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At least 1,189 records · Page 66Linked to original sources

PubMed1992

High-performance liquid chromatography of histamine and 1-methylhistamine with on-column fluorescence derivatization.

An on-column fluorometric derivatization method was developed for the determination of histamine and 1-methylhistamine (HMs) by high-performance liquid chromatography. The system for the derivatization consisted only of a commercially available single-plunger pump and a reversed-phase C18 column supported on synthetic polymer with a mobile phase of acetonitrile and alkaline borate buffer solution containing o-phthalaldehyde as a derivatization reagent. It required no additional reaction system as for a post-column derivatization method. Injected HMs might be derivatized to a fluorophore on the inlet site of the high-performance liquid chromatographic column, followed by chromatography on the same column. Optimization of the on-column reaction conditions resulted in a simple and sensitive analytical method for the determination of HMs with excellent reproducibility and linearity of 0.05-5 micrograms/ml of both HMs. Application of this method to the determination of HMs in food samples resulted in a limit of quantification of 0.05 mg/100 g and in a greater than 95% overall mean recovery at a fortification of 0.1 mg/g of both HMs. This method was furthermore applicable to the determination of histamine released from rat peritoneal mast cells.

Animals↗

PubMed1995

Evaluation of solid-phase extraction procedures in peptide analysis.

Solid-phase extraction (SPE) procedures for peptide isolation and fractionation, based on non-polar and ionic interactions, were evaluated using small synthetic peptides and casein enzymatic hydrolysates. SPE based on hydrophobic phases is a useful, efficient and rapid procedure for peptide extraction and concentration. It allows a successful peptide fractionation using eluents that contain an increasing content of acetonitrile in the presence of trifluoroacetic acid. Differences regarding selectivity are observed between sorbents. Non-polar interaction with C18 sorbents is adequate for the isolation of very polar and hydrophobic peptides. CN sorbents are only adequate for very hydrophobic peptides. PH, CH, C8 and C2 sorbents are useful for isolating and fractionating hydrophobic and very non-polar peptides, but generally not for very polar peptides. Ionic solid-phase extraction using Accell Plus cartridges of QMA (quaternary methylammonium) and CM (carboxymethyl) are very useful for the fractionation of peptide mixtures into basic, acidic and neutral pools of peptides. It can be concluded that SPE using these procedures is a useful tool for the isolation and fractionation of peptides from biological and food samples.

Amino Acid Sequence↗

PubMed1995

Use of toxic equivalency factors for risk assessment for dioxins and related compounds.

TCDD is the most toxic member of a class of polyhalogenated aromatic hydrocarbons that are structurally related, have a similar mechanism of action, and cause the same spectrum of responses. Because of the need to assess the risk from complex mixtures of these chemicals, the international community has adopted an interim approach that assigns relative potency factors to this family of chemicals, based on a comparison with the potency of TCDD. Each chemical that fits the criteria for this class is assigned a toxic equivalency factor, TEF, which is some fraction of that of TCDD. The total toxic equivalency of a mixture, TEQ, is the sum of the weighted potency of each compound in the mixture. Although there may be some variability between different responses in the determination of a TEF value for a compound, endpoint-specific TEFs are usually very similar. There may also be some species differences in TEFs. Again, if pharmacokinetic factors are taken into account, they are usually relatively minor. TEFs based on intake values may also exhibit some differences when compared to those based on target tissue concentrations. Using scientific judgment and a broad data base, interim TEF values have been recommended for PCDDs, PCDFs, and dioxin-like PCBs. Using such values, the TEF approach has been successful at predicting the toxicity of real world mixtures. Ongoing studies from our laboratory have validated the approach for synthetic mixtures that approximate congener ratios found in food samples. Whether non-additive interactions occur with nondioxin-like compounds found in environmentally relevant concentrations remain to be determined.

Animals↗

PubMed1987

Monoclonal antibody for the detection of Clostridium botulinum type A toxin.

