應用高效能液相層析三段四極柱雙重質譜儀對高脂飼料餵養與streptozotocin處理的大鼠進行尿液生物標記物分析之代謝體學研究   全文下載

Metabonomic Study with a High Performance Liquid Chromatography Coupling to a Triple Quadruple Mass Spectrometer to Identify Biomarkers from Urine of High-fat Fed and Streptozotocin Treated Rats

本研究利用高效率液相層析三段四極柱雙重質譜儀進行動物尿液的代 謝體學研究。 Wistar 大鼠（每組5 隻）分別餵以標準飼料或高脂飼料14 週 後，收集其尿液樣品進行小分子代謝物之掃瞄分析。利用統計上之主成分分 析，可以由動物尿液小分子代謝物的掃瞄結果，明顯的區分動物在接受不同 餵養條件下的代謝表現，且其代謝表現差異的情形，亦可歸因於不同負載強 度的小分子代謝物。負載強度愈大的小分子代謝物，愈能說明不同餵養條件 所造成的差異現象。經由雙重質譜分析，發現負載強度較大的小分子代謝物 中，包括了葡萄糖、丙二酸，以及其它克氏循環（Krebs cycle）的代謝物， 顯示在本研究中的不同餵養條件，主要是造成了動物在糖生成與能量合成的 代謝途徑上產生差異。由於代謝體學可以協助驗證基因體學與蛋白質體學的 研究結果，是探討營養、藥物、毒物甚至環境對生物基因表現影響的重要研 究手段之一，本研究成功地利用高效率液相層析三段四極柱雙重質譜儀執行 代謝體學研究，期待未來也可以應用到保健食品功效的驗證與個人化食品的 開發。

High performance liquid chromatography coupled with an electrospray ionization (ESI) tandem mass (MS/MS) detector was employed to scan the urinary metabolites of Wistar rats (n=5 each group) fed with either control diet or high fat diet with a single dose of streptozotocin (STZ) for 14 weeks. Principle component analysis (PCA) of the LC-MS data was performed for pattern re-cognition and it clearly showed that the dosed animals displayed different metabolic responses even though they received the same treatment. Components responsible for the separation were identified with MS/MS. Glucose and malonic acid were among some other components in Krebs cycle to show high loading strength in PCA. It was therefore postulated that the high fat diets have driven the animals toward the metabolic flux to gluconeogenesis and then energy production through Krebs cycle. Excess energy was also stored through fatty acid biosynthesis with the observation of high loading strength of malonic acid. This experiment demonstrated that a triple quadruple mass spectrophotometer is capable of the metabonomic study on diet modulation of animals. Since metabonomics is a congruous part of nutrigenomics that can provide information on nutritional status, toxicity and gene function of an organism, it is hoped that this technique be extended to study the efficacy of functional foods and the development of personalized foods.