Multiplexed quantitation of endogenous estrogens and estrogen metabolites in human peritoneal fluid

Xia Xu, Essam El Dine R. Othman, Haleem J. Issaq, Daniela Hornung, Ayman Al-Hendy, Timothy D. Veenstra

Research output: Contribution to journalArticle

5 Citations (Scopus)

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 μL of peritoneal fluid, yet can quantify 13 distinct EM. Calibration curves for each EM were linear over a 103-fold concentration range and the lower LOQ was 50 fg on-column. For 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.

Original languageEnglish (US)
Pages (from-to)2706-2713
Number of pages8
JournalELECTROPHORESIS
Volume29
Issue number12
DOIs
StatePublished - Jun 1 2008

Fingerprint

Ascitic Fluid
Metabolites
Estrogens
Fluids
Charcoal
Cerebral Palsy
Bioactivity
Calibration
Assays
Health

Keywords

  • Capillary liquid chromatography
  • Electrospray ionization-tandem mass spectrometry
  • Endogenous estrogens and estrogen metabolites
  • Human peritoneal fluid
  • Selected reaction monitoring

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Xu, X., Othman, E. E. D. R., Issaq, H. J., Hornung, D., Al-Hendy, A., & Veenstra, T. D. (2008). Multiplexed quantitation of endogenous estrogens and estrogen metabolites in human peritoneal fluid. ELECTROPHORESIS, 29(12), 2706-2713. https://doi.org/10.1002/elps.200700837

Multiplexed quantitation of endogenous estrogens and estrogen metabolites in human peritoneal fluid. / Xu, Xia; Othman, Essam El Dine R.; Issaq, Haleem J.; Hornung, Daniela; Al-Hendy, Ayman; Veenstra, Timothy D.

In: ELECTROPHORESIS, Vol. 29, No. 12, 01.06.2008, p. 2706-2713.

Research output: Contribution to journalArticle

Xu, X, Othman, EEDR, Issaq, HJ, Hornung, D, Al-Hendy, A & Veenstra, TD 2008, 'Multiplexed quantitation of endogenous estrogens and estrogen metabolites in human peritoneal fluid', ELECTROPHORESIS, vol. 29, no. 12, pp. 2706-2713. https://doi.org/10.1002/elps.200700837
Xu, Xia ; Othman, Essam El Dine R. ; Issaq, Haleem J. ; Hornung, Daniela ; Al-Hendy, Ayman ; Veenstra, Timothy D. / Multiplexed quantitation of endogenous estrogens and estrogen metabolites in human peritoneal fluid. In: ELECTROPHORESIS. 2008 ; Vol. 29, No. 12. pp. 2706-2713.
@article{0d37fcda00f94d07a6a828df1bef530d,
title = "Multiplexed quantitation of endogenous estrogens and estrogen metabolites in human peritoneal fluid",
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 μL of peritoneal fluid, yet can quantify 13 distinct EM. Calibration curves for each EM were linear over a 103-fold concentration range and the lower LOQ was 50 fg on-column. For 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 = "Capillary liquid chromatography, Electrospray ionization-tandem mass spectrometry, Endogenous estrogens and estrogen metabolites, Human peritoneal fluid, Selected reaction monitoring",
author = "Xia Xu and Othman, {Essam El Dine R.} and Issaq, {Haleem J.} and Daniela Hornung and Ayman Al-Hendy and Veenstra, {Timothy D.}",
year = "2008",
month = "6",
day = "1",
doi = "10.1002/elps.200700837",
language = "English (US)",
volume = "29",
pages = "2706--2713",
journal = "Electrophoresis",
issn = "0173-0835",
publisher = "Wiley-VCH Verlag",
number = "12",

}

TY - JOUR

T1 - Multiplexed quantitation of endogenous estrogens and estrogen metabolites in human peritoneal fluid

AU - Xu, Xia

AU - Othman, Essam El Dine R.

AU - Issaq, Haleem J.

AU - Hornung, Daniela

AU - Al-Hendy, Ayman

AU - Veenstra, Timothy D.

PY - 2008/6/1

Y1 - 2008/6/1

N2 - 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 μL of peritoneal fluid, yet can quantify 13 distinct EM. Calibration curves for each EM were linear over a 103-fold concentration range and the lower LOQ was 50 fg on-column. For 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.

AB - 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 μL of peritoneal fluid, yet can quantify 13 distinct EM. Calibration curves for each EM were linear over a 103-fold concentration range and the lower LOQ was 50 fg on-column. For 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.

KW - Capillary liquid chromatography

KW - Electrospray ionization-tandem mass spectrometry

KW - Endogenous estrogens and estrogen metabolites

KW - Human peritoneal fluid

KW - Selected reaction monitoring

UR - http://www.scopus.com/inward/record.url?scp=46249112359&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=46249112359&partnerID=8YFLogxK

U2 - 10.1002/elps.200700837

DO - 10.1002/elps.200700837

M3 - Article

VL - 29

SP - 2706

EP - 2713

JO - Electrophoresis

JF - Electrophoresis

SN - 0173-0835

IS - 12

ER -