Faculty Collaboration Database

Liquid chromatographic-mass spectrometric determination of cyclooxygenase metabolites of arachidonic acid in cultured cells. J Chromatogr B Analyt Technol Biomed Life Sci 2003 Feb 25;785(1):135-45

Date

01/22/2003

Pubmed ID

12535846

DOI

10.1016/s1570-0232(02)00906-6

Scopus ID

2-s2.0-0037465106 (requires institutional sign-in at Scopus site)   44 Citations

Abstract

A liquid chromatographic-electrospray ionization-mass spectrometric (LC-ESI-MS) technique was developed to simultaneously determine the cyclooxygenase metabolites of arachidonic acid (6-keto-PGF(1alpha), PGD(2), PGE(2), PGF(2alpha), and PGJ(2)) produced by cultured cells. Samples were separated on a C(18) column with water-acetonitrile mobile phase, ionized by electrospray, and detected in the positive mode. Selected ion monitoring (SIM) of m/z 353, 335, 335, 319, and 317 were used for quantifying 6-keto-PGF(1alpha), PGD(2), PGE(2), PGF(2alpha), and PGJ(2), respectively. Prostaglandins were detected at concentrations as low as 1 pg (S/N=3) on the column. The method was used to determine the production of PGs from bovine coronary artery endothelial cells (ECs) and human prostate cancer cells (PC-3) with different degree of invasiveness. Bradykinin (10(-6) M) stimulated a marked increase in the production of 6-keto-PGF(1alpha), PGE(2), and PGF(2alpha) and a small increase of PGD(2) by ECs. 6-Keto-PGF(1alpha) was the major metabolite in these cells. The production of PGE(2) was threefold higher and PGD(2) was twofold higher in PC-3-S (invasive) cells than in PC-3-U (non-invasive) cells.

Author List

Nithipatikom K, Laabs ND, Isbell MA, Campbell WB

Author

William B. Campbell PhD Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin

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

Animals
Arachidonic Acid
Cattle
Cells, Cultured
Chromatography, Liquid
Coronary Vessels
Humans
Male
Prostaglandin-Endoperoxide Synthases
Prostaglandins
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization
Tumor Cells, Cultured