臺灣博碩士論文加值系統

我們發展出簡單、連續性、自動化的分析方法，線上偵測MDA-TBA複合物。此方法包含了使用微透析取樣技術以及線上衍生並配合高效能液相層析儀進行分析。透析液的流速為1μl/min直接線上與TBA混合，在85℃環境下加熱30分鐘，之後由線上自動注射器將樣品送入高效能液相層析儀中，並藉由分光光度計進行偵測。層析條件包含甲醇-磷酸緩衝液（Ph6.8）（45:55,v/v）藉由逆相層析管柱進行分析，移動相流速為1.2ml/min。線性範圍由0.051 μM到2.43 μM(相關係數為0.999)，以TEP標準液所測到的平均微透析探針的回收率為18.4 ± 1.0 %，intra-assay與inter-assay精密度（％RSD）範圍為1.1 % 到 10.5 %之間。偵測極限為signal/noise為3時，其偵測極限為0.6p mol。因此我們發展出更為靈敏的MDA-TBA複合物的分析方法。另外，我們將此法應用來偵測不飽和脂肪酸的過氧化變化，以亞麻油酸於37℃水浴，再給予過氧化氫與金屬，所產生hydrojyl radical而造成亞麻油酸過氧化，測定其時間對MDA-TBA複合物對應關係。

A simple, continuous and fully automatic method was developed for on-line monitoring malondialdehyde (MDA) as its 2-thiobarbituric acid (TBA) complex. This method involved the use of microdialysis sampling, on-line derivatization and on-line high performance liquid chromatography analysis. Dialysate perfused from a microdialysis probe at 1 μl min-1 was on-line reacted with TBA at 85 ℃ for 30 min and automatic injected into a HPLC system equipped with a spectrophotometric detector. The chromatographic conditions consisted of a mobile phase of methanol-phosphate buffer (pH 6.8) (45:55, v/v) pumped through a reversed-phase column at a flow rate of 1.2 ml/min. For standards ranging from 0.051 μM to 2.43 μM, the response was linear (correlation coefficient = 0.999). The average in-vitro dialysate recovery (%) of MDA-TBA adduct in tetraethoxypropane standards was 18.4 ± 1.0 %. The intra-assay and inter-assay precision (% R.S.D.) ranged from 1.1 % to 10.5 %. The detection limit was 0.6 pmole for analyte at a signal to noise ration of three. Thus, a more sensitive quantitation of MDA-TBA adduct present in the assay system can be achieved. In addition, this method was applied to monitor the MDA-TBA adduct levels after treatment with H2O2 and metals to linoleic acid.

目 錄
第一章 前言
1.1自由基與脂質過氧化作用……………………...……………..…...………1
1.2鎂(Magnesium, Mg)的介紹……………………..………..……..…...….….9
1.3錳(manganese, Mn)的介紹………….…………………..…..……….…11
1.4 脂肪酸…………………………………….……..……….……..…...…….14
1.5研究目的與重點……………………………..………………….…...…….15
第二章 微透析取樣技術的介紹
2.1 微透析設計概念……………………………..…………………..…….….16
2.2 微透析取樣技術的特色…………………..………….…………...………18
2.3 微透析取樣技術在分析上的考量…………………..…………….….…..19
2.4微透析在體內研究的應用…………………..…………………..….……..20
2.5 微透析在基礎醫學與臨床醫學上的應用…………………..……………22
第三章 儀器與分析原理
3.1 儀器連線技術…………………..………………………………...………24
3.2 微透析取樣分析的原理…………………..………………….…..………26
3.3 高效能液相層析法…………………..……………………….…..………28
第四章 實驗部分
4.1 實驗材料及方法………………………….…..…………………..………31
4.2 分析條件…………………..……………….……………………..………36
4.3 分析步驟…………………..………………………………….…..………37
第五章 結果與討論…………………………………………….…...……40
第六章 結論………………………………………………….…………….47
第七章 參考文獻………………………………………..….……………..49
第八章 圖與表……………….………………………………

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