論小分子代謝產物鑑定流程建立與其應用於益生菌共生發酵液之開發

Taipei Medical University Institutional Repository > 醫學科技學院 > 醫學檢驗暨生物技術學系所 > 博碩士論文 > Item 987654321/63439

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題名:	論小分子代謝產物鑑定流程建立與其應用於益生菌共生發酵液之開發 The establishment of small molecule metabolite identification process and its application in the development of probiotic symbiotic fermentation broth
作者:	莊朝傑 Chuang, Jau-Jye
貢獻者:	醫學檢驗暨生物技術學系碩士在職專班林榮俊
關鍵詞:	酵素結合免疫吸附分析法;超高效能液相層析串聯質譜儀;超高效能液相層析軌道阱質譜儀;動物實驗;血清素;多巴胺 ELISA;UHPLC tandem mass spectrometry;UHPLC orbitrap mass spectrometry;animal experiment;Serotonin;Dopamine
日期:	2023-07-03
上傳時間:	2024-01-22 13:23:12 (UTC+8)
摘要:	背景：益生菌以其改善腸菌群、調節免疫功能和抑制肥胖等多種對人體有益的作用而受到關注。本研究選用佰研生化科技股份有限公司提供的乳酸菌共生發酵液進行實驗，旨在探索其中對人體有益的成分，並通過細胞實驗和動物實驗模型驗證攝入該發酵液對代謝的影響。方法：首先，利用細胞株CaCO2進行體外細胞實驗，並使用ELISA kit定量分析血清素的含量。隨後，通過動物模型進行體內試驗，模擬攝入發酵液對人體的影響，並監測體重和血液生化指標的變化。同時，開發並建立了液相層析串聯質譜（LC-MS/MS）的分析方法，以檢測動物實驗中代謝物的改變。結果：佰研生化科技提供的乳酸菌共生發酵液（GA6）在體外細胞實驗中促進了CaCO2細胞的血清素分泌，在動物實驗中也增加了血清素的分泌。此外，我們成功建立了糞便檢體中多巴胺與血清素的LC-MS/MS分析方法，並通過方法確效驗證了該分析方法的準確度和精密度。 Background: Probiotics have gained attention for their diverse beneficial effects on human health, including improving gut microbiota, regulating immune function, and suppressing obesity. In this study, we employed a lactobacillus symbiotic fermentation liquid provided by Sagittarius Life Science Co., Ltd. to investigate its beneficial components and evaluate the metabolic impact of its consumption through cellular and animal experimental models. Methods: Firstly, we conducted in vitro cell experiments using the CaCO2 cell line and quantitatively measured serotonin levels using an ELISA kit. Subsequently, we performed in vivo experiments using an animal model to simulate the effects of consuming the fermentation liquid, monitoring changes in body weight and blood biochemical markers. Additionally, we developed and established a liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis method to detect alterations in metabolites during the animal experiments. Results: The lactobacillus symbiotic fermentation liquid (GA6) provided by Sagittarius Life Science Co., Ltd. significantly enhanced serotonin secretion in CaCO2 cells during the in vitro cell experiments. Consistently, it also led to increased serotonin secretion in the animal experiments. Moreover, we successfully developed an LC-MS/MS analysis method for dopamine and serotonin in fecal samples and validated the accuracy and precision of this analytical approach through method validation experiments.
描述:	碩士指導教授:林榮俊委員:葉添順委員:梁有志委員:林榮俊
資料類型:	thesis
顯示於類別:	[醫學檢驗暨生物技術學系所] 博碩士論文

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