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WO2007013717A1 - Oligosaccharide de chitosane, soluble dans l'eau et de grande qualite, composition comportant cet oligosaccharide et procede de preparation de ladite composition - Google Patents

Oligosaccharide de chitosane, soluble dans l'eau et de grande qualite, composition comportant cet oligosaccharide et procede de preparation de ladite composition Download PDF

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Publication number
WO2007013717A1
WO2007013717A1 PCT/KR2005/003598 KR2005003598W WO2007013717A1 WO 2007013717 A1 WO2007013717 A1 WO 2007013717A1 KR 2005003598 W KR2005003598 W KR 2005003598W WO 2007013717 A1 WO2007013717 A1 WO 2007013717A1
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Prior art keywords
chitosan oligosaccharide
molecular weight
soluble chitosan
water
quality water
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PCT/KR2005/003598
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English (en)
Inventor
Jae Woon Nah
Teok Rae Jung
Hwan-Jeong Jeong
Su Young Chae
Chang Yong Choi
Mi Kyeong Jang
Original Assignee
Jae Woon Nah
Teok Rae Jung
Hwan-Jeong Jeong
Su Young Chae
Chang Yong Choi
Mi Kyeong Jang
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Application filed by Jae Woon Nah, Teok Rae Jung, Hwan-Jeong Jeong, Su Young Chae, Chang Yong Choi, Mi Kyeong Jang filed Critical Jae Woon Nah
Publication of WO2007013717A1 publication Critical patent/WO2007013717A1/fr

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
    • C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
    • C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS
    • A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
    • A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
    • A21D2/14—Organic oxygen compounds
    • A21D2/18—Carbohydrates
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS
    • A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
    • A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
    • A21D2/34—Animal material
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G9/00—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor
    • A23G9/32—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
    • A23L2/02—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof containing fruit or vegetable juices
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
    • A23L2/52—Adding ingredients
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L23/00—Soups; Sauces; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
    • A23L25/30—Mashed or comminuted products, e.g. pulp, pastes, meal, powders; Products made therefrom, e.g. blocks, flakes, snacks; Liquid or semi-liquid products
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/60—Salad dressings; Mayonnaise; Ketchup
    • A23L27/63—Ketchup
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/275—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of animal origin, e.g. chitin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/14—Ultrafiltration; Microfiltration
    • B01D61/145—Ultrafiltration
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to high-quality water-soluble chitosan oligosaccharide, a composition comprising the same and a method for the preparation thereof.
  • Chitin and chitosan are polysaccharides found in nature and have been highlighted as novel functional materials.
  • Chitin is a polysaccharide composed of lots of ⁇ f-acetylglucosamine monomers
  • Chitosan is prepared by the deacetylation of chitin.
  • Chitosan is a very useful biomaterial derived from shellfishes such as shrimps and crabs, and its chemical name is (l ⁇ 4)2-amino-2-deoxy- ⁇ -D-glucosamine.
  • Chitosan has various bioactivities such as reduction of blood-cholesterol level, inhibition of fat intake, anticancer, immunity enhancement, antidiabetic, wound-healing and antibacterial activity.
  • chitosan is bound to mucous membranes of oral, nasal and intestinal cavities, and thereby accelerates drug absorption, and researches on the application thereof as an absorbefacient for protein drugs or nonabsorbable drugs in oral and various formulations have been extensively carried out. Further, there were records that chitosan had been taken as foods or medicines for the treatment of diseases in Korea, China and Japan from ancient times.
  • chitosan oligosaccharide obtained by enzymatic hydrolysis, acid hydrolysis, hydrolysis, oxidizing disintegration, etc.
  • it has improved solubility and low viscosity in aqueous solution due to low molecular weight and thereby may be used for various applications as biomaterials.
  • un- uniformity occurred in disintegration process leads to broad molecular weight distribution, and thereby raises several problems of reproducibility in practical applications.
  • chemically modified chitosan it has a drawback of deterioration in the characteristics of chitosan itself.
  • Japanese Laid-open Patent Publication No. sho 54-148090 discloses a method for preparing low molecular weight chitosan having a predetermined range of molecular weight by treating chitosan obtained through deacetylation of natural chitin in 0.007 to 0.35% hydrogen peroxide solution of pH 6 to 12.
