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Statements

Subject Item
n2:RIV%2F00216208%3A11160%2F14%3A10282183%21RIV15-MSM-11160___
rdf:type
skos:Concept n20:Vysledek
rdfs:seeAlso
http://www.sciencedirect.com/science/article/pii/S073170851400137X
dcterms:description
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.
dcterms:title
Sample preparation and UHPLC-FD analysis of pteridines in human urine Sample preparation and UHPLC-FD analysis of pteridines in human urine
skos:prefLabel
Sample preparation and UHPLC-FD analysis of pteridines in human urine Sample preparation and UHPLC-FD analysis of pteridines in human urine
skos:notation
RIV/00216208:11160/14:10282183!RIV15-MSM-11160___
n5:aktivita
n13:I n13:S n13:P
n5:aktivity
I, P(EE2.3.20.0235), S
n5:cisloPeriodika
July
n5:dodaniDat
n19:2015
n5:domaciTvurceVysledku
n9:4906268 n9:8491836 n9:7555350
n5:druhVysledku
n12:J
n5:duvernostUdaju
n17:S
n5:entitaPredkladatele
n16:predkladatel
n5:idSjednocenehoVysledku
43768
n5:idVysledku
RIV/00216208:11160/14:10282183
n5:jazykVysledku
n11:eng
n5:klicovaSlova
Dihydroforms; Biopterin; Neopterin; SPE; UHPLC-FD
n5:klicoveSlovo
n7:SPE n7:Dihydroforms n7:Neopterin n7:Biopterin n7:UHPLC-FD
n5:kodStatuVydavatele
NL - Nizozemsko
n5:kontrolniKodProRIV
[492DEED4149B]
n5:nazevZdroje
Journal of Pharmaceutical and Biomedical Analysis
n5:obor
n6:CB
n5:pocetDomacichTvurcuVysledku
3
n5:pocetTvurcuVysledku
3
n5:projekt
n10:EE2.3.20.0235
n5:rokUplatneniVysledku
n19:2014
n5:svazekPeriodika
95
n5:tvurceVysledku
Nováková, Lucie Solich, Petr Tomšíková, Helena
n5:wos
000336558600035
s:issn
0731-7085
s:numberOfPages
8
n15:doi
10.1016/j.jpba.2014.03.012
n14:organizacniJednotka
11160