胡芦巴甾体皂苷的提取精制及其半乳甘露聚糖的利用研究
|
摘要
运用现代科学技术和研究方法,从中药中提取有活性的的化学成分,可以使人们用上高效无毒副作用或毒副作用小的天然药物,充分发挥中草药的优越性。胡芦巴是一种有着广泛应用的传统中药。胡芦巴中富含有活性成分——甾体皂苷,甾体皂苷具有很强的生物活性,如毒杀多种癌细胞,降血脂,抗白细胞等作用。
本论文对胡芦巴甾体皂苷的不同提取工艺进行了优选。主要优化纤维素酶解法提取工艺和超声辅助提取工艺,并进行对比研究,筛选一种胡芦巴甾体皂苷得率较高的工艺。最后选择超声辅助提取的方法提取胡芦巴甾体皂苷,最佳工艺参数：乙醇浓度为50%,料液比为1：20,超声时间为40 min,超声功率为60 W,此时,结果甾体皂苷提取率的平均值为5.89%。
研究了大孔吸附树脂精制胡芦巴甾体皂苷的最佳条件,使用大孔吸附树脂的型号为ADS-7,上柱液浓度为1.20 mg/mL左右,树脂柱高径比为20,上柱吸附流速为1.5mL/min,解吸液为80%的乙醇溶液,解吸流速2 mL/min,选择解吸剂用量为250 mL,解吸率可达到97.80%,树脂经过多次使用后其吸附量稳定在最大吸附量的77%。
胡芦巴半乳甘露聚糖具有显著的降血糖,调血脂和减肥的功效,但其溶解度很小,本论文对其开发利用进行了更进一步的研究。酸水解胡芦巴半乳甘露聚糖制备易溶于水的膳食纤维,随着水解时间的延长,半乳甘露聚糖的黏均相对分子质量变小,由原来的5.5×105可降低到9.9x103,并且溶解度增大。
水解过的胡芦巴半乳甘露聚糖可以用来制备微球,制备的微球可以成为一种潜在的药物载体。
Fenugreek is a kind of traditional Chinese herbs which has a long history. Optimum extracting techniques of steroid saponins from fenugreek seeds were studied. Enzymatic hydrolysis extraction method and ultrasonic extraction technique were studied comparatively. The best extracting technology was screened out. Macro-porous adsorptive resin was used on the steroid saponins purification. A kind of freely soluble dietary fiber was produced by acid hydrolyzing. The micro spheres were prepared by the freely soluble dietary fiber.
The optimum method of extracting steroid saponins from fenugreek seeds by ultrasonic extraction technique was studied. The orthogonal design was adopted to optimize the alcohol extraction process. The contents of steroid saponins from fenugreek seeds were determined by colorimetric method. The order of the factors which affected the extraction was:concentration of ethanol>solid-liquid ratio>extracting time>ultrasonic power. The results showed that the yield of steroid saponins was 5.89% at the conditions of 50% ethanol solution,1:20 solid-liquid ratio and 40 min for extraction.
Taking the ADS-7 macro-porous adsorptive resin on steroid saponins adsorptive capacity and desorption rate as measuring index, all sorts of factors which effected the refining impact were reviewed in static and dynamic state. The adsorption-desorption optimum conditions are as follows:resin adsorbent temperature is room temperature, the steroid saponins concentration in loading solution is 1.20 mg/mL around, desorption solution concentration is 80% ethanol solution, the ratio height to diameter of resin column is 20, flow rate of loading solution is 1.5 mL/min, the dosage of desorption solution is 250 mL and flow rate of desorption solution is 2 mL/min.
Freely soluble dietary fiber was produced by acid hydrolyzing. The viscosity-average molecular weight was definitely decreased. The main chain of the galactomannan is degraded during hydrolyzing, and the whole molecular is sheared into many fragments, which induced the decrease of its viscosity-average molecular weight from 5.5×105 to 9.9×103. There is no obviously difference between FT-IR spectra of original fenugreek galactomannan and products.
FT-IR spectra of fenugreek galactomannan micro sphere showed the association happened between-OH bonds of fenugreek galactomannan, partial hydrogen bond was instead by crosslinked bond.
本论文对胡芦巴甾体皂苷的不同提取工艺进行了优选。主要优化纤维素酶解法提取工艺和超声辅助提取工艺,并进行对比研究,筛选一种胡芦巴甾体皂苷得率较高的工艺。最后选择超声辅助提取的方法提取胡芦巴甾体皂苷,最佳工艺参数：乙醇浓度为50%,料液比为1：20,超声时间为40 min,超声功率为60 W,此时,结果甾体皂苷提取率的平均值为5.89%。
研究了大孔吸附树脂精制胡芦巴甾体皂苷的最佳条件,使用大孔吸附树脂的型号为ADS-7,上柱液浓度为1.20 mg/mL左右,树脂柱高径比为20,上柱吸附流速为1.5mL/min,解吸液为80%的乙醇溶液,解吸流速2 mL/min,选择解吸剂用量为250 mL,解吸率可达到97.80%,树脂经过多次使用后其吸附量稳定在最大吸附量的77%。
胡芦巴半乳甘露聚糖具有显著的降血糖,调血脂和减肥的功效,但其溶解度很小,本论文对其开发利用进行了更进一步的研究。酸水解胡芦巴半乳甘露聚糖制备易溶于水的膳食纤维,随着水解时间的延长,半乳甘露聚糖的黏均相对分子质量变小,由原来的5.5×105可降低到9.9x103,并且溶解度增大。
水解过的胡芦巴半乳甘露聚糖可以用来制备微球,制备的微球可以成为一种潜在的药物载体。
Fenugreek is a kind of traditional Chinese herbs which has a long history. Optimum extracting techniques of steroid saponins from fenugreek seeds were studied. Enzymatic hydrolysis extraction method and ultrasonic extraction technique were studied comparatively. The best extracting technology was screened out. Macro-porous adsorptive resin was used on the steroid saponins purification. A kind of freely soluble dietary fiber was produced by acid hydrolyzing. The micro spheres were prepared by the freely soluble dietary fiber.
