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Biomedical subjects

C Gardana

Publications and source records attributed to C Gardana.

At least 19 recordsLinked to original sources
PubMed2003

LC-APCI-MS/MS analysis of urinary 8-hydroxy-2'-deoxyguanosine.

8-Hydroxy-2'-deoxyguanosine (8OHdG) is regarded as an important biomarker of oxidative DNA damage and it may be estimated by using different techniques in various biological matrices, most notably DNA and urine. In the case of DNA, artifactual oxidation may take place during the isolation of DNA, its hydrolysis and possible derivatization (as for GC-MS), invalidating the measurement of 8OHdG. Therefore, the direct analysis of 8OHdG excreted into urine was preferred. Interferences from the urine matrix were excluded by applying LC-APCI-MS/MS in the multiple reaction monitoring (MRM) mode. The abundant fragment ion at m/z 168 arising from 8OHdG was monitored in the urine sample of volunteers supplemented with tomato concentrate for different times. The procedure allowed the detection of levels of 8OHdG as low as 1 ng/ml in urine sample.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed2003

Polyphenol pattern and antioxidant activity of different tomato lines and cultivars.

BACKGROUND/AIMS: Besides antioxidant vitamins and minerals, fruits and vegetables contain flavonoids and related phenolics. The biological activities of these polyphenols have become well known in recent years evidencing their beneficial effects on human health. In this context, the characterization of the flavonoids present in tomatoes is of great interest. Thus the polyphenol pattern (including flavonols, flavanones and cinnamate derivatives), lycopene and beta-carotene concentrations and the total antioxidant activity (TAA) of the phenolic fraction from different tomato lines and cultivars have been determined. METHODS: The characterization was obtained by means of spectrophotometry and HPLC analyses. RESULTS: Mean values for single flavonoids were 0.68 +/- 0.16 for naringenin, 0.74 +/- 0.12 for rutin and 0.32 +/- 0.06 for a rutin-pentoside. Mean total polyphenol content was 13.15 +/- 1.15 mg/100 g and mean TAA value was 1.3 +/- 0.10 mmol/g. The obtained TAA values resulted in good accordance with the total polyphenol content (R(2) = 0.7928). The main phenolic acids were chlorogenic (mean +/- SE 0.20 +/- 0.03) and caffeic acid (mean +/- SE 0.03 +/- 0.01). Mean levels of lycopene and beta-carotene were 5.38 +/- 0.90 and 1.18 +/- 0.40 mg/100 g, respectively. CONCLUSIONS: Almost all the lines characterised by low carotenoid content produce high levels of polyphenols, and consequently have the most powerful antioxidant potential.

Antioxidants↗
PubMed2002

Analytical methods for quality control of propolis.

Propolis is a resinous substance collected by honeybees from leaf buds and cracks in the bark of various plants, and it is composed of 50% resin (composed of flavonoids and related phenolic acids), 30% wax, 10% essential oils, 5% pollen and 5% various organic compounds. Propolis cannot be used as raw material, and it must be purified by extraction with solvents. This process should remove the inert material and preserve the polyphenolic fraction, which is considered to contribute more to the observed healing effects than the other propolis constituents. Therefore, the assay of propolis polyphenols is of interest, and this paper describes the results obtained in the analysis of propolis by means of a gradient HPLC or mass spectrometry. HPLC in the gradient mode and coupled with photodiode array detection remains the method of choice for the assay of most relevant components of propolis. Direct analysis by APCI-IT-MS represents a valuable alternative to obtain typical fingerprints of propolis and a reliable identification of a large number of propolis components.

Chemistry, Pharmaceutical↗
PubMed2001

Liquid chromatography/atmospheric pressure chemical ionization mass spectrometry of terpene lactones in plasma of volunteers dosed with Ginkgo biloba L. extracts.

Liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (LC/APCI-ITMS) was applied to evaluate the levels of ginkgolides A and B and bilobalide in plasma of volunteers after administration of Ginkgo biloba extracts in free (Ginkgoselect) or phospholipid complex (Ginkgoselect Phytosome) forms, providing 9.6 mg of total terpene lactones. The maximum plasma concentrations, C(max), of total ginkgolides A, B and bilobalide were 85.0 and 181.8 microg/mL for Ginkgoselect and Ginkgoselect Phytosome, respectively. The C(max) values were reached at 120 min for the free form and at 180--240 min for the phospholipid complex form. In both cases, the mean elimination half-life of each terpene lactone was in the range 120--180 min. Due to its sensitivity (about 1 ng/mL) and specificity, LC/APCI-ITMS proved to be a very powerful tool for pharmacokinetic studies of these phytochemicals.

Adult↗
PubMed2001

Plasma levels of caffeic acid and antioxidant status after red wine intake.

The aim of this study was to evaluate the bioavailability of caffeic acid and the modification of plasma antioxidant status following red wine intake. Five healthy male volunteers consumed 100, 200, and 300 mL of red wine corresponding to approximately 0.9, 1.8, and 2.7 mg of caffeic acid, respectively. Plasma samples collected at different times (0-300 min) were evaluated for their content of caffeic acid and their total antioxidant status. Both these parameters, i.e., plasma concentration of caffeic acid and antioxidant potential, were dose-dependent and the C(max) was reached at about 60 min after red wine intake. The results indicate that caffeic acid is bioavailable and it may be correlated with the antioxidant potential of plasma.

Adult↗
PubMed2000

Polyphenol content and total antioxidant activity of vini novelli (young red wines).

Eight commercial Italian vini novelli (red wines prepared by carbonic maceration and supposed to be consumed within three months from their wine-making) were evaluated for their total antioxidant activity. The wines had an average total phenol content (1605.4 +/- 337.4 mg/L gallic acid equivalents) lower than that of wines prepared by traditional maceration and consumable after aging (2057. 3 +/- 524.0 mg/L gallic acid equivalents). The average flavanol content (424.7 +/- 121.3 mg/L catechin equivalents) and the total antioxidant activity (16.8 +/- 3.8 mmol/L Trolox equivalents) of vini novelli were higher than the corresponding values (382.7 +/- 174.5 mg/L catechin equivalents and 12.3 +/- 3.3 mmol/L Trolox equivalents) found for aged wines. Three couples of experimental wines were prepared from the same grapes by traditional or carbonic maceration. These wines showed a different phenolic pattern, anthocyanins being more abundant in vini novelli. However, the average total antioxidant activities of the wines were similar, suggesting that aging (and not the wine-making technique) is the main factor influencing the antioxidant activity of red wines.

Antioxidants↗
PubMed1999

Liquid chromatography/electrospray ionization mass spectrometric characterization of flavonol glycosides in tomato extracts and human plasma.

Flavonoids continue to attract wide attention as possible very useful agents for combating free radical pathologies, i.e. the pathological states associated with free radical overproduction. Commonly used methods for the analysis of plant flavonoids include high performance liquid chromatography (HPLC) and capillary electrophoresis (CE). On the other hand, the soft-ionization approach based on electrospray ionization (ESI-MS) permits highly selective analysis of complex matrices. In this work, we examined firstly the ESI-MS behaviour of representative aglycones and glycosides of flavonols, flavones and isoflavones with the aim of suggesting a possible relationship between structure and mass spectra. Using HPLC coupled to a diode array detector (DAD) for on-line UV spectra acquisition, and in parallel to ESI-MS for mass spectra (LC/DAD-ESI-MS), we have developed methodology to observe flavonols directly in tomato puree extract. In this way, it has been possible to detect intact flavonol glycosides in tomato extracts and to characterize a flavonol trisaccharide. For the first time, using LC/ESI-MS, it has been possible to detect intact flavonol glycosides in plasma of healthy volunteers and to provide further evidence on the absorption of flavonoid glycosides after consumption of common vegetables like tomatoes.

Calibration↗
PubMed1998

Catechin metabolites after intake of green tea infusions.