  • Japanese Laid-open Patent Publication No. hei 01-256395 discloses a method for preparing low molecular weight chitosan by treating chitosan obtained through deacetylation of natural chitin by an enzyme, wherein enzyme reaction is carried out by continuously adding acid to a reaction system while controlling its addition rate and by keeping pH of the system at a substantially constant level in the range of 3 to 9. The method produces D-glucosamine-free low molecular weight chitosan having a molecular weight ranging from 340 to 50,000 Da.
  • Japanese Laid-open Patent Publication No. hei 02-200196 discloses a method for preparing low molecular weight chitosan by treating chitosan with at least one of neutral protease and lipase having decomposing ability of chitosan.
  • chitosan is decomposed by acid hydrolysis using hydrochloric acid, phosphoric acid, and nitric acid, etc.; hydrogen peroxide treatment; or enzyme treatment under acidic solution. Then, for the separation of oligosaccharide, methanol (CH OH) is repeatedly treated several times to separate into a soluble part and an insoluble part, and each oligosaccharide is separated by changing ethanol concentration, and subsequently treated by chromatography or ion-exchange resin column.
  • CH OH methanol
  • An object of the present invention is to provide high-quality water-soluble chitosan oligosaccharide having an average molecular weight of 1,000 to 11,000 Da and a molecular weight distribution of 1 to 1.5.
  • Another object of the present invention is to provide a method for the preparation of high-quality water-soluble chitosan oligosaccharide.
  • Another object of the present invention is to provide a composition containing high- quality water-soluble chitosan oligosaccharide.
  • the present invention provides high-quality water-soluble chitosan oligosaccharide having an average molecular weight of 1,000 to 11,000 Da and a molecular weight distribution of 1 to 1.5.
  • the present invention provides a method for the preparation of high-quality water-soluble chitosan oligosaccharide. [24] Additionally, the present invention provides a composition containing the high- quality water-soluble chitosan oligosaccharide.
  • the present invention provides high-quality water-soluble chitosan oligosaccharide having an average molecular weight of 1,000 to 11,000 Da and a molecular weight distribution of 1 to 1.5, preferably an average molecular weight of 1,000 to 1,700 Da and a molecular weight distribution of 1.3 to 1.5, an average molecular weight of 2,500 to 3,500 Da and a molecular weight distribution of 1.2 to 1.3, an average molecular weight of 5,500 to 6,700 Da and a molecular weight distribution of 1.15 to 1.2, and an average molecular weight of 8,500 to 11,000 Da and a molecular weight distribution of 1.0 to 1.15.
  • high-quality water-soluble chitosan oligosaccharide according to the present invention has an intrinsic viscosity of 0.020 to 0.250 g/D-sec, a degree of deacetylation of 90 to 95%, a moisture content of 1.0 to 3.0%, and a inorganic content of O to l%.
  • the molecular weight and its distribution may be determined by gel permeation chromatography coupled with light scattering detector, and the molecular weight of high-quality water-soluble chitosan oligosaccharide according to the present invention is in the range of 1,000 to 11,000 Da.
  • the molecular weight to be absorbed into the human body is below 10,000 Da.
  • a molecular weight distribution closer to 1 means that the chitosan oligosaccharide is successfully separated, and has a uniform range of molecular weight. Accordingly, the molecular weight distribution of high-quality water-soluble chitosan oligosaccharide ranging from 1 to 1.5 may be thought to be uniform in molecular weight.
  • the intrinsic viscosity of high-quality water-soluble chitosan oligosaccharide according to the present invention is in the range of 0.020 to 0.250 g/ D -sec, which may be determined by a rheometer. In consideration of the relationship with molecular weight, viscosity increases as molecular weight increases. Mark- Houwink constant is calculated by Mark-Houwink equation using molecular weight and intrinsic viscosity, and molecular weight may be obtained by intrinsic viscosity.
  • the degree of deacetylation of high-quality water-soluble chitosan oligosaccharide according to the present invention is high in the range of 90 to 95%, which may be determined by first derivative UV spectroscopic method.
  • the degree of deacetylation is a criterion for the purity of chitosan. Chitin is converted into chitosan by deacetylation, and thus many kinds of chitosans may be obtained according to the conditions of deacetylation.
  • High degree of deacetylation means that the degree of conversion from chitin into chitosan is high, and that the properties of chitosan are more strongly expressed than those of chitin.
