SEPARATION OF URINARY PURINES AND PYRIMIDINES BY ISOTACHOPHORESIS - USEFULNESS IN SCREENING FOR INBORN-ERRORS OF PURINE AND PYRIMIDINE METABOLISM

A new electrophoretic technique-analytical isotachophoresis-was investigated for the separation of urinary purines and pyrimidines. A standard mixture of bases and nucleosides was separated in 45 min and the individual components were readily detectable at the 100 pmol level. The most suitable procedure involved the use of a 43 cm capillary and a buffer system in which the leading electrolyte was 2.5 mM Tris-HCl, pH 7.9, and the terminating electrolyte 20 mM β-alanine, pH 10.6. Using this system urine samples from patients with six different enzyme defects were analyzed. Profiles recorded at 254 and 280 nm could distinguish characteristic ultraviolet absorbing compounds from five of the defects; the exception was adenosine deaminase deficiency.* * Since this article went to press, it has been possible to identify deoxyadenosine as adenine using a repeat scan following acid hydrolysis Isotachophoresis was found to be a simple, rapid, and reproducible technique requiring only a few μl of urine, potentially useful as a preliminary screening test for disorders of purine and pyrimidine metabolism. Other possible uses include the monitoring of allopurinol metabolites in gouty patients. © 1979.