The optimum method of extracting steroid saponins from fenugreek seeds by ultrasonic extraction technique was studied. The orthogonal design was adopted to optimize the alcohol extraction process. The contents of steroid saponins from fenugreek seeds were determined by colorimetric method. The order of the factors which affected the extraction was:concentration of ethanol>solid-liquid ratio>extracting time>ultrasonic power. The results showed that the yield of steroid saponins was 5.89% at the conditions of 50% ethanol solution,1:20 solid-liquid ratio and 40 min for extraction.
Taking the ADS-7 macro-porous adsorptive resin on steroid saponins adsorptive capacity and desorption rate as measuring index, all sorts of factors which effected the refining impact were reviewed in static and dynamic state. The adsorption-desorption optimum conditions are as follows:resin adsorbent temperature is room temperature, the steroid saponins concentration in loading solution is 1.20 mg/mL around, desorption solution concentration is 80% ethanol solution, the ratio height to diameter of resin column is 20, flow rate of loading solution is 1.5 mL/min, the dosage of desorption solution is 250 mL and flow rate of desorption solution is 2 mL/min.
Freely soluble dietary fiber was produced by acid hydrolyzing. The viscosity-average molecular weight was definitely decreased. The main chain of the galactomannan is degraded during hydrolyzing, and the whole molecular is sheared into many fragments, which induced the decrease of its viscosity-average molecular weight from 5.5×105 to 9.9×103. There is no obviously difference between FT-IR spectra of original fenugreek galactomannan and products.
FT-IR spectra of fenugreek galactomannan micro sphere showed the association happened between-OH bonds of fenugreek galactomannan, partial hydrogen bond was instead by crosslinked bond.
引文
[1]南往.胡芦巴[EB/OL].百度百科,2008-2-2.
[2]荆宇,赵余庆,胡芦巴化学成分和药理作用研究进展[J].中草药,2003,34(12)：1146-1149.
[3]杨永利,孙双彦,崔德锋,等.胡芦巴种子化学成分的研究[J].西北师范大学学报,2003,39(1)：64-65.
[4]周维善,庄治平主编.,甾体化学进展.,第一版,北京：科学出版社,2002.
[5]Yoshikawa M. Medicinal food stuffs Ⅳ. Fenugreek seed(1):Structures of trigoneosides Ⅰa, Ⅰb, Ⅱa, Ⅱb, Ⅲa, and Ⅲb, new furostanol saponins from the seeds of Indian Trigonell a foenum-graecumL[J]. Chem Pharm Bull,1997,45(1):81-87.
[6]Yoshikawa M. Medicinal food stuffs Ⅳ. Fenugreek seed(1):Structures of six new furostanol saponins trigoneosides Ⅳa, Ⅴa, Ⅴb, Ⅶb, Ⅶb and Ⅷb fromm the seeds of Trigonell a foenum-graecumL[J]. Heterocycles,1998,47 (1):397-405.
[7]Murakami T, Kishi A, Matsuda H, et al. Medicinal food stuffs. ⅩⅦ. fenugreek seed(3):Structures of new furostanol-type steroidsaponins, trigoneosides Xa, Ⅹa, Ⅺb, Ⅻa, Ⅻb and ⅩⅢ fromm the seeds of Trigonell a foenum-graecumL[J]. Chem Pharm Bull,2000,48(7):994-1000.
[8]Shang MY, Cai SQ, Kadda S, et al. Structure identification of trigoneoside VIII. [J]. Acta Pharm Sin (药学学报),2001,36(11):836-839.
[9]Shang M Y, Cai S Q, Han J, et al. Studies on flavone of Trigonella foenum-graecum L. [J]. China J Chin Mater Med (中国中药杂志),1998,23(10):614-616.
[10]Yingmei H, Sansei N, Yukari N, et al. Flavonol glycosides from the stems of Trigonella foenum-graecum L. [J]. phyto-chemistry,2001,58:577-580.
[11]Shang M Y, Tezuka Y, Cai S Q, etal. Studies on triter-penoids from common fenugreek (Trigonella foenum-grae-cumL.). [J]. Chin tradit Herb Drugs(中草药), 1998,29(10):655-657.
[12]Shang M Y, Kadota S, Wang X, et al Studies on lipo so luble components of common fenugreek seeds (T rigonella foenum-graecum L.) [J].Chin Trad it Herb Drugs(中草药),1999,30(4):256-257.
[13]Shang M Y, Cai S Q, LinW H, et al. Studies on chemical constituents of Trigonella foenum-graecum L. [J]. China J Chin MaterM ed(中国中药杂志),2002,27(4): 277-279.
[14]Fernando P L, Martha R M, Carlo s A C, et al. So to lone production by hairy root cultures of Trigonella foenum-graecum L. in airlift with mesh bioreactors [J]. J Agr Food Chem,2001,49:6012-6019.
[15]缪月秋,梅秋红,顾龚平.胡芦巴中性杂多糖酶解工艺优化及其产物对青春双歧杆菌增殖作用的研究[J].安徽农业大学学报,2006,33(2)：238-242.
