Determination of trans-arachidonic acid isomers in human blood plasma

Chaza M. Zghibeh; V. Raj Gopal; Candace D. Poff; J R Falck; Michael Balazy

doi:10.1016/j.ab.2004.04.030

Determination of trans-arachidonic acid isomers in human blood plasma

Chaza M. Zghibeh, V. Raj Gopal, Candace D. Poff, J R Falck, Michael Balazy

Biochemistry

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

41 Scopus citations

Abstract

Endogenous trans fatty acids originate from diet, but recent studies also suggest that cis-trans isomerization of fatty acids is possible by nitrogen dioxide radical, a product of NO and nitrite oxidation. We developed a method for quantitative analysis of four trans-arachidonic acids (TAA) in human plasma using isotopic dilution gas chromatography/mass spectrometry (GC/MS) with deuterium-labeled internal standard. Esterification of the plasma fatty acid extract with pentafluorobenzyl (PFB) bromide followed by high-performance liquid chromatography purification yielded a fairly pure fraction containing TAA-PFB esters that was analyzed by GC/MS. Partial separation of the TAA isomers was obtained on various GC columns. Comparison of the retention time with the synthetic standards revealed that all four TAA isomers are present in human plasma. The mean concentration of TAA in human plasma was 20.2ng/ml. The levels of isomers were 12.48±1.28, 2.75±0.39, and 4.99±0.74ng/ml for 5E-AA + 11E-AA, 8E-AA, and 14E-AA, respectively. The identification of TAA in plasma suggests that isomerization of arachidonic acid occurs in vivo. Our method allows distinguishing between the dietary and the NO₂- dependent mechanisms of trans fatty acid formation and will be useful in defining the role of TAA as an in vivo marker of nitrooxidative stress in clinical and experimental settings.

Original language	English (US)
Pages (from-to)	137-144
Number of pages	8
Journal	Analytical biochemistry
Volume	332
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2004.04.030
State	Published - Sep 1 2004

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

Access to Document

10.1016/j.ab.2004.04.030

Cite this

@article{289ce1b47fe044adae60e1f5e1bd4b2d,

title = "Determination of trans-arachidonic acid isomers in human blood plasma",

abstract = "Endogenous trans fatty acids originate from diet, but recent studies also suggest that cis-trans isomerization of fatty acids is possible by nitrogen dioxide radical, a product of NO and nitrite oxidation. We developed a method for quantitative analysis of four trans-arachidonic acids (TAA) in human plasma using isotopic dilution gas chromatography/mass spectrometry (GC/MS) with deuterium-labeled internal standard. Esterification of the plasma fatty acid extract with pentafluorobenzyl (PFB) bromide followed by high-performance liquid chromatography purification yielded a fairly pure fraction containing TAA-PFB esters that was analyzed by GC/MS. Partial separation of the TAA isomers was obtained on various GC columns. Comparison of the retention time with the synthetic standards revealed that all four TAA isomers are present in human plasma. The mean concentration of TAA in human plasma was 20.2ng/ml. The levels of isomers were 12.48±1.28, 2.75±0.39, and 4.99±0.74ng/ml for 5E-AA + 11E-AA, 8E-AA, and 14E-AA, respectively. The identification of TAA in plasma suggests that isomerization of arachidonic acid occurs in vivo. Our method allows distinguishing between the dietary and the NO2- dependent mechanisms of trans fatty acid formation and will be useful in defining the role of TAA as an in vivo marker of nitrooxidative stress in clinical and experimental settings.",

author = "Zghibeh, {Chaza M.} and {Raj Gopal}, V. and Poff, {Candace D.} and Falck, {J R} and Michael Balazy",

note = "Funding Information: We thank Dr. Maria Aguero-Rosenfeld and the staff at the Clinical Laboratory of Westchester Medical Center (Valhalla, NY) for expert phlebotomy and clinical chemistry assays and Dr. Jordi L{\'o}pez-F{\'e}rnandez for help with preliminary GC/MS analysis. This work was supported by Grants from NIH (5R01 GM62453, 5R01 GM31278), Philip Morris, Inc. USA, and Robert A. Welch Foundation.",

year = "2004",

month = sep,

day = "1",

doi = "10.1016/j.ab.2004.04.030",

language = "English (US)",

volume = "332",

pages = "137--144",

journal = "Analytical biochemistry",

issn = "0003-2697",

publisher = "Academic Press Inc.",

number = "1",

}

TY - JOUR

T1 - Determination of trans-arachidonic acid isomers in human blood plasma

AU - Zghibeh, Chaza M.

