臺灣博碩士論文加值系統

紅景天是一種生長在歐洲及亞洲高海拔的多用途植物，傳統醫
學長久以來以其根莖做為增強耐力的強壯劑、抗發炎、預防高山症。目前已成為重要的健康輔助食品之一，由於具有強力抗氧化能力，多用於抗老化、增強記憶力、抗疲勞、提升工作能力等。
使用超臨界流體CO2當溶媒，乙醇為共同助萃劑，探討於不同條件下，從中國傳統藥材紅景天中，萃取指標成分紅景天苷之最佳條件。利用HPLC檢測抽出物，換算個別條件下紅景天苷含量之多寡。經實驗得知，以添加乙醇為共萃劑、溫度40 ℃、壓力340 Bar，抽取時間靜態40分鐘，動態40分鐘，為最佳之超臨界萃取條件。
依此條件萃取紅景天藥材，將萃取物分成三批，各混合三種不同比例(zero-low-high/0-30-50%)之Avicel PH102和Chitosan造粒，利用擠壓搓圓技術，製備圓粒。經體外溶離度試驗之研究，分析溶離曲線，結果以含30 % Chitosan之圓粒製劑處方，可獲致最理想之緩釋效果。
本研究得到紅景天每次服用劑量的科學定量與評估方法，能夠進一步開發一種方便服用，並能達到維持體內藥物濃度恆定的紅景天長效緩釋劑型。

Rhodiola rosea L. is a kind of multipurpose plant at altitudes in Europe and Asia. It has been a long time since were used in the traditional medical system, as an adaptogen, anti-inflammation, altitude sickness prevention. With the strongly anti-oxidative property, high activity of anti-fatigue and great efficacy for improving physical working capacity, today it has become an important component of dietary supplements.
In order to find out the best extract condition we extracted the salidroside ingredients from Rhodiola rosea with CO2 supercritical fluid extraction in Chinese Traditional Medicine. The HPLC was applied to the determination of salidroside in all different experimental conditions. In our experiments we got the best one if it was under the condition with extractor pressure in 340 bar, extractor temperature 40℃, ethanol for a co-solvent, and the extraction time 80 minutes. Using the best extraction condition for an extract. To prepared the Rhodiola pellets by extrusion-spheronisation. The chitosan load levels were zero-low-high/0-30-50%. Type of filler recipient was Avicel (PH102). Then to investigate the effect of pellets on the drug released upon dissolution test in vitro. The result showed that the pellets which contained 30 % low load level of chitosan were more suitable for a Rhodiola rosea sustained release pellets.
In this study we obtained a scientific qualitative method of Rhodiola dosage. It can be an expedient oral dosage form with long-lasting released effect that will be able to maintain the concentration of Rhodiola in blood.

目 錄
圖次目錄-------------------------------------------------Ⅰ
表次目錄-------------------------------------------------Ⅲ
中文摘要-------------------------------------------------IV
英文摘要-------------------------------------------------Ⅴ
壹、 緒論-------------------------------------------------1
一、 紅景天藥材簡介----------------------------------1
(一) 紅景天的藥理活性----------------------------4
(二) 紅景天於古書典籍的記載以及近代研究情形之探討--------------------------------------------7
(三) 從紅景天植物中分離出的化學成---------------11
二、 超臨界流體萃取技術-----------------------------13
(一) 超臨界流體的定義及性質---------------------14
(二) 超臨界流體萃取混合物的條件-----------------16
(三) 超臨界流體的溶解力-------------------------18
(四) 超臨界流體技術的應用-----------------------20
三、 圓粒製劑擠壓搓圓技術---------------------------22
(一) 擠壓搓圓方法其製程可分為五個步驟-----------22
(二) 藥物圓粒劑型（pellets）之優勢--------------23
（三）藥物釋出機轉評估----------------------------24
（四）賦形劑--------------------------------------26
貳、 研究動機與目的--------------------------------------28
叁、 實驗材料與研究方法----------------------------------30
一、實驗材料-----------------------------------------30
二、化學試劑-----------------------------------------31
三、儀器設備-----------------------------------------32
四、實驗方法與步驟-----------------------------------34
(一) 藥材鑑別：組織鏡檢方法-------------------------34
(二) 藥材檢品製備方法-------------------------------35
(三) 市售藥材成分鑑別-------------------------------35
(四) 市售粉末及濃縮製劑指標成分分析-----------------35
(五) 超臨界流體萃取法實驗---------------------------36
A. 純CO2抽提法(non co-solvent extraction)-------36
B. 共萃劑萃取法(Co-solvent extraction )----------37
(六) 高效液相層析法(HPLC)分析方法-------------------38
1. 建立標準品檢量線--------------------------38
2. 超臨界流體萃取物之指標成分含量分析--------39
3. 以methanol直接抽提法抽提樣本之定量分析---39
五、緩釋圓粒劑型製備方法------------------------------40
(一) 紅景天藥材有效成分之超臨界抽提-------------40
(二) 萃取物之處理-------------------------------41
(三) 圓粒劑型之製備-----------------------------42
1. 以紅景天超臨界流體萃取液製作圓粒製劑的方法-42
2. 體外溶離度實驗步驟-------------------------43
3. 圓粒劑型之HPLC定量分析實驗步驟------------44
4. 藥物成分溶離累積量之計算-------------------45
肆、 結果與討論------------------------------------------46
一、紅景天藥材外觀及組織切片鏡檢結果-----------------46
1.外部型態---------------------------------------46
2.組織切片---------------------------------------47
二、市售藥材成分鑑別---------------------------------49
三、市售粉末及濃縮製劑指標成分分析-------------------50
四、超臨界流體萃取實驗結果---------------------------51
五、高速液相色層分析方法(HPLC)之結果-----------------54
(一) 利用HPLC方法製作檢量線----------------------54
(二) 紅景天苷分析方法確效實驗----------------------56
六、紅景天緩釋劑型圓粒製劑開發實驗結果之探討---------66
(一) 紅景天有效成分萃取----------------------------66
(二) 紅景天擠壓搓圓緩釋劑型開發結果----------------68
伍、 結論------------------------------------------------76
陸、 參考文獻--------------------------------------------78

