Biomedical subjects

Tommaso R I Cataldi

Publications and source records attributed to Tommaso R I Cataldi.

14 recordsLinked to original sources

PubMed2006

Occurrence of N-acyl-L-homoserine lactones in extracts of some Gram-negative bacteria evaluated by gas chromatography-mass spectrometry.

Acylated homoserine lactones (AHLs) are self-generated signal molecules that mediate population density-dependent gene expression (quorum sensing) in a variety of Gram-negative bacteria. These signal molecules diffuse from bacterial cells and accumulate in the medium as a function of cell growth. In selected foods AHLs contribute to product spoilage. As different bacterial species produce AHL analogs that differ in length of the N-acyl chain, ranging from 4 to 14 carbons and in the substitution at the C-3 position of the side chain (i.e., oxo or hydroxyl group), the suitability and applicability of a gas chromatography-mass spectrometry direct method for characterizing trace amounts of AHLs was evaluated using N-heptanoyl-homoserine lactone as internal standard. Crude cell-free supernatants of bacterial cultures of Aeromonas hydrophila, Aeromonas salmonicida, Pseudomonas aeruginosa, Pseudomonas fluorescens, Yersinia enterocolitica, and Serratia liquefaciens were screened for AHL production in selected ion monitoring mode, using the prominent fragment at m/z 143. The observed profiles of distinguishable N-acyl-homoserine lactones occurring in bacterial extracts were compared and discussed. The presence of a labile 3-oxo-hexanoylhomoserine lactone was evidenced but serious difficulties arose in estimating its concentration as thermal degradation occurs during the gas chromatographic separation. Its electron impact mass spectra was, however, given and interpreted.

4-Butyrolactone↗

PubMed2006

Pulsed electrochemical detection of orotic acid by an activated potential waveform at a gold working electrode following anion-exchange chromatography.

The application of activated pulsed amperometric detection (APAD) for the determination of orotic acid (OrA) in real samples at a gold working electrode in alkaline solutions, in combination with anion-exchange chromatography, is reported. Such an activated potential waveform was designed with an initial step that involves the formation of redox active species (e.g., adsorbed AuOH/AuO), which in turn is halted upon lowering the applied potential at the detection value while the adsorbed gold hydroxide/oxide species are still catalytically active. A direct comparison between the activated potential waveform and the more commonly used pulsed amperometric detection showed roughly a 20-fold increase in sensitivity. The chromatographic separation of OrA was accomplished by using a microbore anion-exchange column eluted with an isocratic mobile phase composed of 100 mM NaOH+40 mM NaNO(3). Orotic acid was determined at the concentration ranges of 0.2-30 microM (r=0.9997) with an absolute detection limit of 80 pg (10 microL injected). The levels of OrA in cows' milk samples evaluated by standard additions, using 5-aminoorotic acid as an internal standard, ranged from 56 to 126 mg/L. Lower levels were found in raw sheep's milk (<20 mg/L). The assay is shown to be very useful in clinical investigations where relatively high levels of OrA in human urine are correlated to metabolic diseases.

Animals↗

PubMed2005

Comparison of silver, gold and modified platinum electrodes for the electrochemical detection of iodide in urine samples following ion chromatography.

