US EPA

4220313 

Journal Article 

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

Bergh, MS; Bogen, IL; Andersen, JM; Øiestad, ÅML; Berg, T 

2018 

Yes 

Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
ISSN: 1570-0232
EISSN: 1873-376X 

Elsevier B.V. 

1072 

161-172 

English 

29175696 

10.1016/j.jchromb.2017.11.016 

WOS:000423637600021 

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. 

UHPLC MS/MS; Ion pair chromatography; Stress hormones; HFBA; Whole blood; Adrenaline