狭叶虎皮楠的化学成分及其生物活性研究
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摘要
虎皮楠科植物为一常绿灌木或小乔木,民间经常用来做创伤药和消炎药,在我国长江以南资源丰富,各地均有栽培。但是长期以来,由于对其认识不够,没有对虎皮楠科植物做深入的研究,直到近年来的研究表明,虎皮楠科植物富含生物碱类,黄酮类等活性化合物,越来越受到重视,很多有价值的成分从虎皮楠科植物中分离出来。本论文主要针对目前还未涉及的狭叶虎皮楠的化学成分进行研究,从中共分离得到了10个单体化合物,并通过红外,紫外,质谱,一维和二维核磁共振谱鉴定了它们的结构,其中有2个为新化合物。
1、采用正相硅胶柱色谱、sephadex LH-20、RP-18和制备型HPLC等方法对狭叶虎皮楠的乙酸乙酯萃取物进行了分离,从中分离得到5个化合物,分别鉴定为:5,7,14-三羟基黄烷(2→O→7,4→8)3′,5′,14′-三羟基黄酮醇(Ⅰ),5,7,14-三羟基黄烷醇(2→O→7,4→8)3′,5′,14′-三羟基黄酮醇(Ⅱ),20(29)-羽扇豆烯-3β,24-二醇(20(29)-lupene-3β,24-diol(Ⅲ),木犀草素(luteolin,Ⅳ),芹菜素(apigenin,Ⅴ),其中化合物Ⅰ,Ⅱ为新化合物。
2、狭叶虎皮楠正丁醇萃取物采用D101型大孔树脂分离,用水、25%、50%、75%和95%的乙醇溶液洗脱为不同极性的组分。25%乙醇洗脱部分通过sephadex LH-20、制备型HPLC分离得到组分5,4′-二羟基黄酮-7-O-β-D-葡萄糖-2′-O-α-(L)-鼠李糖苷(Naringin,Ⅵ)和7,4′-二羟基异黄酮-8-C-β-D-葡萄糖苷(Puerarin,Ⅶ);75%乙醇洗脱部分通过RP-18分离得到化合物对羟基苯甲酸(p-Hydroxybenzoic acid,Ⅷ)和对羟基肉桂酸(p-Hydroxycinnamic acid,Ⅸ)。
3、采用正相硅胶柱色谱等方法对狭叶虎皮楠的石油醚提取物进行了分离,从中分离得到了β-谷甾醇(β-Sitosterol,Ⅹ)。
4、狭叶虎皮楠黄酮类化合物的生物活性研究
研究发现狭叶虎皮楠黄酮类化合物具有较强的抗氧化能力,且皆强于BHT。应用清除DPPH自由基方法评价揭示了不同黄酮类化合物单体抗氧化能力大小依次为:Ⅳ＞Ⅴ＞Ⅱ＞Ⅰ＞Ⅶ＞Ⅵ,并对黄酮类化合物的结构与其抗氧化活性的关系进行了探讨。
利用MTT法测定了新化合物体外抗肿瘤活性,化合物Ⅰ和Ⅱ对人乳腺癌Bcap37细胞的IC_(50)值分别为25.3±2.1μg/mL和22.8±1.4μg/mL,表明其均有不同程度的抑制肿瘤细胞生长的生物活性。
此外,论文还对近年来虎皮楠科植物的化学成分研究进行了综述,本研究的结果为进一步阐明狭叶虎皮楠中的化学成分及其生物活性奠定了一定的基础。
The shrub Daphniphyllum is widely distributed in southern China, which has been used in folk medicine for the healing of wounds and as an antiinflammatory remedy.There were no deep study on the Daphniphytlum for a long time,because of its insufficient understanding. Daphniphyllum plants have been the subject of continued and growing interest,due to the range of biological activities shown by many members of this family.No any phytochemical investigation has been done on the species of D.angustifolium Hutch.previously.On our searching for plant-derived bioactivities compounds,the stems of D.angustifolium Hutch.were chemically studied.10 compounds were isolated,purified from this species.Their structures were determined by IR,UV,MS,1D and 2D NMR analysis,respectively,all of them are first isolated from this plants,and two of them are new compounds.