The electrochemical (EC) detection of iodide at gold, silver and platinum electrodes under similar experimental conditions was evaluated. To achieve optimal amperometric detection, the electrode sensitivity, selectivity, and stability was compared. Isocratic separation of iodide was attained by ion chromatography (IC) using an anion-exchange column with nitrate as an eluent ion (25 mM HNO(3) + 50 mM NaNO(3)). Although the Ag electrode showed the highest selectivity due to the relatively low applied potential (+0.10 V versus Ag|AgCl), it requires continuous surface polishing upon injection of standard solutions or real samples; in addition, the chromatographic peak of iodide exhibited a pronounced dip-tailing. The limit of detection (LoD) of iodide was estimated to be 3.5 microg/L (S/N=3) with an injection volume of 50 microL. Likewise, pulsed electrochemical detection at the silver electrode did not demonstrate the expected results in terms of peak shape and low detection limit. Using the same chromatographic conditions, iodide detection at the Au electrode (E(app)= +0.80 V versus Ag|AgCl) exhibited a regular peak shape accompanied by a sensitivity comparable to the silver one. Yet, upon continuous injections the signal intensity displayed a progressive lowering up to ca. 40% in 6h. Best results in terms of signal stability, peak shape and analytical response were obtained with a modified platinum electrode which allowed to achieve a LoD of 0.5 microg/L (S/N=3). The present IC-EC detection method using a modified Pt electrode (E(app)= +0.85 V versus Ag|AgCl) was successfully applied to determine low contents of iodide in human urine with solid phase extraction as pretreatment. Such a developed method correlated very well with the reference colorimetric method in urine (r=0.95273), and it is specifically suggested when the iodide content is relatively low, i.e., <20 microg/L.

Chromatography, Liquid↗

PubMed2005

Sensitive quantification of iodide by ion-exchange chromatography with electrochemical detection at a modified platinum electrode.

A rapid and very sensitive method for the accurate determination of free iodide in real samples is described. The method is based on anion-exchange chromatographic separation coupled with amperometric detection at a modified platinum electrode under constant applied potential (+0.85 V vs. Ag AgCl). An experimental setup with an in-line and very effective method of electrode modification is proposed using an amperometric thin-layer cross-flow detector and a flowing solution 300 mg/L of iodide; the working electrode is polarised to the limiting current for oxidation of iodide to iodine in acidic solutions with the consequent formation of an iodine-based film. The results indicated that the modified electrode exhibits high analytical response for iodide electrooxidation with good stability and long-life. The signal intensity of daily experimental sessions (8 h), during which standards and real samples were repeatedly injected, exhibits a moderate lowering (i.e. < 6%). Using a mixture of 25 mM HNO3 and 50 mM NaNO3 as an eluent phase in ion-exchange chromatography, the detection limit of iodide was estimated to be 0.5 microg/L (S/N = 3) with an injection volume of 50 microL. This method was applied successfully to quantify the iodide content of milk samples and in wastewaters as well as trace amounts in common vegetables and solutions containing high chloride levels.

Journal Article↗

PubMed2005

A pulsed potential waveform displaying enhanced detection capabilities towards sulfur-containing compounds at a gold working electrode.

Pulsed electrochemical detection of sulfur-containing compounds was successfully investigated by applying a four-step potential waveform at a gold working electrode. This potential waveform called APAD, which stands for activated pulsed amperometric detection, is composed of an activation potential step added to a conventional three-step potential waveform. A key advantage of the APAD at the Au electrode is the ability to enhance sensitivity through the use of a short potential pulse (E(ACT) = +750 mV versus Ag/AgCl and tACT approximately 90 ms) during which the formation of redox active species (presumably OH*) are able to efficiently oxidize organosulfur compounds. The APAD waveform parameters were optimized to maximize the signal-to-noise ratio (S/N) and successfully applied for the sensitive detection of lipoic acid, biotin, iminobiotin, methionine, cystine, cysteine, homocysteine, homocystine, N-acetylcysteine and glutathione, following their separations by high-performance anion-exchange chromatography (HPAEC) using alkaline mobile phases. The detection limits (S/N = 3, 10 microL injected) ranged from 0.3 for cysteine (400 pg) to 0.02 micromol/L for biotin (50 pg) and methionine (30 pg). The response of sulfur-, amine- and alcohol-based compounds was compared by using four selected pulsed potential waveforms. It was found that the APAD exhibits excellent sensitivity for thiocompounds outperforming all other pulsed potential waveforms. Ratios of the peak areas for APAD and the six-step potential integrated waveform increased from 3.2 +/- 0.4 to 13.5 +/- 0.6 for lipoic acid and biotin, respectively.