[16]李德亮,崔元臣,宋丽萍等.田箐胶在麦草造纸中的应用研究[J].中国造纸,1999,18(6)：46-48.
[17]Mirzaeva M R, Rakhmanberdyeva R K, Rakhimov D A. Structure of galactomannan from seeds of Gleditsia macracantha[J]. Chem Nat Compd,2000,35(5):505-507.
[18]Smirnova N I, Shcherbukhin V D. Galactomannan from caspia seeds[J]. Prikl.Biokhim. Mikrobiol,1998,24(5):653-659.
[19]Davison R L. Handbook of Water-soluble Galactomannan and Resins[J]. US,1980.
[20]Kooiman P. Structures of the galactomannans from seeds of Annona Muricata、 Cocos nucifera and sophoria japonica[J]. Carbonhyrate Research,1975,20(2): 329-337.
[21]Olivier N E, Taravel F. Nuclear magnetic resonance evidence of the guar borate interaction[J]. Carbohyarate Research,1987,16(6):1-11.
[22]Shimahara H, Suzuki H, Sugiyama N, et al. Isolation and characterization of olifosaccharides from an enzymic hdrolysate of konjac glucomannan[J]. Agric Biol Chem,1975,39(2):293-229.
[23]郭文松,刘达雯,周训清,等.甾体皂苷和苷元的研制[J].贵州化工,1996,(2)：14-17.
[24]Chen Y Y, Yuang C. The blood glucose reducing of fenugreek seeds abstracts in the alloxan-diabetic S. D. rats[J]. Zhen jiang Med Coll (镇江医学院学报),2001, 11(12):163-166.
[25]Madar Z, Shomer I J, Polysaccharides composition of gel fraction derived from fenugreek and its effect on starch digestion and bile acid absorption in rats[J]. A gric food.Chem,1990,38:1535～1539.
[26]Shani J, Glodschmied A, Joseph B, et al. Hypoglycemic effect of Trigonella foenum graecum and luminus term is seed and their major alkoloid in alloxan-diabetic and normal rats[J]. Arch. Int Pharmacodyn Ther,1974,210:27-36.
[27]Ribes G Y, Sauvaire Y, Baccou J, et al. Antidiabetic effect of subfractions from fenugreek seeds in diabetic dogs[J]. Proc Soc Exp Bio Med,1986,182:159-166.
[28]Amin Ryiad M, Abdul-Ghani Abdul-Salam S, Soleiman Mohammad S. Effect of fenugreek seeds and liupine seeds on the development of the experimental diabetes in rats[J]. Planta Med,1988:286-290.
[29]Sauvaire Y, Ribes G Y, Baccou J, et al. Implication of steroid saponin and sapogenins in the hypocholestero lemic effect of fenugreek[J]. Lipids,1991,26: 191-197.
[30]Shang M Y, Cai S Q, Wang X, et al. Analysis of aminoacid of Trigonella foenum-graecum L. [J]. Chin Med Mater(中药材),1998,21(4)：188-189.
[31]Suja P R, Anuradha C V, iswanathan P. et al. Gastropro-tective effect of fenugreek seeds(Trigonella foenum-graecum L.) on experimental gastricul cerinrats[J]. JE thnophar-macol,2002,81(3):393-397.
[32]Sur P, Das M, Comes A, et al. Trigonella foenum-graecum L. (fenugreek)seed extract as an antineo plastic agennt. [J]. Ohy-tother Res,2001,15(3):257-259.
[33]Zhu B L, Ban Y H, Duan J A. Protective effect of fenugreek seed on acute chemical liver injury[J]. Chun J Ind Med(中国工业医学杂志),2000,13(1)：19-21.
[34]Li L L, Mao X M, Wang X F, et al. The memory facilitation of TFGS and its protective effect against cerebral is chemia[J]. Chin Pharmacol Bull(中国药理学通报),2001,17(1)：92-94.
[35]Weder J K, Haussner K. Inhibitors of human and bovine trypsin and chy-motrypsin in fenugreek(Trigonella foenum-graecum L.) seeds[J]. Z Lebensm Unters Forsch, 1991,192(6):535-540.
[36]Panda S, Tahiliani P, Kar A. Inhibition of triiodothyronine production by fenugreek seed extract in mice and rats[J]. Pharmacol Res,1999,40(5):405-409.
[37]SHEN R L. The research progress of dietary firbre [J]. Shanxi Food Industry,2002, (2):21-24.
[38]TIAN X H. Function characteristic and application of the dietary fibre[J]. Food Research And Development,2002, (03):56-57.
[39]何锦风,郝利民,论膳食纤维[J].食品发酵工业,1999,23(5)：63-72.
[40]刘丹,常伟勤.复合膳食纤维对高胆固醇血症小鼠降脂作用的研究[J].北华大学学报(自然科学版),2006,(01)：35-37.
[41]吴杰.魔芋精粉与几种常用膳食纤维降血脂效果比较及机理探讨[J].营养学报,1993,15(2)：168.
[42]Cummings John H. Dietary carbohydrates and health:do we still need the fibre concept[J]. Clinical nutrition supplements,2004,1(2):5-7.
[43]Heaton K W. Dietary fibre and energy intake[J]. The lancet,1974,303(7853): 368-369.
[44]Riccardi G, et al. Dietary fibber in treatment of diabetes:myth or reality [J]. Digestive and Liver Disease,2002,34(2):140-144.
[45]欧仕益,高孔荣.膳食纤维抑制抑制膳后血糖升高的机理探讨[J].食品科学,1998,19(3)：48-52,
[46]邢小燕,张波.高膳食纤维有利于预防2型糖尿病—新疆克拉玛依市和山西忻州农村饮食状况调查[J].中日友好医院学报,2002,16(3)：134-137.