圖目錄

圖一、紅景天Rhodiola rosea L. (Crassulaceae)之植株----------3
圖二、紅景天Rhodiola rosea L. (Crassulaceae)之藥材--------------3
圖三、紅景天五種重要活性成分及其結構式--------------------11
圖四、超臨界流體萃取儀（Supercritical Fluid extraction；SFE）------13
圖五、純物質的三相點、臨界點與超臨界狀態的關係-------------14
圖六、紅景天根組織詳圖------------------------------------47
圖七、紅景天根組織全圖------------------------------------48
圖八、紅景天根組織詳圖(100倍)----------------------------48
圖九、紅景天市售藥材萃取液--------------------------------53
圖十、紅景天SFE co-solvent萃取法所得萃取液----------------53
圖十一、紅景天SFE non co-solvent萃取法所得萃取液------------53
圖十二、紅景天苷(Salidroside)標準品之HPLC 檢量線-----------56
圖十三、Salidroside標準品之HPLC層析圖譜-------------------60
圖十四、紅景天超臨界流體萃取物之HPLC層析圖譜-------------61
圖十五、紅景天以甲醇萃取之HPLC層析圖譜-------------------61
圖十六、以SFE萃取條件為30 ℃ / 270psi / NON CO-SOLVENT
之層析圖譜---------------------------------------63
圖十七、以SFE萃取條件為30 ℃ / 270psi / CO-SOLVENT之層
析圖譜-------------------------------------------63
圖十八、以SFE萃取條件為40 ℃ / 340psi / CO-SOLVENT之層
析圖譜-------------------------------------------64
圖十九、紅景天SFE萃取物濾液之HPLC層析圖譜---------------67
圖二十、紅景天SFE 萃取物沉澱物之HPLC層析圖譜-------------67
圖二十一、chitosan在三種不同配方下製成2 mm的pellets後之溶離釋出%曲線圖----------------------------------73
圖二十二、圓柱型擠壓搓圓機--------------------------------74
圖二十三、正擠壓出之紅景天棒狀物--------------------------74
圖二十四、擠壓完成圖--------------------------------------74
圖二十五、於圓盤搓圓機中搓圓------------------------------74
圖二十六、紅景天緩釋圓粒製劑製備完成圖--------------------75

表目錄

表一、已知紅景天化學成分分類表----------------------------12
表二、為超臨界流體與氣體、液體的物理性質比較表------------15
表三、為常用之超臨界流體其臨界溫度、壓力與密度之關係------17
表四、紅景天圓粒製劑處方內容------------------------------43
表五、市售藥材成分鑑別比較表-----------------------------49
表六、市售紅景天粉末、濃縮藥粉調查分析結果----------------50
表七、紅景天藥材以超臨界流體萃取實驗，所得結果-------------52
表八、紅景天苷於不同濃度條件之HPLC層析數據---------------55
表九、紅景天苷標準品同日間精密度(Inter-day precision)再現性之確效分析實驗(n = 3)-------------------------------58
表十、紅景天苷標準品異日間精密度(Intra-day precision)再現性之確效分析實驗(n = 3)-------------------------------59
表十一、紅景天藥材以超臨界流體萃取法，在不同的萃取條件下所抽取之紅景天苷產率比較表-----------------------------65
表十二、不同配方紅景天圓粒製劑，其溶離度(%)實驗結果-------69
表十三、以紅景天圓粒製劑配方1所得之溶離實驗結果----------70
表十四、以紅景天圓粒製劑配方I1所得之溶離實驗結果---------71
表十五、以紅景天圓粒製劑配方II1所得之溶離實驗結果--------72

