臺灣博碩士論文加值系統

中文摘要
人參能夠減輕在術後或是藥物治療所造成的疲倦。根據先前研究發現在台灣的乳癌病患服用乳癌藥物tamoxifen時也會同時服用人參，此外，並還沒有任何文獻指出tamoxifen與人參是否具有交互作用。因此，本篇研究目的為建立一個快速且靈敏度好的超高效能液相層析串聯質譜儀 (UHPLC-MS/MS)分析方法，並將此分析方法用於定量tamoxifen與其代謝物，來探討人參對於tamoxifen與其代謝物的藥物動力學，並觀察兩者間是否有交互作用。
根據本試驗所建立的方法，tamoxifen與其代謝物會經由固定相的逆向層析管柱C18 column, 100mm x 2.1mm, 2µm，移動相所使用的有機相與水相分別為乙腈和10 mM的碳酸氫銨溶液，兩者的比例為90 :10，並利用多重反應監測模式偵測質譜的條件，結果分別為tamoxifen，m/z 372.20 → 72.30 (〔M + H〕+), N-desmethyltamoxifen，m/z 358.30 → 58.00 (〔M + H〕+), 4-hydroxytamoxifen，m/z 388.10 → 72.20 (〔M + H〕+), endoxifen，m/z 374.10 → 58.10 (〔M + H〕+) and ondansetron，294.00 → 170 (〔M + H〕+)，確立各物質的質譜條件。
將建立好的條件用來進行tamoxifen的藥物動力學分析，tamoxifen的AUC分別為1.31 ± 0.21, 1.33 ± 0.15和1.19 ± 0.14 hr µg/mL，結果顯示tamoxifen在有餵食人參與沒人參的大鼠組別中，在統計上並沒有顯著性的差異，然而在半衰期與平均滯留時間在人參組都有被延長的趨勢，此外，4-hydroxytamoxifen的AUC也在高劑量的人參組有顯著性上升的現象，而N-desmethyltamoxifen則是隨著人參劑量的提高，AUC的數值就跟著下降，說明tamoxifen與參鬚萃取物之間存在著交互作用，此結果也可給予臨床上關於tamoxifen與人參間交互作用的資訊。

Ginseng has not only been used as easing treatment-related fatigue, but has been co-administered with tamoxifen in Taiwan as well. Furthermore, no article related with the herb-drug interaction of ginseng and tamoxifen. Our investigation was to develop a fast and sensitive UHPLC-MS/MS method for the quantification of tamoxifen and its three main metabolites and pharmacokinetics study in rats after ginseng was administrated. Based on our validated method, tamoxifen and its metabolites were chromatographically separated on the reverse phase C18 endcapped column, and eluted on a isocratic program of acetonitrile-10 mM ammonium bicarbonate (90:10, v:v). The multiple reaction monitoring of m/z 372.20 → 72.30, m/z 358.30 → 58.00, m/z 388.10 → 72.20, m/z 374.10 → 58.10 and 294.00 → 170 were used to quantify the level of tamoxifen, N-desmethyltamoxifen , 4-hydroxytamoxifen, endoxifen and ondansetron, respectively. After oral administration 10 mg/kg of tamoxifen and low, medium, high dose of ginseng root extract, the pharmacokinetic results revealed that the area under the concentration versus time curve (AUC) of tamoxifen combined with the dose of ginseng root hair extract of 0.3 g/kg, 1g/kg, 3 g/kg were 1.31 ± 0.21, 1.33 ± 0.15 and 1.19 ± 0.14 hr µg/mL, respectively. The results demonstrated that AUC was no significant difference between administrating with or without ginseng. However, the half-life and mean retention time in ginseng group, tamoxifen were significant prolonged in rats. Besides, the AUC of 4-hydroxytamoxifen in high dose ginseng group increased significantly. The AUC of N-desmethyltamoxifen decreased with the increasing dose of ginseng. According to above results, Tamoxifen and its metabolites with ginseng root hair extract showed the herb-drug interaction between them. These results may provide some constructive information for clinical application about the interaction of tamoxifen and ginseng.

