臺灣博碩士論文加值系統

由於抗生素濫用造成醫院內的細菌有極高比例的抗藥情形，研發新的抑菌物質以取代目前使用的抗生素已成為當務之急。本研究以五十八種民俗上報導具有相關抑菌活性或在藥理方面有消炎、止痛等功效之中草藥為材料，採用 95% 乙醇萃取抑菌成分，探討其對目前台灣院內主要的抗藥菌綠膿桿菌之抑菌活性。研究結果發現在五十八種中草藥中篩選出二十三種具有抑菌活性，挑選其中較具廣效性的十六種中草藥，測試其粗萃物的最低抑菌濃度 (Minimum Inhibition Concentration，MIC)。結果顯示桂枝、烏梅及牡丹皮的抑菌活性較佳，其 MIC 值約為 1-10 mg/mL，大黃、貝母、山豆根、栝蔞皮及丹參抑菌活性中等，其 MIC 值約為 15-30 mg/mL，而香附、山慈菇、威靈仙、連翹、黃柏、木香、雷丸及石菖蒲的抑菌效果較不理想，其 MIC 值均大於 30 mg/mL。乙醇粗萃物再經由不同極性的溶劑分配萃取後，測試各區分物之活性，結果發現活性區分物之 MIC 值皆有下降的現象。以兩倍 MIC 值的藥物濃度測試各活性區分物的殺菌曲線，顯示試驗菌株可在八小時內死亡。在抗藥性測試方面，各活性區分物於四分之ㄧ倍 MIC 值藥物濃度下連續投藥十天後，試驗菌株並沒有產生抗藥性。此外，由紙錠擴散法 (disc diffusion method) 試驗藥物合併效應，發現桂枝的 Hexane 活性區分物對於 Gentamycin 及 Streptomycin 具有協同作用的效果。同時藉由薄層色層層析初步分離有效成分，再經由生物自析術確認有效成分的活性區帶，並利用紫外線掃描光譜鑑定有效成分之吸收波長。本研究大量篩選具抑菌活性之中草藥，藉由溶劑分配萃取與生物自析術分離並確認有效成分，可供往後中草藥抑菌活性物質提純的參考，純化的有效成分具有成為天然抗生素的潛力。

In the recent years, a large number of antibiotics were misapplied in Taiwan that caused very high ratios of the drug-resistant pathogens in hospitals. Thus, there is an urgent need for new effective anti-microorganism agents to replace those currently in use. In the present study, we screened 58 Chinese herbal medicines that were reported to have antibacterial activities, anti-inflammatory or analgesia in pharmaceutical efficacy to investigate the antibacterial activities of alcoholic extracts against the nosocomial drug-resistant Pseudomonas aeruginosa in Taiwan.The results of bioassay indicated that 23 out of the 58 Chinese herbs showed obvious antimicrobial activity against the test isolates. Among of the 23 herbs, 16 plant extracts that showed broad spectrum of antimicrobial activity against the test organism were selected to determine the minimum inhibitory concentration (MIC). The MIC results from Guizhi, Mume Frutus and Cortex Moutan revealed better antibacterial activity ranged between 1 to 10 mg/mL, Raidx Et Rhizoma Rhei, Bulbus Fritillariae Thunbergii, Radix Sophorae Tonkinensis, and Radix Salviae Miltiorrhizae extracts showed MIC of 15 to 30 mg/mL, whereas for Cyperi Rhizoma, Cremastra appendiculata, Radix Clemafidis Chinesis, Frutus Forsythiae, Cortex Phellodendri, Radix Aucklandiae, Omphala, and Rhizoma Acori Graminei it ranged higher than 30 mg/mL. The crude alcohol extracts were then partition extraction by different solvents based on their polarity. Each fraction was also tested for its antibacterial activity. The MIC value for each active fraction showed significant lower than the crude extract. The results from time-kill studies showed that the test strains were killed within 8 hours when the isolates were exposed to two times of MIC concentration. In the resistance test, exposure of P. aeruginosa to a sub-MIC level of the extract for ten consecutive subcultures did not induce resistance to the active compound. Moreover, combination of the hexane fraction of Guizhi with gentamycin or streptomycin showed a synergism effect by the disc diffusion method. Each of the active partition fractions was also separated and purified by thin layer chromatography (TLC). The active compound on TLC plate was examined by TLC-bioautography and identified by UV/VIS. In this study, we used a bioassay-directed scheme to isolate and partially identify the active antibacterial compounds from Chinese herbal medicines. The present works not only provides extraction reference to active compound from herbs, but also indicates the active substances from Chinese herbs have the potential to be developed into the native antibiotics.

