臺灣博碩士論文加值系統

貓爪藤廣泛使用在治療發炎性的疾病。貓爪藤成分中的羥&;#21554;&;#21722;生物鹼被視為最具生物活性之指標成分。本篇研究目的為建立一個微波輔助萃取方法，並結合高效能液相層析-光二極陣列檢測器及高效能液相層析質譜儀，用於快速萃取、定性以及定量貓爪藤成分中的主要指標成分羥&;#21554;&;#21722;生物鹼，包括rhynchophylline, pteropodine, isomitraphylline 及 isopteropodine。分析方法建立及確效後，接著開發貓爪藤固體分散劑型，藉由劑型開發而增加羥&;#21554;&;#21722;生物鹼的溶解度及累積溶離釋出率。
分析條件探討包括固定相選擇、移動相pH值、流速。微波輔助萃取以實驗設計法中的田口法設計實驗變因，微波輔助萃取優化參數如微波功率、萃取時間、萃取物與溶媒比例及離子液體種類。而固體分散劑型之賦形劑則會選出最適當的界面活性劑以及載體。分析方法優化結果，使用的管柱為Monolithic RP-18e (100 &;#180; 4.6 mm)，移動相為 pH 4.0的0.01 M醋酸銨溶液以及甲醇:乙&;#33096; (1:1, v/v)，流速設定為1.5 mL/min，偵測波長為245 nm，分析時間20分鐘，此分析方法具有良好線性關係 (r2 ≧ 0.9999)，高準確度 (99.6-100.6%)、精密度(RSD ≦ 1.8 %)及回收率 (≧ 98.3%)。微波輔助萃取實驗結果顯示以微波功率為900瓦，加熱時間60秒，而萃取物與溶媒比例為1:40 (w/v)，萃取溶媒則是使用1-hexyl-3-methylimidazolium bromide，可萃取出最高含量之羥&;#21554;&;#21722;生物鹼，萃取結果為傳統萃取之430%。固體分散劑型所使用的載體為聚羥亞烴188，而界面活性劑使用十二烷基硫酸鈉或1-甲基-2-?m咯烷酮，固體分散劑型之成品相較於對照組之溶解度提高290%，而在溶離試驗，累積溶離釋出百分比提高200%。
本研究開發之分析方法成功地應用於分析貓爪藤市售品及貓爪藤固體分散製劑。微波輔助萃取方法也能有效率地萃取出貓爪藤之有效成分羥&;#21554;&;#21722;生物鹼，而固體分散劑型的體外試驗結果顯示此固體分散劑型能提高溶解度及溶離釋出率。

Uncaria tomentosa (cat’s claw) is widely used for the treatment of inflammatory diseases. Oxindole alkaloids are regarded as the most important components responsible for the biological activities attributed to the plant. A rapid method was established using microwave-assisted extraction (MAE) combined with HPLC–PDA and HPLC-MS for the extraction, identification, and quantification of oxindole alkaloids (rhynchophylline, pteropodine, isomitraphylline, and isopteropodine) in U. tomentosa. After method established and validated, we developed a delivery system for enhancing the solubility and dissolution of oxindole alkaloids by solid dispersion technique.
We examined the effects of HPLC parameters, such as flow rate and pH of mobile phase. Orthogonal array design method was used to estimate optimum the MAE extraction conditions. The microwave parameters including irradiation power, extraction time, solid–liquid ratio and kinds of ionic liquids were optimized. Then, searching for a carrier and a surfactant to be the excipient for formulation of solid dispersion. The compounds were separated on a Monolithic Rp-18e column (4.6 × 100 mm), mobile phase containing 10 mM ammonium acetate solution (pH 4.0) and a mixture of methanol and acetonitrile (1:1, v/v), flow rate was 1.5 mL/min. Detection wavelength was set at 245 nm whthin 20 minutes. The HPLC method showed satisfactory linearity (r2 &;#61619; 0.9999), accuracy (99.6-100.6%), precision (RSD &;#61603; 1.8 %), and recovery (&;#61619; 98.3%). Based on oxindole alkaloids content, the maximum predicted oxindole alkaloids under the optimised conditions (900W irradiation power, 60 s extraction time, the ratio of solid to liquid was 1:40, and1-hexyl-3-methylimidazolium bromide as ionic liquid). The oxindole alkaloids content was 430% compared to traditional decoction method. The optimized formulations of U. tomentosa solid dispersion were successfully prepared within N- methylpyrrolidone to be the surfactant and poloxamer 188 to be the carrier. The solubility improvement reached 290% and cumulative dissolution percentage of the oxindole alkaloids was improved approximately 200% compared with the control group.
Finally, the new method was applied to investigate the content of oxindole alkaloids in U. tomentosa of commercial products and solid dispersion. MAE method effectively extract the oxindole alkaloids in U. tomentosa. And the result of in vitro studies showed that U. tomentosa solid dispersion enhanced the solubility and dissolution.

