臺灣博碩士論文加值系統

Tranexamic acid 是皮膚美白化妝品配方的重要試劑，及抑制分解纖維蛋白凝塊的抗血紅蛋白藥物，早期具有止血抗炎的藥理效果。
Tranexamic acid 對紫外光、可見光波長 190~800 nm 之吸光性非常弱，以致於無法以紫外光、可見光偵測器直接予以檢測之。本實驗以 FMOC ( 9-Fluorenylmethyl chloroformate )衍生試劑，在鹼性之緩衝水溶液中與 Tranexamic acid 之胺基反應成衍生物，並建立一套新的HPLC分析方法來檢測水膠面膜中Tranexamic acid的美白成分；移動相為10mM NH4OAc 緩衝水溶液pH 6.7/ 甲醇比例為30 / 70，撘配ODS Hypersil 250×4.6 mm(5 μm)的層析管柱，偵測波長為268 nm，流速為1.0 mL/min，注射量為20 μL，此分析方法可在9分鐘內完成。
在分析方法確效方面，Tranexamic acid的線性迴歸係數R2 = 0.9999，精密度實驗的相對標準偏差( RSD )介於0.01 % ~ 0.31 %之間，準確度實驗的回收率則介於98.0 % ~ 100.0 %。
在經皮穿透試驗方面，將自製的 Tranexamic acid 水膠面膜在 Nylon 濾膜及豬皮進行穿透性實驗，Tranexamic acid 在 3 hr 後的釋離率分別為 17.35 % 和 6.53 %，而相對標準偏差( RSD)分別在 5.71 % 和 8.60 % 以下，穿透試驗的結果可以當作藥物經皮使用的參考依據。

將自製的 Tranexamic acid 水膠面膜在室溫( 25±2℃)及加速( 40±2℃)的安定性試驗條件下進行6個月實驗，室溫試驗的Tranexamic acid含量衰退至94.92 %，而加速試驗的Tranexamic acid含量衰退至90.72 %。

Tranexamic acid is an important reagent in cosmetic skin-whitening formulation, and is an antifibrinolytic drug that inhibits breakdown of fibrin clots, early in the pharmacological of anti-inflammatory effect with bleeding.
The molecule of tranexamic acid has very week absorption at the ultraviolet and visible wavelength from 190 to 800 nm, so that it is impossible to directly detect tranexamic acid with the UV-Vis detector.
In this research, the derivative reagent of FMOC (9-fluorenylmethyl chloroformate) was to use to react with the amino functional group of tranexamic acid at the alkaline condition and to form the UV detectable compound. A HPLC method was developed to determine the whitening ingredient of tranexamic acid in hydrogel mask. The mobile phases were prepared with 10mM NH4OAc buffer solution adjusted at pH 6.7 and methanol with the ratio of 30 to 70. Chromatographic methods was performed with a HPLC system equipped with ODS Hypersil 250×4.6mm(5 μm) column with UV/Vis absorption set at 268 nm, flow rate set at 1.0 mL/min and an injection of 20 μL as well as each runtime about nine minutes.
In the result of analytical validation, the linear regression coefficient (R2) was 0.9999, the relative standard deviations(RSD) of precision experiment were between 0.01 %~0.31 %, and the recoveries of the accuracy experiment were between 98.0 %~100.0 %.
In the tests of transdermal penetration, the hydrogel masks of tranexamic acid were placed in nylon membrane and pig skin to do penetrating experiments, the release rates of tranexamic acid for three hours for nylon membrane and pig skin were 17.35 % and 6.53 %, respectively. The relative standard deviations(RSD) of penetration tests for nylon membrane and pig skin were 5.71 % and 8.60 % below, respectively. The results of penetration tests can be used as the reference of transdermal medicine.
In this experiment, the tranexamic acid hydrogel mask was placed in the room temperature condition(25±2℃) and the accelerated condition (40±2℃) for six-month stability tests, the content of tranexamic acid was declined to 94.92 % in the room temperature test and was declined to 90.72 % in the accelerated test.

