Liquid chromatographic determination of the adenosine receptor agonist CGS 21680 in blood using on-line solid-phase extraction on a phenylboronic acid support and fluorescence detection

Publication Type:

Journal Article

Authors:

Irth, H.; Cleton, A.; Mathot, R.A.; Danhof, M.; AP, I.J.; Tjaden, U.R.; van der Greef, J.

Source:

J Chromatogr B Biomed Appl, Volume 658, Number 1, p.207-12 (1994)

ISBN:

1572-6495 (Print)

Keywords:

Acetates; Adenosine/*analogs; &; derivatives/blood/pharmacokinetics; Animals; *Boronic Acids; Chromatography; High Pressure Liquid/*methods/statistics; &; numerical data; Drug Monitoring; Hydrogen-Ion Concentration; Male; Phenethylamines/*blood/pharma

Abstract:

An analytical method is described for the selective determination of A1 or A2 adenosine receptor agonists in blood. By implementing solid-phase extraction using immobilized-phenylboronic acid (PBA) in sample pretreatment, all adenosine derivatives are retained via their intact cis-diol group. On-line desorption of the analytes from the PBA support to the C18 analytical column is performed by injection of a small plug of perchloric acid. Fluorescence and UV detection are employed for the different adenosine derivatives. The method is applied to the determination of 2-[p-(2-carboxyethyl)phenylethylamino]-5'-N- ethylcarboxyamidoadenosine (CGS 21680, I) in blood using fluorescence detection. The only off-line sample handling step is the extraction of blood with ethyl acetate and subsequent evaporation of the extraction solvent. The detection limit of the method was 0.25 ng (signal-to-noise ratio 3:1) and the determination limit for I in blood (pretreatment of 100 microliters) was 5 ng/ml. The method was validated and used to study the pharmacokinetics of I in rats.