Sensitive determination of anandamide in rat brain utilizing a coupled-column HPLC with fluorimetric detection.
作者： Yoshiko Arai , Takeshi Fukushima , Mika Shirao , Xiangjing Yang , Kazuhiro Imai
DOI: 10.1002/(SICI)1099-0801(200004)14:2<118::AID-BMC936>3.0.CO;2-O
关键词:
摘要: A fluorimetric determination method for N-arachidonoylethanolamine (anandamide) was developed using a precolumn fluorescence derivatization followed by coupled-column high-performance liquid chromatography (HPLC). Anandamide extracted from the rat brain tissue derivatized with 4-N-chloroformylmethyl-N-methylamino-7-N,N-dimethylaminosulfonyl-2,1,3-benzoxadiazole (DBD-COCl), purified solid-phase extraction (Empore™), and assayed HPLC. The HPLC consisted of phenyl (100 × 4.6 mm i.d.) octadecylsilica columns (250 × 4.6 mm i.d.), both connected six-port valve. concentration anandamide in 3.37 ± 0.73 pmol/g 6.47 3.57% intra- inter-day precisions, respectively. Using this method, we investigated alteration 30 min after administration (2 mg/kg, i.p.) to rats pretreated or without phenylmethylsulfonyl fluoride (PMSF; 30 mg/kg, i.p.), an inhibitor amidohydrolase. In PMSF, approx. 16-fold higher than that PMSF (p < 0.01). Copyright © 2000 John Wiley & Sons, Ltd.
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