Green tea contains relatively large amounts of catechins, that have been recognized to be efficient free-radical scavengers. In spite of a largely described antioxidant effect, the metabolic fate of catechins in humans has been scarcely studied. An infusion of green tea (about 400 mg of catechins) was given to healthy volunteers; plasma and urine samples were collected for 5 h and 2 days, respectively. Epigallocatechin gallate and epicatechin gallate were detected in plasma samples, reaching the maximum concentration (2 microM) at 2 h. Urine samples collected at 6-48 h contained detectable amounts of final catechin metabolites, including 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, 3-methoxy-4-hydroxy-hippuric acid and 3-methoxy-4-hydroxybenzoic acid (vanillic acid). The total content of these metabolites averaged 60 mg. The levels of free plasma catechins account only partly for the increased (approximately +20%) total radical-trapping antioxidant parameter (TRAP) detected after green tea intake. Catechin conjugates (glucuronide and sulphate) and metabolites may add further contribution and explain the measured TRAP increase.

Antioxidants↗
PubMed1998

Relationship between rate and extent of catechin absorption and plasma antioxidant status.

Flavonoids are described to exert a large array of biological activities, which are mostly ascribed to their radical-scavenging, metal chelating and enzyme modulation ability. Most of these evidences have been obtained by in vitro studies on individual compounds and at doses largely exceeding those dietary. Little is known about a possible relationship between rate and extent of the absorption and modifications of plasma antioxidants. To elucidate this aspect, human volunteers were supplemented with single doses of green tea catechins in free (Greenselect) or phospholipid complex form (Greenselect Phytosome) equivalent to 400 mg epigallocatechingallate (EGCg). EGCg was chosen as biomarker for green tea catechin absorption, and its time course plasma concentration was correlated to the subsequent percent variations of plasma ascorbate, total glutathione, alpha-tocopherol, beta-carotene and Total Radical Antioxidant Parameter (TRAP). Green tea catechins were absorbed more extensively when administered as phospholipid complex rather than as free catechins. Single dose intake of both forms of catechins produced a transient decrease (10-20%) of plasma ascorbate and total glutathione and an increase of plasma TRAP (16-19%). These variations were consistent with the plasmatic levels of EGCg, ascorbate and total glutathione.

Adult↗
PubMed1997

Identification of Gingko biloba flavonol metabolites after oral administration to humans.

An extract of Ginkgo biloba leaves (EGb) was given to healthy volunteers. Urine samples were collected for 3 days, and blood samples were withdrawn every 30 min for 5 h. The samples were purified through SPE C18 cartridges and analyzed by reversed-phase LC-diode array detection for the presence of EGb metabolites. Only urine samples contained detectable amounts of substituted benzoic acids, i.e., 4-hydroxybenzoic acid conjugate, 4-hydroxyhippuric acid, 3-methoxy-4-hydroxyhippuric acid, 3,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, hippuric acid and 3-methoxy-4-hydroxybenzoic acid (vanillic acid). In contrast to rats no phenylacetic acid or phenylpropionic acid derivatives were found in urine, thus indicating that in humans a more extensive metabolism takes place. As for rats the metabolites found in human urines accounted for less than 30% of the flavonoids given. The same procedure was applied to blood samples, and no metabolites could be detected.

Benzoates↗
PubMed1997

Assay of soluble guanylate cyclase activity by isocratic high-performance liquid chromatography.

A HPLC method alternative to labelled or unlabelled procedures was developed for the assay of guanylate cyclase (GC) activity. The substrate (GTP) and the product (cGMP) of the enzymatic reaction were separated in the isocratic mode on a muBondapak C18 column. The activity of GC was linearly dependent on the amount of cGMP produced in the presence of sodium nitroprusside. This approach was applied to follow the purification of GC from bovine lung and to evaluate its stability in different storage conditions.

Animals↗
PubMed1996

Analysis of Stevia glycosides by capillary electrophoresis.

The determination of diterpene glycosides from Stevia rebaudiana leaves using capillary electrophoresis is described. Analyses were performed on fused silica capillaries with 20 mM sodium tetraborate buffer, pH 8.3, and 30 mM sodium dodecyl sulfate. The effect of the organic solvent injected with the sample solution on the electrophoretic solution has been confirmed, and an absolute amount of 1.6 nL per injected sample was optimal. Rebaudioside A and steviolbioside were isolated by semipreparative high performance liquid chromatography (HPLC), and their structure was assessed by mass spectrometry.

