Evaluation of a novel colorimetric assay for free oxygen radicals as marker of oxidative stress.
BACKGROUND
Free oxygen radicals play an important role in the pathogenesis of many diseases including cardiovascular disease, diabetes, hypertension, cancer and aging. Several methods were developed for the direct or indirect measurement of oxygen free radical and its byproducts. The free oxygen radicals monitor (FORM) is a novel point-of-care system for the rapid measurement of free oxygen radicals (FORT) in blood. We have carefully evaluated the use of this assay for batch analysis of plasma samples in a research environment with respect to factors affecting its performance, including storage temperature and duration.


METHODS
We determined the effect of storage, hemolysis, variability and reproducibility of the FORT in blood and plasma.


RESULTS
Plasma FORT correlated significantly with hsCRP (P<0.0001) and CHOL/HDL ratio (P<0.02). While hsCRP results have shown a greater range of assay variability (27.82%-53.92%), FORT measurements in the same samples have less assay variability (5.63%-9.61%). Collected whole blood can be kept on ice for up to 7 h prior to plasma isolation without affecting FORT values. Storage of plasma has no effect on FORT when stored at 4 degrees C for up to 3 weeks (R(2)=0.685). Comparable values can be obtained in samples stored for up to 3 months at -80 degrees C (R(2)=0.5888) but not at -20 degrees C.


CONCLUSIONS
The day to day variability of FORT, as assessed by multiple measures in a group of controls over time, is minimal. FORT assay is stable when stored at -80 degrees C for a couple of months or at 4 degrees C for a few weeks. FORT correlation with hsCRP and other lipid markers provides an interesting insight and a novel link between oxidative stress, inflammation and lipid metabolism.
Garelnabi
Mahdi O
MO
Division of Cardiothoracic Surgery, Ohio State University Medical Center, 460 W 12th Avenue, Room 350C, Columbus, OH 43210, USA. garelnabi@gmail.com
Brown
W Virgil
WV
Le
Ngoc-Anh
NA
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002097:C-Reactive Protein / Q000378:metabolism; D003124:Colorimetry / Q000379:methods*; D006801:Humans; D006861:Hydrogen Peroxide / Q000097:blood; D008875:Middle Aged; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000097:blood*
D017382:Reactive Oxygen Species; D002097:C-Reactive Protein; D006861:Hydrogen Peroxide