Quantitative stereochemical analysis of subnanogram amounts of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid by sequential chiral phase liquid chromatography and stable isotope dilution mass spectrometry

John Turk; W. Thomas Stump; Bryan A. Wolf; Richard A. Easom; Michael L. McDaniel

doi:10.1016/0003-2697(88)90059-0

Quantitative stereochemical analysis of subnanogram amounts of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid by sequential chiral phase liquid chromatography and stable isotope dilution mass spectrometry

John Turk
, W. Thomas Stump
, Bryan A. Wolf
, Richard A. Easom
, Michael L. McDaniel

Division of Endocrinology, Metabolism & Lipid Research

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

The two enantiomers of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid (12-HETE) are products of different biosynthetic pathways and have distinct biologic actions. Conventional methods of stereochemical analysis of 12-HETE require multimicrogram amounts of material and cannot be applied to systems where the availability of tissue is limited and only trace quantities of 12-HETE are generated. We have developed a method capable of measuring subnanogram amounts of 12-HETE enantiomers which involves addition of racemic, ¹⁸O₂-labeled 12-HETE as an internal standard, chiral phase HPLC of the pentafluorobenzyl ester derivative of 12-HETE, and stable isotope dilution gas chromatographic-negative ion chemical ionization mass spectrometric quantitation of the resolved stereoisomers. This method has been employed to determine the stereochemical composition of 12-HETE produced by isolated pancreatic islets.

Original language	English
Pages (from-to)	580-588
Number of pages	9
Journal	Analytical Biochemistry
Volume	174
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(88)90059-0
State	Published - Nov 1 1988

Keywords

eicosanoids
stereochemistry

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10.1016/0003-2697(88)90059-0

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@article{f27474fb5a524d7eaadd79ab9ca75497,

title = "Quantitative stereochemical analysis of subnanogram amounts of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid by sequential chiral phase liquid chromatography and stable isotope dilution mass spectrometry",

abstract = "The two enantiomers of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid (12-HETE) are products of different biosynthetic pathways and have distinct biologic actions. Conventional methods of stereochemical analysis of 12-HETE require multimicrogram amounts of material and cannot be applied to systems where the availability of tissue is limited and only trace quantities of 12-HETE are generated. We have developed a method capable of measuring subnanogram amounts of 12-HETE enantiomers which involves addition of racemic, 18O2-labeled 12-HETE as an internal standard, chiral phase HPLC of the pentafluorobenzyl ester derivative of 12-HETE, and stable isotope dilution gas chromatographic-negative ion chemical ionization mass spectrometric quantitation of the resolved stereoisomers. This method has been employed to determine the stereochemical composition of 12-HETE produced by isolated pancreatic islets.",

keywords = "eicosanoids, stereochemistry",

author = "John Turk and Stump, \{W. Thomas\} and Wolf, \{Bryan A.\} and Easom, \{Richard A.\} and McDaniel, \{Michael L.\}",

note = "Funding Information: {\textquoteright} This work was supported in part by National Insti-tutes of Health Grants K04-DK-01553 (to JT), ROl-DK-34388 (to JT), ROl-DK-06181 (to MM), and RR-00954 (to the Washington University Mass Spectrometry Facility). {\textquoteleft}To whom correspondence should be addressed at Box 8 118, Washington University School of Medicine. 660 S. Euclid Ave., St. Louis. MO 63 110.",

year = "1988",

month = nov,

day = "1",

doi = "10.1016/0003-2697(88)90059-0",

language = "English",

volume = "174",

pages = "580--588",

journal = "Analytical Biochemistry",

issn = "0003-2697",

number = "2",

}

Quantitative stereochemical analysis of subnanogram amounts of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid by sequential chiral phase liquid chromatography and stable isotope dilution mass spectrometry. / Turk, John; Stump, W. Thomas; Wolf, Bryan A. et al.
In: Analytical Biochemistry, Vol. 174, No. 2, 01.11.1988, p. 580-588.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Quantitative stereochemical analysis of subnanogram amounts of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid by sequential chiral phase liquid chromatography and stable isotope dilution mass spectrometry

AU - Turk, John

AU - Stump, W. Thomas

AU - Wolf, Bryan A.

AU - Easom, Richard A.

AU - McDaniel, Michael L.

N1 - Funding Information: ’ This work was supported in part by National Insti-tutes of Health Grants K04-DK-01553 (to JT), ROl-DK-34388 (to JT), ROl-DK-06181 (to MM), and RR-00954 (to the Washington University Mass Spectrometry Facility). ‘To whom correspondence should be addressed at Box 8 118, Washington University School of Medicine. 660 S. Euclid Ave., St. Louis. MO 63 110.

PY - 1988/11/1

Y1 - 1988/11/1

N2 - The two enantiomers of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid (12-HETE) are products of different biosynthetic pathways and have distinct biologic actions. Conventional methods of stereochemical analysis of 12-HETE require multimicrogram amounts of material and cannot be applied to systems where the availability of tissue is limited and only trace quantities of 12-HETE are generated. We have developed a method capable of measuring subnanogram amounts of 12-HETE enantiomers which involves addition of racemic, 18O2-labeled 12-HETE as an internal standard, chiral phase HPLC of the pentafluorobenzyl ester derivative of 12-HETE, and stable isotope dilution gas chromatographic-negative ion chemical ionization mass spectrometric quantitation of the resolved stereoisomers. This method has been employed to determine the stereochemical composition of 12-HETE produced by isolated pancreatic islets.

AB - The two enantiomers of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid (12-HETE) are products of different biosynthetic pathways and have distinct biologic actions. Conventional methods of stereochemical analysis of 12-HETE require multimicrogram amounts of material and cannot be applied to systems where the availability of tissue is limited and only trace quantities of 12-HETE are generated. We have developed a method capable of measuring subnanogram amounts of 12-HETE enantiomers which involves addition of racemic, 18O2-labeled 12-HETE as an internal standard, chiral phase HPLC of the pentafluorobenzyl ester derivative of 12-HETE, and stable isotope dilution gas chromatographic-negative ion chemical ionization mass spectrometric quantitation of the resolved stereoisomers. This method has been employed to determine the stereochemical composition of 12-HETE produced by isolated pancreatic islets.

KW - eicosanoids

KW - stereochemistry

UR - https://www.scopus.com/pages/publications/0023761540

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DO - 10.1016/0003-2697(88)90059-0

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AN - SCOPUS:0023761540

SN - 0003-2697

VL - 174

SP - 580

EP - 588

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

Quantitative stereochemical analysis of subnanogram amounts of 12-hydroxy-(5,8,10,14)-eicosatetraenoic acid by sequential chiral phase liquid chromatography and stable isotope dilution mass spectrometry

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