[47]韦晖.膳食纤维的研究与开发.纤维素科学与技术,2001,9(1)：56-63,
[48]阂锐,何云海,姚晓敏.开发人体第七营养素膳食纤维[J].中国食物与营养,1999, (3)：18-21.
[49]金其荣.低聚搪及根霉低聚糖生理功能[J].食品科学,1994,(1)：5-8.
[50]梁静谊,将建新,朱莉伟,等.皂荚皂苷提取工艺研究[J].中国野生植物资源,2003,22(6)：58-61.
[51]胡湘春,鱼红闪.穿山龙薯蓣皂苷的分离提纯[J].大连轻工业学院学报,2004,23(3)：25-29.
[52]康阿龙.胡芦巴素新药的开发研究[D].西北大学硕士学位论文,2001.
[53]杨献文,李苏梅,等.麦冬总皂苷提取工艺研究[J].中草药,2002,33(12)：1076-1078.
[54]吴红,梁恒,刘永红,等.山茱萸总皂苷的提取分离及含量测定[J].第四军医大学学报,2003,24(5)：430-432.
[55]秦民坚,吴蓉,黄芸,等.匙羹藤叶总皂苷提取工艺研究[J].现代中药研究与实践,2003,17(3)：39-41.
[56]李成,邱智东,朱彩凤,等.两头尖总皂苷的精制[J].延边大学医学学报,2003,26(1)：7-9.
[57]郭孝武.一种提取中草药成分的方法--超声提取法[J].天然产物研究与开发,1998,11(3)：37-40.
[58]曾里,夏之宁.超声波和微波对中药提取的促进和影响[J].化学研究与应用,2006,14(3)：245-249.
[59]Mason T J. Chemistry with ultrasound (Critical reports on applied chemistry:V28) [M]. London and New York, Elsevier Science Publishers Lid,1990:4-6.
[60]Fu H, Xiao X Y, Zhang N P, et al. Determination and chromatogram recognition of main chemical constituents in Radix Scutellariae of different resources[J]. Chin J Pham Anal,2003,23:33-38.
[61]怀斯曼,酶技术生物手册,1989：.389.
[62]张彩霞,佟少山,王育红等.纤维素酶在穿山龙提取中的应用[J].基层中药杂志.2000,14(2)：32-35.
[63]马桔云,吕芳,千喜水等.纤维素酶用于中药穿心莲提取的初步研究[J].黑龙江医药.2000,13(1)：16-19.
[64]马桔云,赵晶岩,姜颖等.纤维素酶在黄连提取工艺中的应用[J].中草药.2000,31(2)：103-105.
[65]邢秀芳,马桔云,于宏芬等.纤维素酶在葛根总黄酮提取工艺中的应用[J].中草药.2001,32(1)：37-40.
[66]薛伟明,张效林,刘袖洞等.酶-膜法药用植物有效成分提取分离过程应用研究[J].西北大学学报,1997,27(6)：494-498.
[67]陈浩,田景振,赵海霞.吸附澄清技术[J].山东中医杂志,2000,19(3)：176-178.
[68]张万新,陈立新,向红连,等.抗感口服液澄清工艺探求[J].中国中药杂志.1999,24(11)：673-674.
[69]董方言.现代实用中药新剂型新技术[M].北京：人民卫生出版社,470-472.
[70]周跃华,吴笑如,徐德生.大孔吸附树脂纯化麦冬总皂苷[J].中草药,2003,34(12)：1089-1091.
[71]石召华,熊富良,李崇明,等.大孔吸附树脂纯化三七总皂苷工艺研究[J].中成药,2004,26(1)：10-11.
[72]陈彦,蔡宝昌.黄芪皂苷纯化工艺研究[J].中成药,2003,25(11)：866-868.
[73]堵年生,刘峻岭.大孔吸附树脂-紫外分光光度法测定肉苁蓉中苯乙醇苷类的含量[J].天然产物研究与开发,1993,5(3)：30-33.
[74]金京玲,任东鲜,金哲诛,等.大孔树脂法提取蒺藜总皂苷[J].延边大学医学学报,1999,22(1)：29-30.
[75]苏瑞强,张泽平,姜佳峰,等.蒺藜粗皂苷提取工艺研[J].山东医药工业,1997,16(5)：37-40.
[76]蔡雄,刘中秋,王培训,等.大孔吸附树脂工艺[J].中成药,2001,23(9)：631-634.
[77]周萍,廖铁文,杨瑛.大孔树脂富集枸骨叶甾体皂苷[J].湖南中医杂志,2002,18(1)：51-55.
[78]杜江,丁宁,贾宪生.大孔吸附树脂在分离纯化中的应用研究[J].中国中药杂志,2001,26(10)：685-688.
[79]金京玲,任东鲜,金哲诛,等.大孔吸附树脂法提取蒺藜甾体皂苷[J].延边大学医学学报,1999,22(1)：29-31.
[80]麻秀萍,蒋朝晖,杨玉琴,等.大孔吸附树脂对银杏叶黄酮的吸附研究[J].中国中药杂志,1997,22(9)：539-542.
[81]刘中秋,赖小平,吴燕红,等.大孔树脂富集橙皮苷的工艺研究[J].中成药,2001,23(8)：550-552.
[82]B rummer, Cui W, Wang Q. Extraction, urification and physicochemical characterization of fenugreek gum[J]. Food Hydrocolloids,2003,9(9):229-236.