目錄
致謝 I
目錄 II
附表目錄 VI
附圖目錄 VII
Abstract VIII
中文摘要 X
第一章 緒論 1
第一節 Tamoxifen之簡介與相關研究 1
一、Tamoxifen簡介 1
二、Tamoxifen的副作用 1
三、Tamoxifen作用機制與代謝途徑 2
四、Tamoxifen研究背景 2
第二節 紅參之簡介與相關研究 3
一、紅參簡介 3
二、紅參之特性與功效 3
第三節 癌症在中醫之研究與人參在西醫上的應用 4
一、中醫對於癌症治療的策略 4
二、人參在西醫之相關研究 5
三、人參與癌症藥物之合併使用 6
四、Tamoxifen與人參合併使用的相關研究探討 6
第四節 超高效能液相層析串聯質譜儀 (UHPLC-MS/MS) 7
一、高效液相層析儀 8
二、偵測裝置 8
第五節 分析方法確效 9
一、選擇性 (selectivity) 10
二、線性關係 (linearity) 10
三、準確性 (accuracy)與精確性 (precision) 10
四、基質效應 (matrix effect)與回收率 (recovery) 10
五、靈敏度 (sensitivity) 11
六、安定性 (stability) 11
第六節 藥物動力學 11
一、概論 11
二、給藥途徑與定義 12
三、藥物動力學模式 14
四、藥物動力學參數 15
第二章 研究動機與目的 17
第三章 實驗材料與研究方法 18
第一節 實驗材料與化學試劑 18
一、化學試藥 18
二、手術器械與實驗儀器 18
三、實驗動物 19
四、應用軟體 20
第二節 紅參鬚根成分萃取 21
第三節 紅參在Tamoxifen及其代謝物上的藥物動力學作用 21
一、超高效能液相層析串聯質譜儀之分析條件 21
二、標準品與藥物劑量配置 20
三、動物模式與實驗程序 20
四、大鼠給藥與採血時間 21
五、生物檢品的前處理與檢量線配置 21
六、分析方法之確效 23
七、數值分析與統計軟體 25
第四章 實驗與結果 27
第一節 超高效能液相層析串聯質譜儀之分析方法確立 27
第二節 分析方法之確效 28
一、標準品之分析方法 28
二、生物檢品之基質效應與萃取回收率 28
三、準確性與精確性之評估 29
四、穩定度試驗之檢測 30
第三節 紅參鬚根萃取物對Tamoxifen及其代謝物在大鼠體內之藥物動力學 30
第四節 紅參萃取物對Tamoxifen在大鼠體內之藥物動力學比較 32
第五章 結果討論 34
第一節 Tamoxifen與其代謝物分析方法的建立 34
第二節 分析方法之確效 34
第三節 生物檢品前處理 35
第四節 紅參鬚根萃取物對Tamoxifen與其代謝物在大鼠體內之藥物動力學 36
第五節 影響Tamoxifen代謝的細胞色素P450與紅參間的探討 37
第六章 結論 40
第七章 參考文獻 41
附表 45
附圖 52


附表目錄
Table.1 The UHPLC-MS/MS conditions for the identification of the constituents 45
Table. 2 Linear ranges, calibration curves, correlation coefficients (r2 ), and detection limits of four constituents in rat plasma using UHPLC-MS/MS 46
Table. 3 Matrix effect and recovery of tamoxifen and its metabolites in rat plasma 47
Table 4. Intra-day precision and accuracy of tamoxifen and its metabolites 48
Table 5. Inter-day precision and accuracy of tamoxifen and its metabolites 49
Table 6. Stability of tamoxifen and its metabolites in rat plasma 50
Table 7. Pharmacokinetic parameters of tamoxifen and its metabolites in rats 51


附圖目錄
Figure 1. The mass spectra spectrometry and structure of (A) tamoxifen (B) endoxifen (C) N-desmethyltamoxifen (D) 4-hydroxytamoxifen (E) ondansetron 52
Figure 2. Representative MRM chromatograms of (A) blank plasma sample；(B) blank plasma sample spiked with tamoxifen (5 ng/mL)；(C) plasma sample at 30 min after administration of tamoxifen (10 mg/kg, p.o.) 53
Figure 3. Representative MRM chromatograms of (A) blank plasma sample；(B) blank plasma sample spiked with 4-hydroxytamoxifen (10 ng/mL)；(C) plasma sample at 4hr after administration of 4-hydroxytamoxifen (10 mg/kg, p.o.) 54
Figure 4. Representative MRM chromatograms of (A) blank plasma sample；(B) blank plasma sample spiked with N-desmethyltamoxifen (50 ng/mL)；(C) plasma sample at 6hr after administration of N-desmethyltamoxifen (10 mg/kg, p.o.) 55
Figure 5. Representative MRM chromatograms of (A) blank plasma sample；(B) blank plasma sample spiked with endoxifen (50 ng/mL)；(C) plasma sample at 6hr after administration of endoxifen (10 mg/kg, p.o.) 56
Figure 6. Representative MRM chromatograms of (A) blank plasma sample；(B) blank plasma sample spiked with ondansetron (10 ng/mL)；(C) plasma sample at 2hr after administration of ondansetron(10 mg/kg, p.o.) 57
Figure 7. Mean plasma Concentration-time curve of tamoxifen in rat blood after tamoxifen administration (10 mg/kg, p.o. ●) alone and tamoxifen with dose of ginseng root hair extract (0.3 g/kg/day for 5 consecutive days, p.o.) (◯), ginseng root hair extract (1g/kg/day for 5 consecutive days, p.o.)(▼) and ginseng root hair extract (3 g/kg/day for 5 consecutive days, p.o.) (△). 58
Figure 8. Mean plasma Concentration-time curve of N-desmethyltamoxifen in rat blood after tamoxifen administration (10 mg/kg, p.o. ●) alone and tamoxifen with dose of ginseng root hair extract (0.3 g/kg/day for 5 consecutive days, p.o.) (◯), ginseng root hair extract (1g/kg/day for 5 consecutive days, p.o.)(▼) and ginseng root hair extract (3 g/kg/day for 5 consecutive days, p.o.) (△). 59
Figure 9. Mean plasma Concentration-time curve of 4-hydroxytamoxifen in rat blood after tamoxifen administration (10 mg/kg, p.o. ●) alone and tamoxifen with dose of ginseng root hair extract (0.3 g/kg/day for 5 consecutive days, p.o.) (◯), ginseng root hair extract (1g/kg/day for 5 consecutive days, p.o.)(▼) and ginseng root hair extract (3 g/kg/day for 5 consecutive days, p.o.) (△) 60

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