Pteridine determination in human serum with special emphasis on HPLC methods with fluorimetric detection

Conjugated and unconjugated pteridines and their derivatives are important cofactors in cellular metabolism. Hence, the amount of unconjugated pteridines in biological fluids has been found to be modified as a result of several disorders. It is necessary to note that while for the control of pteridines in urine samples there are numerous reference data, the literature referred to for the analysis of these analytes in serum/plasma is scarce. In biological fluids, pteridines can exist in different oxidation states, and these compounds can be classified into two groups according to: (a) oxidized or aromatic pteridines and (b) reduced pteridines. Oxidized pteridines yield a strong fluorescence signal, whereas reduced pteridines present a low quantum yield of fluorescence. In order to enable the analysis of the reduced forms, several preoxidation procedures to generate aromatic rings have been established. Also, stabilization of the reduced forms by the addition of reducing agents has been widely reported. The objective of this paper is to show possibilities and different approaches in the analysis of pteridines in serum samples. We have mainly focused on the description of the current situation in the application of high-performance chromatography methods with fluorimetric detection