Automated analytical systems for drug development studies. V. A system for enzyme kinetic studies

Two similar automated analytical systems using liquid chromatography (LC) and microdialysis as an on-line sampling technique were applied to studies of enzyme kinetics. 2′,3′,5′-Triacetyl-6-azauridine (azaribine) with porcine liver esterase (PLE) and N-acetylphenylalanyl-3,5-diiodotyrosine (AcFY...

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Bibliographic Details
Published in:	Journal of pharmaceutical and biomedical analysis Vol. 14; no. 12; pp. 1691 - 1698
Main Authors:	Zhou, J., Shearer, E.C., Hong, J., Riley, C.M., Schowen, R.L.
Format:	Journal Article Conference Proceeding
Language:	English
Published:	Amsterdam Elsevier B.V 01-09-1996 Elsevier Science
Subjects:	2′,3′,5′-triacetyl-6-azauridine Acetylphenylalanyl-3,5-diiodotyrosine Analysis Animals Automation Azauridine - analogs & derivatives Azauridine - metabolism Biological and medical sciences Chromatography, High Pressure Liquid - methods Dipeptides - metabolism Drug Design Enzyme kinetics Esterases - metabolism General pharmacology Hydrolysis Liver - enzymology Medical sciences Microdialysis Pepsin A - metabolism Pharmacology. Drug treatments Porcine liver esterase Reproducibility of Results Reversed-phase chromatography Swine Automation Microdialysis 2′,3′,5′-triacetyl-6-azauridine Acetylphenylalanyl-3,5-diiodotyrosine Enzyme kinetics Porcine liver esterase Reversed-phase chromatography Drug Enzyme Pepsin A HPLC chromatography Esterases Enzymatic activity Analytical method Development Hydrolases Aspartic endopeptidases Kinetics Proteinases
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Summary:	Two similar automated analytical systems using liquid chromatography (LC) and microdialysis as an on-line sampling technique were applied to studies of enzyme kinetics. 2′,3′,5′-Triacetyl-6-azauridine (azaribine) with porcine liver esterase (PLE) and N-acetylphenylalanyl-3,5-diiodotyrosine (AcFY') with pepsin were used as model compounds. The microdialysis sampling technique permitted the rapid separation of low molecular weight analytes from macromolecules, thus simultaneously achieving clean-up of the samples and quenching of the reaction. The combination of rapid LC analysis and microdialysis sampling provided selectivity and automation. The systems are rugged and give reproducible results in agreement with those from manual sampling methods.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0731-7085 1873-264X
DOI:	10.1016/0731-7085(96)01792-X