中文摘要 I
英文摘要 III
誌謝 V
目錄 VI
表目錄 IX
圖目錄 X
第一章 緒論 1
第二章 文獻回顧 5
2-1 綠膿桿菌 5
2-2 細菌抗藥性 8
2-3 傳統溶劑萃取植物的有效成分 13
2-4 植物內常見之有效成分 14
2-5 中草藥之抑菌成分 20
2-6 抗微生物藥劑的體外試驗 22
第三章 研究目的及動機 27
第四章 實驗材料與方法 28
4-1 實驗材料 28
4-1-1 中草藥 28
4-1-2 菌株 28
4-1-3 培養基 28
4-1-4 藥品 29
4-1-5 儀器設備 29
4-2 實驗方法 29
4-2-1 實驗架構 29
4-2-2 菌株篩選分離 31
4-2-3 氧化酶試驗 31
4-2-4 菌株保存及培養 32
4-2-5 菌株抗生素敏感試驗 32
4-2-6 中草藥萃取 33
4-2-7 中草藥抑菌活性測試 33
4-2-8 中草藥粗萃物之 MIC 測定 33
4-2-9 分配萃取 34
4-2-10 分配萃取各活性區分之抑菌測試及活性區分物之 MIC 測定 36
4-2-11 殺菌曲線 36
4-2-12 抗藥性測試 36
4-2-13 活性區分物與抗生素之合併效應 37
4-2-14 薄層色層層析 37
4-2-15 紫外線光譜掃描 38
第五章 實驗結果 39
5-1 抑菌活性測試 39
5-1-1 氧化酶試驗 39
5-1-2 抗生素敏感試驗 39
5-1-3 中草藥粗萃物抑菌活性試驗 40
5-1-4 中草藥粗萃物之 MIC 測定 40
5-1-5 中草藥粗萃物之分配萃取及活性區分物 41
5-1-6 中草藥活性區分物之 MIC 測定 41
5-1-7 殺菌曲線 41
5-1-8 中草藥有效成分與抗生素之合併效應 41
5-1-9 抗藥性測試 42
5-2 有效成分之純化分離及鑑定 42
5-2-1 各活性區分薄層層析及 TLC 生物活性試驗 42
5-2-2 紫外線掃描光譜 43
第六章 討論 44
參考文獻 74
附錄 85

1張上淳，陳美智，胡幼圃 (2000)，台灣人用抗生素與動物用抗生素使用量之調查研究，感染控制雜誌，第 13 期第 6 卷，334-345。

2溫譽玲 (2003)，藥草粗莖物及區分物應用於新鮮豬肉微生物控制之研究，國立中興大學食品科學研究所，碩士論文。

3陳盈潔 (2002)，槴子萃取液之抑菌及抗氧化之探討，屏東科技大學食品科學研究所，碩士論文。

4徐翊庭 (2004)，中藥萃取物之抑菌活性及皮膚美白效果之評估，中原大學醫學工程研究所，碩士論文。

5蔡淑雯 (2001)，不同品種竹筍萃取液抑菌性之探討，國立屏東科技大學食品科學系，碩士論文。

6李昇憲 (2005)，以中草藥萃取物研發天然抗真菌試劑及其在抑制植物致病菌之應用，元培科學技術學院生物技術研究所，碩士論文。

7Xu, H.X. and Lee, S.F. (2001) Activity of plant flavonoids against antibiotic-resistant bacteria. Phytother. Res. 15(1), 39-43.

