叶下珠中叶下珠素的提取纯化工艺研究
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摘要
本研究以叶下珠为研究对象,建立了叶下珠中叶下珠素的高效液相色谱法(HPLC)检测方法,同时采用HPLC测定不同产地和不同部位的叶下珠药材中叶下珠素含量;对叶下珠中叶下珠素的提取与分离纯化工艺进行了较深入而系统的研究,确定了工艺流程和分离纯化条件,同时对纯化的叶下珠素单体进行了质谱鉴定。主要研究内容如下:
HPLC定量检测叶下珠素的含量。确定液相色谱条件为:色谱柱C_(18)(250mm×4.6mm,5um),检测波长230nm,柱温30℃,进样量20μL,流速0.8mL/min,以甲醇—水(80:20)为流动相等梯度洗脱。在此液相条件下,叶下珠样品液中五种组分得到较好地分离。叶下珠素在2.72μg/mL—136μg/mL浓度范围内线性关系良好。
HPLC测定不同产地和(广西,南宁)不同部位叶下珠药材中叶下珠素含量。结果表明,海南产叶下珠中叶下珠素含量最高为0.664%,广西次之(0.565%),广东,湛江为(0.178%),其他产地叶下珠素含量均较低。不同部位以叶中含量最高(1.04%),果实(0.03%)、茎(0.02%)、枝(0.018%)、根(0.01%)中含量都很低。
叶下珠中叶下珠素的提取工艺研究。乙醇超声法提取,进行单因素试验和正交试验,确定最佳提取工艺参数。结果表明,影响叶下珠素提取效率的各因素主次顺序是:乙醇浓度＞液料比＞提取时间,提取温度的影响高度显著。最佳提取工艺参数为:乙醇浓度95%,液料比25,提取时间20min,重复提取两次,叶下珠素含量为0.687%。
叶下珠素的纯化工艺研究。比较HPD100、HPD100A、HPD700等七种大孔树脂对叶下珠素的吸附分离特性,确定选择HPD100大孔树脂。较优纯化条件为:上样流速为1BV/h,选择80%乙醇为洗脱溶剂,洗脱流速为2BV/h,洗脱液用量约为9BV,在该条件下叶下珠素纯度从9.47%提高到55.2%
通过硅胶柱层析纯化叶下珠素粗产品得到纯度为83%,选择石油醚对叶下珠素粗品进行重结晶得到了纯度为96.79%。
对纯化的叶下珠素单体进行了质谱鉴定,结果表明纯化所得晶体与叶下珠素标准品质谱图基本一致。
Phyllanthus urinaria L.as the raw material,established the HPLC detects the content of phyllanthin by quantitative an alysis,the content of phyllanthin in different areas and parts of phyllanthus urinaria L.were measured;Deep and systematical researches had done to the extraction、isolation and purification of phyllanthin from phyllanthus nurinaria L.,isolation and purification factors were studied and established;The purification of phyllanthin was identified by mass spectra.The main research content is as folows:
The HPLC detects the content of phyllanthin by quantitative analysis.The conditi ons of deter mining liquid phase chromatogram are as folows:chromatogr am column C_(18)(250mmx4.6mm,),the wavelength of detector at 230nm,tempera ture at 30℃,sample size20μL,rate at 0.8mL/min,met hanol:water(80:20v/v)as mobile Phase.In this case,at least 5 components alcohol extactive can be separated well from impurities.Good linear relatio nships between the peak area values and concentrations have beenfound.The relationship of the phyllanthin is good in the range of 2.72μg/mL-136μg/mL.
The content of phyllanthin in different areas and parts of phyllanthus urinaria L.were measured by HPLC.The results showed that the content of phyllanthin in phyllanthus urinaria L.from Hainan was 0.664%,Guangxi and Guangdong zhanjiang were 0.565%and 0.178%,the content of phyllanthin in leaf,fruit, steam,root and branch were 1.04%,0.03%,0.02%,0.018%,0.01%.
