Quantitative analysis of 11-dehydrocorticosterone and corticosterone for preclinical studies by liquid chromatography/triple quadrupole mass spectrometry.
Rapid Commun Mass Spectrom
; 34 Suppl 4: e8610, 2020 Sep.
Article
em En
| MEDLINE
| ID: mdl-31677354
ABSTRACT
RATIONALE The activity of the glucocorticoid activating enzyme 11ß-hydroxysteroid dehydrogenase type-1 (11ßHSD1) is altered in diseases such as obesity, inflammation and psychiatric disorders. In rodents 11ßHSD1 converts inert 11-dehydrocorticosterone (11-DHC) into the active form, corticosterone (CORT). A sensitive, specific liquid chromatography/tandem mass spectrometry method was sought to simultaneously quantify total 11-DHC and total and free CORT in murine plasma for simple assessment of 11ßHSD1 activity in murine models. METHODS:
Mass spectrometry parameters were optimised and a method for the chromatographic separation of CORT and 11-DHC was developed. Murine plasma was prepared by 101 chloroform liquid-liquid extraction (LLE) for analysis. Limits of quantitation (LOQs), linearity and other method criteria were assessed, according to bioanalytical method validation guidelines.RESULTS:
Reliable separation of 11-DHC and CORT was achieved using an ACE Excel 2 C18-AR (2.1 × 150 mm; 2 µm) fused core column at 25°C, with an acidified water/acetonitrile gradient over 10 min. Analytes were detected by multiple reaction monitoring after positive electrospray ionisation (m/z 345.1.1 â 121.2, m/z 347.1 â 121.1 for 11-DHC and CORT, respectively). The LOQs were 0.25 and 0.20 ng/mL for 11-DHC and CORT, respectively.CONCLUSIONS:
This LC/MS method is suitable for the reliable analysis of 11-DHC and CORT following simple LLE of murine plasma, bringing preclinical analysis in line with recommendations for clinical endocrinology and biochemistry.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Corticosterona
/
Cromatografia Líquida
/
Espectrometria de Massas em Tandem
Tipo de estudo:
Guideline
/
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Rapid Commun Mass Spectrom
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Reino Unido