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in the range of 19.1~29.7% as well against the mutagenicity of NPYR 目 錄 頁次 第一章 緒言……………………………………………………… 1 第二章 文獻回顧………………………………………………… 3 一、 影響 亞硝胺生成的因素………………………………… 3 (一) 硝酸鹽及亞硝酸鹽 含量………………………………… 3 (二) pH 值……………………………………………….……. 5 (三) 二級胺種 類……………………………………………… 5 (四) 加工方式及食品組成 份………………………………… 7 (五) 微生物作 用……………………………………………… 7 二、 食品中亞硝胺的含 量…………………………………… 10 (一) 醃肉中的亞硝胺含 量…………………………………… 10 (二) 魚類製品的亞硝胺含 量………………………………… 12 (三) 脫脂乳粉及黃豆製品的亞硝胺含 量…………………… 14 (四) 蔬菜的亞硝胺含 量……………………………………… 14 (五) 酒精飲料的亞硝胺含 量………………………………… 14 三、 亞硝胺的毒 性…………………………………………… 17 (一) 急性 毒…………………………………………………… 17 (二) 致癌性 毒………………………………………………… 18 四、 食品中抑制亞硝胺 突變的成分………………………… 20 (一) 維生素 C………………………………………………… 20 (二) 維生素 E……………………………………………….... 22 (三) 酚類化合 論文 目次
素……………………………………………… 25 (五) 其他化合 物……………………………………………… 25 (六) 微生 物…………………………………………………… 26 五、 乳酸菌簡 介……………………………………………… 27 六、 乳酸菌與健 康…………………………………………… 27 (一) 構成益生菌的要 件……………………………………… 30 (二) 腸道菌與人類關 係……………………………………… 30 (三) 益生菌對人體的益 處…………………………………… 34 第三章 材料與方 法……………………………………………… 40 一、 實驗材 料………………………………………………… 40 (一) 試驗菌 株………………………………………………… 40 (二) 亞硝胺種 類……………………………………………… 40 (三) 培養 基…………………………………………………… 40 (四) 化學藥 品………………………………………………… 40 (五) 市售含乳酸菌發酵 乳..…………………………………. 41 二、 儀器設 備………………………………………………… 42 三、 實驗方法及步 驟………………………………………… 42 (一) 乳酸菌培養液製 備……………………………………… 42 (二) 不同 pH 值乳酸菌溶液的製 備…………………………. 42 (三) 不同 pH 值緩衝溶液的製 備……………………………. 43 (四) 亞硝胺標準品的製 備…………………………………… 43 (五) Ames test 試驗原
之培養及製備……….... 44 (七) S. typhimuriumTA98 及 TA100 基因型態之確 認…….... 45 (八) 亞硝胺的致突變性試驗………………………..………. 49 (九) 乳酸菌的致突變性試驗………………………………… 50 (十) 乳酸菌 對亞硝胺的抗致突變性試驗.…………………... 51 (十一) 市售含乳酸菌發 酵乳抑制亞硝胺致突變性調查….…... 52 六、 統計分 析………………………………………………… 52 第四章 結果與討 論……………………………………………… 53 一、 試驗菌株基因型態之 確認……………………………… 53 二、 亞硝胺濃度對 S. typhimurium 致 突變性的影響………. 53 三、 乳酸菌對 S. typhimurium 生長的影 響…………………. 64 四、 乳酸菌培養液對亞硝胺之抗致突變 性………………… 69 五、 乳酸菌上層液對亞硝胺之抗致突變 性………………… 71 六、 乳酸菌菌體對亞硝胺之抗致突變 性…………………… 71 七、 市售含乳酸菌發酵乳抑制亞硝胺致突變性調 查……… 74 第五章 結論……………………………………………………… 78 參考文獻 …………………………………………………………… 80 附錄 一、 市售含乳酸菌發酵乳樣品……………………………… 92 圖 目 錄 頁 次 圖一、 亞硝胺生成機制之通式……………………………………. 4 圖 二、 大腸桿菌懸浮液和米麴菌菌叢中 DMNA 和亞硝酸鹽生成的情 形………………………………………………………. 11 圖三、 改變抑制劑 和亞硝酸鹽的莫耳濃度比例對二甲胺生成 NDMA 的影 響……………………………………………… 24 圖四、 不同部位消化道之 腸道菌分布情形………………………. 32 圖五、 腸道菌隨人類年齡變化之 改變情形…………………….… 33 圖六、 雙叉乳酸桿菌培養液之 pH 調整 對大腸桿菌生長的影響…….. 36 表 目 錄 頁次 表一、 各種二級胺之 pKa 值及最適於亞硝化之 pH 值……….. 6 表二、 不同加工方式對培根中 nitrosopyrrolidine 生成量的影響 8 表三、 餵食無菌老鼠及普通老鼠含脯胺 酸及亞硝酸鈉的飲用水後，排泄物中 NPRO 的存在情 形……………………. 9 表四、 傳統豬肉培根及仿培根產品之揮發性亞硝 胺含量…….. 13 表五、 黃豆製品中 NDMA 的含 量……………………………… 15 表六、 各式酒精性飲料中 N- nitrosodimethylamine 的含量……. 16 表七、 亞硝胺對 BD 大白鼠之半數致 死劑量及其目標器官….. 19 表八、 亞硝基化合物致癌之目標器 官………………………….. 21 表九、 由 Dadih 發酵乳分離之乳酸菌對 N- nitroso-dimethyl-amine(NDEA)致突變性的抑制情形…………………….. 28 表 十、 Bifidobacterium longum 胞內萃取液去除亞硝胺的百分 比………………………………………………………….. 29 表十一、 乳酸 菌對病原菌的抑制情形…………………………..… 35 表十二、 安氏試驗 (Ames Test)系統之 TA 菌株基因型…………. 46 表十三、 Salmonella typhimurium TA98 及 TA100 之基因型態檢測 54 表十四、 NDMA Salmonella typhimurium TA98 之致突變性…. 56 表十五、 NDEA Salmonella typhimurium TA98 之致突變性….. 57 表十六、 NPYR Salmonella typhimurium TA98 之致突變性…… 58 表十七、 NDMA Salmonella typhimurium TA100 之致突變性… 60 表十八、 NDEA Salmonella typhimurium TA100 之致突變性…. 61 表十九、 NPYR
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