8Mahasneh, A.M. and El-Oqlah, A.A. (1999) Antimicrobial activity of extracts of herbal plants used in the traditional medicine of Jordan. J. Ethnopharmacol. 64(3), 271-276.

9Duffy, C.F. and Power, R.F. (2001) Antioxidant and antimicrobial properties of some Chinese plant extracts. Int. J. Antimicrob. Agents. 7(6), 527-529.

10Penna, C., Marino, S., Vivot, E., Cruanes, M.C., de D Munoz, J., Cruanes, J., Ferraro, G., Gutkind, G. and Martino, V. (2001) Antimicrobial activity of Argentine plants used in the treatment of infectious diseases. Isolation of active compounds from Sebastiania brasiliensis. J. Ethnopharmacol. 77(1), 37-40.

11Iauk. L., Lo Bue, A.M., Milazzo, I., Rapisarda, A. and Blandino, G. (2003) Antibacterial activity of medicinal plant extracts against periodontopathic bacteria. Phytother. Res. 17(6), 599-604.

12Cateni, F., Zilic, J., Falsone, G., Scialino, G. and Banfi, E. (2003) New cerebrosides from Euphorbia peplis L.: antimicrobial activity evaluation. Bioorg. Med. Chem. Lett. 13(24), 4345-4350.

13Seidel, V. and Taylor, P.W. (2004) In vitro activity of extracts and constituents of Pelagonium against rapidly growing mycobacteria. Int J. Antimicrob. Agents. 23(6), 613-619.

14Lopez, A., Hudson, J.B. and Towers, G.H. (2001) Antiviral and antimicrobial activities of Colombian medicinal plants. J. Ethnopharmacol. 77(2-3), 189-196.

15Barbour, E.K., Al, Sharif. M., Sagherian, V.K., Habre, A.N., Talhouk, R.S. and Talhouk, S.N. (2004) Screening of selected indigenous plants of Lebanon for antimicrobial activity. J. Ethnopharmacol. 93(1), 1-7.

16de Souza, G.C., Haas, A.P., von Poser, G.L., Schapoval, E.E. and Elisabetsky, E. (2004) Ethnopharmacological studies of antimicrobial remedies in the south of Brazil. J. Ethnopharmacol. 90(1), 135-143.

17Sato, Y., Suzaki, S., Nishikawa, T., Kihara, M., Shibata, H. and Higuti, T. (2000) Phytochemical flavones isolated from Scutellaria barbata and antibacterial activity against methicillin-resistant Staphylococcus aureus. J. Ethnopharmacol. 72(3), 483-488.

18Ahmad, I. and Beg, A.Z. (2001), Antimicrobial and phytochemical studies on 45 Indian medicinal plants against multi-drug resistant human pathogens. J. Ethnopharmacol. 74(2), 113-123.

19Wiart, C., Mogana, S., Khalifah, S., Mahan, M., Ismail, S., Buckle, M., Narayana, A.K. and Sulaiman, M. (2004) Antimicrobial screening of plants used for traditional medicine in the state of Perak, Peninsular Malaysia. Fitoterapia. 75(1), 68-73.

20張讚昌，邱南昌，尤封陵，施玟玲，朱敬儀，謝雅如，王瑜琦，李文珍 (2003)，實用微生物學，華格那出版社，16-25-16-28。

21丁明哲 (1996)，醫用微生物學，合記圖書出版社，333-339。

22Rusin, P.A., Rose, J.B., Haas, C.N. and Gerba, C.P. (1997) Risk assessment of opportunistic bacterial pathogens in drinking water. Rev Environ. Contam. Toxicol. 152, 57-83.

