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Biomedical subjects

G Lercker

Publications and source records attributed to G Lercker.

At least 19 recordsLinked to original sources
PubMed2005

Evaluation of the antioxidant capacity of individual phenolic compounds in virgin olive oil.

Virgin olive oil has a high resistance to oxidative deterioration due to its tryacylglycerol composition low in polyunsaturated fatty acids and due to the presence of a group of phenolic antioxidants composed mainly of polyphenols and tocopherols. We isolated several phenolic compounds of extra virgin olive oil (phenyl-ethyl alcohols, lignans, and secoiridoids) by semipreparative high-performance liquid chromatography (HPLC) and identified them using ultraviolet, atmospheric pressure chemical ionization, and electrospray ionization MS detection. The purity of these extracts was confirmed by analytical HPLC using two different gradients. Finally, the antioxidant capacity of the isolated compounds was evaluated by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl radical, by accelerated oxidation in a lipid model system (OSI, oxidative stability instrument), and by an electrochemical method.

Antioxidants↗
PubMed2004

Calculation of the molar absorptivity of polyphenols by using liquid chromatography with diode array detection: the case of carnosic acid.

Antioxidant activity of vegetable extracts is related to the nature and the amount of active components, mainly polyphenols; therefore, a correct quantification of these molecules should be required to define their concentration in such kind of vegetable extracts. A fast and accurate method to calculate molar absorption coefficients (epsilon), by using HPLC, has been tested on standard polyphenols and caffeine, and should be widely adapted for standardless quantitative analysis. Molar absorptivity (epsilon) of carnosic acid (CA) was determined from 200 to 300 nm, by the proposed method and those values were compared to tert-butyl-hydroxytoluene (BHT) ones for further comparative quantification.

Abietanes↗
PubMed2004

Magnetic resonance spectroscopy and chromatographic methods identify altered lipid composition in human renal neoplasms.

We report on the characterization of the lipid obtained from cortical and medullary normal human kidney tissue, benign renal neoplasms (oncocytoma) and 2 different types of malignant renal neoplasms (chromophobic cell carcinoma and clear cell carcinoma). The total lipid fractions were analyzed by 13C magnetic resonance spectroscopy and thin-layer chromatography, whereas the composition of the total fatty acids and the content of total cholesterol were determined by gas chromatography. alpha-Tocopherol was detected and quantified by high-performance liquid chromatography. The spectroscopic and chromatographic analysis revealed significant differences in the renal tissues examined. It was confirmed that cholesteryl esters (mainly oleate) are typical of clear cell renal carcinomas. Their potential role as prognostic and diagnostic factors is discussed, with particular emphasis on its capability to indicate the tumor diffusion in healthy renal parenchyma. alpha-Tocopherol is prevalent in clear cell carcinoma and it is present in nearly the same low amounts in cortex, medulla and chromophobic cell renal carcinoma. Q10 coenzyme and dolichols were detected by thin-layer chromatography and they are present in significant amounts in the cortex and the benign oncocytoma. Great variations were found in the distribution of saturated and unsaturated fatty acids, especially in the docosapentaenoic, docosahexaenoic and arachidonic acids and the corresponding omega-6/omega-3 fatty acids ratio.

Chromatography↗
PubMed2001

Pressurized liquid extraction of lipids for the determination of oxysterols in egg-containing food.

Pressurized liquid extraction (PLE, ASE) was compared with the Folch procedure (a solid-liquid extraction with chloroform/methanol 2:1, v/v) for the lipid extraction of egg-containing food; the accuracy of PLE for the quantitative determination of oxysterols in whole egg powder was evaluated. Samples of spray-dried whole egg, an Italian vanilla cake (Pandoro) and egg noodles were used. Two different extraction solvents (chloroform/methanol 2:1, v/v, and hexane/isopropanol 3:2, v/v) were tested at different extraction temperatures and pressures (60 degrees C at 15 MPa, 100 degrees C at 15 MPa, 120 degrees C at 20 MPa). No significant differences in the lipid recovery of the egg powder sample using PLE were found. However, PLE of the vanilla cake and egg noodles with the chloroform/methanol mixture was not selective enough and led to the extraction of a non-lipid fraction, including nitrogen-containing compounds. In the same samples, the pressurized hexane/isopropanol mixture gave a better recovery result, comparable to that obtained using the Folch method. Cholesterol oxidation products of the Folch extract and the pressurized liquid extract of spray dried egg powder (obtained with hexane/isopropanol 3:2, v/v, at 60 degrees C and 15 MPa) were determined by gas chromatography. PLE performed under these conditions is suitable to replace the Folch extraction, because the differences between the two methods tested were not statistically significant. Moreover, PLE shows important advantages, since the analysis time was shortened by a factor of 10, the solvent costs were reduced by 80% and the use of chlorinated solvents was avoided.