Acetylcysteine↗

PubMed2005

Analysis of tomato glycoalkaloids by liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

Steroidal glycoalkaloids (SGAs) extracted from tomato leaves and berries (Lycopersicon esculentum Mill.) were separated and identified using optimized reversed-phase liquid chromatography with electrospray ionization (ESI) and ion trap mass spectrometry (ITMS). The ESI source polarity and chromatographic conditions were evaluated. The ESI spectra contain valuable information, which includes the mass of SGAs, the mass of the aglycones, and several characteristic fragment ions. Cleavage at the interglycosidic bonds proximal to the aglycones is the most prominent process in the ESI process. A protonated molecule, [M+H]+, accompanied by a mixed adduct ion, [M+H+Na]2+, was observed for alpha-tomatine (i.e., m/z 1034.7 and 528.9) and dehydrotomatine (i.e., m/z 1032.6 and 527.9) in positive ion mode spectra. The structures of these tomato glycoalkaloids were confirmed using tandem mass spectrometry. The identification of a new alpha-tomatine isomer glycoalkaloid, named filotomatine (MW 1033), which shares a common tetrasaccharide structure (i.e., lycotretraose) with alpha-tomatine and dehydrotomatine, and soladulcidine as an aglycone, is described for the first time. It occurs in significant amounts in the extracts of wild tomato foliage. Multistage mass spectrometry both of the protonated molecules and of the doubly charged ions was used for detailed structural elucidation of SGAs. Key fragmentations and regularities in fragmentation pathways are described and the fragmentation mechanisms involved are proposed.

Chromatography, High Pressure Liquid↗

PubMed2004

Determination of carnosine in feed and meat by high-performance anion-exchange chromatography with integrated pulsed amperometric detection.

Carnosine (beta-alanyl-L-histidine) is a dipeptide regarded as an important molecular marker of the presence of processed animal proteins including meat and bone meal in animal feed. For its identification and quantification a sensitive and selective method by high-performance anion-exchange chromatography coupled with integrated pulsed amperometric detection (HPAEC-IPAD) was developed. The assay is based on isocratic elution with 100 mM NaOH as the mobile phase. Interferences of real matrices were efficiently removed; carnosine could be determined at the concentration ranges 0.1-100 microM with a rather low detection limit of 0.23 ng. Unlike feeds for dogs and cats, no carnosine peak was observed in all examined feeds for ruminants. The good analytical characteristics allowed camosine determination down to 5 microg/g of feed.

Animal Feed↗

PubMed2003

Improved determination of taurine by high-performance anion-exchange chromatography with integrated pulsed amperometric detection (HPAEC-IPAD).

The advantages of the high selectivity of high-performance anion-exchange chromatography (HPAEC) and the sensitive response of taurine at a gold electrode with integrated pulsed amperometric detection (IPAD) have been combined, in order to establish a new analytical method for its determination in real matrices. Potential-time settings of the potential waveform were optimized in order to get the highest amperometric response. The separation of taurine in milk samples was achieved using an alkaline eluent (100 mM NaOH) containing 1 mM Ba(OAc)(2) and a column temperature of 15 degrees C. The inherent merits of using a barium-modified eluent, in terms of taurine separation and detection, are demonstrated. The enhancement in sensitivity under these experimental conditions makes it suitable for taurine determination in milk. Indeed, this method allows high recovery of taurine and satisfies the necessary requirements with respect to accuracy, repeatability and sensitivity with a detection limit of 50 nmol/L, which corresponds to 2.5 pmol. The taurine content in milk samples of some common mammals was evaluated, including human milk. In goat's milk, the taurine content ranged from 46 to 91 mg/L, whereas human and buffalo milk samples exhibited an average content of 18 mg/L and 23 mg/L, respectively.

Animals↗

PubMed2003

Determination of free proline and monosaccharides in wine samples by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD).