23Appelbaum, P.C. and Bowen, A.J. (1978) Opportunistic infection of chronic skin ulcers with Pseudomonas putrefaciens. B.r J. Dermatol. 98(2), 229-231.

24Plotnikova, J.M., Rahme, L.G. and Ausubel, F.M. (2000) Pathogenesis of the human opportunistic pathogen Pseudomonas aeruginosa PA14 in Arabidopsis. Plant Physiol. 124, 1766-1774.

25Prince, A. (1992) Adhesins and receptors of Pseudomonas aeruginosa associated with infection of the respiratory tract. Microb. Pathog. 13(4), 251-260.

26Franzetti, F., Cernuschi, M., Esposito, R. and Moroni, M. (1992) Pseudomonas infections in patients with AIDS and AIDS-related complex. J. Intern. Med. 231(4), 437-443.

27Fichtenbaum, C.J., Woeltje, K.F. and Powderly, W.G. (1994) Serious Pseudomonas aeruginosa infections in patients infected with human immunodeficiency virus: a case-control study. Clin. Infect. Dis. 19(3), 417-422.

28Mendelson, M.H., Gurtman, A., Szabo, S., Neibart, E., Meyers, B.R., Policar, M., Cheung, T.W., Lillienfeld, D., Hammer, G. and Reddy, S. (1994) Pseudomonas aeruginosa bacteremia in patients with AIDS. Clin. Infect. Dis. 18(6), 886-895.

29Dropulic, L.K., Leslie, J.M., Eldred, L.J., Zenilman, J. and Sears, C.L. (1995) Clinical manifestations and risk factors of Pseudomonas aeruginosa infection in patients with AIDS. J. Infect. Dis. 171(4), 930-937.

30Govan, J.R. and Deretic, V. (1996) Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol. Rev. 60(3), 539-574.

31Oliver, A., Canton, R., Campo, P., Baquero, F. and Blazquez, J. (2000) High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. Science. 288(5469), 1251-1254.

32蔡文城 (1996)，實用臨床微生物診斷學第八版，九州圖書文物股份有限公司，738-1207。

33Brodsky, M.H. and Nixon, M.C. (1973) Rapid method for detection of Pseudomonas aeruginosa on MacConkey agar under ultraviolet light. Appl. Microbiol. 26(2), 219-220.

34Baron, E.J. (2001) Rapid identification of bacteria and yeast: summary of a National Committee for Clinical Laboratory Standards proposed guideline. Clin. Infect. Dis. 33, 220-225.

35黃彥智 (2003)，影響動物實驗的外在因素-微生物感染，中華民國實驗動物學會第十六期會訊。

36陳淑惠，趙玉玲，張玉雯 (2003)，南部某區域醫院院內感染調查分析，感染控制雜誌，第 13 卷第 6 期，346-353。

37張上淳，蔡佳倫，王振泰，黃高彬，呂學重，莊銀清，王立信，王復德，劉有增，劉清泉，周明淵，王敦正，陳國東 (2004)，台灣醫學中心與區域醫院 1999 -2002 年院內感染之概況，感染控制雜誌，第 14 卷第 1 期，1-11。

38賴珠煌 (2003)，細菌抗藥性與乙內醯酶，國立中正大學化學暨生物化學研究所，碩士論文。

39Hsueh, P.R., Chen, M.L., Sun, C.C., Chen, W.H., Pan, H.J., Yang, L.S., Chang, S.C., Ho, S.W., Lee, C.Y., Hsieh, W.C. and Luh, K.T. (2002) Antimicrobial Drug Resistance in Pathogens Causing Nosocomial Infections at a University Hospital in Taiwan, 1981-1999. Emerg. Infect. Dis. 8, 63-68.

