Rapid and simple determination of histamine-N-methyl transferase activity by high-performance liquid chromatography with UV detection.
A rapid, simple and low-cost assay method of histamine-N-methyltransferase activity was developed. Methylhistamine, which was separated from the enzymatic reaction system on reversed-phase high-performance liquid chromatography using an ion-paired chromatographic technique, was detected spectrophotometrically at 226 nm. The mobile phase used for the separation of methylhistamine was 0.05M NH4H2PO4 (pH 3.0) containing 2 mM of sodium octanesulfonate. The new assay technique could detect methylhistamine as an enzyme activity product of histamine-N-methyltransferase in the brain and kidney of rats. Chloropheniramine maleate, an antihistamine, activated the histamine-N-methyltransferase. Whether neurotransmitter or neuromodulator, the role of histamine in the brain has not yet been made clear. Therefore, the present method could be applicable for the enzymatic investigation of histamine metabolism in central nervous system or inflammatory reactions.
Inoue
H
H
Department of Plastic and Reconstructive Surgery, St. Marianna University School of Medicine, 2-16-1 Sugao, Miyamae, Kawasaki 216-8511, Japan. h2inoue@marianna-u.ac.jp
Hisamatsu
K I
KI
Yamauchi
M
M
Kumagai
N
N
D000818:Animals; D001921:Brain / Q000201:enzymology; D002744:Chlorpheniramine / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007692:Diatomaceous Earth; D004789:Enzyme Activation / Q000187:drug effects; D006634:Histamine H1 Antagonists / Q000494:pharmacology; D006637:Histamine N-Methyltransferase / Q000032:analysis*; D018836:Inflammation Mediators / Q000032:analysis; D007668:Kidney / Q000201:enzymology; D051381:Rats; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet
D006634:Histamine H1 Antagonists; D018836:Inflammation Mediators; D012997:Solvents; D002744:Chlorpheniramine; D007692:Diatomaceous Earth; C018540:Extrelut; D006637:Histamine N-Methyltransferase