Quantitative analysis of 11-dehydrocorticosterone and corticosterone for preclinical studies by liquid chromatography/triple quadrupole mass spectrometry
© 2019 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd..
RATIONALE: The activity of the glucocorticoid activating enzyme 11β-hydroxysteroid dehydrogenase type-1 (11βHSD1) is altered in diseases such as obesity, inflammation and psychiatric disorders. In rodents 11βHSD1 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 11βHSD1 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 10:1 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 ➔ 121.2, m/z 347.1 ➔ 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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:34 Suppl 4 |
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| Enthalten in: |
Rapid communications in mass spectrometry : RCM - 34 Suppl 4(2020) vom: 14. Sept., Seite e8610 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Verma, Manu [VerfasserIn] |
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| Links: |
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| Themen: |
11-dehydrocorticosterone |
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| Anmerkungen: |
Date Completed 23.07.2021 Date Revised 05.10.2022 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1002/rcm.8610 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 500 | |a Date Revised 05.10.2022 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2019 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd. | ||
| 520 | |a RATIONALE: The activity of the glucocorticoid activating enzyme 11β-hydroxysteroid dehydrogenase type-1 (11βHSD1) is altered in diseases such as obesity, inflammation and psychiatric disorders. In rodents 11βHSD1 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 11βHSD1 activity in murine models | ||
| 520 | |a 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 10:1 chloroform liquid-liquid extraction (LLE) for analysis. Limits of quantitation (LOQs), linearity and other method criteria were assessed, according to bioanalytical method validation guidelines | ||
| 520 | |a 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 ➔ 121.2, m/z 347.1 ➔ 121.1 for 11-DHC and CORT, respectively). The LOQs were 0.25 and 0.20 ng/mL for 11-DHC and CORT, respectively | ||
| 520 | |a 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 7 | |a 11-dehydrocorticosterone |2 NLM | |
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| 650 | 7 | |a Corticosterone |2 NLM | |
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| 700 | 1 | |a Sooy, Karen |e verfasserin |4 aut | |
| 700 | 1 | |a Just, George |e verfasserin |4 aut | |
| 700 | 1 | |a Nixon, Mark |e verfasserin |4 aut | |
| 700 | 1 | |a Morgan, Ruth |e verfasserin |4 aut | |
| 700 | 1 | |a Andrew, Ruth |e verfasserin |4 aut | |
| 700 | 1 | |a Chapman, Karen E |e verfasserin |4 aut | |
| 700 | 1 | |a Homer, Natalie Z M |e verfasserin |4 aut | |
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