40Hsueh, P.R., Tseng, S.P., Teng, L.J. and Ho, S.W. (2005) Pan-drug-resistant Pseudomonas aeruginosa causing nosocomial infection at a university hospital in Taiwan. Clin. Microbiol. Infect. 11, 670-673.

41趙廷坤，凌保東，周歧新 (2004)，銅綠假單胞菌主動外排系統研究進展，四川生理科學雜誌，第 26 卷第 3 期。

42劉蓉，周黎明，王浴生 (2004)，銅綠假單胞菌對 β-內酰胺類抗生素耐藥機制，四川生理科學雜誌，第 26 卷第 4 期。

43Hancock, R.E. and Carey, A.M. (1979) Outer membrane of Pseudomonas aeruginosa: heat-2-mercaptoethanol-modifiable proteins. J. Bacteriol. 140(3), 902-910.

44Li, X.Z., Nikaido, H. and Poole, K. (1995) Role of mexA-mexB-oprM in antibiotic efflux in Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 39(9), 1948-1953.

45Nikaido, H., Nikaido, K. and Harayama, S. (1991) Identification and characterization of porins in Pseudomonas aeruginosa. J. Biol. Chem. 266(2), 770-779.

46Livermore, D.M. (2002) Multiple mechanisms of antimicrobial resistance in Pseudomonas aeruginosa: our worst nightmare? Clin. Infect. Dis. 34, 634-640.

47Schweizer, H.P. (2003) Efflux as a mechanism of resistance to antimicrobials in Pseudomonas aeruginosa nd related bacteria: unanswered questions. Genet. Mol. Res. 2(1), 48-62

48盧建宏 (2001)，以傳統溶劑及超臨界二氧化碳萃取薑辣味成分之研究，國立成功大學化學研究所，碩士論文。

49許芳華 (2001)，黃岑抑菌性質及抑菌成分分離純化之研究，國立中興大學食品科學研究所，碩士論文。

50朱建儒 (2002)，探討通氣量對於樟芝醱酵生產生物鹼之影響，國立中央大學化學工程與材料工程研究所，碩士論文。

51Kavitha, D., Shilpa, P.N. and Devaraj, S.N. (2004) Antibacterial and antidiarrhoeal effects of alkaloids of Holarrhena antidysenterica WALL. Indian J. Exp. Biol. 42(6), 589-594.

52Tanaka, J.C., Silva, C.C., Oliveira, A.J., Nakamura, C.V. and Dias Filho, B.P. (2006) Antibacterial activity of indole alkaloids from Aspidosperma ramiflorum. Braz. J. Med. Biol. Res. 39(3), 387-391.

53郝小燕，彭琳，葉蘭，黃能慧，沈月毛 (2004)，飛龍掌血生物總鹼抗炎鎮痛作用的研究，中西醫結合學報，第 2 卷第 6 期，450-452。

54劉翠哲，佟繼銘，張麗敏 (2006)，白屈菜總生物鹼對豚鼠的平喘作用，中國醫院藥學雜誌，第 26 卷第 1 期，27-29。

55喻晶，胡文淑 (1996)，甲基蓮心鹼對大鼠血壓，血小板聚集和血栓形成的影響，中國藥理學通報，第 12 卷第 6 期，533-535。

56李振濤，朱奇，紀宇，李愛秀 (2005)，植物生物鹼抗腫瘤作用機制的研究進展，藥學進展，第 29 期第 5 卷，193-197。

57袁珊琴，趙毅民，尤勇 (2004)，西蒙1號蕃薯毛狀根生物鹼成分的研究，藥學學報，第 39 卷第 10 期，818-820。

58Rubia, L.B. and Gomez, R. (1977) TLC sensitivity of six modifications of Dragendorff''s reagent. J. Pharm. Sci. 66(11), 1656-1657

59唐菁吟 (2003)，篩選富含類黃酮蔬果探討其免疫調節及抗發炎作用，國立中興大學食品科學系，碩士論文。

60黃恔瑋 (2003)，植物化學物質之抗氧化力及其抗突變性，中國文化大學生活應用科學研究所，碩士論文。

61Alam, S. (2004) Synthesis, antibacterial and antifungal activity of some derivatives of 2-phenyl-chromen-4-one. J. Chem. Sci. 116(6), 325-331.