A sensitive and selective analytical method for the simultaneous separation and quantitative determination of proline and free monosaccharides in wine samples by high-performance anion-exchange chromatography coupled with pulsed amperometric detection is described. Under optimized experimental conditions, a complete separation was obtained in less than 30 min, using an isocratic elution with 10 mM NaOH and 1 mM Ba(OAc)(2). No postcolumn addition of strong bases to the eluent for enhancing detection sensitivity was needed. Upon 25-fold sample dilution and purification to avoid interference of tannins, pigments, and phenolic compounds, the fingerprinting of common monosaccharides (i.e., arabinose, glucose, fructose, galactose, and xylose) and proline in wines, musts, and vinegars can be easily accomplished. The method allows high recovery and satisfies the necessary requirements for accuracy, repeatability, and sensitivity. Values obtained for proline content ranged from 470 to 1190 mg/L in "Aglianico" red wines (mean value, 870 +/- 192 mg/L, n = 21) and from 168 to 286 mg/L in white wines (mean value, 208 +/- 32 mg/L, n = 11). Lower levels were found in musts of red and white grapes, 550 and 87 mg/L, respectively. The lowest content of proline, ca. 10 mg/L, was found both in white and red vinegars.

Acetic Acid↗

PubMed2003

Evaluation of glycoalkaloids in tubers of genetically modified virus Y-resistant potato plants (var. Désirée) by non-aqueous capillary electrophoresis coupled with electrospray ionization mass spectrometry (NACE-ESI-MS).

The glycoalkaloid content of transgenic potatoes was evaluated by an optimised method based on non-aqueous capillary electrophoresis coupled on-line with electrospray ionization-mass spectrometry (NACE-ESI-MS). The potato material consisted of tubers from a conventional cv. Désirée and from three lines of modified plants resistant, intermediate and susceptible to infection by potato virus Y (PVY). The main glycoalkaloids were confirmed to be alpha-solanine and alpha-chaconine with parent ion masses m/z 852 and 868, respectively. In addition, an unknown minor peak at m/z 850.6 was found both in conventional (control) and susceptible line potato tubers. Such a compound exhibited an MS(2) spectrum with fragments ions at 704 and 396 m/z derived by loss of two ions, i.e. m/z 146 and 307, most likely corresponding to a rhamnose unit and a [glucose-(rhamnose)(2)] moiety, respectively. Up to 30-80-fold higher concentrations of total glycoalkaloids were found in the peel compared to flesh samples of all tubers examined. TGA content was nearly doubled in peel samples of resistant compared to control lines, and these levels were lower than the limit recommended for food safety, i.e. 20-60 mg of TGA per 100 g fresh weight. Moreover, it was established that tubers produced by virus-resistant clones are substantially equivalent in glycoalkaloid contents to those produced by conventional potato varieties.

Electrophoresis, Capillary↗

PubMed2002

Assay of riboflavin in sample wines by capillary zone electrophoresis and laser-induced fluorescence detection.

To routinely assay the concentration of riboflavin (RF) in wines, a rapid and sensitive method was developed and evaluated. The method is based on a simple sample preparation, capillary zone electrophoretic separation and laser-induced fluorescence detection (CZE-LIF). Sample preparation required only dilution and filtration. Under optimized conditions, the limit of detection of riboflavin was 0.5 micro g/L, using a hydrodynamic sample introduction of 10 s at 54 mbar. The method was fully validated: the recovery of RF in wines was >95%. The concentrations of RF within the three sample types of Italian wines investigated here ranged from 69 to 151 micro g/L with a mean value (+/-SD) of 112 +/- 25 micro g/L, from 74 to 193 micro g/L with a mean value of 115 +/- 45 micro g/L, and from 156 to 292 micro g/L with a mean value of 226 +/- 40 micro g/L, for white, rosé, and red wines, respectively. Such an accurate and highly sensitive CZE-LIF method represents a powerful improvement over previous methods in terms of sensitivity, simplicity, and efficiency. It is well suited to satisfy the demands for accurate and sensitive detection with minimal sample preparation and cleanup.

Electrophoresis, Capillary↗

PubMed2002

Determination of glycoalkaloids and relative aglycones by nonaqueous capillary electrophoresis coupled with electrospray ionization-ion trap mass spectrometry.