Hybridomas producing monoclonal antibodies against type A Hall strain Clostridium botulinum toxin were generated by fusing mouse myeloma cell line P3-X63-Ag8.653 with spleen cells of Balb/c mice immunized with C. botulinum type A toxoid. The monoclonal antibody from one hybridoma, identified as No. 424, was selected from 61 others for its high antibody titre. This monoclonal antibody was used in a double-sandwich enzyme-linked immunosorbent assay (ELISA) system to detect type A toxin in culture fluids and in foods. The monoclonal antibody did not react with either C. botulinum toxin types B, C, D, E and F or with other clostridial species tested. This particular monoclonal antibody (No. 424) did not neutralize type A toxin in the mouse bioassay procedure but detected approximately 10 mouse lethal doses of type A toxin/ml culture fluid. Monoclonal antibody and rabbit antitoxin to type A C. botulinum toxin were useful in a double-sandwich ELISA for the rapid and specific detection of type A toxic fluids in culture and in food samples.

Animals↗

PubMed1995

Modifications to a carbon paste glucose-sensing enzyme electrode and a reduction in the electrochemical interference from L-ascorbate.

A glucose-sensing enzyme electrode was prepared by incorporating polyethylene glycol-modified glucose oxidase, horseradish peroxidase and 1,1'-dimethylferrocene into a carbon paste. The modification of glucose oxidase with polyethylene glycol was effective for increasing the enzyme activity in the carbon paste owing to the enhanced affinity of the polyethylene glycolmodified enzyme for the hydrophobic carbon paste matrix. In contrast, however, the enzyme activity of the polymer-modified peroxidase was lower than the unmodified peroxidase in the carbon paste matrix because of a severe loss of the enzyme activity during the modification with polyethylene glycol. Hence, the enzyme pair of polyethylene glycol-modified glucose oxidase and unmodified peroxidase was used for preparing the enzyme electrode. The reductive current response of the electrode to glucose was recorded at -0.2 V vs. Ag/AgCl. After the addition of glucose (100 microM), the current increased immediately and reached a plateau (delta = -0.12 microA) within 30 s. The current response was linear up to a glucose concentration of 500 microM and the detection limit was 20 microM (S/N = 5). Interference from ascorbate was very small: the current response to 1 mM glucose (-1.1 microA) was slightly reduced to -0.9 microA when 1 mM ascorbate was added to the glucose-containing solution. In biological and food samples, the concentration of ascorbate is generally quite low compared with the glucose concentration. The interference from ascorbate could actually be ignored for the purpose of determining glucose in soft drinks.

Ascorbic Acid↗

PubMed2004

Optimisation of the composition of a screen-printed acrylate polymer enzyme layer with respect to an improved selectivity and stability of enzyme electrodes.

Glucose oxidase (GOD) was immobilized on screen-printed platinum electrodes by entrapment in a screen printable paste polymerized by irradiation with UV-light. The influences of different additives, in particular polymers and graphite, on the sensitivity and stability of the sensor and the permeability of the enzyme layer for a possible electrochemical interferent were investigated. The chosen additives were Gafquat 755N, poly-L-lysine, bovine serum albumin (BSA), sodium dodecylsulfate (SDS), polyethylene glycol (PEG), Nafion and graphite. All additives led to increases of glucose signals, i.e. improved the sensitivity of glucose detection with Gafquat 755N, poly-L-lysine, SDS and graphite showing the strongest influences with increases by a factor 4, 6.5, 5 and 10, respectively. Ascorbic acid was used as a model interferent showing the influence of the enzyme layer composition on the selectivity of the sensor. The addition of Gafquat 755N or poly-L-lysine led to higher signals not only for glucose, but also for ascorbic acid. SDS addition already reduced the influence of ascorbic acid, which was almost completely eliminated when Nafion (5%) and PEG (10%) were added. A comparable beneficial effect on the selectivity of the sensors was also observed for the addition of 0.5% graphite. Thus, the enzyme electrodes with PEG, Nafion or graphite as additives in the enzyme layer were applied to glucose determinations in food samples and samples obtained from E. coli cultivations.

Acrylates↗

PubMed2004

Oligonucleotide-modified screen-printed gold electrodes for enzyme-amplified sensing of nucleic acids.

