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Biomedical subjects

K A Kovar

Publications and source records attributed to K A Kovar.

At least 19 recordsLinked to original sources

Analysis and stability of Hyperici oleum.

Hyperici Oleum (St. John's wort oil) used in wound healing contains no hypericin. By using the sunlight maceration method described in the supplement to DAB 6 (EB 6), lipophilic breakdown products of this compound are obtained which lend the oil its red colour. Hyperforin, which is responsible for the oil's therapeutic activity could, for the first time, be identified and quantitatively determined by TLC and HPLC after solid-phase extraction. The stability of hyperforin is limited; sufficient shelf-life could only be achieved by hot maceration of dried flowers with eutanol G and storage in the absence of air. By gradient HPLC further polar hyperforin analogues were detected in those St. John's wort oils in which hyperforin had decomposed. At the same time flavonoids and xanthones could be identified. A procedure for the quantitative determination of flavonoids in St. John's wort was validated. The action of light during preparation of the oil led to a rise in the content of flavonoids.

Flavonoids

An enzymatic assay for the colorimetric and fluorimetric determination of uric acid in sera.

The methods described in this paper are based on the uricase catalyzed oxidation of uric acid to allantoine and hydrogen peroxide. By making use of the catalytic activity of peroxidase the generated H2O2 is measured either spectrophotometrically with 3-methyl-benzothiazoline-2-one hydrazone (MBTH) and 3-dimethylaminobenzoic acid (DMAB) (M1) or fluorimetrically with tyramine (M2) or L-tyrosine (M3). The methods are simple, sensitive and selective. The procedures developed can be rapidly and readily performed on patient serum samples without deproteinization using 100 microliters and 5 microliters for colorimetric and fluorimetric assay, respectively.

Colorimetry

Determination of histamine and its metabolic products in the pig gastric mucosa.

This paper describes analytical techniques for quantitative and qualitative determination of histamine and its metabolites (N tau-methylhistamine, N alpha-methylhistamine and N alpha, N alpha-dimethylhistamine) in the pig gastric mucosa. These metabolites and N alpha-acetylhistamine, imidazolyl-4-acetic acid and N tau-methylimidazolylacetic acid were synthesized as reference compounds and analyzed by using the dansylation technique. TLC of dansylated mucosal extracts in various solvents (in situ fluorescence measurements) in combination with TLC/IR and TLC/MS transfer technique demonstrated the presence of histamine. The only metabolite was N tau-methylhistamine.

Animals

Methylation of histamine in the gastric mucosa.

Methylation of histamine in the gastric mucosa of various species has been studied in vitro and in vivo. When gastric mucosal homogenates of rat, guinea pig, cat, dog, and pig were incubated with 14C-histamine in the presence of S-adenosyl methionine Ntau-methylhistamine was formed as the only methylated histamine derivative. Excessive concentration of Ntau-methylhistamine inhibited the formation of Ntau-methylhistamine. Pretreatment of guinea pigs with prednisolone, aminoguanidine, or pentagastrin neither influenced in vitro the formation of Ntau-methylhistamine significantly nor shifted the methylation reaction towards the side chain. Pretreatment of guinea pigs with the histamine H2-receptor blocker cimetidine significantly enhanced the formation of Ntau-methylhistamine in vitro. In pooled samples of guinea pig gastric mucosa histamine and Ntau-methylhistamine were found. Under none of these circumstances was Nalpha-methyl-or Nalpha,Nalpha-dimethylhistamine found. It is concluded that in rat (to a small extent), guinea pig, cat, dog, and pig gastric mucosa the formation of Ntau-methylhistamine is the only methylating pathway of histamine. In cases where other methylated histamine derivates (Nalpha-methyl- and/or Nalpha,Nalpha-dimethylhistamine) are found they must origin in sources other than the gastric mucosa.

Animals