ASSAY FOR TRYPTOPHAN-HYDROXYLASE ACTIVITY IN RAT-BRAIN BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUORESCENCE DETECTION

Authors
IIZUKA R ISHIDA J YOSHITAKE T NAKAMURA M YAMAGUCHI M
Citation
R. Iizuka et al., ASSAY FOR TRYPTOPHAN-HYDROXYLASE ACTIVITY IN RAT-BRAIN BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUORESCENCE DETECTION, Biological & pharmaceutical bulletin, 19(5), 1996, pp. 762-764
Citations number
10
Categorie Soggetti
Pharmacology & Pharmacy
Journal title
Biological & pharmaceutical bulletin → ACNP
ISSN journal
09186158
Volume
19
Issue
5
Year of publication
1996
Pages
762 - 764
Database
ISI
SICI code
0918-6158(1996)19:5<762:AFTAIR>2.0.ZU;2-T
Abstract
A highly sensitive and selective liquid chromatographic (HPLC) method with post-column fluorescence detection has been developed for the det ermination of tryptophan 5-monooxygenase activity in rat brain tissue homogenate. 5-Hydroxytryptophan, formed enzymatically from tryptophan (incubation time, 20 min), is extracted with perchloric acid and deter mined by HPLC. Detection is performed fluorometrically after post-colu mn derivatization by a reaction with benzylamine in the presence of po tassium hexacyanoferrate (III), The lower limit of detection for 5-hyd roxytryptophan formed enzymatically is 100 fmol at a signal-to-noise r atio of three.