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微透析-HPLC-ECD聯(lián)用在線測(cè)定煙堿暴露大鼠腦內(nèi)單胺神經(jīng)遞質(zhì)的動(dòng)態(tài)變化

發(fā)布時(shí)間:2018-07-31 18:23
【摘要】:為探討煙堿依賴的形成機(jī)理,實(shí)時(shí)評(píng)價(jià)煙堿引起的單胺神經(jīng)遞質(zhì)變化效應(yīng),采用在線微透析和高效液相色譜-電化學(xué)檢測(cè)器(HPLC-ECD)聯(lián)用方法測(cè)定了大鼠腦紋狀體中多巴胺(DA)、3,4-二羥基苯乙酸(DOPAC)、高香草酸(HVA)、5-羥色胺(5-HT)和5-羥吲哚乙酸(5-HIAA)的含量。以2.0μL/min灌流,每15 min收集1次透析液并自動(dòng)注入五氟苯基柱,在梯度洗脫條件下分離檢測(cè)。結(jié)果表明:方法的線性范圍為0.2~500.0 ng/m L,日內(nèi)和日間精密度均≤9.7%,加標(biāo)回收率為86.2%~105.9%,體外透析探針回收率為25.3%~28.3%。以2.0 mg/kg煙堿進(jìn)行腹腔注射后,大鼠紋狀體中DA和5-HT的釋放增加但未見持續(xù)增強(qiáng),而HVA和5-HIAA的量明顯升高。該方法簡(jiǎn)便快速,靈敏度高,選擇性好,適合于煙堿作用下活體動(dòng)物腦內(nèi)單胺類神經(jīng)遞質(zhì)變化的動(dòng)態(tài)分析。
[Abstract]:In order to investigate the formation mechanism of nicotine dependence and evaluate the effect of monoamine neurotransmitters induced by nicotine in real time. On-line microdialysis and high performance liquid chromatography-electrochemical detector (HPLC-ECD) were used to determine the contents of (HVA) 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in rat striatum. Dialysate was collected every 15 min and injected into pentafluorophenyl column at 2.0 渭 L/min for separation and detection under gradient elution. The results showed that the linear range of the method was 0.2n 500.0 ng/m / L, the precision between day and day was 鈮,

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