Chromatography, Gas↗
PubMed2000

Chromatographic analysis of unsaponifiable compounds of olive oils and fat-containing foods.

The analysis of the "minor components" present in food lipids is usually hampered by the large diversity of compounds found in this fraction. High-purity degree reagents and solvents, good collection techniques and highly sensitive analysis are required in order to accurately identify and quantify these components. Chromatographic techniques have proven to be particularly suitable for these determinations, especially capillary gas chromatography. This study reports several analytical cases of the main classes of components of the unsaponifiable matter obtained from olive oils or food matrices.

Chromatography↗
PubMed2000

Mass spectral fragmentations of cholesterol acetate oxidation products.

In this work, electron-impact mass spectroscopy (EI-MS) was employed on a wide range of sterol compounds in order to study their behaviour with regard to their functional groups. In particular, some specific mechanisms of fragmentation occurring in these substrates (i.e. retro-Diels-Alder reaction, neutral molecules eliminations, specific hydrogen migrations) were investigated. Loss of the alkyl side chain and of the D ring were observed in all cases. Finally, a classification of sterols on the basis of characteristic mass spectral fragments is suggested, and further applications to substrates with functional groups on positions other than the A and B rings is proposed.

Animals↗
PubMed2000

The protective role of different green tea extracts after oxidative damage is related to their catechin composition.

The antioxidant activities of three different green tea extracts were investigated and compared by two different methods. By the first method, which evaluated the direct protective effect of the green tea extracts on lipid peroxidation, the extracts were added, at different concentrations, to a lipid model system, made by refined peanut oil, freshly submitted to a further bleaching and subjected to forced oxidation at 98 degrees C, by an oxidative stability instrument. By the second method, the effectiveness of the same extracts was checked in cultures of neonatal rat cardiomyocytes exposed to a free radical-generating system by evaluating conjugated diene production and lactate dehydrogenase release. All of the extracts revealed a strong antioxidant activity by both the methods, and a particular effectiveness was demonstrated by the extracts having higher amounts of (-)-epigallocathechin-3-gallate and (-)-epigallocathechin, as analyzed by reverse-phase HPLC analysis.

Animals↗
PubMed2000

Determination of furan fatty acids in extra virgin olive oil.

The presence of 4 different furan fatty acids (F-acids) was detected in 18 samples of transmethylated monovarietal extra virgin olive oil: methyl 10,13-epoxy-11,12-dimethyloctadeca-10,12-dienoate [diMeF(9,5)], methyl 12,15-epoxy-13,14-dimethyleicosa-12,14-dienoate [diMeF(11,5)] and both olefinic derivatives of diMeF(11,5) with one unsaturation on the side chains conjugated with the furan ring. Transmethylated oils were analyzed by normal phase high-performance liquid chromatography coupled on-line with capillary gas chromatography. After the gas chromatographic separation step, a more selective detection of F-acids was achieved by using a photoionization detector mounted in series with a flame ionization detector. The concentration of F-acids ranged between 50 ppb (detection limit of the method) and 2.1 ppm in the oil. The olefinic derivatives of diMeF(11,5) acids detected were not artifacts created during the sample preparation or during the chromatographic analysis.

Chromatography, Gas↗
PubMed1995

Determination of polycyclic aromatic hydrocarbons in water and water-based alcoholic beverages.

This paper proposes a simple HPLC method for the determination of polycyclic aromatic hydrocarbons (PAHs) in water, wine and beer. Samples were purified by PAH collection in solid-phase extraction (SPE) and analysed by reversed-phase HPLC (Supelcosil LC-PAH column from Supelco). For the beer sample, recoveries amounted to 28% for naphthalene and varied from 57% to 103% for the other PAHs; results are quantitative starting from fluoranthene (FI, the seventh component eluted). Almost all the beer and wine samples showed the presence of benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF), benzo(a)pyrene (BaP), benz(ghi)perylene (BghiP) and indeno(1,2,3-cd)pyrene (IP), and in some cases there were traces of FI, benzo(a)anthracene (BaA) and dibenz(ah) anthracene (DBahA). Total contents of PAHs ranged from trace amounts to 0.72 ppb. Traces of BbF, BkF, BaP, BghiP and IP were also found in the wine samples.