The extraction technics of phyllanthin was extracted from Phyllanthus urinaria L.by ethanol ultrasonic extraction,through oneway and orthogonal experiment the optimum parameters of extracting phyllanthin had been determined. It was showed that the main factors influencing extraction efficiency were as follows:the ethanol concentration＞the ethanol amount＞extraction times.The optim um parameters was:the concentration of ethanol is 95%,the multiple of ethanol to material is 25,extraction time is 20min,by two extractions,the content of phyllanthin was 0.687%.
The purifying technology of phyllanthin.The study compared adsorption properties of seven macroporous resins,includingHPD100,HPD100A,HPD700, D101、D201、NKA-Ⅱ、NKA-9、HPD 100 resin was determined as the best sorbent.The optimum purification condition o btained was:the driedre sina sre ference,the flow rate on absorption is 1BV/h,9BV80%ethanol,the flow rate on desorption is 2.0BV/h.Under these conditions,the purity of phyllanthin has greatly improved from 9.47%to 55.2%,
Through silica gel chromatographic column to purify phyllanthin from overall phyllanthin,the purify of phyllanthin obtained was 83%.Phyllanthin was purifid from overall phyllanthin to obtain pure phyllanthin by crystal and recrystal means.its purify was up to 96.79%.
Purification of phyllanthin under-the-MS identification,the results showed phyllanthin was purified from phyllanthus urinaria L.and standards of quality spectrum basically the same.
HPLC定量检测叶下珠素的含量。确定液相色谱条件为:色谱柱C_(18)(250mm×4.6mm,5um),检测波长230nm,柱温30℃,进样量20μL,流速0.8mL/min,以甲醇—水(80:20)为流动相等梯度洗脱。在此液相条件下,叶下珠样品液中五种组分得到较好地分离。叶下珠素在2.72μg/mL—136μg/mL浓度范围内线性关系良好。
HPLC测定不同产地和(广西,南宁)不同部位叶下珠药材中叶下珠素含量。结果表明,海南产叶下珠中叶下珠素含量最高为0.664%,广西次之(0.565%),广东,湛江为(0.178%),其他产地叶下珠素含量均较低。不同部位以叶中含量最高(1.04%),果实(0.03%)、茎(0.02%)、枝(0.018%)、根(0.01%)中含量都很低。
叶下珠中叶下珠素的提取工艺研究。乙醇超声法提取,进行单因素试验和正交试验,确定最佳提取工艺参数。结果表明,影响叶下珠素提取效率的各因素主次顺序是:乙醇浓度＞液料比＞提取时间,提取温度的影响高度显著。最佳提取工艺参数为:乙醇浓度95%,液料比25,提取时间20min,重复提取两次,叶下珠素含量为0.687%。
叶下珠素的纯化工艺研究。比较HPD100、HPD100A、HPD700等七种大孔树脂对叶下珠素的吸附分离特性,确定选择HPD100大孔树脂。较优纯化条件为:上样流速为1BV/h,选择80%乙醇为洗脱溶剂,洗脱流速为2BV/h,洗脱液用量约为9BV,在该条件下叶下珠素纯度从9.47%提高到55.2%
通过硅胶柱层析纯化叶下珠素粗产品得到纯度为83%,选择石油醚对叶下珠素粗品进行重结晶得到了纯度为96.79%。
对纯化的叶下珠素单体进行了质谱鉴定,结果表明纯化所得晶体与叶下珠素标准品质谱图基本一致。
Phyllanthus urinaria L.as the raw material,established the HPLC detects the content of phyllanthin by quantitative an alysis,the content of phyllanthin in different areas and parts of phyllanthus urinaria L.were measured;Deep and systematical researches had done to the extraction、isolation and purification of phyllanthin from phyllanthus nurinaria L.,isolation and purification factors were studied and established;The purification of phyllanthin was identified by mass spectra.The main research content is as folows:
The HPLC detects the content of phyllanthin by quantitative analysis.The conditi ons of deter mining liquid phase chromatogram are as folows:chromatogr am column C_(18)(250mmx4.6mm,),the wavelength of detector at 230nm,tempera ture at 30℃,sample size20μL,rate at 0.8mL/min,met hanol:water(80:20v/v)as mobile Phase.In this case,at least 5 components alcohol extactive can be separated well from impurities.Good linear relatio nships between the peak area values and concentrations have beenfound.The relationship of the phyllanthin is good in the range of 2.72μg/mL-136μg/mL.
