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Clinical Chemistry 36: 435-440, 1990;
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Clinical Chemistry, Vol 36, 435-440, Copyright © 1990 by American Association for Clinical Chemistry

Ultraviolet-absorbing organic anions in uremic serum separated by capillary zone electrophoresis, and quantification of hippuric acid

AC Schoots, TP Verheggen, PM De Vries and FM Everaerts
Faculty of Chemical Engineering, Eindhoven University of Technology, The Netherlands.

Organic anions accumulated in blood serum of patients with chronic renal failure were separated by a novel technique: closed-system capillary zone electrophoresis (CZE) in a pH6 carrier-electrolyte system. Hippuric acid (HA), p-hydroxyhippuric acid, and uric acid were identified by their co-elution with standards prepared in ultrafiltered normal serum and by comparison with the corresponding ultraviolet- detected peaks positively identified in the HPLC analyses. Analysis time for the entire profile is 8 min. Repeatabilities (CVs) of CZE migration times and peak areas of the three acids in serum samples were about 0.7% and 6%, respectively. We quantified HA in 10 ultrafiltered uremic serum samples and compared results with those by a previously described HPLC procedure. The very good agreement further supports the identification of hippuric acid. Accuracy and precision of the CZE method were similar to those for the HPLC gradient-elution method, but analysis time for HA (8 min) is much less than by HPLC (90 min). Our technique is very suitable for selective, rapid analysis for (ultraviolet-absorbing) anionic constituents in ultrafiltered uremic serum, without any sample pretreatment.




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Copyright © 1990 by the American Association for Clinical Chemistry.