1.The chemical constituent in the ethyl acetate extract of D. angustifolium Hutch.was separated by chromatography on silica gel, RP-18,Sephadex LH-20 and Pre-HPLC to yield 5 compounds:5,7, 14-trihydroxyflavane(2→0→7,4→8)3',5',14'-trihydroxyflavonol(Ⅰ), 5,7,14-trihydroxyflavane(2→O→7,4→8)3',5',14'-trihydroxyflavonol (Ⅱ),20(29)-lupene-3β,24-diol(Ⅲ),luteolin(Ⅳ),apigenin(Ⅴ). CompoundsⅠandⅡwere new compounds.
2.The chemical constituent in the n-butanol extract of D. angustifolium Hutch.was isolated by D101 macroporous adsorption resin eluted with water,25%,50%,75%and 95%ethanol to obtain the components with different polarity.The fraction of 25%ethanol was isolated by sephadex LH-20,Pre-HPLC to obtain compound 5,4'-dihydroxy-isofavone-7-O-β-D-Glc-2'-O-α-(L)-Rhamnoglucoside(Ⅵ) and 7,4'-Dihydroxy-isofavone-8-C-β-D-glucopyranoside(Ⅶ).The fraction of 75%ethanol was isolated by RP-18 to obtain compound p-Hydroxybenzoic acid(Ⅷ) and p-Hydroxycinnamic acid(Ⅸ).
3.The chemical constituent in the petroleum ether extract of D. angustifolium Hutch.was separated by repeated chromatography on silica gel,to obstain compoundβ-Sitosterol(Ⅹ).
4.Biological activity of daphniphyllum flavonoids.
The research discovered the daphniphyllum flavonoids have strong antioxidant activity.Their antioxidant activity is stronger than BHT's. DPPH radical-scavenging appraisal indicated that the order of antioxidant activity of different daphniphyllum flavonoids was as follows:Ⅳ＞Ⅴ＞Ⅱ＞Ⅰ＞Ⅶ＞Ⅵ.The structure-activity relationship of Flavonoids antioxidants was discussed.
In vitro test(MTT method),new compounds(Ⅰ),and(Ⅱ) showed moderate anti-tumor activities against human mammary carcinoma (Bcap37),human hepatocellular carcinoma(SMMC7721).
In addition,the advances in isolation and bioactivities of components from D.angustifolium Hutch.were reviewed.The results of present study will be useful to further phytochemical investigation on D. angustifolium Hutch.
1、采用正相硅胶柱色谱、sephadex LH-20、RP-18和制备型HPLC等方法对狭叶虎皮楠的乙酸乙酯萃取物进行了分离,从中分离得到5个化合物,分别鉴定为:5,7,14-三羟基黄烷(2→O→7,4→8)3′,5′,14′-三羟基黄酮醇(Ⅰ),5,7,14-三羟基黄烷醇(2→O→7,4→8)3′,5′,14′-三羟基黄酮醇(Ⅱ),20(29)-羽扇豆烯-3β,24-二醇(20(29)-lupene-3β,24-diol(Ⅲ),木犀草素(luteolin,Ⅳ),芹菜素(apigenin,Ⅴ),其中化合物Ⅰ,Ⅱ为新化合物。
2、狭叶虎皮楠正丁醇萃取物采用D101型大孔树脂分离,用水、25%、50%、75%和95%的乙醇溶液洗脱为不同极性的组分。25%乙醇洗脱部分通过sephadex LH-20、制备型HPLC分离得到组分5,4′-二羟基黄酮-7-O-β-D-葡萄糖-2′-O-α-(L)-鼠李糖苷(Naringin,Ⅵ)和7,4′-二羟基异黄酮-8-C-β-D-葡萄糖苷(Puerarin,Ⅶ);75%乙醇洗脱部分通过RP-18分离得到化合物对羟基苯甲酸(p-Hydroxybenzoic acid,Ⅷ)和对羟基肉桂酸(p-Hydroxycinnamic acid,Ⅸ)。
3、采用正相硅胶柱色谱等方法对狭叶虎皮楠的石油醚提取物进行了分离,从中分离得到了β-谷甾醇(β-Sitosterol,Ⅹ)。
4、狭叶虎皮楠黄酮类化合物的生物活性研究
研究发现狭叶虎皮楠黄酮类化合物具有较强的抗氧化能力,且皆强于BHT。应用清除DPPH自由基方法评价揭示了不同黄酮类化合物单体抗氧化能力大小依次为:Ⅳ＞Ⅴ＞Ⅱ＞Ⅰ＞Ⅶ＞Ⅵ,并对黄酮类化合物的结构与其抗氧化活性的关系进行了探讨。
利用MTT法测定了新化合物体外抗肿瘤活性,化合物Ⅰ和Ⅱ对人乳腺癌Bcap37细胞的IC_(50)值分别为25.3±2.1μg/mL和22.8±1.4μg/mL,表明其均有不同程度的抑制肿瘤细胞生长的生物活性。
此外,论文还对近年来虎皮楠科植物的化学成分研究进行了综述,本研究的结果为进一步阐明狭叶虎皮楠中的化学成分及其生物活性奠定了一定的基础。
The shrub Daphniphyllum is widely distributed in southern China, which has been used in folk medicine for the healing of wounds and as an antiinflammatory remedy.There were no deep study on the Daphniphytlum for a long time,because of its insufficient understanding. Daphniphyllum plants have been the subject of continued and growing interest,due to the range of biological activities shown by many members of this family.No any phytochemical investigation has been done on the species of D.angustifolium Hutch.previously.On our searching for plant-derived bioactivities compounds,the stems of D.angustifolium Hutch.were chemically studied.10 compounds were isolated,purified from this species.Their structures were determined by IR,UV,MS,1D and 2D NMR analysis,respectively,all of them are first isolated from this plants,and two of them are new compounds.