目錄 I
表次目錄 IV
圖次目錄 V
摘要 VII
Abstract VIII
縮寫表 X
壹、 緒論 1
一、 貓爪藤基本概述 1
(一) 貓爪藤介紹 1
(二) 貓爪藤藥理活性介紹 1
(三) 貓爪藤之活性成分 2
二、 文獻回顧 4
三、 微波輔助萃取法 8
(一) 微波原理介紹 8
(二) 微波輔助萃取應用 10
(三) 離子液體定義及其應用 11
四、 實驗設計法 14
五、 固體分散劑型 17
(一) 固體分散劑型概述 17
(二) 固體分散劑型成分釋出機轉 18
(三) 固體分散劑型製備方法 18
(四) 固體分散劑型常用載體介紹 22
六、 研究目的 25
貳、 實驗材&;#63934;與儀器設備 27
一、 試藥與試劑 27
(一) 標準品 27
(二) 貓爪藤粗萃物及市售品 27
(三) 微波輔助萃取溶媒 27
(四) 固體分散劑型之載體 28
(五) 固體分散劑型之界面活性劑 28
(六) 溶離試驗試劑 29
(七) 溶媒 29
二、 實驗儀器設備 30
三、 軟體 31
參、 實驗方法 33
一、 貓爪藤分析方法開發 33
(一) 建立高效能液相層析-光二極陣列檢測器 (HPLC-PDA)之分析方法 33
(二) 建立高效能液相層析質譜儀 (HPLC-MS)之分析方法 34
二、 微波輔助萃取法 34
(一) 微波輔助萃取法條件優化 34
(二) 傳統萃取法 36
三、 高效能液相層析-光二極陣列檢測器 (HPLC-PDA)之分析條件確效 38
(一) 檢量線之配置 39
(二) 同日、&;#63842;日精密度與準確度 39
(三) 回收率 40
四、 市售貓爪藤定量分析 42
(一) 超音波震盪法 42
(二) 微波輔助萃取法 42
五、 固體分散劑型製備 43
(一) 乙醇萃取物萃取方法 43
(二) 賦形劑篩選 43
(三) 固體分散劑型之配方 44
六、 製劑成品評估 46
(一) 溶解度試驗 46
(二) 溶離試驗 46
(三) 安定性試驗 48
七、 統計方法 48
肆、 實驗結果與討論 50
一、 貓爪藤分析方法開發 50
(一) 高效能液相層析-光二極陣列檢測器 (HPLC-PDA)之分析方法 50
(二) 高效能液相層析質譜儀 (HPLC-MS)之分析方法 53
二、 微波輔助萃取 61
(一) 微波輔助萃取法優化結果 61
(二) 傳統萃取法結果 62
三、 高效能液相層析-光二極陣列檢測器 (HPLC-PDA)之分析條件確效 73
(一) 檢量線之配置 73
(二) 同日、&;#63842;日精密度與準確度 74
(三) 回收率 74
四、 市售貓爪藤定量分析 78
(一) 超音波震盪法 78
(二) 微波輔助萃取法 78
五、 固體分散劑型製備 79
(一) 乙醇萃取物萃取方法 79
(二) 賦形劑篩選 79
(三) 固體分散劑型 82
六、 製劑成品評估 85
(一) 溶解度試驗 85
(二) 溶離試驗 85
(三) 安定性試驗 87
伍、 結論 105
陸、 參考文獻 107

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