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研究生:李致廷
研究生(外文):Chih-Ting Li
論文名稱:文蛤殼及蜆殼水溶性萃取物抗氧化活性及煅燒粉末殺菌作用探討
論文名稱(外文):Studies on Antioxidative Activity of Water Extracts and Bactericidal Effects of Calcinated Powders of Hard Clam and Freshwater Clam Shells
指導教授:黃登福黃登福引用關係
指導教授(外文):Deng-Fwu Hwang
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:中文
論文頁數:77
中文關鍵詞:文蛤抗氧化活性煅燒殺菌作用
外文關鍵詞:Hard clamsFreshwater clamsAntioxidant activitiesCalcinationBactericidal effects
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二枚貝殼約由96% ~ 98% 之無機物 (碳酸鈣為主) 以及2% ~ 3% 之有機物組成,本研究目的為探討台灣最主要養殖之海水貝文蛤 (Meretrix lusoria) 和淡水貝蜆 (Corbicula fluminea) 殼廢棄物之水溶性有機物和無機物利用。第一部分將文蛤殼及蜆殼經過清洗後,以蒸餾水萃取 (130℃,1.5 atm,1小時),所得水溶性物質即為文蛤殼水溶性萃取物 (hard clam shell water extract; HSE) 和蜆殼水溶性萃取物 (freshwater clam shell water extract; FSE)。HSE和FSE之蛋白質含量分別約為17.37% 和12.43%,小於6 kDa之胜肽含量分別約為11.78% 和9.11%,且不含醣類以及多酚類,兩種萃取液之pH值皆為弱鹼性。蛋白質電泳結果顯示,HSE和FSE的蛋白質分子量大約為10 kDa。FSE和HSE之總胺基酸種類皆以甘胺酸 (glycine) 為最高,天門冬胺酸 (aspartic acid)、酪胺酸 (tyrosine) 以及苯丙胺酸 (phenylalanine) 等含量亦偏高。抗氧化試驗結果顯示,FSE之DPPH (1,1-diphenyl-2-picrylhydrazyl) 自由基清除能力較HSE佳,其半抑制濃度 (half maximal inhibitory concentration; IC50) 為124.70 μg/ml。HSE之螯合亞鐵離子能力較FSE佳,其IC50分別為0.25與1.36 mg/ml。還原力結果顯示FSE之還原力較HSE佳,FSE之還原力約為HSE之5倍。HSE和FSE之ABTS+ (2,2-azino-bis (3- ethylbenthiazoline sulfonic acid)) 清除能力於低濃度下無顯著差異,高濃度時HSE高於FSE。以人類皮膚纖維母細胞CCD966SK作為細胞模式探討其抗氧化力,結果發現FSE在不具細胞毒性之濃度下,可減緩過氧化氫造成之細胞死亡,反之HSE則無作用。
第二部分將萃取剩餘之粉末經高溫1,050℃煅燒24小時,其回收率約為45%,且煅燒文蛤殼粉末 (calcinated hard clam shell powder; CHSP) 和煅燒臺灣蜆殼粉末 (calcinated freshwater clam shell powder; CFSP) 中的主要結構從碳酸鈣轉變為氧化鈣,另外兩者之粒徑直徑均比未煅燒之粒徑小。CHSP和CFSP之溶液或懸浮液,pH值皆為強鹼性 (11.5-12.4),且隨著貯存時間到達第6天,CHSP和CFSP之pH值並無顯著改變。以0.1% 之CHSP或CHSP處理變形桿菌 (Proteus vulgaris) 和腸道沙門氏菌 (Salmonella enterica) 3分鐘後,可殺滅約7 log CFU/ml之菌數。0.5% 之CFSP及CHSP可殺滅7 log CFU/ml之頭狀葡萄球菌 (Staphylococcus capitis),1% 之CFSP及CFSP約可殺滅6.25 ~ 7.42 log CFU/ml之李斯特單核球菌 (Listeria monocytogenes),但未經煅燒之文蛤殼或蜆殼之粉末不具顯著殺菌能力。綜合上述,文蛤殼及蜆殼水溶性萃取物具有抗氧化能力,可做為皮膚保護材料之應用,而剩餘之粉末經高溫煅燒後,則具有殺菌能力,可做為食品清洗劑之應用。

