高纯低聚果糖分离纯化技术的研究
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摘要
低聚果糖具有促进人体肠道内双歧杆菌生长的显著作用,并有保护肝脏,提高机体免疫力等生理功能,广泛应用于食品和保健品行业。但大规模生产的低聚果糖产品为多种组分的混合物,纯度不高限制了公众对其生理功能的认同感。由于技术条件限制,国内市场紧缺高纯度的低聚糖产品,这使得纯化低聚果糖具有极其现实的意义。
本课题初步研究了对低聚果糖糖浆的脱色效果方法。结果表明70℃,pH=3.8,活性炭用量为1.4%,脱色50min时活性炭对低聚果糖具有很好的脱色效果。新生态碳酸钙法用于低聚果糖脱色效果不理想。
同时研究了利用膜分离法将脱色后的低聚果糖进行两级纳滤和一次反渗透,并将其分离为90%以上纯度和50%以上纯度的低聚果糖产物Ⅰ和产物Ⅱ以及葡萄糖和蔗糖的混合产物Ⅲ。产物Ⅰ虽然产率并不高,但却为后续凝胶层析分离低聚果糖单一组分提供了原料来源,同时也为Bio-gel P-2凝胶层析分离奠定了良好的分离基础。
利用聚丙烯酰胺凝胶P-2,当流速为0.12mL·min~(-1),120×1cm I.D,进样量为0.5mL时,以高纯度低聚果糖为原料初步得到蔗果三糖、蔗果四糖以及蔗果五糖的单一组分,原料纯度越高,单一组分纯度也越高。葡聚糖凝胶LH-20在纯水作为流动相时分离低聚果糖单一组分不理想。
运用KS与CS系列凝胶糖柱,以纯水为流动相,三柱串联方法能够有效分离低聚果糖,本研究为低聚糖类物质提供了新型、环保节能的分离方法。
Fructooligosaccharide (FOS) exhibit a stimulating effect on the growth of the beneficial Bifidobacteria in animal intestines and protect liver, improve immunity etc, it's currently used in the so-called health foods and feedstuff and so on.Because of the short of high-purity FOS, ti's very usefull for purifying FOS. However, large-scale production of FOS is mixture of multiplicate components, it's low-purity, restricts the public's sense of identification with their physiological functions. Due to technical constraints, high-grade products of oligosaccharides is short in the domestic market, which makes FOS purification is extremely practical significance.
This experiment preliminary studied on decolourization of FOS syrup. The experiments show that the decolourization effect was the best when the amount of activated carbon is 1.4 percent, decolorization temperature is 70℃, pH is 3.8 and bleaching time is 40 min. fresh lime carbonate is not effective for the decolourization of FOS.
FOS is purified by NF after decolourization. It is purified by twice NF process and once RO, then we can get three products: productⅠ's purity is above 90%, productⅡ's purity is above 90%, productⅢis the mixture of sucrose and glucose. productⅠis not high yielded, but it is the raw material for the separation of FOS and it is useful for the separation with Bio-gel P-2.
We can get 1-kestose, nystose and fructofuranosylnystose by separation with Bio-gel P-2 when the flow rate is 0.12mL·min~(-1),column longth is 120×1 cm I.D ,injection volumn is 0.5mL.They will have high purity when the raw material is more pure.It's non-ideal for separation with sephadex LH-20 when the mobile phase is water.
It's effective for the separation with KS & CS series sugar column when the mobile phase is water and with three connected columns. This method is new, and it's good for environmental protection.
本课题初步研究了对低聚果糖糖浆的脱色效果方法。结果表明70℃,pH=3.8,活性炭用量为1.4%,脱色50min时活性炭对低聚果糖具有很好的脱色效果。新生态碳酸钙法用于低聚果糖脱色效果不理想。
同时研究了利用膜分离法将脱色后的低聚果糖进行两级纳滤和一次反渗透,并将其分离为90%以上纯度和50%以上纯度的低聚果糖产物Ⅰ和产物Ⅱ以及葡萄糖和蔗糖的混合产物Ⅲ。产物Ⅰ虽然产率并不高,但却为后续凝胶层析分离低聚果糖单一组分提供了原料来源,同时也为Bio-gel P-2凝胶层析分离奠定了良好的分离基础。
利用聚丙烯酰胺凝胶P-2,当流速为0.12mL·min~(-1),120×1cm I.D,进样量为0.5mL时,以高纯度低聚果糖为原料初步得到蔗果三糖、蔗果四糖以及蔗果五糖的单一组分,原料纯度越高,单一组分纯度也越高。葡聚糖凝胶LH-20在纯水作为流动相时分离低聚果糖单一组分不理想。
运用KS与CS系列凝胶糖柱,以纯水为流动相,三柱串联方法能够有效分离低聚果糖,本研究为低聚糖类物质提供了新型、环保节能的分离方法。
Fructooligosaccharide (FOS) exhibit a stimulating effect on the growth of the beneficial Bifidobacteria in animal intestines and protect liver, improve immunity etc, it's currently used in the so-called health foods and feedstuff and so on.Because of the short of high-purity FOS, ti's very usefull for purifying FOS. However, large-scale production of FOS is mixture of multiplicate components, it's low-purity, restricts the public's sense of identification with their physiological functions. Due to technical constraints, high-grade products of oligosaccharides is short in the domestic market, which makes FOS purification is extremely practical significance.
This experiment preliminary studied on decolourization of FOS syrup. The experiments show that the decolourization effect was the best when the amount of activated carbon is 1.4 percent, decolorization temperature is 70℃, pH is 3.8 and bleaching time is 40 min. fresh lime carbonate is not effective for the decolourization of FOS.
