Liquid chromatography-mass spectrometry-based in vitro metabolic profiling reveals altered enzyme expressions in eicosanoid metabolism

Su Hyeon Lee, Eung Ju Kim, Dong Hyoung Lee, Won Yong Lee, Bong Chul Chung, Hong Seog Seo, Man Ho Choi

Research output: Contribution to journalArticle

Abstract

Background: Eicosanoids are metabolites of arachidonic acid that are rapidly biosynthesized and degraded during inflammation, and their metabolic changes reveal altered enzyme expression following drug treatment. We developed an eicosanoid profiling method and evaluated their changes on drug treatment. Methods: Simultaneous quantitative profiling of 32 eicosanoids in liver S9 fractions obtained from rabbits with carrageenan-induced inflammation was performed and validated by liquid chromatography-mass spectrometry coupled to anion-exchange solid-phase purification. Results: The limit of quantification for the devised method ranged from 0.5 to 20.0 ng/mg protein, and calibration linearity was achieved (R2>0.99). The precision (% CV) and accuracy (% bias) ranged from 4.7 to 10.3% and 88.4 to 110.9%, respectively, and overall recoveries ranged from 58.0 to 105.3%. Our method was then applied and showed that epitestosterone treatment reduced the levels of all eicosanoids that were generated by cyclooxygenases and lipoxygenases. Conclusions: Quantitative eicosanoid profiling combined with in vitro metabolic assays may be useful for evaluating metabolic changes affected by drugs during eicosanoid metabolism.

Original languageEnglish
Pages (from-to)342-352
Number of pages11
JournalAnnals of Laboratory Medicine
Volume36
Issue number4
DOIs
Publication statusPublished - 2016 Jul 1

Fingerprint

Eicosanoids
Liquid chromatography
Metabolism
Liquid Chromatography
Mass spectrometry
Mass Spectrometry
Enzymes
Drug therapy
Epitestosterone
Lipoxygenases
Pharmaceutical Preparations
Inflammation
Carrageenan
Prostaglandin-Endoperoxide Synthases
Metabolites
Arachidonic Acid
Liver
Calibration
Purification
Anions

Keywords

  • Arachidonic acid
  • Eicosanoids
  • Epitestosterone
  • Liquid chromatography-mass spectrometry
  • Liver S9 fraction

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Liquid chromatography-mass spectrometry-based in vitro metabolic profiling reveals altered enzyme expressions in eicosanoid metabolism. / Lee, Su Hyeon; Kim, Eung Ju; Lee, Dong Hyoung; Lee, Won Yong; Chung, Bong Chul; Seo, Hong Seog; Choi, Man Ho.

In: Annals of Laboratory Medicine, Vol. 36, No. 4, 01.07.2016, p. 342-352.

Research output: Contribution to journalArticle

@article{9ec0daaae3d74828a9bf4b6da33e7042,
title = "Liquid chromatography-mass spectrometry-based in vitro metabolic profiling reveals altered enzyme expressions in eicosanoid metabolism",
abstract = "Background: Eicosanoids are metabolites of arachidonic acid that are rapidly biosynthesized and degraded during inflammation, and their metabolic changes reveal altered enzyme expression following drug treatment. We developed an eicosanoid profiling method and evaluated their changes on drug treatment. Methods: Simultaneous quantitative profiling of 32 eicosanoids in liver S9 fractions obtained from rabbits with carrageenan-induced inflammation was performed and validated by liquid chromatography-mass spectrometry coupled to anion-exchange solid-phase purification. Results: The limit of quantification for the devised method ranged from 0.5 to 20.0 ng/mg protein, and calibration linearity was achieved (R2>0.99). The precision ({\%} CV) and accuracy ({\%} bias) ranged from 4.7 to 10.3{\%} and 88.4 to 110.9{\%}, respectively, and overall recoveries ranged from 58.0 to 105.3{\%}. Our method was then applied and showed that epitestosterone treatment reduced the levels of all eicosanoids that were generated by cyclooxygenases and lipoxygenases. Conclusions: Quantitative eicosanoid profiling combined with in vitro metabolic assays may be useful for evaluating metabolic changes affected by drugs during eicosanoid metabolism.",
keywords = "Arachidonic acid, Eicosanoids, Epitestosterone, Liquid chromatography-mass spectrometry, Liver S9 fraction",
author = "Lee, {Su Hyeon} and Kim, {Eung Ju} and Lee, {Dong Hyoung} and Lee, {Won Yong} and Chung, {Bong Chul} and Seo, {Hong Seog} and Choi, {Man Ho}",
year = "2016",
month = "7",
day = "1",
doi = "10.3343/alm.2016.36.4.342",
language = "English",
volume = "36",
pages = "342--352",
journal = "Annals of Laboratory Medicine",
issn = "2234-3806",
publisher = "Seoul National University",
number = "4",

