Malondialdehyde Kit Evaluated for Determining Plasma and Lipoprotein Fractions that React with Thiobarbituric Acid

Abstract

The determination of thiobarbituric acid reactants (TBARs) is a widely used method for investigating overall lipid peroxidation. An assay kit that could be used with plasma and lipid fractions would facilitate standardization of the method. The results reported here indicate that the malondialdehyde (MDA) kit manufactured by Sobioda (Grenoble, France) complies with criteria of good analytical practices. The detection limit was 0.11 mumol of MDA per liter of plasma. The within-run (CV = 1.8-3.3%) and between-run (CV = 3.3-4.4%) precisions were acceptable. The analytical recovery of MDA after supplementing human plasma samples with tetraethoxypropane standards varied from 88% to 100%. The mean (SD) lipoperoxide concentration determined in 32 healthy adults, ages 20-40 years, was 2.51 (0.25) mumol/L. No significant sex-related difference was noted: 2.57 (0.28) in men vs 2.44 (0.20) mumol/L in women. Applying the method to lipid fractions showed that lipoprotein fractions contain relatively little MDA: 0.07 (0.03) mumol/L of plasma for very-low-density lipoproteins and 0.19 (0.10) mumol/L for low-density lipoproteins.

Protective effects of ascorbic acid against the genetic and epigenetic alterations induced by 3,5‐dimethylaminophenol in AA8 cells

“…Lipid peroxidation (LP) in nuclear and cytoplasmic extracts was quantified by measuring the concentration of TBARS by a spectrofluorometric assay using a 'TBARS assay kit' at an exitation wavelength of 532 nm and emission wavelength of 553 nm with the same endpoints by Richard et al (1992). Quantification was achieved by parallel measurements of a standard curve of known TBARS concentrations, and the results were expressed as nmol g À1 protein.…”

Selenium supplementation decreases nuclear factor‐kappa B activity in peripheral blood mononuclear cells from type 2 diabetic patients

“…This measure was performed using the Sobioda TBARS kit (Grenoble, France) as described by Richard et al . [12], in which the fluorescence of thiobarbituric acid (TBA) reactants in plasma is recorded. The reagents in the kit consisted of TBA, perchloric acid and a calibration solution consisting of 1,1,3,3-tetraethoxypropane (TEP), 20 mmol L − 1 , in ethanol.…”

Vitamin E Decreases Urine Lipid Peroxidation Products in Young Healthy Human Volunteers under Normal Conditions

“…An important advantage of fluorometry relies on its great increase in sensitivity in relation to HPLC with absorbance detection (Iwata & Kikugawa 1987;Knight er al. 1987;Wong et al 1987;Richard et al 1992). The lower sensitivity of the measurement of thiobarbituric acid reactive substances by spectrophotometry leads to the need of concentrating human urine previously to detection of diethylthiobarbituric acid reactive substances by HPLC with absorbance detection at 532 nm (Guichandart et al 1994).…”

“…Furthermore, the fluorescence measurement of thiobarbituric acid reactive substances eliminates interferences from various substances (Okawa et al 1979;Bird & Draper 1984). It has been demonstrated that the direct fluorescence measurement of thiobarbituric acid reactive substances in human plasma highly correlates with thiobarbituric acid reactive substances measured in the same plasma samples by HPLC with absorbance detection at 532 nm (r=0.8; P<O.OOOl; Richard et al 1992).…”

“…Since the measurement of urinary thiobarbituric acid reactive substances by spectrophotometry alone can be inaccurate due to the presence of interfering substances, point measurements and excitation and emission spectral studies were also performed by fluorometry. Fluorescent measurement of thiobarbituric acid reactive substances, originally developed by Yagi (1976) for plasma eliminates the interference of various substances occurring in spectrophotometric measurements (Conti et al 1991;Yin 1995), increases the sensitivity over that of spectrophotometry, detects various different lipid peroxidation products further increasing sensitivity over that obtained specifically measuring a single product, and recently showed a high correlation with HPLC measurements of thiobarbituric acid reactive substances in human plasma (Richard et al 1992). The fluorescent method can be preferable for large scale clinical trials in humans due to its higher sensitivity, simplicity, speed of analysis and capacity for simultaneously processing a large amount of samples, when compared to other methods like the measurement of thiobarbituric acid products by HPLC with absorbance detection at 532 nm.…”