Abidov M., Grachev S., Seifulla RD., Ziegenfuss TN., Extract of Rhodiola rosea radix reduces the level of c-reactive protein and creatinine kinase in the blood. Bull Exp Biol Med. 2004 ; 138 (1) : 63 - 64.

Abidov M., Crandal F., Grachev S., Seifulla RD., Ziegenfuss TN., Effect of extracts from Rhodiola rosea and Rhodiola crenulata (Cressulaceae) root on ATP content in mitochondria of skeletal muscles. Bull Exp Biol Med. 2003 ; 136 (6) : 585 - 587.

Afanas’ev S.A., Krylatov A.V., Lasukova T.V., Lishmanov Y.B.,
Role of inducible stress proteins in cardioprotective effects of
Rhodiola rosea. Biokhimiya. 1996 ; 61 : 1779 - 1784.

Azizov AP., and Seifulla RD., The effect of elton, leveton, fitoton and adapton on the work capacity of experimental animals. Eksp Klin Farmakol. 1998 ; 61 : 61 - 63.

Bocharova O.A., Matveev B.P., Baryshnikov A.I., Figurin K.M.,
Serebriakova R.V., Bodrova N.B., The effect of a Rhodiola rosea extract on the incidence of recurrences of a superficial bladder cancer. Urol Nefrol. 1995 ; 2 : 46 - 47.

Bocharova O.A., Serebriakova R.V., Bodrova N.B., Preventive effect of
Rhodiola rosea in spontaneous liver carcinogenesis in a mice model of
high-tumor strain. Vestn Ross Akad Med Nauk. 1994 ; 5 : 41 - 43.

De B. K., eEijnde B.O., Ramaekers M., Hespel P., Acute Rhodiola rosea intake can improve endurance exercise performance. Int Nutr exerc Metab. 2004；14 (3) : 298- 307.

Dement’eva L.A., and laramenko K.V., Effect of a Rhodiola extract on the tumor process in an experiment. Vapr Onkol. 1987；33 (7) : 57 - 60

Felt O., Buri P., Gurny R., Chitosan: A unique polysaccharide for drug delivery. Drug Dev Ind Pharm. 1998 ; 24: 979 -/993.

Furmanowa M., Hartwich M., Alfermann A.W., Kozminski W., Olejnik M., Rosavin as a product of glycosylation by Rhodiola rosea (rose root) cell cultures. Plant Cell Tissue Organ Cult. 1999a ; 56 : 105 - 110.

Furmanowa, M., Kedzia, B., Hartwich, M., Kozlowski, J., Krajewska-
Patan, A., Mscisz, A., Jankowiak, J., Phytochemical and pharmacological properties of Rhodiola rosea L. Herba Pol. 1999b ; 45 (2) : 108 - 113.

Illum, L., Chitosan and its use as a pharmaceutical excipient. Pharm Res. 1998 ; 15 : 1326 -/1331.

Jittima C., Fridrun P., Michael J.N., The influence of chitosan and sodium alginate and formulation variables on the formation and drug release from pellets prepared by extrusion/spheronisation. Int J Pharma. 2004 ; 275: 41 - 60.

Korsmeyer R.W., Gurny R., Doelker E., Buri, P. and Peppas, N. A., Mechanisms of solute release from porous hydrophilic polymer. Int J Pharm. 1983 ; 15：25-35.

Kristmundsdo´ ttir T., Ingvarsdo´ ttir K., Saemundsdo´ ttir G., Chitosan matrix tablets: the influence of excipients on drug release. Drug Dev Ind Pharm. 1995 ; 21 : 1591 -/1598.

Lee P.I., Kinetics of drug release from hydrogel matrices. J Control Release, 1985；2：277-288.
Lishmanov I.B., Trifonova Z.V., Tsibin A.N., Maslova L.V., Dement’eva L.A., Plasma beta-endorphin and stress hormones in stress and adaptation. Bull Exp Biol Med 1987 ; 103 : 422 - 424.