An electrochemical genosensor for the detection of specific sequences of DNA has been developed using disposable screen-printed gold electrodes. Screen-printed gold electrodes were firstly modified with a mixed monolayer of a 25-mer thiol-tethered DNA probe and a spacer thiol, 6-mercapto-1-hexanol (MCH). The DNA probe sequence was internal to the sequence of the 35S promoter, which sequence is inserted in the genome of GMOs regulating the transgene expression. An enzyme-amplified detection scheme, based on the coupling of a streptavidin-alkaline phosphatase conjugate and biotinylated target sequences was then applied. The enzyme catalysed the hydrolysis of the electroinactive alpha-naphthyl phosphate to alpha-naphthol; this product is electroactive and has been detected by means of differential pulse voltammetry. The assay was, firstly, characterised using synthetic oligonucleotides. Relevant parameters, such as the probe concentration and the immobilisation time, the use of the MCH and different enzymatic conjugates, were investigated and optimised. The genosensor response was found to be linearly related to the target concentration between 0 and 25 nmol/L; the detection limit was 0.25 nmol/L. The analytical procedure was then applied for the detection of the 35S promoter sequence, which was amplified from the pBI121 plasmid by polymerase chain reaction (PCR). Hybridisation conditions (i.e., hybridisation buffer and hybridisation time) were further optimised. The selectivity of the assay was confirmed using biotinylated non-complementary amplicons and PCR blanks. The results showed that the genosensor enabled sensitive (detection limit: 1 nmol/L) and specific detection of GMO-related sequences, thus providing a useful tool for the screening analysis of bioengineered food samples.

Alkaline Phosphatase↗

PubMed2005

Congener-specific concentrations and carbon stable isotope ratios (delta13C) of two technical toxaphene products (Toxaphene and Melipax).

In this study we compared the contribution of individual congeners and the ratios of stable carbon isotopes of two technical toxaphene products. The former US-American product Toxaphene was from 1978 and the East-German product Melipax from 1979. Both technical products showed the known complexity in GC/ECD measurements. Contributions of 24 peaks to each of the technical products were determined by gas chromatography in combination high resolution electron capture negative ion mass spectrometry (GC/ECNI-HRMS). The percentages of the compounds studied in the technical mixtures ranged from approximately 0.05% to approximately 2.5% but showed some individual differences. 2,2,5,5,8,9,9,10,10-nonachlorobornane (B9-1025 or P-62) was identified as a major congener in both mixtures. 2-Endo,3-exo,5-endo,6-exo,8,8,10,10-octachlorobornane (B8-1413 or P26) and 2-endo,3-exo,5-endo,6-exo,8,8,9,10,10-nonachlorobornane (B9-1679 or P-50) were found at similar concentration in both technical products. Identical amounts of Melipax or Toxaphene were combusted to CO2 in an element analyzer and their delta13C values were determined relative to the international standard Vienna PeeDee belemnite (VPDB). The mean delta13C values of both products varied by 2.8% (determined at two different locations) which is roughly one order of magnitude more than the precision obtained in repetitive analyses of the individual products. Thus, both investigated products could be unequivocally distinguished by stable isotope ratio mass spectrometry (IRMS). IRMS analyses may thus be a suitable tool for tracing back toxaphene residues in environmental and food samples to the one or both of the products.

Carbon Isotopes↗

PubMed2005

Determination of acrylamide and methacrylamide by normal phase high performance liquid chromatography and UV detection.

A method using normal phase high performance liquid chromatography (NP-HPLC) with UV detection was developed for the analysis of acrylamide and methacrylamide. The method relies on the chromatographic separation of these analytes on a polar HPLC column designed for the separation of organic acids. Identification of acrylamide and methacrylamide is approached dually, that is directly in their protonated forms and as their hydrolysis products acrylic and methacrylic acid respectively, for confirmation. Detection and quantification is performed at 200 nm. The method is simple allowing for clear resolution of the target peaks from any interfering substances. Detection limits of 10 microg L(-1) were obtained for both analytes with the inter- and intra-day RSD for standard analysis lying below 1.0%. Use of acetonitrile in the elution solvent lowers detection limits and retention times, without impairing resolution of peaks. The method was applied for the determination of acrylamide and methacrylamide in spiked food samples without native acrylamide yielding recoveries between 95 and 103%. Finally, commercial samples of french and roasted fries, cookies, cocoa and coffee were analyzed to assess applicability of the method towards acrylamide, giving results similar with those reported in the literature.

Acrylamide↗

PubMed2005

Novel gas chromatography-mass spectrometry database for automatic identification and quantification of micropollutants.

