1-脫氧野尻黴素 (1-Deoxynojirimycin，DNJ)是一種在桑樹中特有但其它植物少見的生物鹼。DNJ是一種α-葡萄糖苷酶抑制劑，可抑制多醣與雙糖的分解，減少人體對糖的吸收，有效降低體內血糖濃度。因此建立一種可靠方法檢測食品 DNJ 的含量是非常重要的。故本研究以9-Fluorenylmethyl chloroformate (FMOC-Cl) 柱前衍生化結合HPLC與螢光或紫外光檢測器來分析DNJ成分，並進行確效評估以及新鮮桑葉與市售桑葉相關產製品中DNJ之含量分析。得知以HPLC結合螢光偵測器分析DNJ之檢量線的線性良好，相關係數R2達到0.99以上，準確度之回收率介於91.11~98.94%之間，重複性 (同日間)之變異係數(Coefficient of variation, CV)介於1.91~2.58%之間，中間精密度 (異日間) 之變異係數(CV)介於3.04~5.55%之間；最低偵測極限 (Limit of detection, LOD)與最低定量極限 (Limit of quantification, LOQ)分別為0.1 μg/mL以及0.3 μg/mL；顯示此方法檢測DNJ之靈敏度與確效性佳。然而，若以HPLC結合紫外光偵測器設定波長263 nm 檢測DNJ衍生物，靈敏度較低LOD只有15 μg/mL。本研究以HPLC結合螢光偵測器檢測新鮮桑葉與相關產品之結果，顯示葉子中DNJ含量(732.90-915.45 μg/g) 明顯較莖部(143.02-338.69 μg/g)為高，相同組織則以新鮮程度愈高含量愈高，愈成熟的愈低；其次，市售桑葉加工品之DNJ含量明顯較桑葚加工品為高。再者，桑葉經萃取、衍生化後之DNJ-FMOC衍生物於室溫下貯存3週後，大概只減少5.4%，顯示此衍生物安定性佳。另外，建議於桑葉茶製造過程中應盡量不要使用太高的烘乾溫度，以免原料中DNJ被熱分解破壞。整體而言，本研究建立一套簡單、靈敏、可靠定量食品中DNJ含量之檢測方法。

1-Deoxynojirimycin (1-Deoxynojirimycin, DNJ) is a unique alkaloid in mulberry but rare in other plants. DNJ is an α-glucosidase inhibitor, which can inhibit the decomposition of polysaccharides and disaccharides, reduce the body's absorption of sugar, and effectively reduce the blood glucose level in the body. Therefore, it is very important to establish a reliable method to detect the content of DNJ in food. This study was to use 9-Fluorenylmethyl chloroformate (FMOC-Cl) pre-column derivatization combined with HPLC and fluorescence or ultraviolet detectors to analyze DNJ components, and to conduct validation evaluation. It is known that the linearity of the calibration line for the analysis of DNJ by HPLC combined with a fluorescent detector is good, the correlation coefficient R2 is above 0.99, the recovery rate of the accuracy is between 91.11~98.94%, the coefficient of variation (Coefficient of variation, CV) of the repeatability ranged from 1.91 to 2.58%, and the coefficient of variation (CV) of the intermediate precision ranged from 3.04 to 5.55%. Limit of detection (LOD) and limit of quantification, LOQ) were 0.1 μg/mL and 0.3 μg/mL, respectively. This results showed that this method has good sensitivity and accuracy in detecting DNJ. However, if the DNJ derivatives are detected by HPLC combined with a UV detector set at a wavelength of 263 nm, the sensitivity is low and the LOD is only 15 μg/mL. In this study, the results of detecting fresh mulberry leaves and related products by HPLC combined with fluorescent detectors showed that the DNJ contents in leaves (732.90-915.45 μg/g) was significantly higher than that in stems (143.02-338.69 μg/g). The DNJ contents of commercially mulberry leaf products were significantly higher than those of mulberry products. Furthermore, the DNJ-FMOC derivative after extraction and derivatization of mulberry leaves was stored at room temperature for 3 weeks, and the decrease percentage was only about 5.4%, showing that the derivative has good stability. In addition, it is suggested that the drying temperature should not be too high during the manufacturing process of mulberry leaf tea, so as to prevent the DNJ in the raw materials from being destroyed by thermal decomposition. Overall, this study established a simple, sensitive and reliable detection method for quantifying DNJ contents in food.

中文摘要 I
英文摘要 III
誌謝 V
目錄 VII
表目錄 IX
圖目錄 XI
壹、緒論 1
貳、文獻探討 3
一、1-脫氧野尻黴素的來源 3
二、1-脫氧野尻黴素的萃取 9
三、1-脫氧野尻黴素的檢測方法 11
參、材料與方法 16
一、試藥與溶劑 16
二、器具及材料 16
三、儀器設備 17
四、桑葉與市售桑葉製品之檢體製備 18
五、試劑配製 18
六、桑葉與桑葉製品之前處理與衍生化 19
七、HPLC參數設定 20
八、樣品中1-脫氧野尻黴素之鑑別及含量測定 20
九、檢驗方法確效評估 21
十、樣品DNJ衍生物溶液之貯存試驗 24
十一、桑葉乾燥前後DNJ含量變化 24
十二、冷藏桑葉之DNJ貯存安定性 24
肆、結果與討論 25
一、HPLC-螢光檢測方法之確效評估 25
二、以HPLC-PDA檢測方法之確效評估 28
三、與國外文獻方法之確效結果比較 29
四、桑葉萃取DNJ衍生物溶液的貯存安定性試驗 31
五、桑葉、桑葚及其相關產製品中1-脫氧野尻黴素之含量 31
六、桑葉乾燥前後DNJ含量變化 33
七、桑葉於冷藏期間DNJ含量的變化 33
伍、結論 57
陸、參考文獻 58

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