  • bioactivities are almost derived from chitosan, not from chitin, it can be estimated that the higher the conversion rate from chitin into chitosan is, that is, the higher the degree of deacetylation is, the higher the expression of the bioactivities is.
  • the moisture content and the inorganic content of high-quality water- soluble chitosan oligosaccharide are in the ranges of 1 to 3% and 0 to 1% respectively, which may be determined by a thermogravimetric analyzer.
  • the components of shellfishes such as calcareous compounds, heavy metals, proteins, etc., may be contained as impurities in chitosan oligosaccharide. Accordingly, it may be estimated that the lower the content of the impurities is, the higher the purity of chitosan is.
  • high-quality water-soluble chitosan oligosaccharide according to the present invention having low inorganic and moisture contents may be estimated to have high purity.
  • the present invention provides a method for the preparation of high- quality water-soluble chitosan oligosaccharide, comprising the steps of:
  • chitosan oligosaccharide is separated according to the molecular weight thereof, using ultrafiltration membranes.
  • Ultrafiltration is a filtration having filtering effect between microfilration and reverse osmosis, and separates low molecular weight materials from a solution of high molecular weight materials.
  • An ultrafiltration membrane may be prepared from cellulose, polyacrilonitrile copolymer, polyamide, polysulfone, polyimide, and polyvinylidenefluoride, etc., and have a pore size of 5 to 100 D. Ultrafiltration is useful for the separation of solute having a molecular weight of 1,000 to 5,000 Da, and may be applied for the separation of solute and solvent having above 100 times differences in the molecular weights therebetween.
  • Water-soluble chitosan oligosaccharide having high molecular weight used in the preparation of high-quality water-soluble chitosan oligosaccharide according to the present invention may be obtained by conventional methods such as enzymatic hydrolysis (Korean Patent No. 291308), and commercialized reagents may also be used.
  • the step (a) may particularly comprise the following sub-steps of:
  • an aqueous solution of high molecular weight water-soluble chitosan oligosaccharide is prepared and then passed through a 30 kDa ultrafiltration membrane to remove components having a molecular weight above 30 kDa.
  • the filtrate containing water-soluble chitosan oligosaccharide with a molecular weight below 30 kDa is passed through a 10 kDa ultrafiltration membrane to separate into water-soluble chitosan oligosaccharide having a molecular weight of 10 to 30 kDa and a filtrate containing water-soluble chitosan oligosaccharide with a molecular weight below 10 kDa.
  • the 10 to 30 kDa water-soluble chitosan oligosaccharide is recovered and freeze- dried. Further, the filtrate containing water-soluble chitosan oligosaccharide with a molecular weight below 10 kDa is passed through a 3 kDa ultrafiltration membrane to separate into water-soluble chitosan oligosaccharide with a molecular weight of 3 to 10 kDa and a filtrate containing water-soluble chitosan oligosaccharide with a molecular weight below 3 kDa. The 3 to 10 kDa water-soluble chitosan oligosaccharide is recovered and freeze-dried.
  • the filtrate containing water-soluble chitosan oligosaccharide with a molecular weight below 3 kDa is passed through a 1 kDa ultrafiltration membrane to separate into water-soluble chitosan oligosaccharide having a molecular weight of 1 to 3 kDa and a filtrate containing water-soluble chitosan oligosaccharide with a molecular weight below 1 kDa.
  • the ultrafiltration may preferably be performed in an ultrafiltration reactor using a cellulose ultrafiltration membrane. Steps of concentration and the ultrafiltration may preferably be repeated 3 times.
  • step (b) impurities in each water-soluble chitosan oligosaccharide separated in the step (a) are removed, and then freeze-drying is performed.
  • the solution of water-soluble chitosan oligosaccharide is passed through a filter, preferably a 0.8 D filter to remove impurities and then freeze-dried. Freeze-drying is preferably performed at -40°C for 24 hrs.
  • the water-soluble chitosan oligosaccharide separated according to molecular weight and freeze-dried in the step (b) is precipitated in an organic solvent, to give powdered water-soluble chitosan oligosaccharide.
  • the organic solvent used in this step is a mixed solvent of acetone and ether, preferably in the ratio of 50:50 by volume.
  • the powdered water-soluble chitosan oligosaccharide obtained in the step (c) is vacuum-dried. Vacuum-drying is preferably performed at 10 to 13 torr for 48 hrs.