62Alam, S. and Mostahar, S. (2005) Studies of Antimicrobial Activity of two Synthetic 2N, 4N, 6N-trioxygenated Flavones. J. Applied. Sci. 5(2), 327-333.

63徐慶，宋芸娟，李麗亞，侯巧燕 (2005)，荔枝核總黃酮的抗鴨乙型肝炎病毒作用，世界華人消化雜誌，第 13 卷第 17 期，2082-2085。

64Wang, W.N., Yang, X.B., Liu, H.Z., Huang, Z.M. and Wu, G.X. (2005) Effect of Oenanthe javanica flavone on human and duck hepatitis B virus infection. Acta. Pharmacol. Sin. 26(5), 587-592.

65黃為瑜 (2002)，紅色鄉土蔬菜中類黃酮抗氧化力及其對淋巴球DNA氧化損傷的保護作用，中國文化大學生活應用科學研究所，碩士論文。

66Park, B.K., Heo, M.Y., Park, H. and Kim, H.P. (2001) Inhibition of TPA-induced cyclooxygenase-2 expression and skin inflammation in mice by wogonin, a plant flavone from Scutellaria radix. Eur. J. Pharmacol. 425(2), 153-157.

67Chi, Y.S., Lim, H., Park, H. and Kim, H.P. (2003) Effects of wogonin, a plant flavone from Scutellaria radix, on skin inflammation: in vivo regulation of inflammation-associated gene expression. Biochem. Pharmacol. 66(7), 1271-1278.

68王彩芳 (2001)，鞣質生理活性研究進展，國外醫學：中醫中藥分冊，第 23 卷第 5 期，278-282。

69王放銀，趙良忠，段林東，余有貴，李新社 (2004)，老鸛草鞣質的提取及其抑菌效果研究，邵陽學院學報：自然科學版，第 1 卷第 3 期，103-105。

70劉延澤，冀春茹 (1991)，高效液相色譜法在中草藥鞣質類研究中的應用，中草藥，第 22 卷第 2 期，89-93。

71關濱，張秋紅，劉秀芳 (2003)，植物中鞣質類化合物的化學研究進展，山東醫藥工業，第 22 卷第 1 期，34-36。

72郭群群，杜桂彩，李榮貴 (2003)，紫蘇葉揮發油抑菌活性研究，食品工業科技，第 24 卷第 9 期，25-27。

73申利群，莫洪波，尹篤林 (2004)，幾種單環單萜化合物的生物活性及其應用，林產化工通訊，第 38 卷第 2 期，30-35。

74周峰，籍保平，李博，李京晶，王成濤，趙磊 (2005)，十二種中藥揮發油及其濾液體外抑菌活性研究，食品科學，第 26 卷第 3 期，50-53。

75靖會，佐建鋒 (2005)，揮發油的藥理研究進展，西北藥學雜誌，第 20 卷第 2 期，3-4。

76孫健，賀銀鳳，田建軍 (2003)，酸馬奶酒中有機酸的抑菌作用，內蒙古農業大學學報 (自然科學版)，第 24 期第 1 卷，94-97。

77武運，王煒，魏東 (2004)，酶和其它蛋白質及有機酸在食品防腐中的研究，新疆農業科學，第 41 卷第 8 期，92-94。

78鄭玉彬，姚立宏 (2004)，美容用中草藥的化學成分及藥理作用，中國美容醫學，第 13 卷第 3 期，292-293。

79陳知本，陳瓊華，黃玉初，蔡亞農 (1990)，大黃的生化學研究XL. 大黃蒽醌衍生物對淋病雙球菌的抑菌作用，中國藥科大學學報，第 21 期第 6 卷，373-374。

