河魨毒素中毒在許多國家都曾發生，因此，河魨毒素的檢測在食品衛生安全方面是相當重要的。由於河魨毒素標準品相當昂貴且不易取得，而河魨毒素經鹼處理加熱後會分解成C9-base，其基本化學結構具兩個相連的6員環及guanidine類似結構，因此，具有河魨毒素結構功能基特性及三度空間，本研究將有機合成河魨毒素鹼水解物C9-base的結構類似物，再以合成的河魨毒素結構類似物作為分子拓印聚合物的模版，製作分子拓印聚合物，以河魨毒素測試其專致性與敏感性。
本研究發現4-Guanidino-3-hydroxybenzoic acid及4-guanidino- 3-methoxy-benzoic acid皆可藉由4-Amino-3-hydroxyl- benzoic acid及4-amino-3-methoxy-benzoic acid與氨基腈反應而獲得。在合成(2-hydroxy-4-(hydroxymethyl)- phenyl)guanidine之過程中，利用不同的胍基化試劑及還原劑測試反應能力，結果發現，胍基化試劑中，以N,N’-di-Z-1H-pyrazole-1-carbamidine最好，而還原劑中則以lithium triethylborohydride可得到較高的產率，其還原力順序為Li(Et)3BH > LiBH4 > LiAlH4 > LiPyrBH3。而在分子拓印聚合物製備方面，以4-guanidinobenzoic acid為模版，於甲醇溶劑中可成功製成分子拓印聚合物。

Tetrodotoxins (TTXs) are potent natural neurotoxins that selectively block the sodium channels of the animals’ neuron cell membrane. It has been found in many marine and terrestrial organism. Since tetrodotoxin poisoning has occurred in many countries, detection of tetrodotoxin is very important for food safety. Molecular imprinting can be designed and tailor-made various polymers so that the polymers have high specificity and selectivity and act as artificial antibody.
TTX standard is very expensive and is not easy to obtain. Its convention determination is based on alkali treatment to the fluorescent compound, C9-base. Therefore, the main objective of this study was to synthesize TTX-mimic derivatives as template and synthesize molecular imprinted polymer that could bind TTX.
4-Guanidino-3-hydroxybenzoic acid and 4-guanidino-3-methoxy- benzoic acid could be obtained by guanidination of 4-Amino-3-hydroxyl- benzoic acid and 4-amino-3-methoxy-benzoic acid with cyanamide. During the synthesis of (2-hydroxy-4-(hydroxymethyl)- phenyl)guanidine, different guanidination reagents and reducing reagents have been tried. Guanidination with N,N’-di-Z-1H-pyrazole-1-carbamidine and reduction with lithium triethylborohydride provided satisfactory yields. The order of reducing reactivity is Li(Et)3BH > LiBH4 > LiAlH4 > LiPyrBH3. Furthermore, molecularly imprinted polymer using 4-guanidinobenzoic acid as template in methanol has been prepared.

中文摘要…………………………………………………………………i
英文摘要…………………………………………………………………ii
目錄……………………………………………………………………iii

