建立細菌代謝色胺酸（Tryptophan）及其生成吲哚（Indole）衍生物之高效液相層析分析法

腸道中的細菌會利用膳食中蛋白質分解的tryptophan（色胺酸），代謝為indole（吲哚）及indolederivatives（吲哚衍生物），為有效及快速偵測與定量tryptophan及細菌代謝生成的吲哚環衍生物（indole ring derivatives），吾等利用high performance liquid chromatography（高效液相層析，HPLC），移動相為0.1%acetic acid（乙酸）混合methanol（甲醇）（60:40），以碳18逆向層析管柱（C18 reversed phase HPLC columns）進行化合物分離。分析時間40分鐘（3.5分鐘後開始出現起始波峰，直至35.5分鐘後結束）。本研究係利用tryptophan及各種吲哚衍生物的標準品建立能偵測並定量細菌代謝之9種吲哚衍生物的HPLC分析方法，續以Bifidobacterium longum subsp. infantis（長雙歧桿菌亞種嬰兒，B. infantis）培養在modified reinforced clostridial medium（改良強化梭菌培養基，MRC medium），然後利用此方法分析菌株於細胞轉化實驗代謝tryptophan的代謝物，可測得並定量indole-3-lactate（吲哚-3-乳酸）、indole-3-carboxaldehyde（吲哚-3-醛）及indole-3-carboxaldehyde（吲哚-3-乙酸）之生成。

關鍵字

色胺酸；高效液相層析法；吲哚衍生物

並列摘要

Certain gut bacteria utilize tryptophan from dietary proteins to metabolize into indole and indole-derivates. To detect and quantitate tryptophan, indole, and indole-related metabolites effectively, a high-performance liquid chromatography (HPLC) method was constructed. This 40-minute method used 0.1% acetic acid and methanol (60:40) as the mobile phase and C18 reverse phase column to separate a total of ten standard compounds. Using Bifidobacterium longum subsp. infantis (B. infantis) grown in modified reinforced clostridial medium (MRC medium), the metabolites of B. infantis were analyzed by our HPLC method and the production of indole-3-lactate could be monitored and quantified.

並列關鍵字

tryptophan ； high performance liquid chromatography ； indole derivatives

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