TY - JOUR
T1 - Preparation and assay of monohydroxy-eicosatetraenoic acids
AU - Boeynaems, J. M.
AU - Brash, A. R.
AU - Oates, J. A.
AU - Hubbard, W. C.
N1 - Funding Information:
This work was supported by NIH Grants GM I5431 and BRSG-RR-05494 and hy PHS Fellowship 09682. J. M. Boeynaems is Fellow of the Fogarty International Center and Aspirant of the Fonds National de la Recherche Scientifique (Belgium). J. A. Oates is the J. and M. Werthan Professor of Investigative Medicine. We are grateful to Dr. D. Taber for the generous gift of octadeuterated arachidonic acid and for helpful suggestions throughout this study.
PY - 1980/5/15
Y1 - 1980/5/15
N2 - 5-, 8-, 9-, 11-, 12-, and 15-hydroxy-eicosatetraenoic acids (HETEs) were prepared from arachidonic acid by reaction with H2O2 in the presence of Cu2+ ions. They were separated by high-performance liquid chromatography on silica gel (μPorasil), using a linear solvent gradient from hexane to chloroform: only the 8- and 9-isomers were not resolved. Multi-milligram quantities of highly purified HETEs could be easily generated by this method, which thus provides a useful tool to study the biological activity of these compounds. Octadeuterated analogs of HETEs prepared from octadeuterated arachidonic acid by this procedure were suitable for use as internal standards in stable isotope dilution assays, by combined gas chromatography and mass spectrometry, with selected ion monitoring. The detection limit of the HETEs was less than 1 ng.
AB - 5-, 8-, 9-, 11-, 12-, and 15-hydroxy-eicosatetraenoic acids (HETEs) were prepared from arachidonic acid by reaction with H2O2 in the presence of Cu2+ ions. They were separated by high-performance liquid chromatography on silica gel (μPorasil), using a linear solvent gradient from hexane to chloroform: only the 8- and 9-isomers were not resolved. Multi-milligram quantities of highly purified HETEs could be easily generated by this method, which thus provides a useful tool to study the biological activity of these compounds. Octadeuterated analogs of HETEs prepared from octadeuterated arachidonic acid by this procedure were suitable for use as internal standards in stable isotope dilution assays, by combined gas chromatography and mass spectrometry, with selected ion monitoring. The detection limit of the HETEs was less than 1 ng.
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U2 - 10.1016/0003-2697(80)90073-1
DO - 10.1016/0003-2697(80)90073-1
M3 - Article
C2 - 7004263
AN - SCOPUS:0019325415
SN - 0003-2697
VL - 104
SP - 259
EP - 267
JO - Analytical biochemistry
JF - Analytical biochemistry
IS - 2
ER -