A high-performance liquid chromatography-electrospray-tandem mass spectrometry analysis of cortisol and metabolites in placental perfusate

Analytical Biochemistry
H M DoddsS M Pond

Abstract

A reversed-phase high-performance liquid chromatography-electrospray-tandem mass spectrometry assay (HPLC-ESI-MS/MS) was developed to quantitate cortisol, cortisone, 20 alpha- and beta-dihydrocortisol, 20 alpha- and beta-dihydrocortisone, tetrahydrocortisol, and tetrahydrocortisone. The technique was used to analyze perfusate from the isolated human placental lobule for cortisol and its metabolites. Analytes were prepared from the perfusion medium using C18 solid-phase extraction cartridges. The internal standard for the analyses was 6 alpha-methylprednisolone. Chromatography was performed on a Novapak C18 column at ambient temperature using 53% methanol and 47% 10 mM ammonium formate buffer (pH 4.0) as mobile phase, at a flow rate of 80 microL/min. A PE-SCIEX API III triple quadrupole instrument was used for mass spectrometric detection. An ionspray (pneumatically assisted electrospray) interface was used in negative and positive ionization mode. The assay was linear over the range 100-2000 micrograms/L for each analyte. The instrumental limit of detection was 50 pg. Assay imprecision at 400 and 800 micrograms/L was < or = 10% (total coefficient of variation). Accuracy ranged between 83.2% for 20 beta-dihydrocortisone to 102.6...Continue Reading

References

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Citations

May 15, 2013·The Journal of Steroid Biochemistry and Molecular Biology·Jonathan J HirstHannah K Palliser
Apr 23, 2013·The Annals of Thoracic Surgery·Juho Keski-NisulaPertti K Suominen
Dec 3, 2014·The Annals of Thoracic Surgery·Juho Keski-NisulaEero Pesonen
May 8, 2016·The Annals of Thoracic Surgery·Juho Keski-NisulaPertti K Suominen
Mar 16, 2006·Scandinavian Journal of Clinical and Laboratory Investigation·U TurpeinenE Hämäläinen
May 12, 2011·Analytical and Bioanalytical Chemistry·Ilse BecueCarlos Van Peteghem
Jun 11, 2005·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Natsuho ItoRalf Paus
Jan 5, 2002·Journal of Chromatography. a·K ShimadaT Higashi
Jun 29, 1999·Analytical Chemistry·D J Anderson

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