1.The chemical constituent in the ethyl acetate extract of D. angustifolium Hutch.was separated by chromatography on silica gel, RP-18,Sephadex LH-20 and Pre-HPLC to yield 5 compounds:5,7, 14-trihydroxyflavane(2→0→7,4→8)3',5',14'-trihydroxyflavonol(Ⅰ), 5,7,14-trihydroxyflavane(2→O→7,4→8)3',5',14'-trihydroxyflavonol (Ⅱ),20(29)-lupene-3β,24-diol(Ⅲ),luteolin(Ⅳ),apigenin(Ⅴ). CompoundsⅠandⅡwere new compounds.
2.The chemical constituent in the n-butanol extract of D. angustifolium Hutch.was isolated by D101 macroporous adsorption resin eluted with water,25%,50%,75%and 95%ethanol to obtain the components with different polarity.The fraction of 25%ethanol was isolated by sephadex LH-20,Pre-HPLC to obtain compound 5,4'-dihydroxy-isofavone-7-O-β-D-Glc-2'-O-α-(L)-Rhamnoglucoside(Ⅵ) and 7,4'-Dihydroxy-isofavone-8-C-β-D-glucopyranoside(Ⅶ).The fraction of 75%ethanol was isolated by RP-18 to obtain compound p-Hydroxybenzoic acid(Ⅷ) and p-Hydroxycinnamic acid(Ⅸ).
3.The chemical constituent in the petroleum ether extract of D. angustifolium Hutch.was separated by repeated chromatography on silica gel,to obstain compoundβ-Sitosterol(Ⅹ).
4.Biological activity of daphniphyllum flavonoids.
The research discovered the daphniphyllum flavonoids have strong antioxidant activity.Their antioxidant activity is stronger than BHT's. DPPH radical-scavenging appraisal indicated that the order of antioxidant activity of different daphniphyllum flavonoids was as follows:Ⅳ＞Ⅴ＞Ⅱ＞Ⅰ＞Ⅶ＞Ⅵ.The structure-activity relationship of Flavonoids antioxidants was discussed.
In vitro test(MTT method),new compounds(Ⅰ),and(Ⅱ) showed moderate anti-tumor activities against human mammary carcinoma (Bcap37),human hepatocellular carcinoma(SMMC7721).
In addition,the advances in isolation and bioactivities of components from D.angustifolium Hutch.were reviewed.The results of present study will be useful to further phytochemical investigation on D. angustifolium Hutch.
引文
[1]彭镇华,中国长江三峡植物大全(上卷)[M].科学出版社,2005:500.
[2]翟汝康,中国高等植物图鉴(补编第二册)[M].科学出版社,1983:185.
[3]郑勉,闵天禄,中国植物志(第四十五卷,第一分册)[M].科学出版社,1980:1.
[4]Zhi Yu SHAO,Da Yuan ZHU,Yue Wei GUO.A New Flavan-3-ol Glucoside from Daphniphyllum oldhamii[J].Chinese Chemical Letters,2003,14(9):926-929.
[5]Gamez E J C,Luyengi L,Lee S.K,et al.Antioxidant flavonoid glycosides from Daphniphyllum calycinum[J].J.Nat.Prod.1998,61:706-709.