中文摘要………………………………………………………………………...I
英文摘要……………………………………………………………………….III
誌謝……………………………………………………………………………..V
目錄……………………………………………………………………………VI
表目錄………………………………………………………………………....IX
圖目錄…………………………………………………………………………XI
第一章 緒論…………………………………………………………..1
1.1 前言………………………………………………………………………...1
1.2 研究目的…………………………………………………………………...3
第二章 文獻整理……………………………………………………..4
2.1 皮膚的構造………………………………………………………………...4
2.2 黑色素的生成與作用機制………………………………………………...9
2.3 什麼是面膜……………………………………………………………….12
2.4 傳明酸( Tranexamic acid )的簡介………………………………………..15
2.5 衍生化反應與應用……………………………………………………….19
2.6 藥物經皮吸收系統……………………………………………………….21
2.7 儀器原理分析…………………………………………………………….26
第三章 實驗………………………………………………………..34
3.1 藥品與試劑………………………………………………………….…….34
3.2 儀器設備…………………………………………………………….…….35
3.3 藥品配置……………………………………………………………….….37
3.4 最佳化衍生條件…………………………………………………….…….38
3.5 分析方法與條件建立……………………………………………….…….38
3.6 最佳化衍生條件之建立…………………………………………….…….42
3.7 衍生物安定性的探討……………………………………………….…….44
3.8 分析方法的確效…………………………………………………….…….44
3.9 經皮穿透試驗……………………………………………………….…….46
3.10 面膜的製備………………………………………………………….…...47
3.11 空白面膜的製備…………………………………………………….…...48
3.12 不同溫度及時間對水膠面膜的影響…………………………………....49
3.13 豬皮前處理……………………………………………………………....49
3.14 面膜中成份萃取法……………………………………………………....50
3.15 成份含量計算……………………………………………………….…...51
第四章 結果與討論…………………………………………….…..52
4.1 分析方法與條件之建立……………………………………………….….52
4.2 最佳化衍生條件之建立……………………………………………….….66
4.3 衍生物安定性的探討………………………………………………….….70
4.4 分析方法的確效……………………………………………………….….71
4.5 面膜成品的經皮穿透試驗………………………………………….…….80
4.6 不同溫度與時間對面膜含量的影響……………………………….…….83
4.7 不同溫度與時間對面膜成品外觀的影響…………………………….….86
第五章 結論…………………………………………………….…..87
參考文獻………………………………………………………….…88













表目錄
表1. 抑制黑色素生成之美白成分………………………………………….11
表2. 常用面膜成分及功能說明…………………………………………….14
表3. 為經由行政院衛生署查詢之相關廠商製作 Tranexamic acid 美白面膜……………………………………………………………………….18
表4. 正相層析與逆相層析之特性比較…………………………………….29
表5. Tranexamic acid水膠面膜的組成份………………………………….48
表6. 不同有機溶劑的比較………………………………………………….52
表7. 不同有機溶劑比例的比較…………………………………………….54
表8. 不同緩衝溶液成分及 pH 值的比較………………………………....57
表9. 不同緩衝溶液 pH 值的比較………………………………………....60
表10.不同衍生化緩衝水溶液 pH 值的比較……………...…………..…...63
表11.最適當的 HPLC 分析條件……………………………………….......65
表12.自製面膜內容物的標準品與樣品滯留時間………………………….71
表13.系統適用性結果數據( n=6 )…………………………………………..73
表14. Tranexamic acid 三重複注射結果…………………………………...74
表15. Tranexamic acid 標準溶液經 HPLC 分析之重複性數據( n=3 )…..76
表16. Tranexamic acid 標準溶液經 HPLC 分析之再現性數據( n=3 ) 1st day………………………………………………………………………76
表17. Tranexamic acid 標準溶液經 HPLC 分析之再現性數據( n=3 ) 2nd day…………………………………………………………………........77
表18. Tranexamic acid 標準溶液經 HPLC 分析之再現性數據( n=3 ) 3rd day……………………………………………………………………....77
表19.Tranexamic acid 的回收率數據………………………………….…...78
表20. Tranexamic acid 標準溶液經 HPLC 之數據( n=10 )…………….…79
表21. HPLC 對 Tranexamic acid 的偵測極限和定量極限…………….….80
表22.面膜成品在 Nylon 濾膜之經皮穿透試驗據數……………………....81
表23.面膜成品在豬皮樣品之經皮穿透試驗據數…………………………..82
表24. Tranexamic acid 在 25℃及40℃的環境下，6個月間的含量變化..83
表25. Tranexamic acid 在 25℃時的安定性數據…………………………..84
表26. Tranexamic acid 在 40℃時的安定性數據…………………….…….85
表27. Tranexamic acid 面膜成品在 25℃及40℃的環境下，6個月間的顏色與外觀變化………………………………………………………….…86