Beer↗
PubMed1995

Incorporation of cholesterol oxidation products into cell lipids and their influence on the proliferation of cultured cardiomyocytes.

We have investigated the incorporation of cholesterol oxidation products into cardiomyocyte lipids and related this to changes in cell proliferation, evaluated by measuring cellular protein content. Primary cultures of neonatal rat ventricular cells were supplemented with scalar concentrations of several cholesterol oxidation products (cholestan-5 alpha, 6 alpha-epoxy-3 beta-ol, 5 alpha-cholestane-3 beta, 5, 6 beta-triol, 5-cholesten-3 beta, 4 beta-diol, 5-cholesten-3 beta-ol-7-one, and 5-cholesten-3-one). Although all the cholesterol oxidation products were incorporated into the cardiomyocyte lipids when added to the medium at a concentration higher than 0.5 microM, the extent of the incorporation of the different cholesterol oxidation products differed, depending on the concentration in the culture medium and on the chemical structure of the compound. The effects of the cholesterol oxidation products on the cellular protein content were also different: 5 alpha-cholestane-3 beta, 5, 6 beta-triol was shown to be the most potent inhibitor of cell proliferation, followed by cholestan-5 alpha, 6 alpha-epoxy-3 beta-ol, 5-cholesten-3 beta, 4 beta-diol and 5-cholesten-3 beta-ol-7-one. 5-Cholesten-3-one did not affect the cellular protein content. The ability of cholesterol oxidation products to inhibit cell proliferation, and their capacity to increase the permeability of the plasma membrane to calcium, could be deleterious for cardiac cells.

Animals↗
PubMed1994

High-performance liquid chromatography separation and light-scattering detection of phospholipids from cooked beef.

A sensitive high-performance liquid chromatography (HPLC) method for the separation and quantitative analysis of major phospholipids (PLs) in biological systems is described. PLs were purified by solid-phase extraction with an amino (NH2) phase. Separation of PLs was carried out on an HPLC silica gel column, with a mobile phase consisting of chloroform, methanol and ammonium hydroxide, and detection was performed with a light-scattering evaporative detector. HPLC analysis of PLs extracted from ground beef cooked under different conditions and capillary gas chromatography of the fatty acid methyl esters showed that cooking treatments did not have a significant effect on the PL composition and fatty acid contents of the single PLs in ground beef.

Animals↗
PubMed1994

Identification of thermal oxidation products of cholesteryl acetate.

The polar products separated by solid-phase extraction from the peroxidation mixture of cholesteryl acetate, were investigated. The oxidation products were identified by comparing GC retention times as well as the mass spectra against those of available or synthesized standards. The main oxidation products where 7 beta-chydroperoxicholesteryl acetate, 7 alpha-hydroperoxicholesteryl acetate, 7-ketocholesteryl acetate, the alpha and beta isomers of 7-hydroxycholesteryl acetate, the alpha- and beta-epoxy isomers in 5,6 position and several derivatives from the loss of groups (especially the acetic and/or hydroxyl groups in the form of acetic acid and water).

Cholesterol Esters↗
PubMed1994

Effect of cholesterol-5 alpha,6 alpha-epoxide supplementation to cultured cardiomyocytes.

In order to evaluate the effect of one of the main oxysterols derived from cholesterol oxidation, cholesterol-5 alpha,6 alpha-epoxide (epox), on cardiac cells, we have supplemented the culture medium of neonatal rat cardiomyocytes with scalar concentrations of epox (0.1-100 microM). While 0.1 microM epox supplementation was ineffective, epox supplementation in the range 1-100 microM determined a reduction in cellular protein level, without affecting cell viability, and a dose-dependent epox incorporation into cardiomyocyte lipids. Furthermore, in the same concentration range of epox supplementation, a gas chromatographic peak unambiguously identified by gas chromatography-mass spectrometry as cholestane-3 beta,5 alpha,6 beta-triol, an hydrolytic metabolite of epox, was detected. The mechanism of cytotoxicity of epox to cardiomyocytes could be due to the insertion of epox itself into cellular lipids, and to its metabolization to the more toxic triol.

