虎杖白藜芦醇、白藜芦醇苷和花生白藜芦醇含量的研究
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摘要
白藜芦醇(resveratrol)是植物受外界刺激而产生的一类低分子量、具有生物活性的植物抗毒素(phytoalexin)。白藜芦醇苷(piceid)是白藜芦醇的葡萄糖苷形式。本文对福建地产虎杖(Polygonum cuspidatum)饮片的白藜芦醇和白藜芦醇苷含量以及花生(Arachis hypogaea)植株的不同组织、不同加工品白藜芦醇的含量进行了研究。
用95％乙醇从虎杖样品中提取白藜芦醇及白藜芦醇苷，采用反相高效液相色谱(HPLC)法检测二者的含量，色谱柱为ODS Hypersil(4.6mm×250.0mm，5μm)，流动相为乙腈／水(20／80)，检测波长为306nm。结果表明，白藜芦醇在0.01168～0.05840mg／mL的浓度范围内峰面积积分值与浓度的相关系数r=0.9999，相关性极显著(α=0.01)；所测定的虎杖饮片白藜芦醇平均含量为0.441％(干重)，平均回收率为98.33％：白藜芦醇苷在0.0216～0.1944mg／mL的浓度范围内峰面积积分值与浓度的相关系数r=0.9999，相关性极显著(α=0.01)；所测定的虎杖饮片白藜芦醇苷平均含量为1.756％(干重)，平均回收率为98.46％。
用95％乙醇提取花生样品中的白藜芦醇经柱层析纯化(硅胶／氧化铝为固定相，80％乙醇为流动相)后，采用HPLC法检测，色谱柱为ODS Hypersil(4.6mm×250.0mm，5μm)，流动相为甲醇／水(45／55)，检测波长为306nm。实验结果表明，白藜芦醇在0.01184～0.05920mg／mL的浓度范围内峰面积积分值与浓度的相关系数r=0.9999，相关性极显著(α=0.01)；花生根白藜芦醇的含量最高，达0.1795mg／g(干重)，平均回收率为97.98％；茎、壳、煮熟花生中白藜芦醇含量低微；叶、种子、烤花生、花生酱未检测出白藜芦醇。
Resveratrol is a kind of phytoalexin, which is low molecular weight, biologically active metabolite produced by plants as defense to various exogenous stimuli. Piceid is the glycoside of resveratrol. Research was done to determine the content of reveratrol and piceid in Polygonum cuspidatum and the content of reveratrol in the different tissues and different processing samples of peanut.
Reveratrol and piceid were extracted from P.cuspidatum with 95% ethanol and the content was determined by HPLC (high performance liquid chromatographic), with ODS Hypersil column (4.6mmx250.0mm, 5um). A mixture of CH3CN/H2O (20/80) was used as the mobile phase and the wavelength for detecting was 306nm. Results indicated that the linear range of resveratrol was 0.01168 ~ 0.05840mg/mL(r=0.9999, a =0.01).The average content of revesratrol was 0.441%, and the average recovery rate was 98.33%; The linear range of piceid was 0.0216~ 0.1944mg/mL(r=0.9999, a =0.01).The average content of piceid was 1.756%, and the average recovery rate was 98.46%.
Reveratrol was extracted from the different tissues and different processing samples of peanut with 95% ethanol. The extract was purified by chromatographic (the mixture of AI2O3 and silica gel as solid phase, 80% ethanol as mobile phase).Then the content was determined by HPLC, with ODS Hypersil column (4.6mmx250.0mm, 5um). A mixture of CH3OH/H2O (45/55) was used as the mobile phase and the wavelength for detecting was 306nm. Result indicated that the linear range was 0.01184~0.05920mg/mL(r=0.9999, a =0.01).The root of Arachis hypogaea contained the highest content of resveratrol (0.1795mg/g). The average recovery rate was 97.98%. The stem, shell and boiled peanut contained very little resveratrol. The leaf, seed, toasted peanut and peanut butter did not contain resveratrol.
用95％乙醇从虎杖样品中提取白藜芦醇及白藜芦醇苷，采用反相高效液相色谱(HPLC)法检测二者的含量，色谱柱为ODS Hypersil(4.6mm×250.0mm，5μm)，流动相为乙腈／水(20／80)，检测波长为306nm。结果表明，白藜芦醇在0.01168～0.05840mg／mL的浓度范围内峰面积积分值与浓度的相关系数r=0.9999，相关性极显著(α=0.01)；所测定的虎杖饮片白藜芦醇平均含量为0.441％(干重)，平均回收率为98.33％：白藜芦醇苷在0.0216～0.1944mg／mL的浓度范围内峰面积积分值与浓度的相关系数r=0.9999，相关性极显著(α=0.01)；所测定的虎杖饮片白藜芦醇苷平均含量为1.756％(干重)，平均回收率为98.46％。
用95％乙醇提取花生样品中的白藜芦醇经柱层析纯化(硅胶／氧化铝为固定相，80％乙醇为流动相)后，采用HPLC法检测，色谱柱为ODS Hypersil(4.6mm×250.0mm，5μm)，流动相为甲醇／水(45／55)，检测波长为306nm。实验结果表明，白藜芦醇在0.01184～0.05920mg／mL的浓度范围内峰面积积分值与浓度的相关系数r=0.9999，相关性极显著(α=0.01)；花生根白藜芦醇的含量最高，达0.1795mg／g(干重)，平均回收率为97.98％；茎、壳、煮熟花生中白藜芦醇含量低微；叶、种子、烤花生、花生酱未检测出白藜芦醇。
Resveratrol is a kind of phytoalexin, which is low molecular weight, biologically active metabolite produced by plants as defense to various exogenous stimuli. Piceid is the glycoside of resveratrol. Research was done to determine the content of reveratrol and piceid in Polygonum cuspidatum and the content of reveratrol in the different tissues and different processing samples of peanut.