The content of phyllanthin in different areas and parts of phyllanthus urinaria L.were measured by HPLC.The results showed that the content of phyllanthin in phyllanthus urinaria L.from Hainan was 0.664%,Guangxi and Guangdong zhanjiang were 0.565%and 0.178%,the content of phyllanthin in leaf,fruit, steam,root and branch were 1.04%,0.03%,0.02%,0.018%,0.01%.
The extraction technics of phyllanthin was extracted from Phyllanthus urinaria L.by ethanol ultrasonic extraction,through oneway and orthogonal experiment the optimum parameters of extracting phyllanthin had been determined. It was showed that the main factors influencing extraction efficiency were as follows:the ethanol concentration＞the ethanol amount＞extraction times.The optim um parameters was:the concentration of ethanol is 95%,the multiple of ethanol to material is 25,extraction time is 20min,by two extractions,the content of phyllanthin was 0.687%.
The purifying technology of phyllanthin.The study compared adsorption properties of seven macroporous resins,includingHPD100,HPD100A,HPD700, D101、D201、NKA-Ⅱ、NKA-9、HPD 100 resin was determined as the best sorbent.The optimum purification condition o btained was:the driedre sina sre ference,the flow rate on absorption is 1BV/h,9BV80%ethanol,the flow rate on desorption is 2.0BV/h.Under these conditions,the purity of phyllanthin has greatly improved from 9.47%to 55.2%,
Through silica gel chromatographic column to purify phyllanthin from overall phyllanthin,the purify of phyllanthin obtained was 83%.Phyllanthin was purifid from overall phyllanthin to obtain pure phyllanthin by crystal and recrystal means.its purify was up to 96.79%.
Purification of phyllanthin under-the-MS identification,the results showed phyllanthin was purified from phyllanthus urinaria L.and standards of quality spectrum basically the same.
引文
[1]全国中草药汇编编写组编.全国中草药汇编[M](上册),北京:人民卫生出版社,1978,263
[2]Thyagarajan S P,Thiruneelakantan K,Subramanian S,etal.In vitro inac tivation of HBsAg by Eclipla alba Hassk and Phllanthus uiruri linn[J].Indian J Med Res,1982,76(Suppl):124-130
[3]牛晓峰,李维凤.陕西叶下珠属植物资源概况[J].西安交通大学学报(医学版),2003,24(2):119-123
[4]李秉滔.中国叶下珠属植物的修订[J].植物分类学报,1987,25(5):371-375
[5]万振先,周国平,易扬华.叶下珠化学成分的研究(Ⅰ)[J].中草药,1994,25(9):455-456
[6]万振先,易扬华,喻庆禄.叶下珠化学成分的研究(Ⅱ)[J].中草药,1997,28(3):134-135
[7]张兰珍,郭亚健,涂光忠,等.叶下珠多酚化合物的分离与鉴定[J].中国中药杂志,2000,25(12):724-725
[8]陈玉武,任丽娟,李克明,等.叶下珠新多酚化合物的分离与鉴定[J].药学学报,1999,34(7):526-529
[9]张兰珍,郭亚健,涂光忠,等.叶下珠新鞣花鞣质的分离与鉴定[J].药学学报,2004,39(2):119-122
[10]Somanabandhu A,Nitayangkura S,Mahidol C,etal.~1H-and ~(13)C - NMR assignm ents of phyllanthin and hypophyllanthin:lignans that enhance cytotoxic resp onses with cultured multidrug-re sistant cells[J].Journal of Natural Products,1993,56:233-239