[6]Zhi Yu SHAO,Da Yuan ZHU,Yue Wei GUO.Two New Triterpene Esters from Daphniphyllum oldhamii[J].Chinese Chemical Letters,2002,13:1181-1184.
[7]Morita H,Yoshida N,Kobayashi J.Daphnezomines A and B,Novel Alkaloids with an Aza-adamantane Core from Daphniphyllum humile[J].J.Org.Chem.1999,64:7208-7212.
[8]Morita H,Yoshida N,Kobayashi J.Daphnezomines C,D and E,New Alkaloids with an N-Oxide Moiety from Daphniphyllum humile[J].Tetrahedron,1999,55:12549-12566.
[9]Yang S P,Zhang H,Zhang C R,et al.Alkaloids from Daphniphyllum Iongeracemosum[J].J.Nat.Prod.2006,69:79-82.
[10]Morita H,Yoshida N,Kobayashi J.Daphnezomines F and G:Novel Alkaloids with 1-Azabicycio undecane Moiety from Daphniphyllum humile[J].J.Org.Chem.2000,65:3558-3562.
[11]Morita H,Yoshida N,Kobayashi J.Daphnezomines H,I,J,and K,New Daphnilactone-type and Yuzurimine-type Alkaloids from Daphniphyllum humile[J].Tetrahedron,2000,56:2641-2646.
[12]Morita H,Kobayashi J.Daphnezomines L,M,N,and O,new alkaloids from Daphniphyllum humile[J].Tetrahedron,2002,58:6637-6641.
[13]Morita H,Takatsu H,Kobayashi J.Daphnezomines P,Q,R and S,new alkaloids from Daphniphyllum humile[J].Tetrahedron 2003,59:3575-3579.
[14] Kobayashi J, Inaba Y, Shiro M, et al. Daphnicyclidins A-H, Novel Hexa- or Pentacyclic Alkaloids from Two Species of Daphniphyllum[J]. J. Am. Chem. Soc. 2001, 123: 11402-11408.
[15] Morita H, Yoshida N, Kobayashi J. Daphnicyclidins J and K, Unique Polycyclic Alkaloids from Daphniphyllum humile[J]. J. Org. Chem. 2002, 67: 2278-2282.
[16] El Bitar H, Nguyen V H, Gramain, et al. New Alkaloids from Daphniphyllum calycinum[J]. Nat. Prod. 2004, 67:1094-1099.
[17] Li L, He H P, Di Y T, et al. Daphnilongerine, an unprecedented fused pentacyclic ring system alkaloid from Daphniphyllum longeracemosum Rosenth[J]. Tetrahedron Lett. 2006, 47:6259-6262.
[18] Zhan Z.-J, Zhang C R, Yue J M. Caldaphnidines A-F, six new Daphniphyllum alkaloids from Daphniphyllum calycinum[J] Tetrahedron, 2005, 61:11038-11045.
[19] Takatsu H, Morita H, Shen Y C, et al. Daphniglaucins D-H and K, New alkaloids from Daphniphyllum glaucescens[J]. Tetrahedron, 2004, 60: 6279-6284.
[20] El Bitar, H.; Nguyen, V. H.; Gramain, A.;et al. Daphcalycinosidines A and B, new iridoid-alkaloids from Daphniphyllum calycinum[J] Tetrahedron Lett. 2004, 45:515-518.
[21] Kobayashi, J.; Takatsu, H.; Shen, Y.-C. et al. Daphniglaucins A and B, novel polycyclic quaternary alkaloids from Daphiniphyllum glaucescens[J] Org.Lett. 2003, 5:1733-1736.
[22] Di Y T, He H P, Liu H Y, et al. Calycilactone A, a novel hexacyclic alkaloid from Daphniphyllum calycillum[J]. Tetrahedron Lett. 2006,47:5329-5331.
[23] Kobayashi J, Inaba Y, Shiro M, et al. Daphnicylidins A-H, novel hexa- or penta-cyclic alkaloids from two species of Daphniphyllum[J]. J. Am. Chem. Soc. 2001, 123, 11402-11408.
[24] Morita, H.; Ishioka, N.; Takatsu, H. et al. Daphmanidins E and F, Alkaloids from Daphniphyllum teijsmannii[J]. J. Nat. Prod. 2006, 69:418-420.
[25] Zhang, H.; Yang, S.-P.; Fan, C.-Q,et al Daphniyunnines A-E, Alkaloids from Daphniphyllum yunnanense[J]. J.Nat. Prod. 2006,69:553-557.
