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Biomedical subjects

C G Chappell

Publications and source records attributed to C G Chappell.

7 recordsLinked to original sources

Size exclusion chromatography/mass spectrometry applied to the analysis of polysaccharides.

A novel method for analysing polysaccharide materials is described which employs size-exclusion chromatography (SEC) followed by detection by on-line electrospray ionisation mass spectrometry (ESI-MS) and off-line matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS). It is demonstrated through SEC/ESI ion trap mass spectrometry that the formation of multiply charged oligomer ions, which bind up to five sodium cations, allows the rapid analysis of polysaccharide ions with molecular weights in excess of 9 kDa. MALDI spectra generated from fractionation of the effluent collected from the same SEC separation are shown to be in good agreement with the ESI spectra with respect to molecular weight distributions and types of ions generated. ESI and MALDI mass spectra of samples obtained from sequential graded ethanol precipitation and SEC fractionation of acid and enzymatically digested arabinoxylan polysaccharides show important structural differences between polysaccharide fragments. In addition, a comparison is made between the mass spectra of native and permethylated SEC-separated fragments of acid and enzymatically treated arabinogalactan. Linkage information of the permethylated arabinogalactan oligomers can be rapidly established through the use of on-line SEC/ESI-MS( n) experiments.

Bacterial Proteins↗

Modified on-column interface for coupled high-performance liquid chromatography-gas chromatography and its application to the determination of levamisole in milk.

A modified on-column interface is reported for the coupling of high-performance liquid chromatography with gas chromatography, incorporating an adapted, commercially available multidimensional gas chromatography switching system. Novel features include cryogenic analyte focusing, total solvent exclusion from the analytical column and independent carrier gas supplies to the analytical GC column and uncoated pre-column. The instrumentation was used for the determination of the veterinary anthelmintic drug levamisole in milk with analyte detection by both flame ionisation and nitrogen-phosphorus detectors. Detection limits for the assay were 2.2 micrograms l-1 and 0.4 micrograms l-1 by flame ionisation and nitrogen-phosphorus detectors, respectively. The assay was applied to a survey of fourteen milk samples from different dairy outlets.

Animals↗

On-line high-performance liquid chromatographic/gas chromatographic/tandem ion trap mass spectrometric determination of levamisole in milk.

On-line high-performance liquid chromatography/high-resolution gas chromatography/tandem mass spectrometry, performed on a quadrupole ion trap, has been used as a rapid method for the quantification of the anthelmintic drug levamisole in raw milk extracts following a simple extraction procedure. Detection at the 0.5 p.p.b. level and a linear response in the 10-0.5 p.p.b. range is demonstrated. Multiple-scan monitoring techniques have been used to acquire chemical ionization tandem mass spectra and electron impact mass spectra in a single chromatographic run.

Animals↗

Prostacyclin synthesis by vascular tissue. Effect of extracellular Ca2+.

Prostacyclin (PGI2) synthesis in vascular tissue is stimulated by free Ca2+. Concentrations in excess of 4 mM result in a sustained release of PGI2, but not other prostanoids, from rat aorta. This effect arises by increased synthesis of PGI2 and not through the inhibition of its metabolism. Physico-chemical examination of serum and plasma extract has indicated that much of the PGI2-stimulating activity reported for human serum derives from the presence of free Ca2+.

Animals↗

Generation of hydroxyeicosatetraenoic acids by human inflammatory cells: analysis by thermospray liquid chromatography-mass spectrometry.

Arachidonic acid was converted to a series of hydroxyeicosatetraenoic acids (HETEs) by mixed human inflammatory cells following stimulation with the calcium ionophore A23187. HETEs were purified by a simple one-step extraction procedure followed by HPLC. The HPLC was coupled to a Finnigan quadrupole mass spectrometer using the now commercially available thermospray liquid chromatography-mass spectrometry interface. The HPLC eluant was monitored 'on line' by the mass spectrometer. Soft ionisation occurs, generating intense molecular ion species in the negative ion mode (M - H-:m/z 319) for each of the isomeric HETEs. The (M + H+ - H2O) ion at m/z 303 is the major species in the positive ion spectra of HETEs. Mass spectra were obtained on-line post-HPLC for HETEs formed by the human cells, and the HPLC-MS profile compared with that obtained from standards; species corresponding to the 11-, 9- and 5-HETEs were observed.

Arachidonic Acid↗

Prostaglandin D2 and histamine release in cold urticaria.

Prostaglandin (PG) D2 and histamine concentrations have been measured in blood draining cold-challenged forearm skin in patients with cold urticaria. Local venous concentrations of both histamine and PGD2 rose in four patients who developed a whealing response. Plasma histamine concentration increased from a mean resting value of 0.24 +/- 0.09 (SD) ng/ml to peak values of 16.9 to 96.6 ng/ml. Resting concentrations of PGD2 were below the limit of detection (5 pg/ml) in three patients and 62 and 27 pg/ml in the fourth. Peak plasma PGD2 concentration after challenge ranged from 166 to 279 pg/ml. Time course of histamine and PGD2 release was similar with peak concentrations at 6 and 10 minutes, respectively. The maximum clinical response occurred between 10 and 20 minutes after challenge. Our findings demonstrate that PGD2 is produced in association with mast cell degranulation in man, but the amount, relative to histamine, is low. Despite its high potency in production of inflammatory effects, PGD2 probably has only minor direct effects in cold urticaria, although it may act to potentiate other mediators.

Adolescent↗

Measurement of prostaglandin D2 and identification of metabolites in human plasma during intravenous infusion.

A stable isotope dilution assay has been developed for the quantification of prostaglandin D2 (PGD2) in plasma. Samples are analysed by capillary column gas chromatography/negative ion chemical ionisation mass spectrometry (GC/NICIMS). The method employs an internal standard of [2H6]PGD2, prepared biosynthetically by incubation of rat peritoneal mast cells with deuterated arachidonic acid. No PGD2 was detected in peripheral venous plasma samples obtained from 10 healthy male volunteers (detection limit = 5 pg ml-1). Following intravenous infusion of PGD2 at increasing incremental infusion rates ranging from 16-256 ng kg-1 min-1, a dose related increase in the plasma concentration of PGD2 was observed. Plasma levels at 128 ng kg-1 min-1 ranged from 724-1444 pg ml-1. The major circulating metabolites of PGD2, during infusion, were identified as 13,14-dihydro-15-oxo-PGF2 alpha and 15-oxo-PGF2 alpha.

Animals↗