[11]Anjaneyulu ASR,Rao K J,Row L R,etal.Crystalline constituents of Euphorbi aceae,12.Isolation and structural elucidation of 3 new lignans from leaves of Phyllanthus niruri linn[J].Tetra hedron,1973,29:1291-1298
[12]Stevenson R,Williams J R.Concerning phyltetralin synthesis of lignan arylt etralin isomers[J].Tetrahedron,1977,33:2913-29 17
[13]Ward R S,Satyanarayana P,Row L R,etal.The case for a revised structure for hypophyllanthin ananalysis of the ~(13)C-NMR spectra of sryltet ralins[J].Tetr ahe dron Lett,1979,32:3043-3046
[14]Huang Y L,Chen C C,Ou J C.Isolintetralin:a new lignan from phylanthus nir uri[J].Planta Med,1992,58:473-474
[15]Huang Y L,Chen C C,Hsu F L,etal.A new lignan from phyllanthus virgat us[J].Journal of Natural Products,1996,59:520-521
[16]Chia C C,Yu C L,Karin C S,etal.Lignans from phyllanthus urinaria[J].phytoc hemistry,2003,63:825-833
[17]Wang X W,Yuan J P,Yai Z C,etal.Carboxylic acids from phyllanthus urinaria [J].Chemistry of natural compounds,2005,41(1):17-21
[18]Syamasundar K V,Singh B,Thakur R S,etal.Antihepatotoxic principles of phy llantus uiruri herbs[J]Ethnoparmacol,1985,14(1):41-43
[19]Blumberg B S,Millman I,Venkateswaran P S,Thyagarajan S P.Hepatitis B virus and he patocellular carcinoma:treatment of HBV carriers with Phyllan thus amarus[J].Cancer Detect,1989,14:195-201
[20]Thyagarajan S P,Subramanina S,Thirunalasundari T,etal.Effect of Phllanthus amarus on chronic carriers of hepatitis B virus[J].Lancet,1988,1:764-766
[21]Syamasundar K V,Singh B,Thakur R S,et al.Antihepatotoxic Principles of Phyllantus Niruri Herbs[J].Ethnoparmacol,1985,14(1):41-44
[22]任进余,秦存林苦味叶下珠、叶下珠的药理作用及其作用机理.国外医学.中医中药分册[M],19 99,12(2):53-61.
[23]纪徐淮,秦一中,汪伟业,等.rhIFN-ab和苦味叶下珠提取物对人肝癌细胞株PLC/PRF/5产生HBsAg的影响[J].新消化病学杂志,1995,3(3):136-138
[24]王昌俊,袁德培,陈伟,等.叶下珠对人肝癌细胞的影响[J].时珍国药研究,1997,8(6):499-450
[25]黄育华,张建军,晏雪生,等.叶下珠对人肝癌细胞Bel-7402诱导分化的影响[J].湖北中医学院学报,2000,2(1):10-12
[26]Blumberg B S,Millman I,Venkateswaran P S,etal.Hepatitis B virus and primary hepatocellular carcinoma:treatment of HBV carriers with Phyllant husamarus[J].Vaccine,1990,8(Suppl):S86-S92
[27]张建军,黄育华,晏雪生,等.叶下珠药物血清对人肝癌细胞株的诱导分化作用的实验研究[J].中国中医药科技,2002,9(5):289-291
[28]郭卫真,邓学龙,董伯振,等.叶下珠属植物体外抗单纯疱疹病毒Ⅱ型的作用[J].广洲中医药大学学报,2000,17(1):54-57
[29]ZuoChunxu,Li Fengqin,Yao Qingqiang.Antiviral dehydroch ebulic acid met hyl and its preparation[J].CA,1996,124:185 53/u
[30]Zuo Chunxu,Li Fengqin,Yao Qingqiang.Antiviral methyl brevifolin carbox ylate and its preparation[J].CA,1996,124:66574/a
[31]马新宇,裴俊俊,叶下珠药理研究综述[J].齐鲁药事2006 25(2):104-105
[32]kumaran,R.Joel Karunakaran.In vitro antioxidant activities of meth anol extracts of five phyllanthus species from India[J].Swiss Society of Food Science and Technology 2007,40:344-352
[33]R.Harish,T.Shivanandappa.Atioxidant activity and hepatoprotective potenti al of phyllanthus niruri[J].Food Chemistry,2006,95:180-185
[34]仲英,左旭,张琴冈等.叶下珠化学成分及其抗乙肝病毒活性的研究[J].中国中药杂志,1998,23(6):363-364.