[26] Di, Y.-T.; He, H.-P.; Lu, Y.et al. Alkaloids from the leaves of Daphniphyllum longeracemosum[J] J. Nat. Prod. 2006, 69:1074-1076.
[27] Yang, S.-P.; Yue, J.-M. Alkaloids from Daphniphyllum longeracemosum[J].Helvetica Chimica Acta.2006,11:2783-2788.
[28]Hiroshi Morita,Hiroshi Takatsu,Ya-Ching Shenb et al.Daphniglaucin C,a novel tetracyclic alkaloid from Daphniphyllum glaucescens[J].Tetrahedron Letters,2004,45:901-904.
[29]Zhi Yu SHAO,Da Yuan ZHU,Yue Wei GUO.Oldhamioside,A New Phenolic Glucoside from Daphniphyllum oldhamii[J].Chinese Chemical Letters,2004,15(1):52-54.
[30]国家中医药管理局《中华本草》编委会,中华木草,第4卷,上海科学技术出版社,上海,1998,865.
[31]隋晓,韩玉谦,乔世伟等.银杏叶活性成分提取与分离的研究[J].中国食品添加剂,1999:25-28.
[32]刘国诠,于兆娄.色谱柱技术[M].第二版,北京:化学工业出版社,2005.101.
[33]袁黎明.制备色谱技术及运用[M].北京:化学工业出版社,2004.101.
[34]黄建明,郭济贤.大孔树脂对草乌生物碱的吸附性能及提纯工艺复旦学报[J].医学报,2002,30(3):267-269.
[35]谭仁祥主编 植物成分分析 科学出版,2002.
[36]Shashi B M,Asish P K,13C-NMR spectra of pentacyclic triterpenoids—a compilation and some salient features[J].Phytochemistry 1994,37:1517-1575.
[37]Hisham A,Kumar G J,Fujimoto Y,et al.20,29-Epoxysalacianone and 6β-hydroxy-salacianone,two lupine triterepenes from salacia beddomei[J].Phytochemistry,1996,3(42):789-792.
[38]Peraza Sanchez S R,Salazar Aguilar N E,Pena Rodriguez L M.A New Triterpene from the Resin of Bursera simaruba[J].Journal of Natural Products,1995,58:271-274.
[39]Markham K R,Ternai B,Stanley R,et al.Carbon-13 NMR studies of flavonoids Ⅲ,naturally occurring flavonoid glycosides and their acylated derivatives[J].Tetrahedron,1978,34(8):1389-1393.
[40]Li S J,Zhao Y Y,Wang B,et al.Separation and identification of the flavonoids from Buddleia of ficinalis Maxim[J].Acta.Pharmaceutica Sinica,1996,31(11):849-853.
[41]Li YM,Wang TZ,Wang ZXStudies on chemical constituents in dried buds of Lonicera similes Hemsl[J].China Journal of Chinese Materia Medica,2001,26(1):45-47.
[42]Muhammad,Zahid,Akbar,et al.New Cycloartane Flavnol Glycosides from Corchorus depresses[J1.Helvetica chimica,2002,85:689-697.
[43]Feng Yin,Lihong Hu,et al.Dammarane-Type Glycosides from Gynostemma pentaphyllunm[J].Journal of Natural Products,2004,67:942-952.
[44]Masayuki Yoshikawa,Shoko Yamaguchi,et al.Stomachic principle in ginger.Ⅲ[J].Chem.pharm.Bull.1994,42(6):1226-1230.
[45]Zhao PP,Li BM,et al.Studies on the method of determination of combined sugar in glycosides[J].Acta Pharmaceutica Sinica,1987,22(1):70-74.
[46]Shunji Aoki,Kouichi Higuchi et al.Myrmekodsides A and B,Novel Mono-O-alkyl-diglycerols Reversing Tumor Cell Morphology of ras-Transformed Cells from a Marine Sponge Myrmekioderma[J].Tetrahedr.on,1999,55:14865-14870.
[47]Wei Guang ma,Masanori,Mizutani,et ai.Saponions from the roots of Panax notoginseng[J].Phytochemistry,1999,52:1133-1139.
[48]Negi P S,Jayaprakasha G K.Antibacterial activity of grapefruit(Citrus paradisi) peel extracts[J].Eur Food Res Technol,2001,213:484-487.
