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Clinical Chemistry 28: 1745-1748, 1982;
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Clinical Chemistry, Vol 28, 1745-1748, Copyright © 1982 by American Association for Clinical Chemistry

Use of alumina, sephadex G10, and ion-exchange columns to purify samples for determination of epinephrine, norepinephrine, dopamine, homovanillic acid, and 5-hydroxyindoleacetic acid in urine

BH Westerink, FJ Bosker and JF O'Hanlon

We investigated the usefulness of small Sephadex columns, prepared in Pasteur pipettes, in purifying samples to be analyzed for catecholamines, homovanillic acid, and 5-hydroxyindoleacetic acid. After free catecholamines in urine samples were purified on alumina followed by Sephadex G10, a reliable and simultaneous quantification of epinephrine, norepinephrine, and dopamine was achieved by using high- performance liquid-chromatography with electrochemical detection. Purification of urine on Bio-Rad prepacked ion-exchange columns followed by Sephadex G10 resulted in a reliable, fast (200 samples processed per week) method for determination of free catecholamines in urine. Analytical recoveries of both methods were between 80 and 95%, with a CV of about 3%. Single purification on Sephadex G10 sufficed to allow simultaneous determination of homovanillic acid and 5- hydroxyindoleacetic acid in urine. Analytical recovery for this method was about 90%, with a CV of about 5%. Sephadex G10 columns, which can be re-used without regeneration for at least one year, appear to have a great potential in clinical chemistry.




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Copyright © 1982 by the American Association for Clinical Chemistry.