Maslova L.V., Kondrat’ev B.I., Maslov L.N., Lishmanov I.B.,The cardioprotective and antiadrenergic activity of an extract of Rhodiola rosea in stress. Eksp Klin Farmakol. 1994 ; 57 : 61- 63.

Markus G., Yurdanur Y., and Ikhlas A., Khan, Analysis of the Marker Compounds of Rhodiola rosea L. (Golden Root) by Reversed Phase High Performance Liquid Chromatography. Chem Pharm Bull. 2001 ; 49 (4) : 465 - 467.
Ohsugi M., Fan W., Hase K., Xiong Q., Tezuka Y., Komatsu K., Namba
T., Saitoh T., Tazawa K., and Kadota S.. Active-oxygen scavenging
activity of traditional nourishing-tonic herbal medicines and active
constituents of Rhodiola sacra. J Ethnopharmacol. 1999 ; 67 (1) : 111 –
119.

Paul W., Sharma C.P., Chitosan, a drug carrier for the 21st century : a review. S.T.P. J Pharm Sci. 2000 ; 10 : 5 -/22.

Peppas N. A., and Sahlin J.J., A simple equation for description of solute relase. III. Coupling of diffusion and relaxatio. J Control Release. 1987a ; 5：23 - 36.

Petkov, V.D., Yonkov, D., Mosharo., A., Kambourova, T., Alova, L., Petkov, V.V., Todorov, I., E.ects of alcohol aqueous extract from Rhodiola rosea L. roots on learning and memory. Acta Physiol Pharmacol Bulg. 1986 ; 12 : 3 - 16.

Razina T.G., Zueva E.P., Amosova E.N., Krylova S.G., Medicinal herb preparations as a means of additional therapy:Experimental oncology evidence. Eksp Klin Farmakol. 2000 ; 63 : 59–61.

Richard P.B. M.D., Patricia L.G. M.D., and Zakir R., Rhodiola rosea: A Phytomedicinal Overview. J Am Botanic Coun. 2002；56 : 40 - 52.

Ritger P. L., and Peppas N. A., A simple equation for description of solute relase. II. Fickian and anomalous release from swellable devices. J Control Release. 1987b ; 5：37 – 42

Rohloff J., Vollatiles from rhizomes of Rhodiola rosea L. Phytochemistry. 2002 ; 59 : 655 – 661

Roberts D.D., Pollien P., Milo C., Solid-phase micro-extraction method development for headspace analysis of volatile .avor compounds. J Agric Food Chem. 2000 ; 48 : 2430 - 2437.

Roberta D.S., Roberta D.B., Carla S., Umberto M., Luigi C., Marina D., In vitro protective effect of Rhodiola rosea extract against hypochlorous acid-induced oxidative damage human erythrocytes. Biofactors. 2004；20：147-159.

Santos H., Veiga F., Pina M., Podczeck F., Sousa J., Physical properties
of chitosan pellets produced by extrusion-spheronization: influence of
formulation variables. Int J Pharm. 2002 ; 246：153 - 169.

Shevtsov V.A., Zholus B.I., Shervarly I., Vol’skij V. B., and Korovin Y. P., Arandomized trial of two different doses of a SHR-5 Rhodiola rosea extract versus placebo and control of capacity for mental work. Phytomedicine. 2003 ; 10 : 95 - 105.

Sokolov, S.Y., Ivashin, V.M., Zapesochnaya, G.G., Kurkin, V.A.,
Shchavlinskii, A.N., 1985. Studies of neurotropic activity of new
compounds isolated from Rhodiola rosea. Khim Farm Zh. 19,
1367- 1371.

Spasov A.A., Wikman G.K., Mandrikov V.B., et al., A double-blind,
placebo-controlled pilot study of the stimulating and adaptogenic effect of
Rhodiola rosea SHR-5 extract on the fatigue of students caused by stress
during an examination period with a repeated low-dose regimen.
Phytomedicine. 2000 ; 7 : 85 - 89.

Tolonen A., Gyorgy Z., Jalonen J., Neubauer P., Hohtola A., LC/MS/MS identification of glycosides produced by biotransformation of cinnamyl alcohol in Rhodiola rosea compact callus aggregates. Biomedical chromatograph. 2004；18 (8) : 550 – 558.

Udintsev S.N., and Shakhov V.P., The role of humoral factors of
regenerating liver in the development of experimental tumors and the
effect of Rhodiola rosea extract on this process. Neoplasma. 1991 ; 38 : 323 - 331.