A novel gas chromatography-mass spectroscopy (GC-MS) database for identification and quantification of micropollutants in environmental and food samples is reported. GC retention times, calibration curves, and mass spectra of nearly 700 chemicals were registered in the database, and the GC retention times of registered chemicals in actual samples were predicted from the retention times of n-alkanes measured before sample analysis. Differences between predicted and actual retention times were less than 3 s, an accuracy that is nearly identical to that obtained by analysis of standard substances. After the retention times were predicted, a calibration file for the GC-MS instrument was created from the predicted retention times, calibration curves, and mass spectra of the registered chemicals. With the resulting calibration file, automated identification of all the chemicals in actual samples was possible without the use of standards, and the identification method was as reliable as conventional methods. When the GC inlet, column, and tuning conditions were adjusted using GC-MS performance check standards, relative standard deviations of 20% or less for determination values could be obtained. More than 90% of the chemicals in the database could be detected at a sensitivity sufficient for all practical purposes (100 pg or less). Because each chemical in the database, to which new substances can easily be added, can be determined in 1 h, micropollutants in samples can be analyzed efficiently and inexpensively.

Calibration↗

PubMed2006

Fast determination of anions on a short coated column.

In this paper, a simple method for the separation and determination of common inorganic anions by fast ion-exchange chromatography, using a modified short (25 mm x 4.6 mm) monolithic column, is reported. Coating the column with a cationic surfactant, cetylpyridinium chloride (CPC), the isocratic separation of some inorganic anions in minutes was possible, by direct or indirect UV detection. The coated column demonstrated excellent stability over time, even at a high flow-rate, giving retention times with an average relative standard deviation of 1.3% for over 10 consecutive runs. The developed column exhibited unusual selectivity for common anions, was successfully applied to the rapid analysis of inorganic anions of food samples, river water and factory waste water samples.

Anion Exchange Resins↗

PubMed2004

Determination of less polar heterocyclic aromatic amines in standardised beef extracts and cooked meat consumed in Austria by liquid chromatography and fluorescence detection.

An analysis method was developed for the determination of trace levels of less polar heterocyclic aromatic amines (HAs) in food samples. The development started from a frequently used sample pre-treatment scheme which was slightly improved to make it applicable with high-performance liquid chromatography (HPLC) with fluorescence detection. The method was applied for the analysis of a standardised beef extract containing 5-15 ng/g of HAs and the results are compared with those of the other participants in the same European project. In addition, the method was used for the analysis of less polar HAs in cooked meat consumed in Austria.

Amines↗

PubMed2004

Separation procedures applicable to lignan analysis.

Lignans are a class of secondary plant metabolites produced by oxidative dimerization of two phenylpropanoid units. They have been found in many plants of Oriental medicine. In consequence of recent knowledge it is held that lignans are responsible for the key pharmacological activities of these plants. This review surveys the chromatographic, electromigration and hyphenated methods so far applied for the separation of lignans in Oriental plants used in phytotherapy as well as for the analyses of these lignans and their metabolites in biological matrices and food samples. In addition, the sample clean-up procedures--solvent extractions and supercritical fluid extractions--are also included.

Chromatography↗

PubMed2005

Real-time RT-PCR for norovirus screening in shellfish.

Real-time RT-PCR, combining amplification and detection of virus-specific amplicons, is a promising tool for norovirus detection in environmental or food samples such as shellfish. We developed a real-time RT-PCR assay based on one-step detection using single primer sets and probes for norovirus genogroups I and II. Seventy and seven RT-PCR units of genogroup I and II reference norovirus strains, respectively, were detected in artificially contaminated oysters. Validation of the new method on 150 archived naturally contaminated shellfish confirmed the utility of the genogroup II primer set to detect a large range of different strains circulating in France since 1995, but genogroup I strains were detected infrequently.

Animals↗

PubMed2006

Comparative evaluation of new TaqMan real-time assays for the detection of hepatitis A virus.

