尿液及組織中大豆異黃酮葡萄糖醛酸結合物代謝物檢測   全文下載

Detection of Conjugated Soy Metabolites in Urinary and Tissue Samples after Methanol Extraction

以甲醇預處理人類，大白鼠及豬隻攝食大豆食物後的尿液樣品後，以液相離子阱質譜儀分析尿液樣品中的大豆異黃酮的?原及結合物。並利用同法處理及分析大白鼠的肝組織。結果顯示，在大白鼠尿液中檢測出染料木黃酮，木質素黃酮，黃豆素黃酮及雌馬酚的葡萄糖醛酸結合物，及木質素黃酮的硫酸結合物。在人尿及豬尿中未檢測出雌馬酚及其代謝物。根據遲滯時間，人尿中的大豆異黃酮代謝物流出序列為7-木質素黃酮-葡萄糖醛酸，7-黃豆素黃酮-葡萄糖醛酸，4’-木質素黃酮-葡萄糖醛酸，7-，及4’-染料木黃酮-葡萄糖醛酸。豬尿的流出序列與人尿相同，但最後流出物為5-染料木黃酮-葡萄糖醛酸。大白鼠尿液中檢測出雌馬酚的葡萄糖醛酸結合物及木質素黃酮的硫酸結合物。大白鼠尿液代謝物流出序列與豬尿相似，但在7-木質素黃酮-葡萄糖醛酸後流出木質素黃酮-硫酸結合物，在7-染料木黃酮-葡萄糖醛酸後流出7-雌馬酚-葡萄糖醛酸及4’-雌馬酚-葡萄糖醛酸。大白鼠肝組織中檢測出 7-，及4’-染料木黃酮-葡萄糖醛酸及其他?原代謝物。以液相電化學檢測器及液相大氣壓化學電噴灑法作定量分析後，結果顯示在大白鼠及豬隻的尿液中可以檢測出黃豆素黃酮的?原及結合物代謝物，在人尿中則只能檢測出黃豆素黃酮的結合物代謝物。

Glucuronide conjugates of soy isoflavone metabolites genistein, daidzein, glycitein and equol in urinary samples from women, female pigs and female rats were pretreated with methanol instead of solid phase extraction or liquid-liquid extraction after enzymatic digestion. The samples were analyzed using an LC-ESI-MS/MS ion trap system. Analysis of rat liver sample using this procedure was also attempted. By comparison the elution sequences with results from previous studies, glucuronide conjugates including 7-gluc-Dai, 7-gluc-Gly, 4’-gluc-Dai, 7-gluc-Gen, then 4’-gluc-Gen were identified sequentially in women urine. However, 5-gluc-Gen was also observed in female porcine urine. In rat urine, sul-Dai was observed after 7-gluc-Dai, while 7-gluc-Equ and 4’-gluc-Equ were observed after 7-gluc-Gen. The metabolite profiles in rat liver tissue samples were found with 7-gluc-Dai and 7- gluc-Gen. However, glucuronide conjugates of equol and sulfate conjugate of daidzein were only found in rat urine. Interestingly, quantification studies performed by LC-APCI/MS and LC-ECD showed that the aglycone and conjugated glycitein were both found in porcine and rat urine, but only conjugated glycitein was found in woman urine.