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Increased plasma malondialdehyde levels in glomerular disease as determined by a fully validated HPLC method
J Templar1, S P Kon, T P Milligan
1Department of Renal Medicine and Transplantation, Royal London Hospital, Whitechapel, UK.
Summary
A new HPLC method accurately measures malondialdehyde (MDA) in plasma, revealing elevated levels in chronic renal failure patients, particularly those with glomerulonephritis, indicating independent oxidative injury.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Nephrology
Background:
- Reactive oxygen species contribute to renal disease pathogenesis.
- Malondialdehyde (MDA) is a marker of lipid peroxidation, typically measured by the less specific TBARS method.
- A more sensitive and specific method for MDA measurement is needed.
Purpose of the Study:
- To develop and validate a high-performance liquid chromatography (HPLC) method for accurate MDA measurement in plasma.
- To apply this method to assess MDA levels in patients with various renal diseases.
Main Methods:
- Plasma samples underwent antioxidant treatment and protein precipitation.
- MDA was derivatized and separated using C18 reverse-phase HPLC.
- Detection of the MDA-(TBA)2 adduct was performed at 532 nm.
Main Results:
- The HPLC assay demonstrated linearity and a low detection limit (0.06 microM).
- MDA levels were significantly higher in patients with end-stage renal failure (ESRF) and chronic renal failure (CRF) compared to controls.
- Elevated MDA levels in CRF patients with glomerulonephritis were observed, independent of creatinine levels.
Conclusions:
- A reliable and sensitive HPLC method for MDA measurement in plasma was established.
- This method allows for the detection of picomole quantities of MDA.
- Significantly increased MDA levels in glomerulonephritis patients suggest oxidative injury independent of renal impairment.