Three novel real-time TaqMan RT-PCR assays targeting the 5'-UTR, the viral protease and the viral polymerase regions of the hepatitis A virus (HAV) were developed, evaluated and compared against a new published 5'-UTR TaqMan assay (JN) and a widely used conventional RT-PCR assay (HAVc). All conventional RT-PCR (HAV, SH-Prot and SH-Poly systems) and TaqMan (SH-Prot, SH-Poly, JN and SH-5U systems) assays evaluated were consistent for the detection of the three different HAV strains (HM-175, HAS-15 and LSH/S) used and reproducible for both RNA duplicates with the exception of two reproducibility discrepancies observed with both 5'-UTR real-time systems (JN and SH-5U). Limits of detection for conventional HAV, SH-Prot and SH-Poly RT-PCR systems were found to be equivalent when tested with serially diluted suspensions of the HM-175 strain. Although the four real-time RT-PCR TaqMan assays evaluated herein produced similar and consistent quantification data down to the level of one genomic equivalent copy with their respectively cloned amplicons, significant and important differences were observed for the detection of HAV genomic RNA. Results showed that the new real-time TaqMan SH-Poly and SH-Prot primer and probe systems were more consistent and sensitive by 5 logs as compared to both 5'-UTR designs (JN and SH-5U) used for the detection of HAV genomic RNA as well as for the detection in cell culture by cytopathic effect. Considering their higher analytical sensitivity, the proposed SH-Poly and SH-Prot amplification systems could therefore represent valuable tools for the detection of HAV in clinical, environmental and food samples.

Base Sequence↗

PubMed2002

Generation of anticalins with specificity for a nonsymmetric phthalic acid ester.

A set of engineered lipocalins, so-called anticalins, that bind benzyl butyl phthalate, a potential pollutant of environmental and food samples or medical plastic ware, has been generated. To this end, the synthesis of a derivative of the target analyte carrying an activatable carboxylate group at the end of an aliphatic spacer arm was established. This compound was covalently coupled to amino-functionalized paramagnetic beads. Using phage display technology three variants were selected from a random library of the bilin-binding protein (BBP), a prototypic lipocalin, which exhibit binding activity toward the nonsymmetric phthalic acid ester. These anticalins (denominated PhtA, PhtB, and PhtC) possess dissociation constants of 9.1, 6.2, and 11.6 microM, respectively. Specificity for the binding of other phthalic acid esters was studied. No cross-reactivity was found for diethyl phthalate, while binding to dibutyl phthalate was observed with higher dissociation constants. Interestingly, two differing types of binding behavior were observed among the three selected anticalins. Sequence comparison of these engineered lipocalins with the wild-type BBP revealed that all of the 16 randomized positions carried an amino acid exchange and that a certain sequence pattern had been selected, thus pointing toward a peculiar mode of structural interaction. Our data suggest that the generation of anticalins may provide an alternative to antibodies for the creation of stable receptor proteins against haptens with bioanalytical relevance.

Animals↗

PubMed2003

Bioaccumulation and damaging action of polymetal industrial dust on laboratory mice Mus musculus alba. I. Analysis of Zn, Cu, Pb, and Cd disposition and mathematical model for Zn and Cd bioaccumulations.

The concentrations of Zn, Cu, Pb, and Cd in the liver, kidneys, spleen, bones, and carcass of laboratory mice BALB/cy were observed in toxicological experiments. Polymetal industrial dust containing these metals was given to experimental animals at 1% concentration mixed with conventional animal food. Samples for analyses were taken on Days 15, 40, 60, 90, and 120 posttreatment. The experimental data clearly support the established antagonistic interactions among cadmium, zinc, copper, and lead. A mathematical model was proposed to study the main tendencies of heavy metal bioaccumulation under conditions of metal interaction and excessive exposure. The experimental results were assessed on the basis of the model. A rate constant of renal excretion greater than that of hepatic excretion was obtained, which agrees with the observed inversion of cadmium kidney/liver ratio in the conditions of very high exposure.

Administration, Oral↗

PubMed2000

Analysis of herbicide residues in cereals, fruits and vegetables.

The determination of herbicide residues in cereals, fruits and vegetables by chromatographic methods is reviewed. The principal chemical groups of herbicides, like phenoxyacids, benzonitriles, ureas, triazines, dinitroanilines, chloroacetamides, carbamates, uracils, glyphosate and bipyridylium compounds, are considered. This review briefly provides some basic information on food sample extraction, clean-up, derivatization and determination of herbicide residues.

Edible Grain↗