80周曾芬，崔蓉，張永生，鄧學瑞，張瑜 (2000)，大蒜、大黃對幽門螺桿菌抑菌作用的實驗研究，中華實用醫學，第 2 卷第 6 期，1-3。

81鄒承淑，邵濟鈞，蘇冬梅，王秋波，王春華，曲雲英 (1990)，青島嶗山地區72種中藥抑菌作用的研究，青島醫學院學報，第 26 卷第 1 期，45-49。

82徐小雲，王雲霞，李智勇 (2003)，威靈仙化學成分和藥理作用研究進展，現代中醫藥 (陜西)，第4期，67-69。

83杜毅，陳瑞，趙麗蓉，班長俊 (2000)，石菖蒲的化學與藥理研究新進展，山西中醫，第 16 卷第 1 期，53-54。

84丁鳳榮，邱世翠，郭麗華，宮照龍，楊新 (2003)，牡丹皮的體外抑菌作用研究，時珍國醫國藥，第 14 卷第 8 期，452-452。

85李方軍 (2004)，牡丹皮化學成分及藥理作用研究進展，安徽醫藥，第 8 卷第 1 期，9-10。

86黃曉暉，陳思東，譚劍斌 (2002)，中藥桂枝提取物殺滅微生物效果研究，廣東藥學院學報，第 17 卷第 4 期，300-301。

87韓愛霞，綦躍花，邱世翠，杜海嶺，董豔軍 (2004)，桂枝體外抑菌作用研究，時珍國醫國藥，第 15 卷第 11 期，743-743。

88孫吉蘭，常雲亭，宋海英，邱世翠，李波清 (2003)，丹參的體外抑菌作用研究，時珍國醫國藥，第 14 卷第 12 期，725-725。

89王桂閩，馬西強 (2003)，丹參在臨床上的應用，中國民間療法，第 11 卷第 12 期，51-52。

90李陽，黎雪如，周婭，王寧萍 (1996)，寧夏貝母總鹼和蛇膽川貝液體外抑菌活性研究，寧夏醫學雜誌，第 18 卷第 3 期，147-148。

91張勇慧，阮漢利，吳繼洲 (2003)，貝母的藥理作用研究概況，醫藥導報，第 22 卷第 11 期，797-799。

92郭志堅，郭書好，何康明，劉慧瓊，潘珊珊 (2002)，黃柏葉中黃酮醇甙含量測定及其抑菌實驗，暨南大學學報：自然科學與醫學版，第 23 卷第 5 期，64-66。

93王躍華，徐文俊，何俊蓉，朱劍宏，孫雁霞 (2004)，川黃柏離體培養及藥用成分的抑菌試驗研究，中國中藥雜誌，第 29 卷第 10 期，1002-1003。

94李峰，賈彥竹 (2004)，黃柏的臨床藥理作用，中醫藥臨床雜誌，第 16 卷第 2 期，191-191。

95許臘英，余鵬，毛維倫，劉芬 (2003)，中藥烏梅的研究進展，湖北中醫學院學報，第 5 卷第 1 期，52-57。

96劉友平，陳鴻平，萬德光，嚴鑄雲，陳林 (2004)，烏梅的研究進展，中藥材，第 27 卷第 6 期，459-462。

97劉圓，白志川，于杰 (2004)，藏藥復方中余甘子和甘草及木香的薄層定性鑒別，西南農業大學學報，第 26 卷第 5 期，625-628。

98林芳蘭 (2002)，綠膿桿菌環境菌株對抗生素、重金屬及農藥抗性之分析，國立中山大學生物科學系研究所，碩士論文。

99Bauer, A.W., Kirby, W.M., Sherris, J.C. and Turck, M. (1966) Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol. 45(4), 493-496.

