Purification and characterization of the human liver cytochromes P-450 involved in debrisoquine 4-hydroxylation and phenacetin O-deethylation, two prototypes for genetic polymorphism in oxidative drug metabolism.

Debrisoquine/sparteine-type polymorphism of drug oxidation. Purification and characterization of two functionally different human liver cytochrome P-450 isozymes involved in impaired hydroxylation of the prototype substrate bufuralol

The principles and applications of fluorescence detection and fluorescence introducing reagents and methods in HPLC are reviewed. The design and requirements for fluorescence detectors, flow cells and excitation sources and the conversion of non-fluorescent compounds into fluorescent products by pre-column and post-column derivatization reactions are discussed. For the applications the emphasis is on drug analysis, where possible in biological fluids (serum, urine, etc.). The last paragraphs are divided in a number of sections in which newly developed and some scarcely used reagents are mentioned shortly; a more complete treatment is given of the reagents and labels most frequently used in the derivatization of certain functional groups. In this discussion the methods of derivatization as well as the selectivity, stability, fluorescence behaviour of the reagents/labels and derivatives and the reaction conditions are included. An up-to-date survey of the applications of fluorescence detection in l...

Fluorescence Detection in High Performance Liquid Chromatography

tribution, and excretion of AT, NT, IMI, or DM1. The assay was usually by spectrophotometry or simple chemical analyses, some of which involved separation of parent drug from its metabolites by chromatography (1-9). None of these assays were sensitive enough for analysis of amounts of drugs likely to be in plasma or blood. Fluorimetric techniques were developed that had adequate sensitivity but were usually nonspecific because of metabolite interference (10-12). More suitable assays were developed in 1967, first by Moody and colleagues (13), who described a more advanced fluorimetric procedure for IMI and DM1, and then by Hammer and Brodie (14), who developed an isotope-derivative assay for NT or DM1. A further advance came h 1970-71 with the first suitable GC techniques for DM1, NT, and AT (15-18). Since then, the majority of published methods have involved GC, at first with the more conventional detectors, flame-ionization or electron-capture, but more recently with either nitrogen-specific detection or mass-fragmentography. Other contemporary methods investigated include thin-layer chromatography-densitometry (TLC-D), HPLC, and radioimmunoassay. In this review, we will discuss the approaches used since 1967, emphasizing specificity, sensitivity, accuracy, precision (reproducibility), expense, convenience, and ease of sample processing. A procedure will be considered specific if drug metabolites do not interfere and if only drugs that have been shown not to interfere with the assay procedure are administered concomitantly. The sensitivity required for the routine analysis of plasma amounts of most tricyclic drugs is ap

Measurement of tricyclic antidepressants. Part I. A review of methodology.

Bufuralol (Ro 34787, Angium®) is a non selective β-adrenoceptor blocking drug with some degree of sympathomimetic action and a longer duration of action than propranolol. Plasma concentrations of bufuralol and 1 -hydroxybufuralol, its main blood derivative which shows similar β-adrenoceptor blocking properties, were determined in healthy volunteers after a 60 mg oral and a 20 mg intravenous dose. Peak plasma concentrations were higher for the parent drug but due to a longer elimination half-life, the metabolite concentrations became higher after a few hours (bufuralol t1/2=2.7±0.9 h, metabolite t1/2=6.1±l.5h) The bioavailability of the tablet tested was 46±15%. The occurrence of side-effects in a subject with abnormal pharmacokinetics of the drug in this study and in a previous study with this drug suggested the possibility of a pharmacogenetic anomaly. Determination of the plasma metabolic ratio in the family of this subject and in a larger population confirmed that aliphatic hydroxylation of bufuralol is under polymorphic control. Phenotyping of our volunteers whith debrisoquine showed the present pharmacogenetic anomaly to be the same as the one reported for debrisoquine alicyclic hydroxylation. The occurrence of side-effects in poor metabolizers as seen with bufuralol illustrates the clinical relevance of the hydroxylation polymorphism. In Switzerland the frequency of poor metabolizers is about 9% as previously reported for Caucasian British subjects.

The genetic control of bufuralol metabolism in man.

Determination of bufuralol and its major metabolites in plasma by high-performance liquid chromatography

Determination of amitriptyline and nortriptyline in human plasma by quantitative thin-layer chromatography.

A specific and sensitive method for the determination of the anticoagulant phenprocoumon in plasma.

Sensitive assay for the tricyclic antidepressant Ro 11-2465 in biological fluids by high-performance liquid chromatography and fluorescence detection

Determination of the imidazo quinazoline derivative Ro 13-6438 in biological fluids by high-performance liquid chromatography

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