Determination Of Malonaldehyde Research Articles

Preliminary Assessment of Stress and Genotoxicity Biomarkers in Bivalve Molluscs from the Gulf of Annaba, Algeria.

The aim of this work is to assess biomarker responses to stresses caused by pollution of the marine environment in the Gulf of Annaba, using the bivalve mollusc Donax trunculus and Mytilus galloprovincialis. Males and females were collected during the summer from site 1 considered to be far from pollution sources (SM1 and SD1), site 2 (SM2 and SD2) exposed to a mixture of contaminants, and site 3 (SM3 and SD3) receiving the untreated sewage. The whole body was used for the determination of malonaldehyde (MDA), reduced glutathione (GSH) and DNA damage. The MDA concentration from site 2 of both species has indicated significant augmentation. The two bivalves from S2 and S3 have recorded significant decrease of GSH level, with the highest number of Comet scores. A significant site and sex effect was measured for the level of GSH and MDA, accompanied with a significant site effect for DNA damage. The genotoxicity seems correlated with the stress biomarkers and dependent on the effluent types.

Improved determination of malonaldehyde by high-performance liquid chromatography with UV detection as 2,3-diaminonaphthalene derivative

A rapid, specific and simple procedure is proposed for the determination of free malonaldehyde (MA) contained in fish tissue. The method is the optimization of the reaction of MA with 2,3-diaminonaphthalene to afford a naphtodiazepinium ion that present a UV absorption at 311nm, useful for MA determination by HPLC with UV detection. The reaction proceeds in the presence of 25% acetonitrile at 37°C in 20min at pH 2 using 2,4-pentanedione as internal standard. The method has been applied to homogenized samples of canned mackerel fillets that were treated with 2,3-diaminonaphthalene in an acidic aqueous:acetonitrile mixture. The produced naphtodiazepinium ion was extracted in acetonitrile by a salting-out homogeneous liquid-liquid extraction. A standard calibration was carried out in the range 0.625-10nmol/g. The reliability of the procedure is demonstrated by linearity (r(2)=0.998), limit of detection (0.16nmol/g), limit of quantification (0.22nmol/g), repeatibility (RSD 5.57%), and intermediate precision (RSD 8.92%).

A rapid, sensitive and solvent-less method for determination of malonaldehyde in meat by stir bar sorptive extraction coupled thermal desorption and gas chromatography/mass spectrometry with in situ derivatization.

The traditional methods for analysis of malonaldehyde (MDA), such as the thiobarbituric acid (TBA) assay, require strong acidity at high temperature for derivatization and lack specificity in analysis. Stir bar sorptive extraction (SBSE) coupled with thermal desorption-gas chromatography/mass spectrometry (TD-GC/MS) with in situ derivatization using pentafluorophenylhydrazine (PFPH) under mild conditions is an emerging technique for MDA analysis. MDA in meat was derivatized with PFPH at pH ~4 for 1 h at room temperature, forming a relative stable derivative of MDA-PFPH. The derivative of MDA-PFPH was simultaneously extracted using SBSE. Then, MDA-PFPH was thermally released and quantitatively analyzed by GC/MS in selected ion monitoring (SIM) mode. The method of SBSE-TD-GC/MS for MDA analysis with in situ derivatization was optimized and validated with good linearity, specificity and limit of detection/quantification (LOD/LOQ). The method was successfully applied for analysis of MDA in raw and cooked meat (pork). The SBSE-TD-GC/MS method was suitable to monitor and analyze MDA in meat samples at trace levels. The simple, sensitive and solvent-less method with moderated in situ derivatization can be applied for analysis of MDA in a wide variety of foods and biological samples.

Nonlinear Four-Way Kinetic-Excitation−Emission Fluorescence Data Processed by a Variant of Parallel Factor Analysis and by a Neural Network Model Achieving the Second-Order Advantage: Malonaldehyde Determination in Olive Oil Samples

Four-way data were obtained by recording the kinetic evolution of excitation-emission fluorescence matrices for the product of the Hantzsch reaction between the analyte malonaldehyde and methylamine. The reaction product, 1,4-disubstituted-1,4-dihydropyridine-3,5-dicarbaldehyde, is a highly fluorescent compound. The nonlinear nature of the kinetic fluorescence data has been demonstrated, and therefore the four-way data were processed with parallel factor analysis combined with a nonlinear pseudounivariate regression, based on a quadratic polynomial fit, and also with a recently introduced neural network methodology, based on the combination of unfolded principal component analysis, residual trilinearization, and radial basis functions. The applied chemometric strategies are not only able to adequately model the nonlinear data but also to successfully determine malonaldehyde in olive oil samples. This is possible since the experimentally recorded four-way data, modeled with the above-mentioned advanced chemometric approaches, permit the achievement of the second-order advantage. This allows us to predict the analyte concentration in a complex background, in spite of the nonlinear behavior and in the presence of uncalibrated interferences. The present work is a new example of the use of higher-order data for the resolution of a complex nonlinear system, successfully employed in the context of food chemical analysis.