[49]Cheng G,Bai Y J,Zhao Y Y,et al.Flavonoids from Ziziphus jujuba Mill var.spinosa[J].Tetrahedron,2000,56:8915-8919.
[50]Xiao YQ,Li L,You X L,et al.A new compound from Gastrodia elata Blume[J].J Asian Nat Prod Res.2002,4(1):73-79
[51]方允中,郑荣梁.自由基生物学的理论与应用[M],北京:科学出版社,2002:7.
[52]郑建仙.功能性食品(第二卷)[M],北京:中国轻工业出版社,1999.50.
[53]Blois M S.Antioxidant determinations by the use of a stable free radical[J].Nature,1958,26:1199-1200.
[54]iIhami G,Dali B,Akc ahan G.Antiradical and antioxidant activity of total anthocyanins from Perilla pankinensis decne[J].Journal of Ethnopharmacology,2005,101(1):287-293
[55]Zheng XL,Sun HX,Liu XL,Chen YX,Qian BC.stilbic acid induced COLO205 cell apoptosis by regulating bcl-2 and bax expression and activating caspase-3.Acta Pharmacol Sin.2004;25(8):1090-1095.
[56]杨力.黄酮醇及其甙的抗氧化活性研究[D].兰州大学,有机化学,2003.
[57]刘莉华,宛晓春,李大样等.黄酮类化合物抗氧活性构效关系的研究进展[J]安徽农业大学学报,2002,29(3):265-270.
[58]张红雨,陈德展.酚类抗氧化剂清除自由基活性的理论表征与应用[J].生物物理学报,2000,16(1):1-8.
[59]白凤梅,蔡同一.类黄酮生物活性及其机理的研究进展[J].食品科学,1999,8:11-13.
[60]Husain S,Cillard J,Cillard P.Hydroxyl radical scavenging activity of flavonoids[J].Phytochemistry,1987,26(9):2487-2491.
[2]翟汝康,中国高等植物图鉴(补编第二册)[M].科学出版社,1983:185.
[3]郑勉,闵天禄,中国植物志(第四十五卷,第一分册)[M].科学出版社,1980:1.
[4]Zhi Yu SHAO,Da Yuan ZHU,Yue Wei GUO.A New Flavan-3-ol Glucoside from Daphniphyllum oldhamii[J].Chinese Chemical Letters,2003,14(9):926-929.
[5]Gamez E J C,Luyengi L,Lee S.K,et al.Antioxidant flavonoid glycosides from Daphniphyllum calycinum[J].J.Nat.Prod.1998,61:706-709.
[6]Zhi Yu SHAO,Da Yuan ZHU,Yue Wei GUO.Two New Triterpene Esters from Daphniphyllum oldhamii[J].Chinese Chemical Letters,2002,13:1181-1184.
[7]Morita H,Yoshida N,Kobayashi J.Daphnezomines A and B,Novel Alkaloids with an Aza-adamantane Core from Daphniphyllum humile[J].J.Org.Chem.1999,64:7208-7212.
[8]Morita H,Yoshida N,Kobayashi J.Daphnezomines C,D and E,New Alkaloids with an N-Oxide Moiety from Daphniphyllum humile[J].Tetrahedron,1999,55:12549-12566.
[9]Yang S P,Zhang H,Zhang C R,et al.Alkaloids from Daphniphyllum Iongeracemosum[J].J.Nat.Prod.2006,69:79-82.
[10]Morita H,Yoshida N,Kobayashi J.Daphnezomines F and G:Novel Alkaloids with 1-Azabicycio undecane Moiety from Daphniphyllum humile[J].J.Org.Chem.2000,65:3558-3562.
[11]Morita H,Yoshida N,Kobayashi J.Daphnezomines H,I,J,and K,New Daphnilactone-type and Yuzurimine-type Alkaloids from Daphniphyllum humile[J].Tetrahedron,2000,56:2641-2646.
[12]Morita H,Kobayashi J.Daphnezomines L,M,N,and O,new alkaloids from Daphniphyllum humile[J].Tetrahedron,2002,58:6637-6641.
[13]Morita H,Takatsu H,Kobayashi J.Daphnezomines P,Q,R and S,new alkaloids from Daphniphyllum humile[J].Tetrahedron 2003,59:3575-3579.
[14] Kobayashi J, Inaba Y, Shiro M, et al. Daphnicyclidins A-H, Novel Hexa- or Pentacyclic Alkaloids from Two Species of Daphniphyllum[J]. J. Am. Chem. Soc. 2001, 123: 11402-11408.