  • the above-prepared high-quality water-soluble chitosan oligosaccharide has an average molecular weight of 1,000 to 11,000 Da, a molecular weight distribution of 1 to 1.5, an intrinsic viscosity of 0.020 to 0.250 g/D-sec, a degree of deacetylation of 90 to 95%, a moisture content of 1.0 to 3.0%, and a inorganic content of 0 to 1%.
  • the high-quality water-soluble chitosan oligosaccharide according to the present invention may be usefully applied in various fields, for example pharmaceutical applications or functional health foods.
  • the present invention further provides a composition including high-quality water- soluble chitosan oligosaccharide.
  • a composition according to the present invention may further comprise one or more active substances having the same or similar function to that of high-quality water- soluble chitosan oligosaccharide.
  • a composition according to the present invention may be prepared by additionally comprising one or more pharmaceutically acceptable carriers suitable for administration.
  • a pharmaceutically acceptable carrier may be saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and their mixture.
  • Other conventional additives such as antioxidant, buffer solution, bacteriostat, etc. may be added, if necessary.
  • a composition may be formulated into pills, capsules, granules, tablets or injection formulations such as aqueous solution, suspension, emulsion, etc. by additionally comprising diluents, dispersing agents, surfactants, binders and lubricants.
  • a composition may preferably be formulated according to diseases or components by appropriate methods known in the art or methods disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18 , 1990).
  • composition according to the present invention may be parenterally administrated
  • a dosage may be controlled according to the patient's condition such as weight, age, sex, health condition, diet, administration time, administration route, excretion ratio, and disease severity, etc.
  • a daily dosage may be in the range from 0.01 to 100 D/D, which may be administrated at one time or divided into several doses.
  • a composition according to the present invention may be a functional health food.
  • high-quality water-soluble chitosan oligosaccharide according to the present invention may be added alone or in an admixture with other food or food ingredients by conventional methods.
  • the amount of effective ingredients to be mixed may be properly controlled according to its purpose (prevention, health or curative treatment).
  • high-quality water-soluble chitosan oligosaccharide according to the present invention may be added in an amount below 100 parts by weight, preferably below 50 parts by weight based the total raw material.
  • the added amount may be below this range. Because of no problem in safety, the effective ingredient may be used in the amount above this range.
  • Foods are not limited to special kinds. Foods, to which chitosan oligosaccharide may be added, are meat, sausage, bread, chocolate, candies, snacks, cookies, pizza, instant noodles, other noodles, chewing gums, dairy products such as ice creams, various kinds of soup, drinking water, tea, drink products, alcoholic beverage and vitamin complex, etc. including all kinds of health foods in conventional means.
  • Health-improving beverage composition according to the present invention may additionally comprise various flavorings or natural carbohydrates.
  • Natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; and polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol and erythritol, etc.
  • natural flavorings such as thaumatin and stevia extracts, or synthetic flavorings such as saccharin and aspartame may be used.
  • a composition according to the present invention may further comprise various kinds of nutritional supplements, vitamins, electrolytes, flavorings, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective- colloid thickners, pH regulators, stabilizers, preservatives, glycerin, alcohols and carbonating agents used in carbonated beverages, etc.
  • a composition according to the present invention may comprise fruit flesh used for natural fruit juices, fruit juice drinks and vegetable drinks. These ingredients may be used alone or in combination thereof. The ratio of these additives may not be important. However it may generally be selected to be in the range of 0.01 to 0.1 parts by weight to 100 parts by weight of the composition.
  • the high-quality water-soluble chitosan oligosaccharide according to the present invention has an average molecular weight of 1,000 to 11,000 Da, a molecular weight distribution of 1 to 1.5, an intrinsic viscosity of 0.020 to 0.250 g/D-sec, a degree of deacetylation of 90 to 95%, a moisture content of 1.0 to 3.0%, and a inorganic content of O to l%.
  • High-quality water-soluble chitosan oligosaccharide according to the present invention separated according to the molecular weight thereof using ultrafiltration membranes has very narrow molecular weight distribution and high purity, and exhibits little cytotoxicity, thereby being usefully applied in various fields, for example pharmaceutical applications or functional health foods.
  • FIG. 1 is a NMR spectrum for fraction 4 passed through an 1 kDa ultrafiltration membrane.
  • FIG. 2 is a NMR spectrum for fraction 3 passed through a 3 kDa ultrafiltration membrane and not passed through an 1 kDa ultrafiltration membrane.