The composition of bivalve shells includes about 96% - 98% inorganic compounds (mainly CaCO3) and 2% - 3% organic compounds. The purpose of this study was to investigate the antioxidative activities of water soluble extracts and the bactericidal effects of calcinated powders from the most abundant freshwater bivalve, freshwater clam (Corbicula fluminea), and seawater bivalve, hard clam (Meretrix lusoria), in Taiwan. In the first part of this study, first of all, the shells of hard clam and freshwater clam were cleaned-up and then crushed to a powder. An equal weight of deionized water was added to the powder and extracted at 130℃ and 1.5 atm for 60 min. The extract solution was filtered and dialyzed. Finally, the solution was lyophilized and employed as the water extract of freshwater clam shells (FSE) and hard clams (HSE). The protein content of HSE and FSE was about 17.37% and 12.43%, respectively. The peptide (&;lt; 6 kDa) content of HSE and FSE was about 11.78% and 9.11%, respectively. Either FSE or FSE didn’t contain carbohydrates and polyphenols. Both HSE and FSE solutions were weak basic. The amino acid composition of FSE and HSE was mainly glycine. Aspartic acid, tyrosine and phenylalanine were also rich. In the antioxidant activity study, the IC50 (half maximal inhibitory concentration) value of DPPH (1,1-diphenyl-2- picrylhydrazyl) scavenging activity of FSE was about 124.70 μg/ml. HSE didn’t have DPPH radical scavenging activity. The IC50 values of Fe2+-inhibition of FSE and HSE were 1.36 and 0.25 mg/ml, respectively. The reducing power of FSE was five times more than HSE. There was no significant difference of ABTS+ scavenging activity between FSE and HSE at low concentrations. In the cell experiments, the cell viability of CCD966SK cells was not affected by FSE or HSE treatment. Furthermore, H2O2-induced oxidative stress in CCD966SK cells was reduced by FSE treatment, but not by HSE treatment.
In the second part of this study, freshwater clam shell powder (FSP) and hard clam shell powder (HSP) was calcinated at 1,050℃ for 24 hr, and the yields of calcinated FSP (CFSP) and calcinated HSP (CHSP) were both about 45%. The main composition of CFSP and CHSP was converted to CaO from CaCO3. On the other side, the particle sizes were also decreased. Both of CFSP and CHSP was strong basic (pH 11.5-12.4). After 6 days storage, the pH of CFSP and CHSP suspended solutions were not changed significantly. Bacterial counts of Proteus vulgaris and Bacillus subtilis were reduced about 7 log CFU/ml by either 0.1% (w/v) CFSP or CHSP for 3 min immersion. Bacterial counts of Staphylococcus capitis were reduced about 7 log CFU/ml by 0.5% (w/v) either CFSP or CHSP for 3 min immersion. Finally, bacterial counts of Listeria monocytogenes were reduced about 6.25-7.42 log CFU/ml by either 1% (w/v) CFSP or CHSP for 3 min immersion. Oppositely, neither HSP nor FSP had no bactericidal effect. Hence, antioxidant activities were found in both HSE and FSE, which can be used for skin protection materials. Bactericidal effects were found in both CFSP and CHSP solutions, which can be used as food cleaners.

摘要 I
Abstract II
壹、文獻整理
一、文蛤與蜆簡介
(一) 文蛤與蜆之分類 2
(二) 文蛤及蜆之生態和養殖現況 2
(三) 文蛤與蜆肌肉之一般組成分 3
二、二枚貝殼廢棄物
(一) 水產廢棄物之種類與利用 4
(二) 二枚貝類殼廢棄物之組成分與結構 4
(三) 二枚貝類殼廢棄物之生理活性及利用 6
三、活性氧及抗氧化簡介
(一) 活性氧種類及來源及造成之傷害 8
(二) 抗氧化系統及抗氧化劑 8
四、煅燒殼粉之殺菌作用
(一) 食品及環境相關病原菌 10
(二) 煅燒牡蠣殼粉之殺菌機制 11
貳、研究內容
第一章、文蛤殼及蜆殼水溶性萃取物抗氧化活性探討
一、前言 13
二、材料與方法 14
三、結果 23
四、討論 27
五、圖表 29
第二章、文蛤殼及蜆殼煅燒粉末殺菌作用探討
一、前言 42
二、材料與方法 43
三、結果 46
四、討論 49
五、圖表 52
參考文獻 69
謝辭 77

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