壹、前言…………………………….……………………………………1
貳、文獻整理……………………………………………………………3
一、河魨毒素相關研究………………………………………………3
(一) 河魨毒素簡介……………….…………………………………3
(二) 河魨毒素化學結構與性質……………………………………3
(三) 河魨毒素中毒案例……….………………………….………5
(四) 河魨毒素檢測方法……………………………………………7
1.鼠生物檢測法.………………………………………………7
2.離體組織法……………………………………………………7
3.組織培養法……………………………………………………8
4.鹼水解物螢光法及紫外光分光光度法………………………8
5.LC-MS儀器檢測法……………………………………… 9
6.免疫學檢測法………………………………………………10
二、胍類化合物的合成…………………………………………………11
(一) 胍類化合物……………………………………………………11
(二) 胍類化合物液相合成之進展…………………………………11
1.利用氨基腈、溴化腈加成製備胍類化合物………………………11
2.利用胺基亞胺基甲磺酸合成胍類化合物…………………………12
3.利用1H-Pyrazole-1-carboxamidine hydrochloride合成胍類化合物………………………………………………………………………13
4.利用thiourea及S-alkylisothioureas合成胍類化合物………14
5.利用胍基化試劑 (Mtr-保護胍、Pmc-保護胍)合成胍類化合物…17
6.利用雙保護triflylguanidines合成胍類化合物…………………18
三、分子拓印技術相關研究…………………………………………20
(一) 分子拓印技術的發展………………………………………20
(二) 分子拓印聚合物的製作流程………………………………20
(三) 共價與非共價鍵結…………………………………………21
(四) 分子拓印聚合物組成要素……………………….………22
1.功能性單體………………………………………….…….……22
2.交聯劑……………………………………………………………25
3.溫度影響……………..…………………………………………28
4.溶劑作用……………………………………………………28
(五) 分子拓印聚合物的應用……………………………………29
1.食品分析方面……………………………………………………29
1.1.紅酒中ochratoxin A含量檢測………………………………31
1.2.Caffeine的分離………………………………………………33
2.分子拓印聚合物在分離鏡相異構物之應用………………35
2.1.胺基酸衍生物Cbz-L-Trp及Fmoc-L-Trp鏡相異構物分離……36
2.2.Nilvadipine鏡相異構物之分離………………………………37
2.3.抗組織胺藥物 d-chlorpheniramine……………………39
叁、材料與方法…………………………………………………………41
一、實驗材料……………………………………………………………41
1.藥品………………………………………………………………41
2.儀器設備…………………………………………………………43
二、實驗方法……………………………………………………………44
(一) 有機合成仿TTX結構類似物……………………………………46
1.有機合成仿TTX結構類似物 (I)、(II)……………………………46
1.1.仿TTX結構類似物 (I)……………………………………………46
1.2.仿TTX結構類似物 (II)……………………………………………47
2.有機合成仿TTX結構類似物 (III)…………………………………47
2.1.在胺基上加保護基 (3)……………………………………………48
2.2.溴化反應 (4)………………………………………………………49
2.3.溴取代反應 (5)……………………………………………………50
2.4.將羰基還原及去trimethylsilanolate (6)……………………50
2.5.去保護基 (III)……………………………………………………50
3.有機合成仿TTX結構類似物 (IV)……………………………………51
3.1.甲酯化反應 (8a)(8b)……………………………………………52
3.2.Guanidinylation…………………………………………………52
3.3.Methyl 4-aminobenzoate還原成醇………………………………54
3.4.Guanidinylation產物還原成醇 (10a)…………………………55
3.5.Guanidinylaton產物還原成醇 (10b)……………………………56
3.6.仿TTX結構類似物 (IV)……………………………………………56
(二) 分子拓印聚合物製作………………………………………57
1.高溫(65℃)熱起始，自由基聚合反應……………………57
2.低溫(4℃)紫外光照射光起始，自由基聚合反應…………58
(三) MIP對河魨毒素專制性及敏感性測試………………………59
肆、結果與討論…………………………………………………………60
(一) 有機合成仿TTX結構類似物……………………………………60
1.有機合成仿TTX結構類似物 (I)、(II)……………………………60
1.1.仿TTX結構類似物 (I)……………………………………………60
1.2.仿TTX結構類似物 (II)……………………………………………60
2.有機合成仿TTX結構類似物 (III)…………………………………61
2.1.在胺基上加保護基 (3)……………………………………………61
2.2.溴化反應 (4)………………………………………………………61
3.有機合成仿TTX結構類似物 (IV)……………………………………62
3.1.甲酯化反應 (8a)(8b)……………………………………………62
3.2.Guanidinylation…………………………………………………62
3.3.Guanidinylaton產物還原成醇……………………………………64
3.4.仿TTX結構類似物 (IV)……………………………………………65
(二) 分子拓印聚合物製作………………………………………66
伍、結論…………………………………………………………………67
陸、參考文獻……………………………………………………………68

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