Direct injection/h.p.l.c. methods for the analysis of drugs in biological samples.

1. Direct injection h.p.l.c. methods for zaprinast, and pantoprazole and its sulphone metabolite were developed. 2. Optimal recovery of pantoprazole and its sulphone metabolite was effected by the absence of transfer losses and the effective adjustment of sample pH on-line. 3. Acetonitrile reduced the recovery of pantoprazole and its sulphone metabolite at acetonitrile concentrations greater than 5% in serum. 4. Direct injection h.p.l.c. methods minimize sample handling losses, reduce human contact with biological samples and are sufficiently accurate and reproducible to be used to support pharmacodynamic and toxicokinetic studies.

Determination of temelastine and a hydroxymethyl-pyridyl metabolite in biological fluid by high-performance liquid chromatography.

A high-performance liquid chromatographic method for the analysis of temelastine (1) and 2-[4-(5-bromo-3-methylpyrid-2-yl)-butylamino]-5-[6-hydroxymethy lpyrid-3- ylmethyl]-pyrimidin-4(1H)-one (1-A) in biological fluid is presented. The method combines the previously reported extraction procedure and new chromatography conditions capable of resolving 1, 1-A, and structurally similar compounds formed by the oxidation of 1. The modified method has been used to measure concentrations of 1 and 1-A in biological fluids taken from the rat and dog, and to look for the presence of 1-A in humans following administration of 1.

Two systems for the automated analysis of drugs in biological fluids using high-performance liquid chromatography.

This paper describes two fully automated assays. One for zaprinast, a cGMP specific phosphodiesterase inhibitor, which uses the Gilson-Advanced Automated Sample Processor combination, and the other for an H+/K+ ATPase inhibitor and its sulphone metabolite, which uses direct injection. Both assays were developed to support pharmacokinetic studies at therapeutic doses in small animals as well as in man. Plasma or serum (20-200 microliters) is placed directly into an autosampler and all subsequent manipulations are performed mechanically.

Sample preparation for the HPLC analysis of drugs in biological fluids.

Sample preparation for the analysis of drugs in biological fluids consists of a number of unit operations that are used for (i) release of the drug from a conjugate or biological matrix; (ii) removal of endogenous compounds that could interfere with the assay; and (iii) techniques for liquid handling. The trends in sample preparation that have occurred over the past 10 years in the authors' laboratory are discussed. In general, there has been a move from the traditional liquid-liquid extraction to methods using bonded-silica which permit rapid throughput and efficient extraction. Automation of sample preparation has seen further gains in productivity; however, the present generation of equipment lack the control and communication systems that are essential for the development of the automated integrated laboratory of the future.

Analysis of drugs from biological fluids using disposable solid phase columns.

The traditional liquid-liquid extraction method for the removal of drug from biological matrix is being superseded by solid phase extraction. This involves the selection of an appropriate sorbent (normal-phase, reversed-phase, ion-exchange etc.), but once this has been achieved the method is quick and simple to operate. Most sample handling losses are avoided so recovery of drug is high and it is easily automated. Disposable columns have several advantages. Samples of 0.05-2.0 ml can be analysed routinely. Several wash stages can be included in a method to provide a specific extraction prior to a quick analysis by high-performance liquid chromatography (HPLC), radioimmunoassay, UV etc. A potential problem is that retention of the drug may involve more than one mechanism. Elution of drug may therefore require a stronger eluting solvent than analytical HPLC systems using the same stationary phase.