[35]Benner R,Weliky K,Hedges J 1.Early diagenesis of mangrove leaves in a tropicalest uary:Molecular-levela nalyses of neutral sugars and lignin-deriv ed phenols[J].Geochimicaet C osmochimica Acta,1990,54(7):1991-2001.
[36]马金强,贾正平,王彬,等.瑞香狼毒总木脂素与长春新碱的体外抗肿瘤 活性比较[J].西北国防医学杂志,2004,25(5):374-377
[37]Schrroder HC,Mertz H,Steffen R,etal.Z.Naturforsch.Sect.C:Biosci,1990,45:1215-1221
[38]Gnabre JN,Ito Y,Ma Y,etal.J.Chroma.A,1996,719(2):353-364
[39]金昔陆,顾静,胡天喜,等.内南五味子木脂素戈米辛J对肝线粒体膜脂质过氧化和超氧阴离子自由基的作用[J].中国药理学通报,2000,16(1)26-30
[40]韩桂秋,中草药中血小板活化因子受体拮抗活性成分的研究[J].自然科学进展国家重点实验室通讯,1995,5(2):161-164
[41]张永忠,李小莉,郭群.辛夷木脂素类成分抗血小板活化因子作用的研究[J].湖北中药杂志,2001,23(10):7-10
[42]A.S.R.Anjaneyulu,K.Jaganmohan Rao,L.R amachandra Row,etal.Crystal line Constituents of Euphorbiaceae-Ⅻ isolation and Structural El ucidation of There New Lignans from the Lea ves of Phyllanthus Niruri Linn.Tetrahedr on,1973,29:1291-1298
[43]Panchagnula Satyanarayana,somepalli Venkateswarlu.Isolation,Structure and Synthesis of New Diarylbutane Lignans from Phyllanthus Niruri,Synthesis of 5 '-Desmethoxy Niranthin and An Antitumour Extractive.Trtrahedron,1991,47(42):8931-8940
[44]Leite DF,Kassuya CA,Mazzuco TL,etal.The cytotoxic effect and the multidrug resistance reversing action of lignans from Phyllanthus amarus[J].Plant Me d,2006,72(15):1353-1358.
[45]Syamsundar KV,Singh B,Thakur RS,etal.Antihepatotoxic principles of Phyll anthus niruri herbs[J]Ethnopharmacol.1985,14(1):41-44.
[46]Venkateswaran P S,Millman I,Blumberg B S,et al.Effects of an extract from phyllanthus urinari on hepatitis B and woodch uck hwpatitis viruses:in vitro and in vivo studies[J].Proc Na ti Acad Sci USA,1987,84:274-279
[47]Candida AL.Kassuya,Aline Silvestre,Octavio Menezes-de-Lima Jr.etal.Antiinfla mmatory and antiallodynic actions of the lignan niranthin isolated from Phyllanthus amarus[J].Europe an Journal of Pharmacology 2006,546:182-188.
[48]Murugaiyah V,Chan KL,.Antihyperuricemic lignans from the leaves of Phyllant hus niruri[J].Planta Med.2006,72(14):126 2-1267.