100NATIONAL COMMITTEE FOR CLINICAL LABORATORY STANDARDS. (1997) Methods for dilution antimicrobial susceptibility test for bacteria that grow aerobically: approved standard. Villanova: NCCLS (M7-A3).

101陳怡心，莊銀清 (2004)，抗生素抑(殺)菌效果的體外測試，感染控制雜誌，第 14 卷第 6 期，365-370。

102Grzybowska, W., Banaszczyk-Rus, M. and Tyski, S. (2005) Comparison of the checkerboard and E-test methods used for the analysis of two antibiotics combination. Med. Dosw. Mikrobiol. 57(1), 65-75.

103Matsuo, K. and Uete, T. (1993) Enhancement of in vitro antimicrobial activity of cefmetazole and cefotiam in combination against methicillin-susceptible and -resistant Staphylococcus aureus studied using checkerboard MIC method and disc diffusion method with discs containing both drugs. Jpn. J. Antibiot. 46(3), 222-233.

104Manga, H.M., Brkic, D., Marie, D.E. and Quetin-Leclercq, J. (2004), In vivo anti-inflammatory activity of Alchornea cordifolia (Schumach. & Thonn.) Mull. Arg. (Euphorbiaceae). J. Ethnopharmacol. 92(2-3), 209-214.

105Xu, H.X. and Lee, S.F. (2004), The antibacterial principle of Caesalpina sappan. Phytother. Res. 8(8), 647-651.

106Yu, H.H., Kim, K.J., Cha, J.D., Kim, H.K., Lee, Y.E., Choi, N.Y. and You, Y.O. (2005), Antimicrobial activity of berberine alone and in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus. J. Med. Food. 8(4), 454-461.

107Kurien, S. and Lorian, V. (1980), Discrepancies between results obtained by agar and Broth techniques in testing of drug combinations. J. Clin. Microbiol. 11(5), 527-529.

108Szabo, B., Lakatos, A., Koszegi, T., Katay, G. and Botz, L. (2005), Thin-layer chromatography-densitometry and liquid chromatography analysis of alkaloids in leaves of Papaver somniferum under stress conditions. J. AOAC. Int. 88(5), 1571-1577.

109Neu, R. (1956), A new reagent for differentiating and determining flavones on paper chromatograms. Naturwissenschaften. 43: 82

110傅素鄉 (2000)，蜂膠中類黃酮物質之測定及其抑菌特性之探討，國立屏東科技大學食品科學系，碩士論文。

111Aqil, F., Khan, M.S., Owais, M. and Ahmad, I. (2005), Effect of certain bioactive plant extracts on clinical isolates of beta-lactamase producing methicillin resistant Staphylococcus aureus. J. Basic Microbiol. 45(2), 106-114.

112Silva, O., Duarte, A., Cabrita, J., Pimentel, M., Diniz, A. and Gomes, E. (1996) Antimicrobial activity of Guinea-Bissau traditional remedies. J. Ethnopharmacol. 50(1), 55-59.

113Daud, A., Gallo, A. and Riera, A.S. (2005) Antimicrobial properties of Phrygilanthus acutifolius. J. Ethnopharmacol. 99(2), 193-197.

114Kubo, I., Fujita, K., Lee, S.H. and Ha, T.J. (2005) Antibacterial activity of polygodial. (2005) Phytother. Res. 19(12), 1013-1017.

115Tadeg, H., Mohammed, E., Asres, K. and Gebre-Mariam, T. (2005) Antimicrobial activities of some selected traditional Ethiopian medicinal plants used in the treatment of skin disorders. J. Ethnopharmacol. 100(1-2), 168-175.

116Monici, M., Mulinacci, N., Baglioni, P., Vincieri, F.F. (1993) Flavone photoreactivity. UV-induced reactions in organic solvents and micellar systems. J. Photochem. Photobiol. B. 20(2-3), 167-72.