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Statements

Subject Item
n2:RIV%2F00027162%3A_____%2F14%3A%230001182%21RIV15-MSM-00027162
rdf:type
skos:Concept n15:Vysledek
dcterms:description
Alkylresorcinols (ARs) are amphiphilic phenolic lipids and their two main metabolites, 3-(3,5-dihydroxyphenyl)-propanoic acid (DHPPA) and 3,5-dihydroxybenzoic acid (DHBA), can be used as biomarkers of whole grain wheat and rye intake. The aim of this study was to develop antibodies against DHBA and DHPPA for use in ELISA analysis. Good calibration curves were obtained for ELISA using alkaline phosphatase (AP) conjugates. The highest sensitivity for DHPPA was found using a reagent combination of anti-DHPPA-BSA and DHPAA-AP in a direct ELISA (IC50 = 1.5 mu mol/L), and for DHBA using a reagent combination of anti-DHBA-OV and DHBA-AP (IC50 = 1.3 mu mol/L). Calibration was conducted in the linear range (0.3-27.4 mu mol/L), with limit of detection (LOD) 0.1 mu mol/L. Intra and inter CVs was in the range of 0.7-7.2% and 5.1-11.5%, respectively, for DHPPA and 1.3-9.4% and 3.5-20%, respectively, for DHBA. Mean recovery was 104% for DHPPA and 102% for DHBA. The ELISA method developed was then used for analysis of 120 urine samples from free-living men and women that had previously been analysed by gas chromatography-mass spectrometry (GC-MS). ELISA produced several-fold higher values than GC-MS. Application of high-resolution Orbitrap mass spectrometry (HR Orbitrap MS) allowed several compounds, including novel putative AR metabolites, to be identified, synthesised and confirmed as compounds with high ELISA cross-reactivity. Alkylresorcinols (ARs) are amphiphilic phenolic lipids and their two main metabolites, 3-(3,5-dihydroxyphenyl)-propanoic acid (DHPPA) and 3,5-dihydroxybenzoic acid (DHBA), can be used as biomarkers of whole grain wheat and rye intake. The aim of this study was to develop antibodies against DHBA and DHPPA for use in ELISA analysis. Good calibration curves were obtained for ELISA using alkaline phosphatase (AP) conjugates. The highest sensitivity for DHPPA was found using a reagent combination of anti-DHPPA-BSA and DHPAA-AP in a direct ELISA (IC50 = 1.5 mu mol/L), and for DHBA using a reagent combination of anti-DHBA-OV and DHBA-AP (IC50 = 1.3 mu mol/L). Calibration was conducted in the linear range (0.3-27.4 mu mol/L), with limit of detection (LOD) 0.1 mu mol/L. Intra and inter CVs was in the range of 0.7-7.2% and 5.1-11.5%, respectively, for DHPPA and 1.3-9.4% and 3.5-20%, respectively, for DHBA. Mean recovery was 104% for DHPPA and 102% for DHBA. The ELISA method developed was then used for analysis of 120 urine samples from free-living men and women that had previously been analysed by gas chromatography-mass spectrometry (GC-MS). ELISA produced several-fold higher values than GC-MS. Application of high-resolution Orbitrap mass spectrometry (HR Orbitrap MS) allowed several compounds, including novel putative AR metabolites, to be identified, synthesised and confirmed as compounds with high ELISA cross-reactivity.
dcterms:title
Development of antibodies for determination of alkylresorcinol metabolites in human urine and elucidation of ELISA cross-reactivity Development of antibodies for determination of alkylresorcinol metabolites in human urine and elucidation of ELISA cross-reactivity
skos:prefLabel
Development of antibodies for determination of alkylresorcinol metabolites in human urine and elucidation of ELISA cross-reactivity Development of antibodies for determination of alkylresorcinol metabolites in human urine and elucidation of ELISA cross-reactivity
skos:notation
RIV/00027162:_____/14:#0001182!RIV15-MSM-00027162
n4:aktivita
n13:P
n4:aktivity
P(LO1218)
n4:cisloPeriodika
NOV
n4:dodaniDat
n5:2015
n4:domaciTvurceVysledku
Wierzbicka, Roksana n18:6844758 n18:3431525
n4:druhVysledku
n12:J
n4:duvernostUdaju
n9:S
n4:entitaPredkladatele
n7:predkladatel
n4:idSjednocenehoVysledku
10968
n4:idVysledku
RIV/00027162:_____/14:#0001182
n4:jazykVysledku
n16:eng
n4:klicovaSlova
Alkylresorcinol; Dietary biomarker; ELISA; Antibodies; High-resolution Orbitrap mass spectrometry
n4:klicoveSlovo
n8:Alkylresorcinol n8:Dietary%20biomarker n8:High-resolution%20Orbitrap%20mass%20spectrometry n8:ELISA n8:Antibodies
n4:kodStatuVydavatele
NL - Nizozemsko
n4:kontrolniKodProRIV
[6D262F8290C4]
n4:nazevZdroje
Journal of Immunological Methods
n4:obor
n17:EI
n4:pocetDomacichTvurcuVysledku
3
n4:pocetTvurcuVysledku
5
n4:projekt
n14:LO1218
n4:rokUplatneniVysledku
n5:2014
n4:svazekPeriodika
413
n4:tvurceVysledku
Landberg, R. Fránek, Milan Kamal-eldin, A. Wierzbicka, Roksana Eyer, Luděk
n4:wos
000345819500002
s:issn
0022-1759
s:numberOfPages
13
n10:doi
10.1016/j.jim.2014.07.007