Liquid-chromatographic determination of amitriptyline and its metabolites in serum, with adsorption onto glass minimized

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Abstract

To study correlations between the concentrations, in serum, of amitriptyline and its most important metabolites with clinical response in patients, we developed a 'high-performance' liquid-chromatographic method for routine determination of amitriptyline, nortriptyline, total 20-hydroxy-amitriptyline, desmethylnortriptyline, and E(trans)- and Z(cis)-10-hydroxynortriptyline. These compounds are extracted from 1 mL of alkalinized serum to hexane/isoamyl alcohol (99/1 by vol). Perazine is the internal standard. To minimize irreversible adsorption of the drugs onto the glassware, 5 μg of maprotiline is added to the organic phase just before evaporation. After a 10-min resolution on a silica column eluted with acetonitrile/methanol NH4OH (1 mol/L), absorbance is measured at 240 nm. Only chlorimipramine, doxepin, procainamide, and N-acetylprocainamide may interfere with assay of the compounds that probably are therapeutically relevant: amitriptyline, nortriptyline, and E-10-hydroxynortriptyline. Uremia, lipemia, and icterus also do not affect the analysis.

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Edelbroek, P. M., De Haas, E. J. M., & De Wolff, F. A. (1982). Liquid-chromatographic determination of amitriptyline and its metabolites in serum, with adsorption onto glass minimized. Clinical Chemistry, 28(10), 2143–2148. https://doi.org/10.1093/clinchem/28.10.2143

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