Determination of adrenaline, noradrenaline and corticosterone in rodent blood by ion pair reversed phase UHPLC–MS/MS

•An ion pair UHPLC–MS/MS method employing HFBA for stress hormone determination.•Simultaneous determination of adrenaline, noradrenaline and corticosterone.•Essential to add HFBA to both mobile phase and reconstitution solvent.•Stress hormone stability investigated using isotope labeled surrogate an...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2018-01, Vol.1072, p.161-172
Main Authors: Bergh, Marianne Skov-Skov, Bogen, Inger Lise, Andersen, Jannike Mørch, Øiestad, Åse Marit Leere, Berg, Thomas
Format: Article
Language:English
Subjects:
Adrenaline
HFBA
Ion pair chromatography
Stress hormones
UHPLC–MS/MS
Whole blood
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Summary:•An ion pair UHPLC–MS/MS method employing HFBA for stress hormone determination.•Simultaneous determination of adrenaline, noradrenaline and corticosterone.•Essential to add HFBA to both mobile phase and reconstitution solvent.•Stress hormone stability investigated using isotope labeled surrogate analytes.•The UHPLC–MS/MS method was used to determine stress hormones in rodent blood. A novel ion pair reversed phase ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS) method for simultaneous determination of the stress hormones adrenaline, noradrenaline and corticosterone in rodent blood was developed and fully validated. Separations were performed on an Acquity HSS T3 column (2.1mm i.d.×100mm, 1.8μm) with gradient elution and a runtime of 5.5min. The retention of adrenaline and noradrenaline was substantially increased by employing the ion pair reagent heptafluorobutyric acid (HFBA). Ion pair reagents are usually added to the mobile phase only, but we demonstrate for the first time that including HFBA to the sample reconstitution solvent as well, has a major impact on the chromatography of these compounds. The stability of adrenaline and corticosterone in rodent blood was investigated using the surrogate analytes adrenaline-d3 and corticosterone-d8. The applicability of the described method was demonstrated by measuring the concentration of stress hormones in rodent blood samples.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2017.11.016