[49]Chia-Chuan Chang,Yu-Chin Lien,Karin C.S,etal.Lignans from Phyllanthus urina ria[J].Phytochemistry 63(2003)825-833
[50]Kadali S.Sagar,Chia-Chuan Chang,Wei-Kung Wang,etal.Preparation and anti- HIV activities of retrojusticidin B analogs and azalignans[J].Bioorganic & Medicinal Chemistry 12(200 4) 4045-4054
[51]Vikneswaran Murugaiyah,Kit-Lam Chan,Determination of four lignans in Phyll anthus niruri L.bya simple high-performance liquid chromatography method with fluorescence detecti on[J].Journal of Chromatography A,1154 (2007) 198-204
[52]Arvind K.Tripathi,Ram K.Verma,Anil K.Gupta,Quantitative determinatio n of phyllanthin and hypophyllanthin in Phyllanthus species by high- perfo rmance thin layer chromatography[J].Phytochem Anal.2006,17(6):394-397
[53]姚庆强,左春旭.叶下珠化学成分的研究[J].药学学报,1993,28(11):829-835
[54]Anti-inflammatory properties of extracts,fractions and lignans isolated from Phyllanthus amarus.[J]Planta Med.2005,71(8):721-726.
[55]Leite DF,Kassuya,The cytotoxic effect and the multidrug resistance revers ing action of lignans from Phyllanthus amarus[J].Planta Med.2006,7 2(15):1353-1358
[2]Thyagarajan S P,Thiruneelakantan K,Subramanian S,etal.In vitro inac tivation of HBsAg by Eclipla alba Hassk and Phllanthus uiruri linn[J].Indian J Med Res,1982,76(Suppl):124-130
[3]牛晓峰,李维凤.陕西叶下珠属植物资源概况[J].西安交通大学学报(医学版),2003,24(2):119-123
[4]李秉滔.中国叶下珠属植物的修订[J].植物分类学报,1987,25(5):371-375
[5]万振先,周国平,易扬华.叶下珠化学成分的研究(Ⅰ)[J].中草药,1994,25(9):455-456
[6]万振先,易扬华,喻庆禄.叶下珠化学成分的研究(Ⅱ)[J].中草药,1997,28(3):134-135
[7]张兰珍,郭亚健,涂光忠,等.叶下珠多酚化合物的分离与鉴定[J].中国中药杂志,2000,25(12):724-725
[8]陈玉武,任丽娟,李克明,等.叶下珠新多酚化合物的分离与鉴定[J].药学学报,1999,34(7):526-529
[9]张兰珍,郭亚健,涂光忠,等.叶下珠新鞣花鞣质的分离与鉴定[J].药学学报,2004,39(2):119-122
[10]Somanabandhu A,Nitayangkura S,Mahidol C,etal.~1H-and ~(13)C - NMR assignm ents of phyllanthin and hypophyllanthin:lignans that enhance cytotoxic resp onses with cultured multidrug-re sistant cells[J].Journal of Natural Products,1993,56:233-239
[11]Anjaneyulu ASR,Rao K J,Row L R,etal.Crystalline constituents of Euphorbi aceae,12.Isolation and structural elucidation of 3 new lignans from leaves of Phyllanthus niruri linn[J].Tetra hedron,1973,29:1291-1298
[12]Stevenson R,Williams J R.Concerning phyltetralin synthesis of lignan arylt etralin isomers[J].Tetrahedron,1977,33:2913-29 17
[13]Ward R S,Satyanarayana P,Row L R,etal.The case for a revised structure for hypophyllanthin ananalysis of the ~(13)C-NMR spectra of sryltet ralins[J].Tetr ahe dron Lett,1979,32:3043-3046
[14]Huang Y L,Chen C C,Ou J C.Isolintetralin:a new lignan from phylanthus nir uri[J].Planta Med,1992,58:473-474
[15]Huang Y L,Chen C C,Hsu F L,etal.A new lignan from phyllanthus virgat us[J].Journal of Natural Products,1996,59:520-521
[16]Chia C C,Yu C L,Karin C S,etal.Lignans from phyllanthus urinaria[J].phytoc hemistry,2003,63:825-833
[17]Wang X W,Yuan J P,Yai Z C,etal.Carboxylic acids from phyllanthus urinaria [J].Chemistry of natural compounds,2005,41(1):17-21
[18]Syamasundar K V,Singh B,Thakur R S,etal.Antihepatotoxic principles of phy llantus uiruri herbs[J]Ethnoparmacol,1985,14(1):41-43
[19]Blumberg B S,Millman I,Venkateswaran P S,Thyagarajan S P.Hepatitis B virus and he patocellular carcinoma:treatment of HBV carriers with Phyllan thus amarus[J].Cancer Detect,1989,14:195-201
[20]Thyagarajan S P,Subramanina S,Thirunalasundari T,etal.Effect of Phllanthus amarus on chronic carriers of hepatitis B virus[J].Lancet,1988,1:764-766
[21]Syamasundar K V,Singh B,Thakur R S,et al.Antihepatotoxic Principles of Phyllantus Niruri Herbs[J].Ethnoparmacol,1985,14(1):41-44
[22]任进余,秦存林苦味叶下珠、叶下珠的药理作用及其作用机理.国外医学.中医中药分册[M],19 99,12(2):53-61.