圖目錄
圗1. 皮膚的構造………………………………………………………………5
圗2. 皮膚表皮層結構圖…………………………………………..…………...6
圖3. 皮膚表皮構造…………………………………………………..………...7
圖4. 黑色素生成的反應途徑………………………………………….……..10
圖5. 美白成分對黑色素之作用方式………………………………….……..12
圖6. 傳明酸( Tranexamic acid )分子結構式…………………………….…..16
圖7. FMOC ( 9-Fluorenylmethyl chloroformate ) 和 Tranexamic acid衍生化之反應式…………………………………………………………….…..20
圖8. 藥物經皮吸收之過程……………………………………………….…..21
圖9. 角質層磚牆結構示意圖………………………………………………...22
圖10.藥物穿透皮膚路徑圖………………………….………………………..24
圖11. HPLC 儀器圖…………………….……………………………………30
圖12.滲透裝置……………………………………….………………………..31
圖13.串聯式滲透裝置………………………………………….……………..32
圖14.恆溫循環水槽………………………………………….………………..32
圖15a. Tranexamic acid 衍生反應前的 UV 吸收圖……………...….…….39
圖15b. Tranexamic acid 衍生反應後的 UV 吸收圖…………………….…40
圖16. 直立式恆溫反應槽示意圖…………………………………………….47
圖17.自製的水膠面膜………………………………………………………..49
圖18. Tranexamic acid 在波長 268 nm，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/乙腈 = 50 / 50 的層析圖…………………………..53
圖19. Tranexamic acid 在波長 268 nm，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 50 / 50 的層析圖….……………………...53
圖20. Tranexamic acid 在波長 268 nm，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 50 / 50 的層析圖………………………....55
圖21. Tranexamic acid 在波長 268 nm，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 40 / 60 的層析圖…………………………55
圖22. Tranexamic acid 在波長 268 nm，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 30 / 70 的層析圖………………………....56
圖23. Tranexamic acid 在波長 268 nm，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 20 / 80 的層析圖……..…………………..56
圖24. Tranexamic acid 在波長 268 nm，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 30 / 70 的層析圖…………………..……..58
圖25. Tranexamic acid 在波長 268 nm，移動相為 10 mM Na2HPO4 緩衝水溶液 ( pH 6 )/ 甲醇 = 30 / 70 的層析圖……………………..……..58
圖26. Tranexamic acid 在波長 268 nm，移動相為 10 mM Na2HPO4 緩衝水溶液 ( pH 7 )/ 甲醇 = 30 / 70 的層析圖…..……………………….59
圖27. Tranexamic acid 在移動相 pH 5.0，波長 268 nm 的層析圖...........60
圖28. Tranexamic acid 在移動相 pH 6.7，波長 268 nm 的層析圖...........61
圖29. Tranexamic acid 在移動相 pH 8.0，波長 268 nm 的層析圖...........61
圖30. Tranexamic acid 在波長268 nm，稀釋溶液為 10mM Na2HPO4 ( pH 9 )，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 30 / 70 的層析圖…………………………………………….……....63
圖31. Tranexamic acid 在波長268 nm，稀釋溶液為10mM Na2HPO4 ( pH 10 )，移動相為 10mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 30 / 70 的層析圖…………………………………………….………64
圖32. Tranexamic acid 在波長268 nm，稀釋溶液為10mMNa2HPO4 ( pH11 )，移動相為 10 mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 30 / 70 的層析圖…………………….………………………………64
圖33. Tranexamic acid 在波長268 nm，稀釋溶液為10mM Na2HPO4 ( pH 12)，移動相為 10mM NH4OAc 緩衝水溶液 ( pH 6.7 )/ 甲醇 = 30 / 70 的層析圖………………………………………………….....65
圖34.不同衍生試劑濃度的比較…………………………………………..…66
圖35.以不同 Vortex 混合時間對衍生物的影響……………………..…….67
圖36.不同反應溫度對衍生物的影響…………………………………….......68
圖37.不同反應時間對衍生物的影響…………………………………..……69
圖38. Tranexamic acid-FMOC 衍生物安定性試驗………………………....70
圖39. Tranexamic acid 標準品滯留時間之層析圖………………………….71
圖40. Tranexamic acid 樣品滯留時間之層析圖………………………….…72
圖41. Tranexamic acid 標準品之 UV 吸收圖( HPLC-PDA )………….…..72
圖42. Tranexamic acid 樣品之 UV 吸收圖( HPLC-PDA )…………….…..73
圖43. Tranexamic acid 標準品之檢量線圖……………………………….…75
圖44.空白面膜樣品萃取溶液衍生後之層圖………………………………...78
圖45.面膜成品在Nylon 濾膜之經皮穿透曲線圖……………………..……81
圖46.面膜成品在豬皮樣品之經皮穿透曲線圖……………………….….….82
圖47.Tranexamic acid 在 25℃及40℃的環境下，6個月間的含量變化曲線圖………………………………………………………………….……84

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