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  • Elevated levels of pteridines can indicate the activation of cellular immune system by certain diseases. No work dealing with the simultaneous determination of urinary neopterin, biopterin and their reduced forms has been published. Therefore, a new SPE-UHPLC-FD method for the analysis of these compounds has been developed. The main emphasis was put on the stability of dihydroforms during the sample processing and storage. As a stabilizing agent, dithiothreitol, at various concentrations, and various pH values (3.8-9.8) of working solutions were tested. Chromatographic separation was performed under HILIC isocratic conditions on BEH Amide column. The method was linear for the calibration standard solutions in the range of 10-10,000 ng/ml (dihydroforms) and 0.5-1000 ng/ml (oxidized forms), and for real samples in the range of 25-1000 ng/ml (dihydroforms) and 1-100 ng/ml (oxidized forms). The development of a new SPE sample preparation method was carried out on different types of sorbents (based on a mixed-mode cation exchange, porous graphitic carbon and a polymer comprising hydrophilic and hydrophobic components). Final validation was performed on a MCAX SPE column. Method accuracy ranged from 76.9 to 121.9%. The intra- and inter-day precision did not exceed 10.7%. The method provided high sensitivity for the use in routine clinical measurements of urine (LLOQ 1 ng/ml for oxidized forms and 25 ng/ml for dihydroforms). Average concentrations of biopterin, neopterin, and dihydrobiopterin found in urine of healthy persons were related to the mol of creatinine (66.8, 142.3, and 257.3 mu mol/mol of creatinine, respectively) which corresponded to the literature data. The concentration of dihydroneopterin obtained using our method was 98.8 mu mol/mol of creatinine.
  • Elevated levels of pteridines can indicate the activation of cellular immune system by certain diseases. No work dealing with the simultaneous determination of urinary neopterin, biopterin and their reduced forms has been published. Therefore, a new SPE-UHPLC-FD method for the analysis of these compounds has been developed. The main emphasis was put on the stability of dihydroforms during the sample processing and storage. As a stabilizing agent, dithiothreitol, at various concentrations, and various pH values (3.8-9.8) of working solutions were tested. Chromatographic separation was performed under HILIC isocratic conditions on BEH Amide column. The method was linear for the calibration standard solutions in the range of 10-10,000 ng/ml (dihydroforms) and 0.5-1000 ng/ml (oxidized forms), and for real samples in the range of 25-1000 ng/ml (dihydroforms) and 1-100 ng/ml (oxidized forms). The development of a new SPE sample preparation method was carried out on different types of sorbents (based on a mixed-mode cation exchange, porous graphitic carbon and a polymer comprising hydrophilic and hydrophobic components). Final validation was performed on a MCAX SPE column. Method accuracy ranged from 76.9 to 121.9%. The intra- and inter-day precision did not exceed 10.7%. The method provided high sensitivity for the use in routine clinical measurements of urine (LLOQ 1 ng/ml for oxidized forms and 25 ng/ml for dihydroforms). Average concentrations of biopterin, neopterin, and dihydrobiopterin found in urine of healthy persons were related to the mol of creatinine (66.8, 142.3, and 257.3 mu mol/mol of creatinine, respectively) which corresponded to the literature data. The concentration of dihydroneopterin obtained using our method was 98.8 mu mol/mol of creatinine. (en)
Title
  • Sample preparation and UHPLC-FD analysis of pteridines in human urine
  • Sample preparation and UHPLC-FD analysis of pteridines in human urine (en)
skos:prefLabel
  • Sample preparation and UHPLC-FD analysis of pteridines in human urine
  • Sample preparation and UHPLC-FD analysis of pteridines in human urine (en)
skos:notation
  • RIV/00216208:11160/14:10282183!RIV15-MSM-11160___
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • I, P(EE2.3.20.0235), S
http://linked.open...iv/cisloPeriodika
  • July
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
http://linked.open.../riv/druhVysledku
http://linked.open...iv/duvernostUdaju
http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 43768
http://linked.open...ai/riv/idVysledku
  • RIV/00216208:11160/14:10282183
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • Dihydroforms; Biopterin; Neopterin; SPE; UHPLC-FD (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • NL - Nizozemsko
http://linked.open...ontrolniKodProRIV
  • [492DEED4149B]
http://linked.open...i/riv/nazevZdroje
  • Journal of Pharmaceutical and Biomedical Analysis
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
http://linked.open...cetTvurcuVysledku
http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 95
http://linked.open...iv/tvurceVysledku
  • Solich, Petr
  • Nováková, Lucie
  • Tomšíková, Helena
http://linked.open...ain/vavai/riv/wos
  • 000336558600035
issn
  • 0731-7085
number of pages
http://bibframe.org/vocab/doi
  • 10.1016/j.jpba.2014.03.012
http://localhost/t...ganizacniJednotka
  • 11160
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