Animals↗
PubMed1992

Improvement of extraction procedure for biogenic amines in foods and their high-performance liquid chromatographic determination.

A high-performance liquid chromatography method is described for the simultaneous determination of the biogenic amines tryptamine, 2-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine in cheese. The optimization of the procedure for the extraction of amines from the matrix is described. The separation of dansyl derivatives of the amines was achieved by reversed-phase liquid chromatography with gradient elution, followed by UV detection at 254 nm. The mobile phase was acetonitrile-0.01 M phosphate buffer (pH 7)-water. Under these conditions, rapid elution of the amines in less than 13 min was obtained. Validation of the method included calibration experiments, addition of standard amines for the determination of amine recoveries and repeatability tests.

Biogenic Amines↗
PubMed1992

Cannabinoid acids analysis.

The cannabinoid pattern of vegetable preparations from Cannabis sativa (hashish, marijuana) allows to recognize the phenotype of the plants, to be used as drug or for fiber. Cannabinoid determination by analytical point of view has represented some problems caused by the complex composition of the hexane extract. Capillary gas chromatography of the hexane extracts of vegetable samples, shows the presence of rather polar constituents that eluted, with noticeable interactions, only on polar phase. The compounds can be methylated by diazomethane and silanized (TMS) by silylating reagents. The methyl and methyl-TMS derivatives are analyzed by high resolution gas chromatography (HRGC) and by gas chromatography-mass spectrometry (GC-MS). The identification of the compounds shows their nature of cannabinoid acids, which the main by quantitative point of view results the cannabidiolic acid (CBDA). It is known that the cannabinoid acids are thermally unstable and are transformed in the corresponding cannabinoids by decarboxilation. This is of interest in forensic analysis with the aim to establish the total amount of THC in the Cannabis preparations, as the active component.

Cannabinoids↗
PubMed1989

Effect of a hyperlipidic diet on lipid composition, fluidity, and (Na+-K+)ATPase activity of rat erythrocyte membranes.

Feeding rats a hyperlipidic diet in which animals were offered daily a variety of high-energy food resulted in a significant increase of serum free fatty acids and a decrease of phospholipids with respect to controls. On the contrary, there were no significant differences in erythrocyte membrane total lipid composition between the two groups. Erythrocyte membranes showed a significant decrease in saturated fatty acid content and a significant increase in (n-6) polyunsaturated fatty acid content; (n-3) polyunsaturated fatty acids significantly decreased. Membrane fluidity, investigated by fluorescence polarization of diphenylhexatriene, significantly increased in the erythrocyte membranes of the experimental group. These results seem compatible with the decreased saturated/unsaturated fatty acid ratio. A significant decrease of (Na+-K+)ATPase activity occurred in erythrocyte membranes of the experimental group rats with respect to the controls.

Animals↗
PubMed1989

Lipid composition, fluidity and enzymatic activities of rat liver plasma and mitochondrial membranes in dietary obese rats.

Total lipids and fatty acid composition were determined in liver plasma and mitochondrial membranes from control and dietary obese rats after 4 weeks of the experimental period. The lipid composition of liver plasma and mitochondrial membranes showed an increase of triacylglycerols in obese rats. The liver plasma membranes showed a decrease of saturated/unsaturated fatty acid ratio and an increase of (n-6) polyunsaturated fatty acids, whereas the (n-3) polyunsaturated acids were decreased. Contrary to what occurs with plasma membranes, few modifications were observed in mitochondrial membranes. Changes of the fatty acid composition of the phospholipid bilayer are of potentially great importance in structural and functional parameters of membrane. Fluidity of liver plasma membranes of dietary obese rats was highly increased, while the mitochondrial ones remained unchanged. These results can be well explained by the decreased saturated/unsaturated fatty acid ratio. A significant decrease of (Na+-K+) ATPase activity (a membrane bound enzyme) was found in plasma membranes of dietary obese rats. Mitochondrial enzymatic activities and oxidative phosphorylation showed few changes except a small, but significant decrease of state 3 respiratory rate. In this study we also determined the fatty acid composition of all the foods offered to animals and their daily intakes in order to discuss their possible influence on changes in structural and functional membrane parameters.

Animals↗