Determination of Malonaldehyde in Bovine Plasma During the Receiving Period and Effects of Phytoderivative Diet Supplementation

Determination of Malonaldehyde in Bovine Plasma During the Receiving Period and Effects of Phytoderivative Diet Supplementation

Improved procedure for the determination of malonaldehyde by gas-chromatography with electron-capture detection as 2,4,6-trichlorophenylhydrazine derivative

A previously described derivatization method using trichlorophenylhydrazine was developed for the estimation of malonaldehyde measured by gas-chromatography (GC) with electron-capture detection. The precision and reliability of the procedure are improved here by the use of methylmalonaldehyde as internal standard and by the introduction of a diverter valve at the end of the capillary column to protect the electron-capture detector, respectively. The method was applied to determine malonaldehyde content in bovine plasma samples.

Specific determination of malonaldehyde by N‐methyl‐2‐phenylindole or thiobarbituric acid

AbstractReactivity of a commercially available test kit (LPO‐586), based on N‐methyl‐2‐phenylindole, toward aldehydes was characterized and compared with that of thiobarbituric acid (TBA). In hydrochloric acid, LPO‐586 produced a violet pigment with malonaldehyde (MA) but not with other tested aldehydes. In methane sulfonic acid, LPO‐586 produced the violet pigment with MA and 4‐hydroxynonenal (HNE), but not with other tested aldehydes. Pigment formation with MA was not inhibited by other aldehydes, but that with HNE was inhibited by alka‐2,4‐dienals. TBA produced a red pigment with MA but not with other tested aldehydes in hydrochloric acid or in acetate with ethylenediaminetetraacetic acid (EDTA). Both the LPO‐586 test in hydrochloric acid and the TBA test in hydrochloric acid or in acetate with EDTA can be used for specific measurement of MA in oxidized lipid samples.

Cyclodextrin enhanced fluorimetric determination of malonaldehyde by the thiobarbituric acid method

The enhancement effects of α-, β-, γ;- and hydroxypropyl-β-cyclodextrins on the fluorescence of a 2:1 thiobarbituric acid/malonaldehyde adduct in acid aqueous solutions have been studied. The best characteristics as a fluorescence enhancement agent showed hydroxypropyl-β-cyclodextrin which bound the adduct sufficiently tightly (K = 180 1 mol −1) mol −1) and caused a five-fold increase in its fluorescence. A kinetic-fluorimetric method of determination of malonaldehyde in the range 0.1–10 μM at room temperature with hydroxypropyl-β-cyclodextrin as the enhancement agent is proposed and applied for the analysis of raw and cooked meat samples.

Determination of Malonaldehyde in the Samples of Cardiac Myocytes by 2.5th Order Differential Voltammetry

The 2.5th order differential polarographic reduction behavior of malonaldehyde (MLD) was studied. A sensitive measurement method of the determination of MLD for biological samples was given by using differential voltammetry with a static mercury drop electrode at a scanning speed of 80mV/s in a solution of 0.1mol/L NH4Cl. The linear response range of current(e″) against the concentration of MLD was 1.0×10−6 to 1.0×10−3 mol L and the detection limit was 1.0 × 10−7mol/L. The contents of MLD in the sample of cardiac myocytes was determined rapidly and simply by means of the method.

Determination of malonaldehyde in Alzheimer's disease: a comparative study of high-performance liquid chromatography and thiobarbituric acid test.

