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Determination of total oncopterin, neopterin and biopterin in human urine by high performance liquid chromatography with solid phase extraction
Author(s) -
Tomandl Josef,
Tallova Jaroslava,
Tomandlova Marie,
Palyza Vladimir
Publication year - 2003
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200301371
Subject(s) - chromatography , solid phase extraction , urine , chemistry , high performance liquid chromatography , elution , extraction (chemistry) , neopterin , biopterin , analyte , biochemistry , medicine , cofactor , tetrahydrobiopterin , enzyme
A new, sensitive method for the determination of oncopterin, biopterin, and neopterin in human urine has been developed using SPE with 6,7‐dimethylpterin as internal standard and gradient HPLC with fluorescence detection. SPE was tested for the pre‐treatment of urine samples on different types of sorbents (strong ion exchange resins, polar adsorbents, and reversed‐phase sorbents). RP‐SPE with subsequent evaporation of eluate has been found to be the most appropriate. The extraction efficiency exceeded 95% for all selected pterins. The extracted pterins were subsequently analyzed on a Purospher RP‐18 RP column. The LOD of oncopterin was 1.43 nmol/L of urine. The intra‐day and inter‐day imprecision at a physiological oncopterin concentration never exceeded 10%. The potential of this method was tested using urine samples of healthy volunteers and cancer patients without methotrexate therapy.