[83]蒋建新,朱莉伟,张卫明等.半湿法分离提取半乳甘露聚糖型植物胶[J].中国野生植物资源,2002,21(4)：10-11.
[84]蒋建新,张卫明,朱莉伟等.烘炒法分离提取半乳甘露聚糖型种子胶[J].中国野生植物资源,2003,22(5)：34-36.
[85]李欣,范明娟,冯连彬等.24种豆科植物种子的半乳甘露聚糖胶的分析[J].植物学报,1980,22(3)：302-304.
[86]Emmanuel B, Alain D, Marie M G G. Study of the solid state hydrolysis of chitosanin[J]. PolymChem,1997,35(9):3181-3191.
[87]Makoto H, Akira L, Fumihiko O. Preparation of low molecula weight chitosanusing Phosphoric acid[J]. CarbohydrPolym,1993,20(6):279-283.
[88]Defave D. Chitinand Chitosan[M]. London:Elsevier APPlied science,1989:415.
[89]Chen R H, et al. Efiects of ultrasonic conditions and storage in acidic lutions on changes in molecular weight and polydispersity of treated chitosan[J]. Carbohydr Res,1997,299(3):287-294.
[90]Barker S A, Foster A B, et al. Isolation and properties of oigosaccharides obtained by controlled fragmentation of chitin[J]. Chemsoc,1958,11(6):2218-2221.
[91]盛以虞,徐开俊,郑风妹,等.壳聚糖在过氧化氢存在下的氧化降解[J].中国药科大学学报,1992,23(3).173-176.
[92]李邦良,高仕瑛,乔新惠等.甲壳低聚糖的制备和分析[J].中国生化药物杂志,1999,20(6)：292-294.
[93]张文清,柴平海,夏纬等.协同氧化制备壳寡糖[J].华东理工大学学报,2000,26(4)：425-428.
[94]Hutadilok N, Mochimasu. The efect of substitution on the hydrolysis of chitosan by anendo chitosanase[J]. CarbohydrRes,1995,68(10):143-147.
[95]郑连英.壳聚糖水解酶的筛选[J].材料研究学报,2000,14(2)：133-135.
[96]缪月秋,梅秋红,顾龚平.胡芦巴中性杂多糖酶解工艺优化及其产物对青春双歧杆菌增殖作用的研究[J].安徽农业大学学报,2006,33(2)：238-242.
[97]平其能.现代药剂学.北京：中国医药科技出版社,2001：652-659.
[98]JanesKA, FresneauMP, MarazuelaA, etal. Chitosan nanopartieles as delivery systems for doxorubicin[J]. Control. Release,2001,73(2-3):255-267.
[99]Risbud MV, Hardikar AA, Bhat SV, et al. PH-sensitive freeze-dried Chitosan-polyvinyl pyrrolidone hydrogels as controlled release system for Antibiotic delivery[J]. Control. Release,2000,68(1):23-30.
[100]Mao HQ, Roy K, Troung-Le VL, et al. Chitosan-DNA nanoparticles as gene carriers: synthesis, characterization and transfection efficiency[J]. Control. Release,2001, 70(3):399-421.
[101]韩哲文.高分子科学实验[M].华东理工大学出版社,2005.
[102]张黎明,赵玉平,王海宽,等.从中药胡芦巴种子提取薯蓣皂甙元工艺的研究[J].天津轻工业学院学报,2003,18(1)：6-8.
[103]卢锦花,胡小玲,岳红.吸附树脂提取分离银杏叶提取物的研究进展[J].化工进展,2001,20(3)：1-4.
[104]张黎明,李春莲.大孔吸附树脂分离纯化山楂叶总黄酮的研究[J].林产化学与工业,2006,26(1)：87-90.
[105]王丽伟,卢拥军,单文文等.瓜尔胶低分子量化降解条件研究[J].油田化学,2006,23(1)：33-35.
[106]肖桂武,曾和平,龙葵多糖的分离、纯化和鉴定[J].药物生物技术,1998,5(3)：157-160.
[2]荆宇,赵余庆,胡芦巴化学成分和药理作用研究进展[J].中草药,2003,34(12)：1146-1149.
[3]杨永利,孙双彦,崔德锋,等.胡芦巴种子化学成分的研究[J].西北师范大学学报,2003,39(1)：64-65.
[4]周维善,庄治平主编.,甾体化学进展.,第一版,北京：科学出版社,2002.
[5]Yoshikawa M. Medicinal food stuffs Ⅳ. Fenugreek seed(1):Structures of trigoneosides Ⅰa, Ⅰb, Ⅱa, Ⅱb, Ⅲa, and Ⅲb, new furostanol saponins from the seeds of Indian Trigonell a foenum-graecumL[J]. Chem Pharm Bull,1997,45(1):81-87.
[6]Yoshikawa M. Medicinal food stuffs Ⅳ. Fenugreek seed(1):Structures of six new furostanol saponins trigoneosides Ⅳa, Ⅴa, Ⅴb, Ⅶb, Ⅶb and Ⅷb fromm the seeds of Trigonell a foenum-graecumL[J]. Heterocycles,1998,47 (1):397-405.
[7]Murakami T, Kishi A, Matsuda H, et al. Medicinal food stuffs. ⅩⅦ. fenugreek seed(3):Structures of new furostanol-type steroidsaponins, trigoneosides Xa, Ⅹa, Ⅺb, Ⅻa, Ⅻb and ⅩⅢ fromm the seeds of Trigonell a foenum-graecumL[J]. Chem Pharm Bull,2000,48(7):994-1000.
[8]Shang MY, Cai SQ, Kadda S, et al. Structure identification of trigoneoside VIII. [J]. Acta Pharm Sin (药学学报),2001,36(11):836-839.