Glycoalkaloids are naturally occurring nitrogen-containing compounds present in many species of the family Solanaceae, including cultivated and wild potatoes (Solanum spp.), tomatoes (Lycopersicon spp.), etc. These compounds have pharmacological and toxicological effects on humans due to their significant anticholinesterase activity and disruption of cell membranes. Herein is reported the development of a capillary electrophoresis (CE) method using nonaqueous (NA) separation solutions in combination with ion trap mass spectrometry (MS and MS/MS) detection for the identification and quantification of glycoalkaloids and their relative aglycones. A mixture 90:10 v/v of MeCN-MeOH containing 50 mM ammonium acetate and 1.2 M acetic acid (applied voltage of 25.5 kV) was selected as a good compromise for the separation and detection of these compounds. The electrospray MS measurements were carried out in the positive ionization mode using a coaxial sheath liquid, methanol-water (1:1) with 1% of acetic acid at a flow rate of 2.5 microL/min. Under optimized experimental conditions, the predominant ion was the protonated molecular ion ([M+H](+)) of solanidine (m/z = 398), tomatidine (m/z = 416), chaconine (m/z = 852), solanine (m/z = 868), and tomatine (m/z = 1034). MS/MS experiments were carried out systematically by changing the relative collisional energy and monitoring the intensities of the fragment ions that were not high enough to allow better quantification than with the mother ions. The method was used for analyzing glycoalkaloids in potato extracts.

Electrophoresis, Capillary↗

PubMedDate not supplied

Ionic content in plant extracts determined by ion chromatography with conductivity detection.

A simple method is described for the determination of the ionic content of vegetable samples by ion chromatography with suppressed conductivity detection. Extracts of leaves of cucumber (Cucumis sativus), leaves and cotyledons of watermelon (Citrullus lanantus), cotyledons of zucchini (Cucurbitapepo), and leaves and roots of olive (Olea europaea) obtained at room temperature yielded chromatographic profiles with substantial differences in the relative contents of Cl-, NO3-, HPO4(2-) and SO4(2-) as well as of Na+, NH4+, K+, Mg2+ and Ca2+. Although NO3-, Cl- and K+ were common to each extracted sample and accounted for most of the ions present, two additional anion peaks (i.e. malate and oxalate) were detected. Among the vegetable tissues investigated, olive roots contained a considerable amount of oxalate (37 mg/g dry weight), while Na+, which is present in very low amount in extracted samples of leaves and cotyledons, represented ca. 30% of the cationic content of olive roots. In all the examined tissue extracts, K+ was the main cation (16-55 mg/g dry weight) and NO3-, Cl- and HPO4(2-) were the main inorganic anions.

Anions↗

PubMedDate not supplied

Surface retention and photochemical reactivity of the diphenylether herbicide oxyfluorfen.

The photochemical behavior of oxyfluorfen [2-chloro-1-(3-etoxy-4-nitrophenoxy)-4-(trifluoromethyl) benzene] on two Greek soils was investigated. Soils were sampled from Nea Malgara and Preveza regions, characterized by a different organic matter content. Soils were spiked with the diphenyl-ether herbicide and irradiation experiments were performed either in the laboratory with a solar simulator (xenon lamp) or outside, under natural sunlight irradiation; other soil samples were kept in the dark to control the retention reaction. Kinetic parameters of both retention and photochemical reactions were calculated using zero-, first- and second- (Langmuir-Hinshelwood) order equations, and best fit was checked through statistical analysis. The soil behaviors were qualitatively similar but quantitatively different, with the soil sampled from the Nea Malgara region much more sorbent as compared with Preveza soil. All studied reactions followed second-order kinetics and photochemical reactions were influenced by retaining capability of the soils. The contributions of the photochemical processes to the global dissipation rates were also calculated. Two main metabolites were identified as 2-chloro-1-(3-ethoxy-4-hydroxyphenoxy)-4-(trifluoromethyl)benzene and 2-chloro-1- (3-hydroxy-4-nitrophenoxy)-4-(trifluoromethyl)benzene.

Environmental Monitoring↗