[15] Morita H, Yoshida N, Kobayashi J. Daphnicyclidins J and K, Unique Polycyclic Alkaloids from Daphniphyllum humile[J]. J. Org. Chem. 2002, 67: 2278-2282.
[16] El Bitar H, Nguyen V H, Gramain, et al. New Alkaloids from Daphniphyllum calycinum[J]. Nat. Prod. 2004, 67:1094-1099.
[17] Li L, He H P, Di Y T, et al. Daphnilongerine, an unprecedented fused pentacyclic ring system alkaloid from Daphniphyllum longeracemosum Rosenth[J]. Tetrahedron Lett. 2006, 47:6259-6262.
[18] Zhan Z.-J, Zhang C R, Yue J M. Caldaphnidines A-F, six new Daphniphyllum alkaloids from Daphniphyllum calycinum[J] Tetrahedron, 2005, 61:11038-11045.
[19] Takatsu H, Morita H, Shen Y C, et al. Daphniglaucins D-H and K, New alkaloids from Daphniphyllum glaucescens[J]. Tetrahedron, 2004, 60: 6279-6284.
[20] El Bitar, H.; Nguyen, V. H.; Gramain, A.;et al. Daphcalycinosidines A and B, new iridoid-alkaloids from Daphniphyllum calycinum[J] Tetrahedron Lett. 2004, 45:515-518.
[21] Kobayashi, J.; Takatsu, H.; Shen, Y.-C. et al. Daphniglaucins A and B, novel polycyclic quaternary alkaloids from Daphiniphyllum glaucescens[J] Org.Lett. 2003, 5:1733-1736.
[22] Di Y T, He H P, Liu H Y, et al. Calycilactone A, a novel hexacyclic alkaloid from Daphniphyllum calycillum[J]. Tetrahedron Lett. 2006,47:5329-5331.
[23] Kobayashi J, Inaba Y, Shiro M, et al. Daphnicylidins A-H, novel hexa- or penta-cyclic alkaloids from two species of Daphniphyllum[J]. J. Am. Chem. Soc. 2001, 123, 11402-11408.
[24] Morita, H.; Ishioka, N.; Takatsu, H. et al. Daphmanidins E and F, Alkaloids from Daphniphyllum teijsmannii[J]. J. Nat. Prod. 2006, 69:418-420.
[25] Zhang, H.; Yang, S.-P.; Fan, C.-Q,et al Daphniyunnines A-E, Alkaloids from Daphniphyllum yunnanense[J]. J.Nat. Prod. 2006,69:553-557.
[26] Di, Y.-T.; He, H.-P.; Lu, Y.et al. Alkaloids from the leaves of Daphniphyllum longeracemosum[J] J. Nat. Prod. 2006, 69:1074-1076.
[27] Yang, S.-P.; Yue, J.-M. Alkaloids from Daphniphyllum longeracemosum[J].Helvetica Chimica Acta.2006,11:2783-2788.
[28]Hiroshi Morita,Hiroshi Takatsu,Ya-Ching Shenb et al.Daphniglaucin C,a novel tetracyclic alkaloid from Daphniphyllum glaucescens[J].Tetrahedron Letters,2004,45:901-904.
[29]Zhi Yu SHAO,Da Yuan ZHU,Yue Wei GUO.Oldhamioside,A New Phenolic Glucoside from Daphniphyllum oldhamii[J].Chinese Chemical Letters,2004,15(1):52-54.
[30]国家中医药管理局《中华本草》编委会,中华木草,第4卷,上海科学技术出版社,上海,1998,865.
[31]隋晓,韩玉谦,乔世伟等.银杏叶活性成分提取与分离的研究[J].中国食品添加剂,1999:25-28.
[32]刘国诠,于兆娄.色谱柱技术[M].第二版,北京:化学工业出版社,2005.101.
[33]袁黎明.制备色谱技术及运用[M].北京:化学工业出版社,2004.101.
[34]黄建明,郭济贤.大孔树脂对草乌生物碱的吸附性能及提纯工艺复旦学报[J].医学报,2002,30(3):267-269.
[35]谭仁祥主编 植物成分分析 科学出版,2002.
[36]Shashi B M,Asish P K,13C-NMR spectra of pentacyclic triterpenoids—a compilation and some salient features[J].Phytochemistry 1994,37:1517-1575.