[9]Shang M Y, Cai S Q, Han J, et al. Studies on flavone of Trigonella foenum-graecum L. [J]. China J Chin Mater Med (中国中药杂志),1998,23(10):614-616.
[10]Yingmei H, Sansei N, Yukari N, et al. Flavonol glycosides from the stems of Trigonella foenum-graecum L. [J]. phyto-chemistry,2001,58:577-580.
[11]Shang M Y, Tezuka Y, Cai S Q, etal. Studies on triter-penoids from common fenugreek (Trigonella foenum-grae-cumL.). [J]. Chin tradit Herb Drugs(中草药), 1998,29(10):655-657.
[12]Shang M Y, Kadota S, Wang X, et al Studies on lipo so luble components of common fenugreek seeds (T rigonella foenum-graecum L.) [J].Chin Trad it Herb Drugs(中草药),1999,30(4):256-257.
[13]Shang M Y, Cai S Q, LinW H, et al. Studies on chemical constituents of Trigonella foenum-graecum L. [J]. China J Chin MaterM ed(中国中药杂志),2002,27(4): 277-279.
[14]Fernando P L, Martha R M, Carlo s A C, et al. So to lone production by hairy root cultures of Trigonella foenum-graecum L. in airlift with mesh bioreactors [J]. J Agr Food Chem,2001,49:6012-6019.
[15]缪月秋,梅秋红,顾龚平.胡芦巴中性杂多糖酶解工艺优化及其产物对青春双歧杆菌增殖作用的研究[J].安徽农业大学学报,2006,33(2)：238-242.
[16]李德亮,崔元臣,宋丽萍等.田箐胶在麦草造纸中的应用研究[J].中国造纸,1999,18(6)：46-48.
[17]Mirzaeva M R, Rakhmanberdyeva R K, Rakhimov D A. Structure of galactomannan from seeds of Gleditsia macracantha[J]. Chem Nat Compd,2000,35(5):505-507.
[18]Smirnova N I, Shcherbukhin V D. Galactomannan from caspia seeds[J]. Prikl.Biokhim. Mikrobiol,1998,24(5):653-659.
[19]Davison R L. Handbook of Water-soluble Galactomannan and Resins[J]. US,1980.
[20]Kooiman P. Structures of the galactomannans from seeds of Annona Muricata、 Cocos nucifera and sophoria japonica[J]. Carbonhyrate Research,1975,20(2): 329-337.
[21]Olivier N E, Taravel F. Nuclear magnetic resonance evidence of the guar borate interaction[J]. Carbohyarate Research,1987,16(6):1-11.
[22]Shimahara H, Suzuki H, Sugiyama N, et al. Isolation and characterization of olifosaccharides from an enzymic hdrolysate of konjac glucomannan[J]. Agric Biol Chem,1975,39(2):293-229.
[23]郭文松,刘达雯,周训清,等.甾体皂苷和苷元的研制[J].贵州化工,1996,(2)：14-17.
[24]Chen Y Y, Yuang C. The blood glucose reducing of fenugreek seeds abstracts in the alloxan-diabetic S. D. rats[J]. Zhen jiang Med Coll (镇江医学院学报),2001, 11(12):163-166.
[25]Madar Z, Shomer I J, Polysaccharides composition of gel fraction derived from fenugreek and its effect on starch digestion and bile acid absorption in rats[J]. A gric food.Chem,1990,38:1535～1539.
[26]Shani J, Glodschmied A, Joseph B, et al. Hypoglycemic effect of Trigonella foenum graecum and luminus term is seed and their major alkoloid in alloxan-diabetic and normal rats[J]. Arch. Int Pharmacodyn Ther,1974,210:27-36.
[27]Ribes G Y, Sauvaire Y, Baccou J, et al. Antidiabetic effect of subfractions from fenugreek seeds in diabetic dogs[J]. Proc Soc Exp Bio Med,1986,182:159-166.
[28]Amin Ryiad M, Abdul-Ghani Abdul-Salam S, Soleiman Mohammad S. Effect of fenugreek seeds and liupine seeds on the development of the experimental diabetes in rats[J]. Planta Med,1988:286-290.
[29]Sauvaire Y, Ribes G Y, Baccou J, et al. Implication of steroid saponin and sapogenins in the hypocholestero lemic effect of fenugreek[J]. Lipids,1991,26: 191-197.
[30]Shang M Y, Cai S Q, Wang X, et al. Analysis of aminoacid of Trigonella foenum-graecum L. [J]. Chin Med Mater(中药材),1998,21(4)：188-189.
[31]Suja P R, Anuradha C V, iswanathan P. et al. Gastropro-tective effect of fenugreek seeds(Trigonella foenum-graecum L.) on experimental gastricul cerinrats[J]. JE thnophar-macol,2002,81(3):393-397.
[32]Sur P, Das M, Comes A, et al. Trigonella foenum-graecum L. (fenugreek)seed extract as an antineo plastic agennt. [J]. Ohy-tother Res,2001,15(3):257-259.
[33]Zhu B L, Ban Y H, Duan J A. Protective effect of fenugreek seed on acute chemical liver injury[J]. Chun J Ind Med(中国工业医学杂志),2000,13(1)：19-21.
[34]Li L L, Mao X M, Wang X F, et al. The memory facilitation of TFGS and its protective effect against cerebral is chemia[J]. Chin Pharmacol Bull(中国药理学通报),2001,17(1)：92-94.
