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Sensitive monitoring of 3-hydroxybutyrate as an indicator of human fasting by capillary electrophoresis in a PAMAMPS coated capillary
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SYSNO ASEP 0558329 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Sensitive monitoring of 3-hydroxybutyrate as an indicator of human fasting by capillary electrophoresis in a PAMAMPS coated capillary Author(s) Tůma, P. (CZ)
Sommerová, B. (CZ)
Koval, Dušan (UOCHB-X) RID, ORCID
Šiklová, M. (CZ)
Koc, M. (CZ)Article number 123582 Source Title Talanta. - : Elsevier - ISSN 0039-9140
Roč. 247, September (2022)Number of pages 7 s. Language eng - English Country NL - Netherlands Keywords clinical analysis ; capillary electrophoresis ; coating ; contactless conductivity detection ; ketogenesis ; stacking OECD category Analytical chemistry Method of publishing Limited access Institutional support UOCHB-X - RVO:61388963 UT WOS 000812357900003 EID SCOPUS 85131460667 DOI 10.1016/j.talanta.2022.123582 Annotation Sensitive electrophoretic determination of 3-hydroxybutyrate (3HB) as an indicator of human ketogenesis is performed in fused silica capillary covalently coated by an anionic copolymer of poly(acrylamide-co-sodium-2-acrylamido-2-methylpropanesulphonate) (PAMAMPS). Baseline separation of 3HB from other components of human serum is achieved in a 20 μm capillary with an effective length of 17 cm covered by 4% PAMAMPS, which generates a cathodic EOF with a mobility of 8.30 ± 0.00 · 10−9 m2/V.s in 80 mM MES/His as background electrolyte. 3HB migrates in counter-current electrophoretic mode against EOF, that effectively improving electrophoretic resolution. Sample pre-treatment is based on adding of 45 μL acetonitrile to 15 μL serum and, after shaking, a 28 mm long zone of supernatant is injected into the capillary, and sharpened after turning on a separation voltage of 20 kV using the technique of large volume sample stacking, where the EOF forces the residual acetonitrile from the capillary. When combined with universal contactless conductivity detection, the achieved LOD and LOQ are 0.43 μM and 1.44 μM, respectively, that are sufficiently low for monitoring the physiological 3HB level. The performed clinical study subsequently showed that serum 3HB increases from a concentration of 71 μM, corresponding to normal food, to level of 1924 μM after 60 h of fasting and returns to the normal physiological concentration 48 h after commencing consumption of high-saccharide food. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434 Year of Publishing 2023 Electronic address https://doi.org/10.1016/j.talanta.2022.123582
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