[37]Hisham A,Kumar G J,Fujimoto Y,et al.20,29-Epoxysalacianone and 6β-hydroxy-salacianone,two lupine triterepenes from salacia beddomei[J].Phytochemistry,1996,3(42):789-792.
[38]Peraza Sanchez S R,Salazar Aguilar N E,Pena Rodriguez L M.A New Triterpene from the Resin of Bursera simaruba[J].Journal of Natural Products,1995,58:271-274.
[39]Markham K R,Ternai B,Stanley R,et al.Carbon-13 NMR studies of flavonoids Ⅲ,naturally occurring flavonoid glycosides and their acylated derivatives[J].Tetrahedron,1978,34(8):1389-1393.
[40]Li S J,Zhao Y Y,Wang B,et al.Separation and identification of the flavonoids from Buddleia of ficinalis Maxim[J].Acta.Pharmaceutica Sinica,1996,31(11):849-853.
[41]Li YM,Wang TZ,Wang ZXStudies on chemical constituents in dried buds of Lonicera similes Hemsl[J].China Journal of Chinese Materia Medica,2001,26(1):45-47.
[42]Muhammad,Zahid,Akbar,et al.New Cycloartane Flavnol Glycosides from Corchorus depresses[J1.Helvetica chimica,2002,85:689-697.
[43]Feng Yin,Lihong Hu,et al.Dammarane-Type Glycosides from Gynostemma pentaphyllunm[J].Journal of Natural Products,2004,67:942-952.
[44]Masayuki Yoshikawa,Shoko Yamaguchi,et al.Stomachic principle in ginger.Ⅲ[J].Chem.pharm.Bull.1994,42(6):1226-1230.
[45]Zhao PP,Li BM,et al.Studies on the method of determination of combined sugar in glycosides[J].Acta Pharmaceutica Sinica,1987,22(1):70-74.
[46]Shunji Aoki,Kouichi Higuchi et al.Myrmekodsides A and B,Novel Mono-O-alkyl-diglycerols Reversing Tumor Cell Morphology of ras-Transformed Cells from a Marine Sponge Myrmekioderma[J].Tetrahedr.on,1999,55:14865-14870.
[47]Wei Guang ma,Masanori,Mizutani,et ai.Saponions from the roots of Panax notoginseng[J].Phytochemistry,1999,52:1133-1139.
[48]Negi P S,Jayaprakasha G K.Antibacterial activity of grapefruit(Citrus paradisi) peel extracts[J].Eur Food Res Technol,2001,213:484-487.
[49]Cheng G,Bai Y J,Zhao Y Y,et al.Flavonoids from Ziziphus jujuba Mill var.spinosa[J].Tetrahedron,2000,56:8915-8919.
[50]Xiao YQ,Li L,You X L,et al.A new compound from Gastrodia elata Blume[J].J Asian Nat Prod Res.2002,4(1):73-79
[51]方允中,郑荣梁.自由基生物学的理论与应用[M],北京:科学出版社,2002:7.
[52]郑建仙.功能性食品(第二卷)[M],北京:中国轻工业出版社,1999.50.
[53]Blois M S.Antioxidant determinations by the use of a stable free radical[J].Nature,1958,26:1199-1200.
[54]iIhami G,Dali B,Akc ahan G.Antiradical and antioxidant activity of total anthocyanins from Perilla pankinensis decne[J].Journal of Ethnopharmacology,2005,101(1):287-293
[55]Zheng XL,Sun HX,Liu XL,Chen YX,Qian BC.stilbic acid induced COLO205 cell apoptosis by regulating bcl-2 and bax expression and activating caspase-3.Acta Pharmacol Sin.2004;25(8):1090-1095.
[56]杨力.黄酮醇及其甙的抗氧化活性研究[D].兰州大学,有机化学,2003.
[57]刘莉华,宛晓春,李大样等.黄酮类化合物抗氧活性构效关系的研究进展[J]安徽农业大学学报,2002,29(3):265-270.
[58]张红雨,陈德展.酚类抗氧化剂清除自由基活性的理论表征与应用[J].生物物理学报,2000,16(1):1-8.
[59]白凤梅,蔡同一.类黄酮生物活性及其机理的研究进展[J].食品科学,1999,8:11-13.
[60]Husain S,Cillard J,Cillard P.Hydroxyl radical scavenging activity of flavonoids[J].Phytochemistry,1987,26(9):2487-2491.