[35]Weder J K, Haussner K. Inhibitors of human and bovine trypsin and chy-motrypsin in fenugreek(Trigonella foenum-graecum L.) seeds[J]. Z Lebensm Unters Forsch, 1991,192(6):535-540.
[36]Panda S, Tahiliani P, Kar A. Inhibition of triiodothyronine production by fenugreek seed extract in mice and rats[J]. Pharmacol Res,1999,40(5):405-409.
[37]SHEN R L. The research progress of dietary firbre [J]. Shanxi Food Industry,2002, (2):21-24.
[38]TIAN X H. Function characteristic and application of the dietary fibre[J]. Food Research And Development,2002, (03):56-57.
[39]何锦风,郝利民,论膳食纤维[J].食品发酵工业,1999,23(5)：63-72.
[40]刘丹,常伟勤.复合膳食纤维对高胆固醇血症小鼠降脂作用的研究[J].北华大学学报(自然科学版),2006,(01)：35-37.
[41]吴杰.魔芋精粉与几种常用膳食纤维降血脂效果比较及机理探讨[J].营养学报,1993,15(2)：168.
[42]Cummings John H. Dietary carbohydrates and health:do we still need the fibre concept[J]. Clinical nutrition supplements,2004,1(2):5-7.
[43]Heaton K W. Dietary fibre and energy intake[J]. The lancet,1974,303(7853): 368-369.
[44]Riccardi G, et al. Dietary fibber in treatment of diabetes:myth or reality [J]. Digestive and Liver Disease,2002,34(2):140-144.
[45]欧仕益,高孔荣.膳食纤维抑制抑制膳后血糖升高的机理探讨[J].食品科学,1998,19(3)：48-52,
[46]邢小燕,张波.高膳食纤维有利于预防2型糖尿病—新疆克拉玛依市和山西忻州农村饮食状况调查[J].中日友好医院学报,2002,16(3)：134-137.
[47]韦晖.膳食纤维的研究与开发.纤维素科学与技术,2001,9(1)：56-63,
[48]阂锐,何云海,姚晓敏.开发人体第七营养素膳食纤维[J].中国食物与营养,1999, (3)：18-21.
[49]金其荣.低聚搪及根霉低聚糖生理功能[J].食品科学,1994,(1)：5-8.
[50]梁静谊,将建新,朱莉伟,等.皂荚皂苷提取工艺研究[J].中国野生植物资源,2003,22(6)：58-61.
[51]胡湘春,鱼红闪.穿山龙薯蓣皂苷的分离提纯[J].大连轻工业学院学报,2004,23(3)：25-29.
[52]康阿龙.胡芦巴素新药的开发研究[D].西北大学硕士学位论文,2001.
[53]杨献文,李苏梅,等.麦冬总皂苷提取工艺研究[J].中草药,2002,33(12)：1076-1078.
[54]吴红,梁恒,刘永红,等.山茱萸总皂苷的提取分离及含量测定[J].第四军医大学学报,2003,24(5)：430-432.
[55]秦民坚,吴蓉,黄芸,等.匙羹藤叶总皂苷提取工艺研究[J].现代中药研究与实践,2003,17(3)：39-41.
[56]李成,邱智东,朱彩凤,等.两头尖总皂苷的精制[J].延边大学医学学报,2003,26(1)：7-9.
[57]郭孝武.一种提取中草药成分的方法--超声提取法[J].天然产物研究与开发,1998,11(3)：37-40.
[58]曾里,夏之宁.超声波和微波对中药提取的促进和影响[J].化学研究与应用,2006,14(3)：245-249.
[59]Mason T J. Chemistry with ultrasound (Critical reports on applied chemistry:V28) [M]. London and New York, Elsevier Science Publishers Lid,1990:4-6.
[60]Fu H, Xiao X Y, Zhang N P, et al. Determination and chromatogram recognition of main chemical constituents in Radix Scutellariae of different resources[J]. Chin J Pham Anal,2003,23:33-38.
[61]怀斯曼,酶技术生物手册,1989：.389.
[62]张彩霞,佟少山,王育红等.纤维素酶在穿山龙提取中的应用[J].基层中药杂志.2000,14(2)：32-35.
[63]马桔云,吕芳,千喜水等.纤维素酶用于中药穿心莲提取的初步研究[J].黑龙江医药.2000,13(1)：16-19.
[64]马桔云,赵晶岩,姜颖等.纤维素酶在黄连提取工艺中的应用[J].中草药.2000,31(2)：103-105.
[65]邢秀芳,马桔云,于宏芬等.纤维素酶在葛根总黄酮提取工艺中的应用[J].中草药.2001,32(1)：37-40.
[66]薛伟明,张效林,刘袖洞等.酶-膜法药用植物有效成分提取分离过程应用研究[J].西北大学学报,1997,27(6)：494-498.
[67]陈浩,田景振,赵海霞.吸附澄清技术[J].山东中医杂志,2000,19(3)：176-178.
[68]张万新,陈立新,向红连,等.抗感口服液澄清工艺探求[J].中国中药杂志.1999,24(11)：673-674.
[69]董方言.现代实用中药新剂型新技术[M].北京：人民卫生出版社,470-472.
[70]周跃华,吴笑如,徐德生.大孔吸附树脂纯化麦冬总皂苷[J].中草药,2003,34(12)：1089-1091.
[71]石召华,熊富良,李崇明,等.大孔吸附树脂纯化三七总皂苷工艺研究[J].中成药,2004,26(1)：10-11.
[72]陈彦,蔡宝昌.黄芪皂苷纯化工艺研究[J].中成药,2003,25(11)：866-868.
