Document Detail

A high-performance liquid chromatographic method for the determination of hypoxanthine, xanthine, uric acid and allantoin in serum.
MedLine Citation:
PMID:  8357939     Owner:  NLM     Status:  MEDLINE    
A method was developed for the simultaneous determination of hypoxanthine, xanthine, uric acid and allantoin based on isocratic reversed-phase chromatography. This HPLC-method additionally allows the direct determination with UV-detection of inosine-5'-phosphate, uridine, thymine, orotic acid, allopurinol and oxipurinol, besides hypoxanthine, xanthine and uric acid in the same chromatographic run. Allantoin elutes in this system near the void volume and a fraction is collected covering the retention time range for this substance. After hydrolysis allantoin is converted to glyoxylate-2,4-dinitrophenylhydrazone, rechromatographed and detected at 360 nm. The coefficient of variation for this method does not exceed 5.0% for a serum concentration of 0.3 mumol/l hypoxanthine and is not greater than 5.3% for a xanthine concentration of 0.3 mumol/l serum. Recoveries were 90-110% for both hypoxanthine and xanthine. The determination of uric acid had an imprecision and inaccuracy not exceeding 1.45% in the concentration range of 103-568 mumol/l. Due to the more complex procedure required for the determination of allantoin, the coefficient of variation between days was 13.6% for a sample containing 0.8 mumol/l allantoin and the recoveries for this analyte were in the range of 86-93%. Reference ranges (mean +/- SD) determined on 171 serum samples from healthy adults were 12.7 +/- 6.6 mumol/l for hypoxanthine, 3.3 +/- 1.4 mumol/l for xanthine, and 15.7 +/- 7.9 mumol/l for allantoin. No significant age or sex dependence was observed. Uric acid concentrations were 320 +/- 55 mumol/l serum for men and 206 +/- 55 mumol/l for women.
R Kock; B Delvoux; H Greiling
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  European journal of clinical chemistry and clinical biochemistry : journal of the Forum of European Clinical Chemistry Societies     Volume:  31     ISSN:  0939-4974     ISO Abbreviation:  Eur J Clin Chem Clin Biochem     Publication Date:  1993 May 
Date Detail:
Created Date:  1993-09-29     Completed Date:  1993-09-29     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  9105775     Medline TA:  Eur J Clin Chem Clin Biochem     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  303-10     Citation Subset:  IM    
Institute of Clinical Chemistry and Pathobiochemistry, Medical Faculty, University of Technology Aachen.
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MeSH Terms
Allantoin / blood*
Chromatography, High Pressure Liquid / methods*
Hypoxanthines / blood*
Middle Aged
Reference Values
Reproducibility of Results
Sensitivity and Specificity
Uric Acid / blood*
Xanthines / blood*
Reg. No./Substance:
0/Hypoxanthines; 0/Xanthines; 68-94-0/Hypoxanthine; 69-89-6/Xanthine; 69-93-2/Uric Acid; 97-59-6/Allantoin

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