Pharmaceutical Sciences Faculty Publications

Multiplexed Quantitation of Endogenous Estrogens and Estrogen Metabolites in Human Peritoneal Fluid

Document Type

Article

Publication Date

6-1-2008

Journal Title

Electrophoresis

ISSN

0173-0835

Volume

29

Issue

12

First Page

2706

Last Page

2713

DOI

10.1002/elps.200700837

PubMed ID

18512681

PubMed Central® ID

PMC4297214

Abstract

Endogenous estrogens and estrogen metabolites (EM) in human peritoneal fluid may play an important role in health and disease, yet little is known regarding their types and levels present in human peritoneal fluid, primarily due to the lack of an analytical method that is capable of directly quantifying their absolute abundances. In this report, we describe the application of a capillary LC-MS/MS method for identifying and quantifying biologically active and total endogenous EM in human peritoneal fluid. The method requires only 50 muL of peritoneal fluid, yet can quantify 13 distinct EM. Calibration curves for each EM were linear over a 10(3)-fold concentration range and the lower LOQ was 50 fg on-column. For a charcoal stripped human peritoneal fluid sample containing 10 pg/mL of each EM, accuracy ranged from 83 to 118%, and intrabatch precision ranged from 0.2 to 4.4% RSD and interbatch precision ranged from 5.5 to 15.5% RSD. The analyses of human female peritoneal fluid shows that at least 10 biologically active and 11 total endogenous EM can be positively identified and quantitatively measured. Many of the biologically active forms are present in high abundance and possess distinct biological activities which warrant further study. Although micellar EKC gave baseline separation of a standard mixture of 10 EM, the LOQs using UV detection were not suitable for the assay of the low level estrogens in biological samples.

Keywords

Ascitic fluid, chromatography, liquid, micellar electrokinetic capillary, endometriosis, estrogens, electrospray ionization, tandem mass spectrometry

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