[73]堵年生,刘峻岭.大孔吸附树脂-紫外分光光度法测定肉苁蓉中苯乙醇苷类的含量[J].天然产物研究与开发,1993,5(3)：30-33.
[74]金京玲,任东鲜,金哲诛,等.大孔树脂法提取蒺藜总皂苷[J].延边大学医学学报,1999,22(1)：29-30.
[75]苏瑞强,张泽平,姜佳峰,等.蒺藜粗皂苷提取工艺研[J].山东医药工业,1997,16(5)：37-40.
[76]蔡雄,刘中秋,王培训,等.大孔吸附树脂工艺[J].中成药,2001,23(9)：631-634.
[77]周萍,廖铁文,杨瑛.大孔树脂富集枸骨叶甾体皂苷[J].湖南中医杂志,2002,18(1)：51-55.
[78]杜江,丁宁,贾宪生.大孔吸附树脂在分离纯化中的应用研究[J].中国中药杂志,2001,26(10)：685-688.
[79]金京玲,任东鲜,金哲诛,等.大孔吸附树脂法提取蒺藜甾体皂苷[J].延边大学医学学报,1999,22(1)：29-31.
[80]麻秀萍,蒋朝晖,杨玉琴,等.大孔吸附树脂对银杏叶黄酮的吸附研究[J].中国中药杂志,1997,22(9)：539-542.
[81]刘中秋,赖小平,吴燕红,等.大孔树脂富集橙皮苷的工艺研究[J].中成药,2001,23(8)：550-552.
[82]B rummer, Cui W, Wang Q. Extraction, urification and physicochemical characterization of fenugreek gum[J]. Food Hydrocolloids,2003,9(9):229-236.
[83]蒋建新,朱莉伟,张卫明等.半湿法分离提取半乳甘露聚糖型植物胶[J].中国野生植物资源,2002,21(4)：10-11.
[84]蒋建新,张卫明,朱莉伟等.烘炒法分离提取半乳甘露聚糖型种子胶[J].中国野生植物资源,2003,22(5)：34-36.
[85]李欣,范明娟,冯连彬等.24种豆科植物种子的半乳甘露聚糖胶的分析[J].植物学报,1980,22(3)：302-304.
[86]Emmanuel B, Alain D, Marie M G G. Study of the solid state hydrolysis of chitosanin[J]. PolymChem,1997,35(9):3181-3191.
[87]Makoto H, Akira L, Fumihiko O. Preparation of low molecula weight chitosanusing Phosphoric acid[J]. CarbohydrPolym,1993,20(6):279-283.
[88]Defave D. Chitinand Chitosan[M]. London:Elsevier APPlied science,1989:415.
[89]Chen R H, et al. Efiects of ultrasonic conditions and storage in acidic lutions on changes in molecular weight and polydispersity of treated chitosan[J]. Carbohydr Res,1997,299(3):287-294.
[90]Barker S A, Foster A B, et al. Isolation and properties of oigosaccharides obtained by controlled fragmentation of chitin[J]. Chemsoc,1958,11(6):2218-2221.
[91]盛以虞,徐开俊,郑风妹,等.壳聚糖在过氧化氢存在下的氧化降解[J].中国药科大学学报,1992,23(3).173-176.
[92]李邦良,高仕瑛,乔新惠等.甲壳低聚糖的制备和分析[J].中国生化药物杂志,1999,20(6)：292-294.
[93]张文清,柴平海,夏纬等.协同氧化制备壳寡糖[J].华东理工大学学报,2000,26(4)：425-428.
[94]Hutadilok N, Mochimasu. The efect of substitution on the hydrolysis of chitosan by anendo chitosanase[J]. CarbohydrRes,1995,68(10):143-147.
[95]郑连英.壳聚糖水解酶的筛选[J].材料研究学报,2000,14(2)：133-135.
[96]缪月秋,梅秋红,顾龚平.胡芦巴中性杂多糖酶解工艺优化及其产物对青春双歧杆菌增殖作用的研究[J].安徽农业大学学报,2006,33(2)：238-242.
[97]平其能.现代药剂学.北京：中国医药科技出版社,2001：652-659.
[98]JanesKA, FresneauMP, MarazuelaA, etal. Chitosan nanopartieles as delivery systems for doxorubicin[J]. Control. Release,2001,73(2-3):255-267.
[99]Risbud MV, Hardikar AA, Bhat SV, et al. PH-sensitive freeze-dried Chitosan-polyvinyl pyrrolidone hydrogels as controlled release system for Antibiotic delivery[J]. Control. Release,2000,68(1):23-30.
[100]Mao HQ, Roy K, Troung-Le VL, et al. Chitosan-DNA nanoparticles as gene carriers: synthesis, characterization and transfection efficiency[J]. Control. Release,2001, 70(3):399-421.
[101]韩哲文.高分子科学实验[M].华东理工大学出版社,2005.
[102]张黎明,赵玉平,王海宽,等.从中药胡芦巴种子提取薯蓣皂甙元工艺的研究[J].天津轻工业学院学报,2003,18(1)：6-8.
[103]卢锦花,胡小玲,岳红.吸附树脂提取分离银杏叶提取物的研究进展[J].化工进展,2001,20(3)：1-4.
[104]张黎明,李春莲.大孔吸附树脂分离纯化山楂叶总黄酮的研究[J].林产化学与工业,2006,26(1)：87-90.
[105]王丽伟,卢拥军,单文文等.瓜尔胶低分子量化降解条件研究[J].油田化学,2006,23(1)：33-35.
[106]肖桂武,曾和平,龙葵多糖的分离、纯化和鉴定[J].药物生物技术,1998,5(3)：157-160.