The concentration of malonaldehyde (MDA) was measured in human erythrocytes obtained from subjects suffering from senile dementia of the Alzheimer type (SDAT), non-demented elderly subjects, and from young controls by two methods: high-performance liquid chromatography (HPLC) and the thiobarbituric acid (TBA) test. The MDA concentration measured by HPLC showed significant differences between SDAT and young control groups (p < 0.01) and between SDAT and non-demented elderly groups (p < 0.01), respectively. Nevertheless, significant differences were not exhibited between young control and non-demented elderly. Moreover, the rate of accumulation of TBA-reactive substances was not significantly different among the three groups. Our results indicate that the HPLC method is highly specific and accurate and distinguishes between true MDA and other aldehydes that may react with TBA. Significant increases in the concentration of MDA of SDAT subjects were found in comparison with the two other groups, indicating that the measurement of MDA in erythrocytes could be used as a marker of oxidative damage in Alzheimer's disease.

Kinetic−Fluorometric Determination of Malonaldehyde Based on the Hantzsch Reaction: Application to Olive Oil Analysis

A kinetic study was completed on the formation of 1,4-disubstituted-1,4-dihydropyridine-3,6 dicarbaldehyde from malonaldehyde and methylamine, via the Hanztsch reaction. The influence of n-hexylaldehyde and acetaldehyde in the reaction has been studied, and it has been concluded that the presence of the aldehydes is not necessary for the reaction. The fluorescence emission of the reaction product was monitored at 470 nm, with excitation at 405 nm, and the effect of instrumental and experimental variables on the reaction was investigated. A pH of 4.0, obtained by a 0.25 M sodium acetate-acetic acid buffer solution, a 10 mM methylamine concentration, a 30% (v/v) 2-propanol content, and a temperature of 75 degrees C were selected for the reaction. A kinetic-fluorometric method to determine malonaldehyde has been proposed, for malonaldehyde concentrations ranging from 0.5 to 2.8 micrograms/mL-1, and a wide study of the possible interferences in the reaction has been performed. The kinetic-fluorometric method was applied to olive oil samples.

Kinetic behaviour of the malonaldehyde-thiobarbituric acid reaction. kinetic-fluorimetric determination of malonaldehyde in human serum

For the first time a fluorimetric-kinetic method for the determination of malonaldehyde (MLD) is proposed. The fluorescence emission of the thiobarbituric acid (TBA)-MLD reaction product has been monitored at 553 nm (excitation wavelength 515 nm), and the instrumental and experimental variables have been studied. A selective and fast kinetic-fluorimetric determination of malonaldehyde is proposed. The sensitivity of the method is very high, the detection limit is 0.30 ng ml −1, the application range is between 1.1 and 50 ng ml −1, and the determination time per sample is 500 s. The kinetic-fluorimetric method has been applied in the determination of two types of pathological human serum: rheumatic and hyperlipemic serum. In both cases, perchloric acid is used to precipitate the proteins.

Kinetic Study of the Malonaldehyde-Azulene Reaction Determination of Malonaldehyde in Human Plasma

The influence of physicochemical conditions in the formation of derivatized malonaldehyde (MLD)-azulene was studied, and kinetic and conventional spectrophotometric data were compared. Conventional and kinetic spectrophotometric procedures were used to study inference of several compounds. An advantageous kinetic method was used to determine MLD in human plasma. The proposed method is faster than the thiobarbituric acid (TBA) method, because 150 s is sufficient to develop the reaction and the kinetic measurements. A comparative study of the results obtained using the proposed kinetic method and those reported in the literature for MLD determination by using the TBA reaction is presented.

Determination of malonaldehyde by coupled high-performance liquid chromatography—spectrofluorimetry after derivatization with luminarin 3

A high-performance liquid chromatographic method with fluorescence detection is described for the specific determination of free malonaldehyde (MDA). Malonaldehyde was labelled with luminarin 3 in acetate buffer (pH 4.0) at room temperature in the dark. An aliquot of the reaction mixture was injected on to an octadecyl-bonded column using acetonitrile—imidazole buffer (30:70, v/v) as the mobile phase. The eluate was monitored with a fluorescence detector at 395 nm (excitation) and 500 nm (emission). A linear calibration graph was established between 7.2 and 90 ng/ml of MDA and the limit of quantification (LOQ) was lower than 7 ng/ml of MDA. The precision was characterized by R.S.D.s of 11% at 7.2 ng/ml and 2% at 90 ng/ml. The structure of the derivative was confirmed as the 5-hydroxy-2-pyrazoline form. UV absorbance and corrected fluorescence spectral data and quantum yields of the luminarin 3 derivative of malonaldehyde are presented.

