关键词: 反相HPLC, 血栓素B₂, 血小板

Abstract: TXB₂ content in platelet were analysed by high-performance liquid chromatography (HPLC) using a pre-column derivatization method,in which a fluorescentreagent,4-bro-momethyl-7-methoxycoumarin (BrMmc) was used.The mothod was applied to studing the effects of Acanthopanax Senticosus (A.S.) extract,a Chinese medicine,on platelet func-tion.The platelet TXB₂ were extracted by a Sep-Pak C₁₈ (waters).In the presence of 18-crown-6 and K₂CO₃,BrMmc reacted with carboxylic group(—COOH) of TXB₂ to form a fluorescent ester (BrMmc-TXB₂),which was seperated on a column (250mm×4.6mm I.D.)packed with Zorbax-ODS (6 μm,Dupont Instruments)and monitored by a fluorescent detector in Ex 345 nm and Em 405 nm.The mobile phase (acetonitrile:water:phosphoric acid;60:40:0.1 V/V),flowed at 1 ml/min at 30℃.The seperation of BrMmc-TXB₂ was completed in 20min.The detection limit was 15 ng using the external standard method.The anti-platelet actions of A.S.in rabbit were studied.The resultes (in vitro) showed that the platelet aggregations induced by AA and ADP were potently inhibited by A.S.in a dose-de-pendent manner and the AA-induced TXB₂ formation in platelet was also reduced.After in-travenous injection of A.S.(120mg/kg),the platelet aggregations induced by AA and ADP were inhibited and the effects lasted 30 and 45 min,respectively.

Key words: RP-HPLC, TXB₂, platelet