Quantitation of monohydroxy fatty acids by high-performance liquid chromatography with fluorescence detection.
A high-performance liquid chromatographic (HPLC) procedure for the separation of hydroxyeicosatetraenoic acids (HETEs) and hydroxyoctadecanoic acids (HODEs) after derivatization of the hydroxy group with 1-anthroylnitrile is described. Anthroyl esters of HETEs were separated from those of HODEs by reversed-phase HPLC. The positional isomers of the HETEs and HODEs were well separated by normal-phase HPLC. The fluorimetric HPLC method has a high sensitivity and naturally occurring HETEs can be quantitatively analyzed at the picomolar level. The amount of 5-HETE in A23187-stimulated polymorphonuclear leukocytes (PMNLs) was determined by the present method. PMNLs produced approximately 150 ng of 5-HETE per 10(7) cells at 5 min stimulation. The amount of 5-HETE determined by fluorimetric detection was consistent with that determined by ultraviolet detection (235 nm).
Metori
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A
School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
Ogamo
A
A
Nakagawa
Y
Y
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis*; D066298:In Vitro Techniques; D008041:Linoleic Acids / Q000032:analysis*; D044243:Linoleic Acids, Conjugated; D009504:Neutrophils / Q000737:chemistry / Q000187:drug effects; D051381:Rats; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D005456:Fluorescent Dyes; D006893:Hydroxyeicosatetraenoic Acids; D008041:Linoleic Acids; D044243:Linoleic Acids, Conjugated; C024347:9-hydroxy-10,12-octadecadienoic acid; D000001:Calcimycin