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Quantification of isotope-labeled and unlabeled folates and folate catabolites in urine samples by stable isotope dilution assay.
Technische Universität München, Germany.
Swedish University of Agricultural Sciences, Uppsala.
German Research Center for Food Chemistry, Leibniz Institute Freising, Germany.
Swedish University of Agricultural Sciences, Uppsala.ORCID-id: 0000-0003-0387-4312
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2013 (Engelska)Ingår i: International Journal for Vitamin and Nutrition Research, ISSN 0300-9831, E-ISSN 1664-2821, Vol. 83, nr 2, s. 112-121Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Dual-label stable isotope dilution assays for the simultaneous quantification of isotopologic folates in clinical samples offer the perspective for differentiating between unlabeled folates from endogenous body pools and administered [13C5]-labeled folates from a test dose when performing bioavailability trials. In contrast to intact folates, this methodology could hitherto not be applied to the quantification of the folate catabolites, p-aminobenzoyl glutamate and p-acetamidobenzoyl glutamate. In this study, [2H4]-p-aminobenzoyl glutamate, [2H4]-p-acetamidobenzoyl glutamate, and unlabeled p-acetamidobenzoyl glutamate were synthesized. The synthesis of the [2H4]-labeled compounds started at unlabeled p-aminobenzoic acid. For the formation of p-acetamidobenzoyl glutamate, p-aminobenzoyl glutamate was acetylated. The new substances were applied successfully in stable isotope dilution assays for the simultaneous quantification of the [13C5]-labeled and unlabeled folate catabolites, p-aminobenzoyl glutamate and p-acetamidobenzoyl glutamate, along with the predominant folate vitamers in urine. The assays were based on clean-up by strong anion exchange followed by liquid chromatography-tandem mass spectrometry detection. Assay sensitivity was sufficient to detect the folate catabolites in physiologic concentrations. The limit of detection was below 0.4 and 0.3 nmol/100 g for p-aminobenzoyl glutamate isotopologues and p-acetamidobenzoyl glutamate isotopologues in urine, respectively. The successful synthesis of [2H4]-p-aminobenzoyl glutamate, [2H4]-p-acetamidobenzoyl glutamate, and unlabeled p-acetamidobenzoyl glutamate and the implementation of these substances in stable isotope dilution assays allows dual-label designs that provide a more detailed insight into human folate metabolism.

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2013. Vol. 83, nr 2, s. 112-121
Nationell ämneskategori
Livsmedelsvetenskap
Forskningsämne
Naturvetenskap
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URN: urn:nbn:se:lnu:diva-51143DOI: 10.1024/0300-9831/a000152PubMedID: 24491884OAI: oai:DiVA.org:lnu-51143DiVA, id: diva2:913526
Tillgänglig från: 2016-03-21 Skapad: 2016-03-21 Senast uppdaterad: 2017-11-30Bibliografiskt granskad

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