  • FIG. 3 is a NMR spectrum for fraction 2 passed through a 10 kDa ultrafiltration membrane and not passed through a 3 kDa ultrafiltration membrane.
  • FIG. 4 is a NMR spectrum for fraction 1 passed through a 30 kDa ultrafiltration membrane and not passed through a 10 kDa ultrafiltration membrane.
  • FIG. 5 is a GPC chromatogram for high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FIG. 6 shows the intrinsic viscosity of fraction 4 of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FIG. 7 shows the intrinsic viscosity of fraction 3 of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FlG. 8 shows the intrinsic viscosity of fraction 2 of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FlG. 9 shows the intrinsic viscosity of fraction 1 of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FlG. 10 shows the relationship between molecular weight and intrinsic viscosity of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FlG. 10 shows the relationship between molecular weight and intrinsic viscosity of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FIG. 11 is a TGA diagram for fraction 4 of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FlG. 12 is a TGA diagram for fraction 3 of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FlG. 13 is a TGA diagram for fraction 2 of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FlG. 14 is a TGA diagram for fraction 1 of high-quality water-soluble chitosan oligosaccharide according to the present invention.
  • FlG. 15 shows the result of cytotoxicity assay of high-quality water-soluble chitosan oligosaccharide according to the present invention. [83]
  • the concentrate was sterilized by high temperature short time method(HTST) and spray-dried to give high molecular weight water-soluble chitosan oligosaccharide.
  • the chitosan oligosaccharide solution passed through a 30 kDa ultrafiltration membrane was separated according to molecular size using a 10 kDa ultrafiltration membrane.
  • the fraction 1 of chitosan oligosaccharide having an average molecular weight of 8,500 to 11,000 Da was obtained by separating the filtrate passed through a 30 kDa membrane and not passed through a 10 kDa membrane.
  • the fraction 2 of water-soluble chitosan oligosaccharide having an average molecular weight of 5,500 to 6,700 Da, the fraction 3 having an average molecular weight of 2,500 to 3,500 Da and the fraction 4 having an average molecular weight of 1,000 to 1,700 Da were obtained by using 3 kDa and 1 kDa ultrafiltration membranes.
  • the ultrafiltration membranes used in these steps were those having MWCO (molecular weight cut-off) of 30 kDa (YM30), 10 kDa (YMlO), 3 kDa (YM3) and 1 kDa (YMl) respectively.
  • the moisture content and the inorganic content of the high-quality water-soluble chitosan oligosaccharide were determined by a TGA (thermogravimetric analyzer)(TGA951, TA Instruments, USA). The determination was performed while heating up to 800 °C, at a heating rate of 10 °C/min, in air flow of 50 D/min.
  • TGA thermogravimetric analyzer
  • the high-quality water-soluble chitosan oligosaccharide contained 1 to 3% moisture and trace amount of ash, indicating that the amount of impurities was very low.
  • each fraction of high-quality water-soluble chitosan oligosaccharide obtained in the above example embodiment was dissolved in PBS 7.4 solution containing 10% FBS to prepare diverse concentrations (6.25, 12.5, 25, 50, 100 and 200 D/D).
  • the chitosan solution was removed.
  • the cells were washed with physiological saline solution and further stabilized for 4 hrs in a fresh culture medium. Subsequently the cells were incubated for 4 hrs in the incubator with cell culture medium containing 0.5 D/D MTT.
  • Relative cell viability was calculated by the following Math Figure 2 using the optical density at 570 D(OD ) of the solution in DMSO. A group containing only cell- free medium was used as a negative control group, and another group without treatment of high-quality water-soluble chitosan oligosaccharide was used as a positive control group.
  • Relative cell viability [ (OD 570 of the test group - OD 570 of the negative control group) / (OD 570 of the positive control group - OD 570 of the negative control group) ]
  • Powder product was prepared by mixing the above ingredients and filling an airtight package therewith.
  • Capsules were prepared by mixing the above ingredients and filling a gelatin capsule by a conventional method.
  • Health-improving meat products, noodle soup or gravies was prepared by adding high-quality water-soluble chitosan oligosaccharide according to the present invention in an amount of 0.1 to 1.0 parts by weight to meat products, soup or gravies.
  • Health-improving ground beef was prepared by adding high-quality water-soluble chitosan oligosaccharide according to the present invention in an amount of 10 parts by weight of to ground beef.
