Determination of 5-hydroperoxyeicosatetraenoic acid produced in rat basophilic leukemia cell line RBL-2H3 by high-performance liquid chromatography with chemiluminescence detection.
A simple and sensitive method, applicable to quantification of 5-hydroperoxyeicosatetraenoic acid (5-HPETE) produced in cells has been developed using high-performance liquid chromatography on a silica gel column with chemiluminescence detection. 5-HPETE was clearly separated from other positional isomers of HPETEs and hydroxyeicosatetraenoic acids with hexane-isopropanol-acetic acid (97:3:0.01, v/v) as the mobile phase. The lower limit of detection was about 100 pg. 5-HPETE produced in 10(7) cells of RBL-2H3 cells stimulated with A23187 was determined as 480+/-30 pg. In the present study, 5-HPETE, which occurs naturally, was detected and quantitated for the first time in intact cells.
Chiba
N
N
Japan Energy Research Center Co. Ltd., Tokyo.
Imai
H
H
Nakagawa
Y
Y
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015471:Leukemia, Basophilic, Acute / Q000378:metabolism* / Q000473:pathology; D007975:Leukotriene B4 / Q000096:biosynthesis / Q000378:metabolism; D015289:Leukotrienes / Q000096:biosynthesis / Q000378:metabolism*; D008163:Luminescent Measurements; D008293:Malates / Q000494:pharmacology; D051381:Rats; D014407:Tumor Cells, Cultured
D015289:Leukotrienes; D008293:Malates; D007975:Leukotriene B4; D000001:Calcimycin; C058705:diethyl malate; C025348:arachidonic acid 5-hydroperoxide