Udintsev S.N., and Schakhov V.P., Decrease of cyclophosphamide
haematotoxicity by Rhodiola rosea root extract in mice with Ehrlich and
Lewis transplantable tumors. Eur J Cancer. 1991 ; 27 : 1182.

Udintsev S.N., Krylova S.G., and Fomina T.I., The enhancement of the
efficacy of adriamycin by using hepatoprotectors of plant origin in
metastases of Ehrlich’s adenocarcinoma to the liver in mice. Vopr Onkol. 1992 ; 38 : 1217 - 1222.

Upadrashta S.M., Katikaneni P.R., Nuessle N.O., Chitosan as a tablet binder. Drug Dev Ind Pharm. 1992 ; 18 : 1701 -/1708.

Wing S. L., Askew E.W., Luetkemeier M., Ryujin L., Kamimori D.T., Grissom G.H., and C. K., Lack of effect of Rhodiola or oxygenated water supplementation on hypoxemia and oxidative stress. Wilderness Environ Med. 2003 ; 14 (1) : 9 - 16.

Xiao H., Tianyou Z., Yun W., Xueli C., Yoichiro I., Seperation of salidroside from Rhodiola crenulata by high-speed counter-current chromatography. J Chromatogr A. 2002 ; 971：237-241.

Yurdamur A., Daneel F., Ehab A., Abourashed, Ikhlas A., Khan, Lotaustralin from Rhodiola rosea roots. Fitoterapia. 2004 ; 75 : 612 - 614.

包文芳, 吳維春, 李葆華, 抗疲勞用藥植物紅景天, 人民軍醫出版社, 北京, 2003；34 – 58。

國家中醫藥管理局＜中華本草＞編委會, 中華本草, 藏藥卷, 上海科學技術出版社, 2002；178 - 180。

龔云, 紅景天與運動疲勞, 西北師範大學學報（自然科學版）, 2001；37 (3) : 110 - 114。

高秋娜, 紅景天的藥理與臨床研究概述, 河南中醫, 2001；21 (2)：76 - 77。

金永日, 雎大員, 于曉風, 呂忠智, 王黎, 紅景天莖葉提取物的抗衰老作用研究, 中國老年學雜誌, 2001；21 (2) : 228 - 229。

明海泉, 夏光成, 張瑞鈞, 紅景天研究發展, 中草藥, 1988；19 (5) : 37 - 41。

武藝, 李愛馥, 陳俊龍, 葛遼海, 馬麗霞, 蔡淑安, 紅景天人參製劑抗皮膚老化掃描電鏡檢查, 中華醫學美容雜誌, 1999；5 (4) : 172 - 175。

李靖, 張蘊琨, 中藥紅景天在競技體育領域中的研究近況, 南京體育學院報, 2002；1 (1)：28 - 31。

戴壽榮, 王蘭金, 汪家春, 潘明達, 紅景天提取物的製備及其抗區氧抗疲勞作用, 海軍醫學雜誌, 1999；20 (1)：44 - 46。

佟有嫦, 孫淑平, 王春泉, 張洪誌, 紅景天等藥物提高運動圓耐力的研究, 航太醫學與醫學工程, 1996；9 (3)：214 - 216 。

陸欽堯, 黎明, 抗衰老的中草藥, 渡假出版社, 台灣, 1995；180 - 181

姜文華, 孟曉婷, 郝利銘, 崔麗, 董智勇, 王淑蘭, 紅景天素抗癡呆效應的實驗研究, 白求恩醫科大學學報, 2001；27 (2)：127 - 129。 陳亞東, 曹秀蘭, 田長有, 王洪生, 張紅英, 高山紅景天對小鼠耐缺氧、抗疲勞及低溫作用的影響, 中國中醫藥科技, 2002；9 (3)：157 - 158。 張慧, 沈俊, 王昊, 汪家春, 紅景天對急性低壓缺氧大鼠內皮素分泌的干預, 航空航天醫藥, 2001；12 (3)：125 - 127。 劉亞玲, 林樹新, 徐明達, 李志超, 陳壁, 張世范, 紅景天對兔燙傷后多器官功能障礙綜合証的治療作用, 第四軍醫大學學報, 2002；23 (10) : 924 - 927。
王莉, 藍曉紅, 李艷麗, 紅景天的現代研究與進展, 中國新藥, 2003；2 (2) : 27 - 28。

李偉, 紅景天屬職務的研究及應用, 首都師範大學學報, 2003；24 (1) : 56 - 59。

周家春, 食品工藝學, 化學工業出版社, 2003；270 - 280。

談駿嵩, 超臨界流體的應用, 科學發展雜誌, 2002；359：12-17。