[23]纪徐淮,秦一中,汪伟业,等.rhIFN-ab和苦味叶下珠提取物对人肝癌细胞株PLC/PRF/5产生HBsAg的影响[J].新消化病学杂志,1995,3(3):136-138
[24]王昌俊,袁德培,陈伟,等.叶下珠对人肝癌细胞的影响[J].时珍国药研究,1997,8(6):499-450
[25]黄育华,张建军,晏雪生,等.叶下珠对人肝癌细胞Bel-7402诱导分化的影响[J].湖北中医学院学报,2000,2(1):10-12
[26]Blumberg B S,Millman I,Venkateswaran P S,etal.Hepatitis B virus and primary hepatocellular carcinoma:treatment of HBV carriers with Phyllant husamarus[J].Vaccine,1990,8(Suppl):S86-S92
[27]张建军,黄育华,晏雪生,等.叶下珠药物血清对人肝癌细胞株的诱导分化作用的实验研究[J].中国中医药科技,2002,9(5):289-291
[28]郭卫真,邓学龙,董伯振,等.叶下珠属植物体外抗单纯疱疹病毒Ⅱ型的作用[J].广洲中医药大学学报,2000,17(1):54-57
[29]ZuoChunxu,Li Fengqin,Yao Qingqiang.Antiviral dehydroch ebulic acid met hyl and its preparation[J].CA,1996,124:185 53/u
[30]Zuo Chunxu,Li Fengqin,Yao Qingqiang.Antiviral methyl brevifolin carbox ylate and its preparation[J].CA,1996,124:66574/a
[31]马新宇,裴俊俊,叶下珠药理研究综述[J].齐鲁药事2006 25(2):104-105
[32]kumaran,R.Joel Karunakaran.In vitro antioxidant activities of meth anol extracts of five phyllanthus species from India[J].Swiss Society of Food Science and Technology 2007,40:344-352
[33]R.Harish,T.Shivanandappa.Atioxidant activity and hepatoprotective potenti al of phyllanthus niruri[J].Food Chemistry,2006,95:180-185
[34]仲英,左旭,张琴冈等.叶下珠化学成分及其抗乙肝病毒活性的研究[J].中国中药杂志,1998,23(6):363-364.
[35]Benner R,Weliky K,Hedges J 1.Early diagenesis of mangrove leaves in a tropicalest uary:Molecular-levela nalyses of neutral sugars and lignin-deriv ed phenols[J].Geochimicaet C osmochimica Acta,1990,54(7):1991-2001.
