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

Abstract

<jats:title>Abstract</jats:title><jats:p>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.</jats:p>

Dilute-and-Shoot-Liquid Chromatography-Quadrupole Time of Flight-Mass Spectrometry for Pteridine Profiling in Human Urine and Its Association with Different Pathologies

“…The use of acidic pH to dissolve pteridines could also cause the cleavage of the double ring, giving rise to other compounds, whereas a basic pH allows the dissolution of the pteridines without affecting their chemical structure. Stabilisers such as dithiothreitol (DTT) and ascorbic acid (AA) were used to slow down the oxidation reaction of pteridines in the presence of air [27,28,35,43]. Individual solutions of the different pterins were prepared at 500 mg L −1 using 2 M NaOH as solvent and DTT and AA were added to each solution at 1 and 4 g L −1 , respectively.…”

“…Individual solutions of the different pterins were prepared at 500 mg L −1 using 2 M NaOH as solvent and DTT and AA were added to each solution at 1 and 4 g L −1 , respectively. Both DTT and AA, added as stabilising agents to preserve the pterins in their native oxidation state [2,27,28,32,43], were supplied by Sigma (St. Louis, MO, USA). Standard solutions of lumazines were prepared at 1000 mg L −1 in Milli-Q water with a few drops of 1 M NaOH.…”

Insights into Molecular Structure of Pterins Suitable for Biomedical Applications

“…The most common analytical methods of pteridine determination are high performance liquid chromatography (HPLC), capillary electrophoresis, and enzyme-linked immunosorbent assay (ELISA) [ 76 , 77 ]. HPLC can be used along with spectrophotometric, fluorescence, electrochemical detection, or mass spectrometry [ 78 ].…”

“…HPLC can be used along with spectrophotometric, fluorescence, electrochemical detection, or mass spectrometry [ 78 ]. The particular biological fluids used for pteridine determination are blood serum [ 77 , 79 ], urine [ 73 ], and cerebrospinal fluid (CSF) [ 78 ].…”

Amino acid-stabilized luminescent gold clusters for sensing pterin and its analogues

“…The changes of the purine concentration ratio are indicative of DNA abnormalities and mutations emerging from certain diseases such as cancer, HIV, epilepsy, etc. 16 The methods of pteridine detection include various chromatography techniques, in particular HPLC 17 and capillary electrophoresis, 18 as well as HPLC-ESI-MS 19 and HPLC-MS/MS 20 techniques, which are time-and labor-consuming and expensive. In this respect, developing new simple, cheap and fast techniques is in great demand.…”