臺灣博碩士論文加值系統

本實驗以液相萃取結合液相層析質譜儀，應用於豆類中激勃素
（gibberellin A3, GA3）及其代謝產物結合性GA3之偵測。由實驗
結果顯示，使用電灑游離法負離子模式對GA3之偵測有較佳的靈敏
度，並以此結果應用豆類中GA3及其代謝產物結合性GA3之偵測。
豆類中GA3分析，以電灑游離法負離子模式之最佳條件，選擇
反應模式為定量方式。此方法對豆類中GA3線性範圍為0.02-20 ng/g
，線性相關係數為0.9970，偵測極限為5 pg/g，精密度為7-11 %。
真實樣品的測定中，可以測得綠豆、黃豆及紅豆中分別含有5.7、
10.6及0.5 ng/g的GA3。在GA3代謝物之偵測方面，使用液相萃取結
合層析質譜術於綠豆的葉、莖及根三部位GA3代謝產物進行分析，
實驗中推測GA3代謝產物經過氣體碰撞後會斷去醣基產生m/z 345離
子或離去中性分子m/z 106，也可能保留GA3的基峰離子，m/z 239。
以m/z 345離子及m/z 239離子之母代離子掃描及m/z 106之中性丟失
掃描，其結果皆無法得到疑似結合性GA3的訊號，推測在綠豆各部位
中GA3 的代謝產物含量太低，造成此方法無法得到結合性GA3的訊號。
由以上結果顯示，液相萃取結合液相層析質譜術於豆類中GA3的
檢測，具有其可行性，未來也可應用此技術於其它各種植物中微量
GA3之定量分析。

This project evaluated solvent extraction combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to determine the trace amount of gibberellic acid (GA3) in pulse. The electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) of triple-quadrupole mass spectrometer were used to compare the ionization efficiency in determination of GA3. GA3 was extracted from pulse by using 20 mL of 80 % methanol in water. Samples were then separated on a RP18 column interfaced with a tandem mass spectrometer. LC-ESI/MS/MS was proved a very selective and sensitive method for determination of trace GA3. The assay was linear over the range of 0.02-20 ng/g with a LOD of 5 pg/g. GA3 detected in Vigna radiata (L.) Wilczek, Glycine max (L.) Merr. and Phaseolus angularis W. F. Wight were 5.7, 10.6 and 0.5 ng/g, respectively. Parent ions and neutral loss scans were used to determine the bond GA3 in Vigna radiata (L.) Wilczek’s leaves, roots and steams. The parent ion spectra of the characteristic ions m/z 239, m/z 345 and neutral loss of 106 Da were used for monitoring the bond GA3. From the results, none bond of GA3 was none detected in Vigna radiata (L.) Wilczek’s leaves, roots and steams.
The proposed method offers not only a low level sensitivity to determine trace amount of GA3 in pulse, but also the ease of analysis with a simple treatment. In view of the simplicity, sensitivity and selectivity, the present method is recommendable for the determination of GA3 in plant for botany.

壹、緒論................................................1
1.1前 言...............................................1
1.2激勃素概述..........................................3
1.3激勃素的作用........................................4
1.4激勃素的結構........................................6
1.5相關法令規章........................................9
1.6激勃素的萃取興分離..................................9
1.7激勃素的檢測分析方法...............................11
1.8質譜儀儀器原理.....................................14
1.9綠豆及其植株.......................................24

貳、實驗部分...........................................27
2.1藥品、實驗器材和儀器設備...........................27
2.2藥品配置...........................................30
2.3探討最佳化條件之操作模式...........................32
2.4分析不同離子源分析激勃素之線性範圍、線性方程式、
線性相關係數及偵測極限.............................34
2.5綠豆中激勃素的萃取.................................34
2.6分析綠豆中激勃素之線性範圍、線性方程式、線性相關
係數及偵測極限.....................................35
2.7分析激勃素之回收率及精密度.........................36
2.8綠豆各部位中激勃素之代謝物偵測................,,...36
2.9真實樣品中激勃素之偵測.............................37

參、結果與討論......................,,.................38
3.1激勃素標準品溶液之液相層析質譜分析.................38
3.2液相層析質譜儀參數之探討...........................45
3.3不同離子源分析激勃素之線性範圍、線性分程式、線性
相關係數及偵測極限.................................68
3.4分析液相萃取結合液相層析質譜儀對激勃素之線性範圍
、線性分程式、線性相關係數及偵測極限...............68
3.5分析液相萃取結合液相層析質譜儀對激勃素之回收率及
精密度.............................................72
3.6真實樣品中激勃素之偵測.............................77
3.7綠豆中不同部位激勃素之偵測.........................77
3.8綠豆中激勃素之代謝產物偵測.........................77

肆、結論...............................................87

伍、參考文獻...........................................89

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