Nephrology Dialysis Transplantation, Vol 14, Issue 4 946-951, Copyright © 1999 by Oxford University Press
J Templar, S Kon, T Milligan, D Newman and M Raftery
Background. Reactive oxygen species and particularly
free radical induced lipid peroxidative tissue damage have been implicated
in the pathogenesis of various renal diseases. Lipid peroxidation is
assessed indirectly by the measurement of secondary products, such as
malondialdehyde (MDA), using the widely employed thiobarbituric acid
reactive substances (TBARS) method. However, this method lacks sensitivity
and specificity. We have therefore developed and validated an HPLC
(high-performance liquid chromatography) method for measurement of MDA and
applied this to a variety of plasma samples in renal patients.
Methods. The optimized method involves antioxidant
treatment of the plasma sample, followed by a protein precipitation step
using trichloroacetic acid, acid hydrolysis and formation of an MDA
thiobarbituric acid complex. The MDA-(TBA)2 adduct is separated from other
interfering compounds by C18 reverse-phase HPLC techniques, with visible
detection at 532 nm. Results. The assay was linear
over the ranges 0.25-1.0 &mgr;M MDA and the detection limit was 0.06
&mgr;M MDA. Within-run precision was <4.5% and between-run
precision was <10.0%. MDA plasma concentrations (mean±SD)
were higher in ESRF diabetic patients (0.32±0.14 &mgr;M,
n=20), non-diabetic ESRF patients
(0.32±0.09 &mgr;M, n=20), and CRF
patients (0.14±0.06 &mgr;M, n=40)
compared to healthy controls (0.11±0.03 &mgr;M,
n=40), (P<0.001,
P<0.001 and P=0.008).
Levels were similar in healthy controls with normal renal function and
transplanted patients (0.12±0.03 &mgr;M MDA,
n=40), (P=NS). No correlation was
observed between MDA and creatinine levels
(r2=0.05,
n=80), which suggests that MDA does not correlate with
the degree of renal impairment. We matched CRF patients with glomerular and
non-glomerular causes of renal failure for creatinine levels and found that
MDA levels were higher in patients with glomerulonephritis
(0.16±0.06 &mgr;M) than in those with CRF from
non-glomerular causes (0.12±0.04 &mgr;M,
P=0.002). Conclusions. We have
introduced a reliable and sensitive HPLC technique to enhance the
specificity of MDA-(TBA)2 measurement, with a significant improvement in
HPLC column life. Using this method, picomole quantities of MDA can be
detected in plasma. We have shown that MDA levels are significantly raised
in patients with CRF due to glomerulonephritis, regardless of serum
creatinine, which suggests that there is oxidative injury independent of
any possible MDA retention due to renal impairment. Keywords:
glomerulonephritis; high performance liquid chromatography;
malondialdehyde; reactive oxygen species; thiobarbituric acid
TECHNICAL REPORT
Increased plasma malondialdehyde levels in glomerular disease as determined by a fully validated HPLC method
Department of Renal Medicine and Transplantation and Department of Clinical Biochemistry, Royal London Hospital, Whitechapel, London E1 1BB, UK; Current address: Southwest Thames Institute for Renal Research, St Helie NHS Trust, Carshalton, Surrey SM5 1AA, UK; Corresponding author
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  D Loru, A Incani, M Deiana, G Corona, A Atzeri, M. Melis, A Rosa, and M. Dessi Protective effect of hydroxytyrosol and tyrosol against oxidative stress in kidney cells Toxicology and Industrial Health, May 1, 2009; 25(4-5): 301 - 310. [Abstract] [PDF]
|
|
|
|
 |
|
 M. F. Walter, R. F. Jacob, R. E. Bjork, B. Jeffers, J. Buch, Y. Mizuno, R. P. Mason, and on behalf of the PREVENT Investigators Circulating Lipid Hydroperoxides Predict Cardiovascular Events in Patients With Stable Coronary Artery Disease: The PREVENT Study J. Am. Coll. Cardiol., March 25, 2008; 51(12): 1196 - 1202. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z Yetkin-Ay, B Cadir, E Uskun, F. Bozkurt, N Delibas, F. Gultepe, and I Ergurhan-Ilhan The periodontal status of indirectly lead-exposed apprentices working in autorepair workshops Toxicology and Industrial Health, November 1, 2007; 23(10): 599 - 606. [Abstract] [PDF]
|
|
|
|
 |
|
 E. Papalambros, F. Sigala, S. Georgopoulos, K. I. Paraskevas, I. Andreadou, X. Menenakos, P. Sigalas, A. L. Papalambros, G. Vourliotakis, A. Giannopoulos, et al. Malondialdehyde as an Indicator of Oxidative Stress During Abdominal Aortic Aneurysm Repair Angiology, September 1, 2007; 58(4): 477 - 482. [Abstract] [PDF]
|
|
|
|
 |
|
 V. B. Tognetti, J. F. Palatnik, M. F. Fillat, M. Melzer, M.-R. Hajirezaei, E. M. Valle, and N. Carrillo Functional Replacement of Ferredoxin by a Cyanobacterial Flavodoxin in Tobacco Confers Broad-Range Stress Tolerance PLANT CELL, August 1, 2006; 18(8): 2035 - 2050. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Goulart, M.C. Batoreu, A.S. Rodrigues, A. Laires, and J. Rueff Lipoperoxidation products and thiol antioxidants in chromium exposed workers Mutagenesis, September 1, 2005; 20(5): 311 - 315. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Vega-Lopez, K.-J. Yeum, J. L Lecker, L. M Ausman, E. J Johnson, S. Devaraj, I. Jialal, and A. H Lichtenstein Plasma antioxidant capacity in response to diets high in soy or animal protein with or without isoflavones Am. J. Clinical Nutrition, January 1, 2005; 81(1): 43 - 49. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. F. Walter, R. F. Jacob, B. Jeffers, M. M. Ghadanfar, G. M. Preston, J. Buch, and R. P. Mason Serum levels of thiobarbituric acid reactive substances predict cardiovascular events in patients with stable coronary artery disease: A longitudinal analysis of the PREVENT study J. Am. Coll. Cardiol., November 16, 2004; 44(10): 1996 - 2002. [Abstract] [Full Text] [PDF]
|
|