[36]马金强,贾正平,王彬,等.瑞香狼毒总木脂素与长春新碱的体外抗肿瘤 活性比较[J].西北国防医学杂志,2004,25(5):374-377
[37]Schrroder HC,Mertz H,Steffen R,etal.Z.Naturforsch.Sect.C:Biosci,1990,45:1215-1221
[38]Gnabre JN,Ito Y,Ma Y,etal.J.Chroma.A,1996,719(2):353-364
[39]金昔陆,顾静,胡天喜,等.内南五味子木脂素戈米辛J对肝线粒体膜脂质过氧化和超氧阴离子自由基的作用[J].中国药理学通报,2000,16(1)26-30
[40]韩桂秋,中草药中血小板活化因子受体拮抗活性成分的研究[J].自然科学进展国家重点实验室通讯,1995,5(2):161-164
[41]张永忠,李小莉,郭群.辛夷木脂素类成分抗血小板活化因子作用的研究[J].湖北中药杂志,2001,23(10):7-10
[42]A.S.R.Anjaneyulu,K.Jaganmohan Rao,L.R amachandra Row,etal.Crystal line Constituents of Euphorbiaceae-Ⅻ isolation and Structural El ucidation of There New Lignans from the Lea ves of Phyllanthus Niruri Linn.Tetrahedr on,1973,29:1291-1298
[43]Panchagnula Satyanarayana,somepalli Venkateswarlu.Isolation,Structure and Synthesis of New Diarylbutane Lignans from Phyllanthus Niruri,Synthesis of 5 '-Desmethoxy Niranthin and An Antitumour Extractive.Trtrahedron,1991,47(42):8931-8940
[44]Leite DF,Kassuya CA,Mazzuco TL,etal.The cytotoxic effect and the multidrug resistance reversing action of lignans from Phyllanthus amarus[J].Plant Me d,2006,72(15):1353-1358.
[45]Syamsundar KV,Singh B,Thakur RS,etal.Antihepatotoxic principles of Phyll anthus niruri herbs[J]Ethnopharmacol.1985,14(1):41-44.
[46]Venkateswaran P S,Millman I,Blumberg B S,et al.Effects of an extract from phyllanthus urinari on hepatitis B and woodch uck hwpatitis viruses:in vitro and in vivo studies[J].Proc Na ti Acad Sci USA,1987,84:274-279
[47]Candida AL.Kassuya,Aline Silvestre,Octavio Menezes-de-Lima Jr.etal.Antiinfla mmatory and antiallodynic actions of the lignan niranthin isolated from Phyllanthus amarus[J].Europe an Journal of Pharmacology 2006,546:182-188.
[48]Murugaiyah V,Chan KL,.Antihyperuricemic lignans from the leaves of Phyllant hus niruri[J].Planta Med.2006,72(14):126 2-1267.
[49]Chia-Chuan Chang,Yu-Chin Lien,Karin C.S,etal.Lignans from Phyllanthus urina ria[J].Phytochemistry 63(2003)825-833
[50]Kadali S.Sagar,Chia-Chuan Chang,Wei-Kung Wang,etal.Preparation and anti- HIV activities of retrojusticidin B analogs and azalignans[J].Bioorganic & Medicinal Chemistry 12(200 4) 4045-4054
[51]Vikneswaran Murugaiyah,Kit-Lam Chan,Determination of four lignans in Phyll anthus niruri L.bya simple high-performance liquid chromatography method with fluorescence detecti on[J].Journal of Chromatography A,1154 (2007) 198-204
[52]Arvind K.Tripathi,Ram K.Verma,Anil K.Gupta,Quantitative determinatio n of phyllanthin and hypophyllanthin in Phyllanthus species by high- perfo rmance thin layer chromatography[J].Phytochem Anal.2006,17(6):394-397
[53]姚庆强,左春旭.叶下珠化学成分的研究[J].药学学报,1993,28(11):829-835
[54]Anti-inflammatory properties of extracts,fractions and lignans isolated from Phyllanthus amarus.[J]Planta Med.2005,71(8):721-726.
[55]Leite DF,Kassuya,The cytotoxic effect and the multidrug resistance revers ing action of lignans from Phyllanthus amarus[J].Planta Med.2006,7 2(15):1353-1358