Determination of malonaldehyde in hyperlipemic sera

Determination of malonaldehyde in hyperlipemic sera

Determination of malonaldehyde in human plasma: elimination of spectral interferences in the 2-thiobarbituric acid reaction.

A selective, derivative spectrophotometric method has been developed for the determination of malonaldehyde (MLD), based on a reaction with 2-thiobarbituric acid (TBA). The proposed method has been applied to the determination of MLD in human plasma. A study to eliminate several spectral interferences is described. A comparative study of the results obtained using the proposed derivative method and a conventional TBA method applied to human plasma is presented and the advantages of the proposed method over the conventional method for the determination of MLD in human plasma are evaluated.

Analysis of free malonaldehyde formed in lipid peroxidation systemsvia a pyrimidine derivative

A high‐performance liquid chromatographic (HPLC) method for the determination of malonaldehyde (MA) in foods and biological samples was developed. MA was derivatized by reaction with urea under acidic conditions to form 2‐hydroxypyrimidine, which was subsequently measured by HPLC. The highest yield (98%) of the product was obtained when 100 nmol of MA was reacted with 60 mmol of urea for 60 min at 100°C. Arachidonic acid, linolenic acid, linoleic acid and oleic acid were oxidized by a FeCl2/H2O2 reagent in aqueous solution. MA formed was determined as 2‐hydroxypyrimidine by HPLC. Arachidonic acid produced the highest level of MA (60 nmol/mg fatty acid), whereas oleic acid did not produce any. The formation levels of MA in microsomes upon enzymatic and nonenzymatic oxidation were 34 nmol/mL and 45 nmol/mL, respectively. Antioxidative activity of α‐tocopherol was also monitored successfully by this HPLC method.

Malonaldehyde determination in foods by ion-pairing high-performance liquid chromatography

A direct method which involved no thiobarbiturate chromogen formation for the analysis of malonaldehyde in foods by ion-pairing high-performance liquid chromatography was developed. After deproteinization and cleaning, diluted samples (20 μl) were injected onto an octadecylsilane column (25 cm × 4·6 mm) i.d., 5 μm) and eluted with 30 m m sodium phosphate buffer, pH 6·5 containing 30% (v/v) ethanol and 0·5 m m tetradecyltrimethylammonium bromide. Detection was accomplished by monitoring absorbance at 267 nm. The lower limit for reliable quantification was 5 pmol per injection. Traces or no malonaldehyde were detected in fresh samples of selected foods. However, after a period of several weeks of refrigerator storage of opened containers, or after chemically induced lipid peroxidation, food samples contained a substantial amount of malonaldehyde.

Determination of malonaldehyde and formaldehyde formed from fatty acid ethyl esters upon microwave and thermal heating

Aqueous fresh and frozen solutions of ethyl arachidonate, ethyl linolenate, and ethyl linoleate were exposed to microwave and thermal heating to induce lipid peroxidation. Malonaldehyde and formaldehyde were derivatized to 1-methylpyrazole with N-methylhydrazine and to thiazolidine with cysteamine, respectively, and were then quantitated with capillary gas chromatography with a nitrogen-phosphorus detector. For both heating methods, the malonaldehyde concentration depended on the degree of unsaturation in the fatty acid. The samples containing arachidonate produced the most malonaldehyde (7.8-11.2 nmol/mg), followed by those containing linolenate (2.9-5.3 nmol/mg) and linoleate (0.7-1.7 nmol/mg). There were no significant differences in the amounts of malonaldehyde formed between the heating methods for both fresh and frozen solutions. Ethyl arachidonate and ethyl linolenate produced about the same amount of formaldehyde (5.8-10.4 nmol/mg), whereas ethyl linoleate produced slightly less (3.2-5.3 nmol/mg) formaldehyde.

Improved fluorometric determination of malonaldehyde

Lipoperoxidation is implicated in various pathological conditions. Malonaldehyde (MDA) is the most commonly used marker of this process. We propose simple modifications to Yagi's fluorometric assay for MDA determinations, to avoid long and tedious manipulations by eliminating the first precipitation and washing steps, analogous to HPLC methods, and to increase both the sensitivity and the specificity of the assay by measuring synchronous fluorescence. The proposed technique is easier, faster, and more sensitive than Yagi's method (Academic Press, 1982: Lipid peroxides in biology and medicine). The results obtained with the novel method correlate with those from the HPLC method described by Therasse and Lemonnier (J Chromatogr Biomed Appl 1987;413:237-41).