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研究生:劉孟翰
研究生(外文):Liu Meng Han
論文名稱:長鏈二元酸鑑定與定量方法之評估研究
論文名稱(外文):The evaluation of identified and quantitative methods of long chain dicarboxylic acid
指導教授:黃維寧
指導教授(外文):Huang Wei Ning
學位類別:碩士
校院名稱:元培科技大學
系所名稱:生物技術研究所
學門:醫藥衛生學門
學類:其他醫藥衛生學類
論文種類:學術論文
畢業學年度:102
語文別:中文
論文頁數:70
中文關鍵詞:長鏈二元酸熱帶假絲酵母
外文關鍵詞:Long chain dicarboxylic acidCandida tropicalis
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  • 被引用被引用:1
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長鏈二元酸(Long chain dicarboxylic acid;DC)為廣泛用於製造香精、聚合物、尼龍、合成樹脂及高級潤滑油之原料,工業上主要由化學合成法生產,但需高溫高壓環境,除工安隱憂外,對環境亦會造成汙染。相較之下,透過微生物醱酵將正烷類分解以生產長鏈二元酸之技術,具有安全、環保的優點。由於醱酵及純化製程直接影響長鏈二元酸產量,因此製程技術的精進有賴於迅速分析與精準定量的方法。本研究將基因改良後的Candida tropicalis酵母菌ATCC20962醱酵獲得之二元酸,利用紫外光 (UV) 吸收光譜、氣相層析結合火焰游離偵測器(GC-FID)、高效液相層析 (HPLC) 結合紫外光偵測器、酸鹼滴定法以及衰減全反射紅外光譜法 (ATR-FTIR)多項技術來進行分析。結果顯示除傳統GC-FID可精準定量二元酸外,HPLC結合紫外光偵測器也可以定量鑑定不同碳鏈長二元酸且具備不需前處理樣品之優點,衰減全反射紅外光譜法方法,除了可以對烴烷類酸定量外,還可快速的鑑定樣品中的不純物成份,但是在不同鏈長及單元、二元酸上是無法以此法加以區別,HPLC結合兩波段UV偵測器則將可以補足以上各不同方法的盲點,以上這些方法應可以作為醱酵與純化二元酸的製程改良中不同階段的分析利器。

Long-chain dicarboxylic acids (Long chain dicarboxylic acid; DC) is widely used in the manufacture flavors , polymers, nylon , synthetic resins and high grade lubricating oil feedstock , industrial production mainly by chemical synthesis , but need high temperature and pressure environment , in addition to industrial safety concerns , but also on the environment caused by pollution. In contrast, the positive through microbial fermentation to produce alkanes decomposition of long- chain dicarboxylic acid technology, safety, environmental protection advantages. Since fermentation and purification processes directly affect the long-chain dicarboxylic acid production , so sophisticated process technology depends on the rapid analysis and accurate quantitative method . In this study, genetically modified yeast Candida tropicalis ATCC20962 after fermentation obtained dicarboxylic acid , the use of ultraviolet (UV) absorption spectroscopy, gas chromatography combined with free flame detector (GC-FID), high performance liquid chromatography (HPLC) combined with ultraviolet detectors, acid-base titration and attenuated total reflection infrared spectroscopy (ATR-FTIR) to analyze a number of techniques . The results showed that in addition to traditional GC-FID can be accurately quantified dicarboxylic acid outside , HPLC UV detector can also be combined with quantitative identification of dicarboxylic acids with different carbon chain length and have the advantages of no sample pre-treatment , attenuated total reflection infrared spectroscopy methods , in addition to a hydrocarbon alkane acid quantitatively , but also rapid identification of impurities in the sample composition , but different chain lengths and unit diacids this method is unable to distinguish between , HPLC combined with two -band UV detector complement will be blind spots of different methods of the above , these methods should be used as dicarboxylic acid fermentation and purification process of the different stages of improvement weapon.
目 錄
頁次
中文摘要 I
英文摘要 II
目錄 III
圖目錄 VI
表目錄 X
第一章 緒論 1
1.1 長鏈二元酸 1
1.2 應用領域 1
1.3 目前主要生產方法 2
1.4 新興生產技術的開發 2
1.5 轉化機制研究 3
1.6 關鍵菌種相關研究 4
1.7 研究目的 5
第二章 材料與方法 6
2.1實驗材料 6
2.1.1 菌源: 6
2.1.2 藥品: 6
2.1.3 儀器: 8
2.2 實驗方法 10
2.2.1培養基的配製 10
2.2.2菌種培養 10
2.2.3 醱酵 11
2.2.4 醱酵液純化及前處理 11
2.2.5中油提供之代檢測醱酵樣品 11
2.2.6 紫外光(UV)分光光度計 12
2.2.7 滴定法 12
2.2.8 霍氏紅外光譜(FTIR) 12
2.2.9 氣相層析(GC) 13
2.2.10 高效能液相層析(HPLC) 13
第三章 結果與討論 15
3.1分析方法探討 15
3.1.1紫外光(UV)分光光度計 15
3.1.2滴定法 15
3.1.3霍氏紅外光譜(FTIR) 16
3.1.4氣相層析(GC) 18
3.1.5高效能液相層析(HPLC) 19
3.2 醱酵條件及純化方法測試 21
第四章 結論 23
圖表 24
參考文獻 62

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