AU - Raj Gopal, V.

AU - Poff, Candace D.

AU - Falck, J R

AU - Balazy, Michael

N1 - Funding Information: We thank Dr. Maria Aguero-Rosenfeld and the staff at the Clinical Laboratory of Westchester Medical Center (Valhalla, NY) for expert phlebotomy and clinical chemistry assays and Dr. Jordi López-Férnandez for help with preliminary GC/MS analysis. This work was supported by Grants from NIH (5R01 GM62453, 5R01 GM31278), Philip Morris, Inc. USA, and Robert A. Welch Foundation.

PY - 2004/9/1

Y1 - 2004/9/1

N2 - Endogenous trans fatty acids originate from diet, but recent studies also suggest that cis-trans isomerization of fatty acids is possible by nitrogen dioxide radical, a product of NO and nitrite oxidation. We developed a method for quantitative analysis of four trans-arachidonic acids (TAA) in human plasma using isotopic dilution gas chromatography/mass spectrometry (GC/MS) with deuterium-labeled internal standard. Esterification of the plasma fatty acid extract with pentafluorobenzyl (PFB) bromide followed by high-performance liquid chromatography purification yielded a fairly pure fraction containing TAA-PFB esters that was analyzed by GC/MS. Partial separation of the TAA isomers was obtained on various GC columns. Comparison of the retention time with the synthetic standards revealed that all four TAA isomers are present in human plasma. The mean concentration of TAA in human plasma was 20.2ng/ml. The levels of isomers were 12.48±1.28, 2.75±0.39, and 4.99±0.74ng/ml for 5E-AA + 11E-AA, 8E-AA, and 14E-AA, respectively. The identification of TAA in plasma suggests that isomerization of arachidonic acid occurs in vivo. Our method allows distinguishing between the dietary and the NO2- dependent mechanisms of trans fatty acid formation and will be useful in defining the role of TAA as an in vivo marker of nitrooxidative stress in clinical and experimental settings.

AB - Endogenous trans fatty acids originate from diet, but recent studies also suggest that cis-trans isomerization of fatty acids is possible by nitrogen dioxide radical, a product of NO and nitrite oxidation. We developed a method for quantitative analysis of four trans-arachidonic acids (TAA) in human plasma using isotopic dilution gas chromatography/mass spectrometry (GC/MS) with deuterium-labeled internal standard. Esterification of the plasma fatty acid extract with pentafluorobenzyl (PFB) bromide followed by high-performance liquid chromatography purification yielded a fairly pure fraction containing TAA-PFB esters that was analyzed by GC/MS. Partial separation of the TAA isomers was obtained on various GC columns. Comparison of the retention time with the synthetic standards revealed that all four TAA isomers are present in human plasma. The mean concentration of TAA in human plasma was 20.2ng/ml. The levels of isomers were 12.48±1.28, 2.75±0.39, and 4.99±0.74ng/ml for 5E-AA + 11E-AA, 8E-AA, and 14E-AA, respectively. The identification of TAA in plasma suggests that isomerization of arachidonic acid occurs in vivo. Our method allows distinguishing between the dietary and the NO2- dependent mechanisms of trans fatty acid formation and will be useful in defining the role of TAA as an in vivo marker of nitrooxidative stress in clinical and experimental settings.

UR - http://www.scopus.com/inward/record.url?scp=4143144321&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4143144321&partnerID=8YFLogxK

U2 - 10.1016/j.ab.2004.04.030

DO - 10.1016/j.ab.2004.04.030

M3 - Article

C2 - 15301958

AN - SCOPUS:4143144321

SN - 0003-2697

VL - 332

SP - 137

EP - 144

JO - Analytical biochemistry

JF - Analytical biochemistry

IS - 1

ER -

Determination of trans-arachidonic acid isomers in human blood plasma

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this