  • Health-improving dairy products were prepared by adding high-quality water- soluble chitosan oligosaccharide according to the present invention in an amount of 0.1 to 1.0 parts by weight to milk and then making butter or ice creams from the milk.
  • High-quality water-soluble chitosan oligosaccharide according to the present invention and subsidiary ingredients such as liquid fructose (0.5%), oligosaccharide (2%), sucrose (2%), salt (0.5%) and water (75%) were uniformly mixed, and then pasteurized.
  • the pasteurized product was packed in small containers such as glass bottles, pet bottles, etc., to give a health-improving beverage.
  • Health-improving vegetable juice was prepared by adding 0.5 g of high-quality water-soluble chitosan oligosaccharide according to the present invention to 1,000 D tomato or carrot juice.
  • Health-improving fruit juice was prepared by adding 0.1 g of high-quality water- soluble chitosan oligosaccharide according to the present invention to 1,000 D apple or grape juice.
  • High-quality water-soluble chitosan oligosaccharide according to the present invention separated according to the molecular weight thereof using ultrafiltration membranes has very narrow molecular weight distribution and high purity, and exhibits little cytotoxicity, thereby being usefully applied in various fields, for example pharmaceutical applications or functional health foods.

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Abstract

La présente invention se rapporte à un oligosaccharide de chitosane, soluble dans l'eau et de grande qualité, à une composition comportant cet oligosaccharide et à un procédé de préparation de ladite composition. Plus particulièrement, l'oligosaccharide de chitosane, soluble dans l'eau, de grande qualité conforme à la présente invention possède un poids moléculaire moyen compris entre 1 000 et 11 000 Da, une répartition de poids moléculaire comprise entre 1 et 1,5, une viscosité intrinsèque comprise entre 0,020 et 0,250 g/D-sec, un degré de désacétylation compris entre 90 et 95 %, une teneur en humidité comprise entre 1,0 et 3,0 %, et une teneur en éléments inorganiques comprise entre 0 et 1 %. Cet oligosaccharide de chitosane, soluble dans l'eau et de grande qualité, peut être obtenu par séparation en fonction de son poids moléculaire au moyen de membranes d'ultrafiltration. L'oligosaccharide de chitosane soluble dans l'eau et de grande qualité de la présente invention présente une répartition de poids moléculaire très étroite ainsi qu'une grande pureté, et il possède une faible cytotoxicité, ceci lui permettant d'être utilisé dans divers domaines, par exemple dans des applications pharmaceutiques ou des aliments santé fonctionnels.
PCT/KR2005/003598 2005-07-27 2005-10-27 Oligosaccharide de chitosane, soluble dans l'eau et de grande qualite, composition comportant cet oligosaccharide et procede de preparation de ladite composition WO2007013717A1 (fr)

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WO2016005519A1 (fr) * 2014-07-10 2016-01-14 Novozymes A/S Composition d'oligosaccharide et son procédé de préparation
US20170035929A1 (en) * 2015-08-07 2017-02-09 Victor Matthew Phillips Flowable hemostatic gel composition and its methods of use
CN109394678A (zh) * 2018-10-15 2019-03-01 安徽徽科生物工程技术有限公司 组织冲洗液及其在医疗领域中的应用
WO2019043740A2 (fr) 2017-01-24 2019-03-07 Jordan University Of Science And Technology Procédé de production et procédé d'utilisation de chitosane soluble à poids moléculaire élevé