}

TY - JOUR

T1 - Liquid chromatography-mass spectrometry-based in vitro metabolic profiling reveals altered enzyme expressions in eicosanoid metabolism

AU - Lee, Su Hyeon

AU - Kim, Eung Ju

AU - Lee, Dong Hyoung

AU - Lee, Won Yong

AU - Chung, Bong Chul

AU - Seo, Hong Seog

AU - Choi, Man Ho

PY - 2016/7/1

Y1 - 2016/7/1

N2 - Background: Eicosanoids are metabolites of arachidonic acid that are rapidly biosynthesized and degraded during inflammation, and their metabolic changes reveal altered enzyme expression following drug treatment. We developed an eicosanoid profiling method and evaluated their changes on drug treatment. Methods: Simultaneous quantitative profiling of 32 eicosanoids in liver S9 fractions obtained from rabbits with carrageenan-induced inflammation was performed and validated by liquid chromatography-mass spectrometry coupled to anion-exchange solid-phase purification. Results: The limit of quantification for the devised method ranged from 0.5 to 20.0 ng/mg protein, and calibration linearity was achieved (R2>0.99). The precision (% CV) and accuracy (% bias) ranged from 4.7 to 10.3% and 88.4 to 110.9%, respectively, and overall recoveries ranged from 58.0 to 105.3%. Our method was then applied and showed that epitestosterone treatment reduced the levels of all eicosanoids that were generated by cyclooxygenases and lipoxygenases. Conclusions: Quantitative eicosanoid profiling combined with in vitro metabolic assays may be useful for evaluating metabolic changes affected by drugs during eicosanoid metabolism.

AB - Background: Eicosanoids are metabolites of arachidonic acid that are rapidly biosynthesized and degraded during inflammation, and their metabolic changes reveal altered enzyme expression following drug treatment. We developed an eicosanoid profiling method and evaluated their changes on drug treatment. Methods: Simultaneous quantitative profiling of 32 eicosanoids in liver S9 fractions obtained from rabbits with carrageenan-induced inflammation was performed and validated by liquid chromatography-mass spectrometry coupled to anion-exchange solid-phase purification. Results: The limit of quantification for the devised method ranged from 0.5 to 20.0 ng/mg protein, and calibration linearity was achieved (R2>0.99). The precision (% CV) and accuracy (% bias) ranged from 4.7 to 10.3% and 88.4 to 110.9%, respectively, and overall recoveries ranged from 58.0 to 105.3%. Our method was then applied and showed that epitestosterone treatment reduced the levels of all eicosanoids that were generated by cyclooxygenases and lipoxygenases. Conclusions: Quantitative eicosanoid profiling combined with in vitro metabolic assays may be useful for evaluating metabolic changes affected by drugs during eicosanoid metabolism.

KW - Arachidonic acid

KW - Eicosanoids

KW - Epitestosterone

KW - Liquid chromatography-mass spectrometry

KW - Liver S9 fraction

UR - http://www.scopus.com/inward/record.url?scp=84969221032&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84969221032&partnerID=8YFLogxK

U2 - 10.3343/alm.2016.36.4.342

DO - 10.3343/alm.2016.36.4.342

M3 - Article

C2 - 27139607

AN - SCOPUS:84969221032

VL - 36

SP - 342

EP - 352

JO - Annals of Laboratory Medicine

JF - Annals of Laboratory Medicine

SN - 2234-3806

IS - 4

ER -