Faculty Collaboration Database

Measurement of steroid synthesis in zona glomerulosa cells by liquid chromatography-electrospray ionization-mass spectrometry: inhibition by nitric oxide. Anal Biochem 2005 Feb 15;337(2):203-10

Date

02/05/2005

Pubmed ID

15691500

DOI

10.1016/j.ab.2004.11.025

Scopus ID

2-s2.0-13244259473 (requires institutional sign-in at Scopus site)   12 Citations

Abstract

A liquid chromatography-electrospray ionization-mass spectrometry method was developed to simultaneously determine the concentrations of aldosterone, corticosterone, cortisol, deoxycorticosterone, pregnenolone, and progesterone in bovine adrenal zona glomerulosa (ZG) cells. Steroids were extracted by liquid-liquid extraction, separated on a reverse-phase C18 column, ionized by electrospray, and detected by single-quadrupole mass spectrometry in a positive ion mode. All steroids formed sodium adducts at high abundance. Factors affecting the formation and signal of sodium adducts were investigated. The limits of detection (S/N=3) using selected ion monitoring are 2 pg for these steroids and 10 pg for pregnenolone. DETA NONOate, a nitric oxide donor, inhibited the basal, angiotensin-II-stimulated, and 25-hydroxycholesterol-stimulated syntheses of these steroids in ZG cells in a concentration-dependent manner. The technique demonstrates the ability to determine the individual steroid in each enzymatic step of aldosterone synthesis and the activity of steroidogenic enzymes in adrenal ZG cells.

Author List

Nithipatikom K, Holmes BB, Isbell MA, Hanke CJ, Gomez-Sanchez CE, Campbell WB

Authors

William B. Campbell PhD Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin
Craig J. Hanke PhD Assistant Dean, Associate Professor in the Medical School Regional Campuses department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Angiotensin II
Animals
Cattle
Chromatography, Liquid
Deuterium
Nitric Oxide
Reference Standards
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization
Steroids
Zona Glomerulosa