US10258065B2 (en) 2014-07-10 2019-04-16 Novozymes A/S Polypeptides having xylanase activity and polynucleotides encoding same
CN110066838A (zh) * 2019-06-12 2019-07-30 山东昌瑞生物科技有限公司 一种高含量液体壳寡糖的生产方法
CN110551780A (zh) * 2019-09-04 2019-12-10 长春工业大学 一种酶解和清洁氧化耦合降解壳聚糖制备壳寡糖的方法
WO2020024053A1 (fr) * 2018-07-31 2020-02-06 The Royal Institution For The Advancement Of Learning/Mcgill University Procédés de production de chitosane
CN111171182A (zh) * 2020-03-02 2020-05-19 江西师范大学 一种用芳香醛修饰壳寡糖制备壳寡糖单体的方法
US10660945B2 (en) 2015-08-07 2020-05-26 Victor Matthew Phillips Flowable hemostatic gel composition and its methods of use
CN112608958A (zh) * 2020-12-17 2021-04-06 黄河三角洲京博化工研究院有限公司 一种壳寡糖的制备方法及减肥片
CN112617108A (zh) * 2021-01-08 2021-04-09 重庆森丰林业科技发展有限责任公司 一种去涩与抗褐变的枸杞叶面条制备方法
CN112972489A (zh) * 2021-03-19 2021-06-18 季华实验室 具有减肥降脂功能的壳聚糖基组合物及制备方法、应用
CN115381022A (zh) * 2022-08-22 2022-11-25 江南大学 一种生鲜面制品的制备及绿色保鲜工艺
CN116003883A (zh) * 2023-02-07 2023-04-25 江西师范大学 一种β-环糊精包被壳寡糖及其制备方法
CN117106431A (zh) * 2023-08-28 2023-11-24 新疆兰德伟业油田服务有限公司 低分子聚合物压裂液

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WO2016005519A1 (fr) * 2014-07-10 2016-01-14 Novozymes A/S Composition d'oligosaccharide et son procédé de préparation
US12185740B2 (en) 2014-07-10 2025-01-07 Novozymes A/S Polypeptides having xylanase activity and polynucleotides encoding same
US11470859B2 (en) 2014-07-10 2022-10-18 Novozymes A/S Polypeptides having xylanase activity and polynucleotides encoding same
US10842172B2 (en) 2014-07-10 2020-11-24 Novozymes A/S Polypeptides having xylanase activity and polynucleotides encoding same
US10258065B2 (en) 2014-07-10 2019-04-16 Novozymes A/S Polypeptides having xylanase activity and polynucleotides encoding same
US10751444B2 (en) * 2015-08-07 2020-08-25 Victor Matthew Phillips Flowable hemostatic gel composition and its methods of use
US20170035929A1 (en) * 2015-08-07 2017-02-09 Victor Matthew Phillips Flowable hemostatic gel composition and its methods of use
US10660945B2 (en) 2015-08-07 2020-05-26 Victor Matthew Phillips Flowable hemostatic gel composition and its methods of use
WO2019043740A2 (fr) 2017-01-24 2019-03-07 Jordan University Of Science And Technology Procédé de production et procédé d'utilisation de chitosane soluble à poids moléculaire élevé
WO2020024053A1 (fr) * 2018-07-31 2020-02-06 The Royal Institution For The Advancement Of Learning/Mcgill University Procédés de production de chitosane
US11597780B2 (en) 2018-07-31 2023-03-07 The Royal Institution For The Advancement Of Learning/Mcgill University Methods for producing chitosan
CN109394678A (zh) * 2018-10-15 2019-03-01 安徽徽科生物工程技术有限公司 组织冲洗液及其在医疗领域中的应用
CN110066838A (zh) * 2019-06-12 2019-07-30 山东昌瑞生物科技有限公司 一种高含量液体壳寡糖的生产方法
CN110551780A (zh) * 2019-09-04 2019-12-10 长春工业大学 一种酶解和清洁氧化耦合降解壳聚糖制备壳寡糖的方法
CN111171182A (zh) * 2020-03-02 2020-05-19 江西师范大学 一种用芳香醛修饰壳寡糖制备壳寡糖单体的方法
CN112608958A (zh) * 2020-12-17 2021-04-06 黄河三角洲京博化工研究院有限公司 一种壳寡糖的制备方法及减肥片
CN112608958B (zh) * 2020-12-17 2022-12-02 黄河三角洲京博化工研究院有限公司 一种壳寡糖的制备方法及减肥片
CN112617108A (zh) * 2021-01-08 2021-04-09 重庆森丰林业科技发展有限责任公司 一种去涩与抗褐变的枸杞叶面条制备方法
CN112972489A (zh) * 2021-03-19 2021-06-18 季华实验室 具有减肥降脂功能的壳聚糖基组合物及制备方法、应用
CN115381022A (zh) * 2022-08-22 2022-11-25 江南大学 一种生鲜面制品的制备及绿色保鲜工艺
CN115381022B (zh) * 2022-08-22 2023-11-07 江南大学 一种生鲜面制品的制备及绿色保鲜工艺
CN116003883A (zh) * 2023-02-07 2023-04-25 江西师范大学 一种β-环糊精包被壳寡糖及其制备方法
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