Metori A, Ogamo A, Nakagawa Y
School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
J Chromatogr. 1993 Dec 22;622(2):147-51. doi: 10.1016/0378-4347(93)80260-b.
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).