Diterpenes↗
PubMed1996

Radical scavenger activity of three flavonoid metabolites studied by inhibition of chemiluminescence in human PMNs.

3,4-Dihydroxyphenylacetic acid, 3-hydroxyphenylacetic acid, and 3-(4-hydroxyphenyl)-propionic acid, metabolites which arise from quercetin glycosides, respectively, from flavones and probably from procyanidins by the human intestinal microflora, have been tested for their effects on oxygen radical production by human PMNs stimulated with FMLP or opsonized zymosan. Oxygen radicals were detected by luminol-augmented chemiluminescence measurements. Furthermore free radical scavenging activity of these metabolites was investigated in a cell-free system in which oxygen radicals were generated by horseradish peroxidase with H2O2 as substrate. 3,4-Dihydroxyphenylacetic acid reduced considerably chemiluminescence in PMNs in an amount which was much more pronounced than those of the other two metabolites. Concentrations of 1 mumol/l showed an inhibition by 84% with FMLP as stimulant and by 15% with opsonized zymosan, indicating that different signal transduction pathways are influenced in PMNs. Using the same conditions the unmetabolized quercetin showed an inhibition of chemiluminescence by 74% (FMLP), resp. 20% (opsonized zymosan). In the cell free system 3,4-dihydroxyphenylacetic acid suppressed much more effectively chemiluminescence than 3-hydroxyphenylacetic acid. In contrast, 3-(4-hydroxyphenyl)-propionic acid led to an increase of chemiluminescence generated in the cell-free system (FMLP and zymosan), i.e. by 30% by 25%, at the highest concentration of 4 mumol/l. In conclusion, flavonoid metabolites differ in their effects on free radical production of PMNs and their radical scavenging potencies.

Flavonoids↗
PubMed1995

Identification of flavonoid metabolites after oral administration to rats of a Ginkgo biloba extract.

An extract of Ginkgo biloba leaves (EGb) was administered by gastric probe to Wistar female rats, and urine and faeces samples were collected for 5 days and whole blood samples were withdrawn every 30 min for 6 h. After purification with SPE C18 cartridges, the samples were analysed by reversed-phase LC-diode array detection (LC-DAD) for residual flavonoid glycosides, aglycones and metabolites. No glycosides or aglycones were detected in urine, faeces or blood and extensive degradation of EGb flavonoids within 24 h was detected. Among the seven different phenylalkyl acids detected by LC-DAD, 3,4-dihydroxyphenylacetic acid (I), hippuric acid (II), 3-hydroxyphenylacetic acid (III), homovanillic acid (IV) and benzoic acid (VII) were directly confirmed by on-line mass spectrometry using an electrospray interface (ES-MS). Peaks V and VI needed to be collected and separately examined and they were found to be 3-(4-hydrophenyl)propionic acid and 3-(3-hydrophenyl)propionic acid, respectively. As further evidence, the identity of metabolites I, II, III, IV, V and VII was confirmed by co-chromatography with authentic standards.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1995

Determination of sunscreen agents by micellar electrokinetic chromatography.

The separation of UV-A and UV-B sunscreens by micellar electrokinetic chromatography has been studied. The optimized method, which involves the presence of an anionic surfactant (sodium dodecyl sulphate) and an organic modifier in the background electrolyte, was applied to determine these sunscreens in cosmetic products. Identification was achieved by "on-line" UV spectra. Recovery was in the range of 88-92% and the lower limit of detection was 0.15 mg ml-1.

Cinnamates↗
PubMed1994

Influence of structure on the behavior of flavonoids in capillary electrophoresis.

In an attempt to elucidate the relation between structure and electrophoretic behavior, the effect of pH, surfactant and buffer on electrophoretic mobilities and resolution of several flavonoids, differing in structure, has been investigated. Ring-B substitution and the presence of a free hydroxy group at C7 of ring A play an important role, whereas O-methoxylation is less significant. Glycosylation also contributes in determining different degrees of complexation with borate, thus affecting the mobilities.

Borates↗