procedure has significant advantages over alternative methods with respect to specificity, sensitivity, simplicity, and speed. The assay enables the detection of iPF2α-III, iPF2α-IV, and iPF2α-VI over a linear dynamic range of 0.1−50 ng/mL in rat urine samples. This range covers the basal levels of these F2-isoprostanes. The limit of quantitation (LOQ) for the standard isoprostanes was about 0.3 ng/mL. The average recoveries ranged from 73 to 115% depending upon the individual F2-isoprostane isomers in rat urine. Additionally, the method was used to determine increases of endogenous urine iPF2α-VI and iPF2α-III in hamsters challenged with either low-fat or high-fat diets.
ISSN:0021-8561
1520-5118
DOI:10.1021/jf101146q