Reveratrol and piceid were extracted from P.cuspidatum with 95% ethanol and the content was determined by HPLC (high performance liquid chromatographic), with ODS Hypersil column (4.6mmx250.0mm, 5um). A mixture of CH3CN/H2O (20/80) was used as the mobile phase and the wavelength for detecting was 306nm. Results indicated that the linear range of resveratrol was 0.01168 ~ 0.05840mg/mL(r=0.9999, a =0.01).The average content of revesratrol was 0.441%, and the average recovery rate was 98.33%; The linear range of piceid was 0.0216~ 0.1944mg/mL(r=0.9999, a =0.01).The average content of piceid was 1.756%, and the average recovery rate was 98.46%.
Reveratrol was extracted from the different tissues and different processing samples of peanut with 95% ethanol. The extract was purified by chromatographic (the mixture of AI2O3 and silica gel as solid phase, 80% ethanol as mobile phase).Then the content was determined by HPLC, with ODS Hypersil column (4.6mmx250.0mm, 5um). A mixture of CH3OH/H2O (45/55) was used as the mobile phase and the wavelength for detecting was 306nm. Result indicated that the linear range was 0.01184~0.05920mg/mL(r=0.9999, a =0.01).The root of Arachis hypogaea contained the highest content of resveratrol (0.1795mg/g). The average recovery rate was 97.98%. The stem, shell and boiled peanut contained very little resveratrol. The leaf, seed, toasted peanut and peanut butter did not contain resveratrol.
引文
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[2] Hain Ruediger, Reif Hans-Joerg., Krause Elvira., Langebartels Ruth Kindl Hel mut., Vornam Barbara., Wiese Wilfried., Schmelzer Elmon., Schreier Peter H. Disease resistance results from foreign phytoalexin expression in a novel plant.Nature, 1993, 361(6408): 153-156
[3] Jang M., Cai L., Udeani G. O., Slowing K. V., Thomas C. F., Beecher C. W. W., Fong H. H. S., Farnsworth N. R., Kinghorn A. D., Mehta R. G., Moon R. C., Pezzuto J. M. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science, 1997, 275: 218-220
[4] Jeandet P., Bessis R., Gautheron B. The production of resveratrol(3,5,4'-trihydroxystilbene) by grape berries in different developmental stages. Am J Enol Vitic, 1991, 42: 41-46
[5] Ingham J.L. 3,5,4,Trihydroxystibene as a phytoalexin from groundnuts (Arachis hypogaea). Phytochemistry, 1976, 15: 1791-1793
[6] Mannila Erkki, Talvitie Antti, Kolehmainen Erkki. Anti-leukaemic compounds derived from stilbenes in picea abies bark. Phytochemistry, 1993, 33(4): 813-816
[7] Iinuma Munekazu, Ohyama Masayoshi, Tanaka Toshiyuki. Six flavonostilbenes and a flavanone in roots of sophora alopecuroides. Phytochemistry, 1995, 38(2): 519-526
[8] Kotha Subbaramaiah, Wen Jing Chung, Pedro Michaluart, Nitin Telang,
Tadashi Tanabe, Hiroyasu Inoue, Meishiang Jang, John M. Pezzuto, Andrew J. Dannenberg. Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary epithelial cells. J Biol Chem, 1998,273: 21875-21882
[9] Martinez J., Moreno JJ. Effect of resveratrol, a natural polyphenolic compound, on reactive oxygen species and prostaglandin production. Biochem Pharmacol,2000, 59(7): 865-870
[10] Shih A., Davis F.B., Lin H.Y., Davis P.J. Resveratrol induces apoptosis in thyroid cancer cell lines via a MAPK-and p53-dependent mechanism. J Clin Endocrinol Metab, 2002, 87(3): 1223-1232
[11] Jang M., Pezzuto J. M. Effect of resveratrol on 12-O-tetradecanoylphorbol-13-acetate-induced oxidative events and gene expression in mouse skin. Cancer Lett, 1998, 134(1): 81-89
[12] Frankel E. N., Parks E. J., Xu R., Schneeman B. O., Davis P.A., German J.B. Effect of n-3 fatty acid-rich fish oil supplementation on the oxidation of low density lipoproteins. Lipids, 1994, 29(4): 233-236
[13] Goldberg D. M., Hahn S. E., Parkes J. G. Beyond alcohol: beverage consumption and cardiovascular mortality. Clinica Chimica Acta, 1995, 237: 155-187
[14] Bertelli A. A., Giovannini L., Giannessi D., Migliori M., Bemini W., Fregoni M., Bertelli A. Antiplatelet activity of synthetic and natural resveratrol in red wine. Int J Tissue React, 1995, 17(1): 1-3
[15] Wang Z., Zou J., Huang Y., Cao K, Xu Y, Wu JM. Effect of resveratrol on platelet aggregation in vivo and vitro. Chin Med J(Engl), 2002, 115(3): 378-380
[16] Zou J., Huang Y., Chen Q., Wei E, Cao K, Wu JM. Effects of resveratrol on
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