FOS is purified by NF after decolourization. It is purified by twice NF process and once RO, then we can get three products: productⅠ's purity is above 90%, productⅡ's purity is above 90%, productⅢis the mixture of sucrose and glucose. productⅠis not high yielded, but it is the raw material for the separation of FOS and it is useful for the separation with Bio-gel P-2.
We can get 1-kestose, nystose and fructofuranosylnystose by separation with Bio-gel P-2 when the flow rate is 0.12mL·min~(-1),column longth is 120×1 cm I.D ,injection volumn is 0.5mL.They will have high purity when the raw material is more pure.It's non-ideal for separation with sephadex LH-20 when the mobile phase is water.
It's effective for the separation with KS & CS series sugar column when the mobile phase is water and with three connected columns. This method is new, and it's good for environmental protection.
引文
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[2]陈常秀,李同树.果糖的研究进展[J].山东饲料,2004,2:14-15
[3]吴文剑,范家恒.低聚果糖的研究进展[J].甘蔗糖业,2004,5:28-36
[4]井明艳,孙建义.天然活性成分-低聚果糖的应用前景[J].饲料研究,2004,1:22-27
[5]田萍,姚武群.果寡糖的研究和应用[J].中国饲料,2003,4:18-19
[6]陈旭东,马秋刚,计成.果寡糖研究进展[J].饲料广角,2003,9:19-23
[5]杨海军,高成林.高纯度低聚果糖的生产及应用[J].中国食品添加剂增刊,2003.142-143
[6]Agency Response Letter.GRAS Notice No.GRN 000044,FDA,2000,11
[7]Press Release of GTC Nutrition,2001,1
[8]C.S.Brazel.M icroencapsulation:efering solutions for the Food Industry.Cereal Foods World.1999,44(6):388-393.
[9]杨玉峰,冯光德,何建等.低聚果糖对动物的作用机理及其应用[J].中国饲料,2001,13:2-3
[10]汪世华,彭利民,张会等.低聚果糖的开发与应用[J].中国乳品工业,2002,30(2):31-34
[11]曹霞,张伟.微生物酶法制备低聚果糖的研究[J].食品科学,2000,21(6):31-35
[12]马莺,孙建华.β-D呋喃果糖苷酶合成低聚果糖的工艺研究[J].食品与发酵工业,2003,29(2):69-71
[13]魏远安,马丽.固定化酶法合成蔗果低聚糖的研究[J].食品与发酵工业,1995.4:12-16
[14]江波,王璋.固定化黑曲霉增殖细胞生产低聚果糖的研究[J].无锡轻工大学学报,1996,15(1):12-18
[15]史锋,江波.连续生产低聚果糖的最佳操作参数[J].无锡轻工大学学报,2000,19(1):1-4
[16]江波,王璋.共固定化生产高含量低聚果糖的研究[J].食品与发酵工业,1996.1:1-7
[17]江波,王璋.双酶法生产高纯度低聚果糖的研究[J].食品科学,1997,18(9):24-27
[18]Sarote Siri sansaneeyakul,Sittwat Lertsiri,Preecha.Product ion of Fructo-Oligosacchalides from Sucrose[J].Nat.Sci.,2000,34:262-269
[19]孙蔚榕,韩亮,鲍元兴.低聚糖的纳滤分离技术[J].无锡轻工大学学报,2002,22(6):574-578
[20]李秀琴,杨星惠,安素欣.低聚果糖的纯化方法[J].河北化工,2003,1:6-7
[21]鲍元兴,孙蔚榕,杨维亚.低聚异麦芽糖的纳滤分离技术和色谱分离技术[J],无锡轻工大学学报.2001,20(7):4-7
[22]魏远安,姚评佳,谢庆武等.蔗果低聚糖的研究和生产应用[J].食品与发酵工业,1998,26(1):48-53
[23]Monika Antosova,Milan Polakovic & Vladimir Bales.Separation of fructooligosaccharides on a cation-exchange HPLC column in silver form with refractometric detection[J].Biotechnology Techniques,1999
[24]C.L'homme,A.Puigserver,A.Biagini,Effect of food-processing on the degradation of fructooligosaccharides in fruit[J],Food Chemistry,2003
[25]M.Bekers,J.Laukevics,D.Upite etc.Fructooligosaccharide and levan producing activity of Zymomonas mobilis extracellular levansucrase[J],Process Biochemistry,2002
[26]P.T.Sangeetha,M.N.Rameshb,S.G.Prapulla a,Fructooligosaccharide production using fructosyl transferase obtained from recycling culture of Aspergillusoryzae CFR 202[J],Process Biochemistry,2005
[27]C.Corradinia,F.Bianchia,D.Matteuzzib etc,High-performance anion-exchange chromatography coupled with pulsed amperometric detection and capillary zone electrophoresis with indirect ultra violet detection as powerful tools to evaluate prebiotic properties of fructooligosaccharides and inulin[J],Journal of Chromatography A,2004
[28]P.T.Sangeetha,M.N.Ramesh,S.G.Prapulla,Maximization of fructooligosaccharide production by two stage continuous process and its scale up[J],Journal of Food Engineering,2005
[29]Dey Chyi Sheul,Po Jang Liol,Shih Tse Chenl etc.Production of fructooligosaccharides in high yield using a mixed enzyme system of β-fructofuranosidase and glucose oxidase[J],Biotechnology Letters,2001
[30]Noureddine Benkeblia,Natsuko Takahashi,Keiji Ueno,Shuichi Onodera.Tetraand penta-fructooligosaccharide(FOS)isomers assessment in onion bulb tissues:effect of temperature and storage time and Norio Shiomi[J],science derect,2005
[31]Katarina Reiffova,Radomira Nemcova,Thin-layer chromatography analysis of fructooligosaccharides in biological samples[J].Journal of Chromatography,2006
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