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Pre-analytical factors affecting ascorbic and uric acid quantification in human plasma.

Angelo Zinellu1, Salvatore Sotgia, Luca Deiana

  • 1Department of Biomedical Sciences, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy. angelozinellu@libero.it

Journal of Biochemical and Biophysical Methods
|March 8, 2006
PubMed
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This study optimized a capillary electrophoresis assay for measuring ascorbic acid (AA) and uric acid (UA). Key findings include stabilizing AA with cysteine and EDTA, recommending EDTA blood collection, and optimizing protein precipitation for accurate results.

Area of Science:

  • Analytical Chemistry
  • Biochemistry

Background:

  • Ascorbic acid (AA) is prone to oxidation during sample preparation.
  • Accurate measurement of AA and uric acid (UA) requires careful consideration of pre-analytical factors.

Purpose of the Study:

  • To optimize a capillary electrophoresis (CE) assay for simultaneous measurement of AA and UA.
  • To investigate pre-analytical factors influencing AA stability and measurement accuracy.

Main Methods:

  • Development of a rapid CE method (<4 min) using a specific capillary and buffer.
  • Evaluation of standard preparation (cysteine, EDTA), blood collection methods (serum vs. EDTA plasma), and protein precipitation techniques.
  • Validation of the optimized CE method against a reference CE assay using Bland-Altman analysis in 32 healthy volunteers.

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Main Results:

  • Standard AA solutions require cysteine and EDTA to prevent oxidation.
  • EDTA plasma collection yields more reliable AA measurements than serum.
  • The choice and amount of precipitating agent significantly impact analyte recovery.
  • The developed CE method demonstrated accuracy comparable to a reference assay.

Conclusions:

  • Optimized pre-analytical steps, including standard preparation and sample collection, are crucial for accurate AA and UA quantification by CE.
  • The new CE assay provides a rapid and accurate method for measuring serum AA and UA.