JP2003183296A - Method for producing glucosamine or (and) chitosan oligomer composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for industrially advantageously producing a glucosamine or (and) a chitosan oligomer composition from a chitin-based material. <P>SOLUTION: The glucosamine or (and) the chitosan oligomer composition is easily produced by reacting the chitin-based material with a concentrated hydrofluoric acid containing 75-95% hydrogen fluoride under a mild condition and dehydrofluorinating the product. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The chitin-based material is an exoskeleton component that covers the body surface of molluscs such as crustaceans (crab, shrimp, krill) and insects, and is used for the tendons of cuttlefish, crab and shrimp. Is also included. In the plant kingdom, it is present in the cell walls of molds, yeasts, mushrooms, green algae and the like. Chitin is a polymer of N-acetylglucosamine and is a biomass comparable to cellulose, which is a structural factor of plants. The deacetylated version of chitin is called chitosan, whose monomer is glucosamine.

The present invention relates to a method for industrially producing a glucosamine or / and chitosan oligomer composition from a chitin-based material, specifically, glucosamine or a salt thereof, a chitosan oligomer or a salt thereof, Specifically, it relates to a method for producing a glucosamine derivative containing a deacetylated chitin oligomer or a salt thereof as a main component.

[0003]

2. Description of the Related Art The inventors of the present invention have conducted many studies on the dissolution and decomposition of polymeric substances in high concentration hydrofluoric acid. For example, concentrated hydrofluoric acid is excellent as a solvent that separates and dissolves linear polyester, Kitano,
Kikuyama et al. (Japanese Patent Laid-Open No. 11-5866).
issue).

Like cellulose, chitin is a substance that is difficult to dissolve in a solvent, and the solvent known so far is concentrated sulfuric acid,
Concentrated phosphoric acid, anhydrous hydrogen fluoride, formic acid anhydride, boron fluoride 1
.About.2 hydrate, a thermally saturated aqueous solution of rhodan lithium and the like.
Chitin has a drawback that it dissolves in these solvents, unlike cellulose, but the reaction solution turns black, which is probably because the glucosamine produced is subjected to aerial oxidation. What is observed when cellulose is dissolved in the above acid is the formation of an ester of the cellulose oligomer and the above acid, and such a situation is also observed in the case of a chitin oligomer or a chitosan oligomer. For example, what is known in the reaction between chitin and anhydrous hydrogen fluoride is that glucosamine is produced as a by-product of a fluorine-containing lower sugar such as 2-acetamido-2-deoxy-α-D-glucopyranosylfluoride. Or (and) the purity of the chitosan oligomer composition decreases, and labor is required to purify the target substance (C. Bosso, J. Defaye et al., Carbohydr.Res., 156, 57.
~ 68, 1986; 186, 177-188, 1898; 261, (2),
267-277, 1994; 327, (1-2), 1-3, 2000).

[0005]

The dissolution and decomposition of chitin with anhydrous hydrogen fluoride occurs much more smoothly than with other hydrogen halides such as hydrogen chloride, and liquefaction can be achieved within a relatively short time. However, when a large amount of chitin is added to a large amount of anhydrous hydrogen fluoride at a time to react at 20 to 25 ° C, chitin is finally hydrolyzed to glucosamine hydrofluoride and acetic acid, and then a part of glucosamine. However, if it is left to stand for a long time due to secondary air oxidation, the reaction mixture becomes a blackish brown to black liquid. In this case, finally, the reaction products are glucosamine hydrofluoride and glucosamine acetate and a black tar-like oxide, and only a small amount of chitosan oligomer hydrofluoride is obtained.

[0006]

[Means for Solving the Problems] 96-1 for chitin-based materials
When 00% hydrogen fluoride is reacted, the fluorine-containing lower sugar is produced as described above, and accordingly, a considerable amount of hydrogen fluoride is wasted. On the other hand, in order to prevent the production of fluorine-containing lower sugars in the preliminary study of the present inventors, 75-9
It was found that 5% concentrated hydrofluoric acid should be used. Here, as a result of intensive studies, the present inventor has developed a production method under a new condition for obtaining a glucosamine or / and chitosan oligomer composition from a chitin-based material. That is, 100 parts by weight of a chitin-based material was dispersed and dissolved in 100 to 2000 parts by weight of concentrated hydrofluoric acid containing 75 to 95% by weight of hydrogen fluoride at −20 to + 20 ° C. for hydrolysis. The method for producing a glucosamine or / and chitosan oligomer composition characterized by dehydrofluorination from a solution has been completed.

The chitin-based material as used herein refers to a single or mixture obtained or collected from crustaceans, insects, molluscs or microorganisms. In addition, dehydrofluoric acid is used to remove insoluble matter from the reaction mixture, and then vacuum distillation, spray drying, freeze drying, neutralization filtration,
It refers to a method for producing a glucosamine or / and chitosan oligomer composition, which is concentrated through a neutralization centrifugation method.

The chitin-based material is a linear polysaccharide in which N-acetyl-D-glucosamine is β (1 → 4) -bonded,
Chitosan is a linear polysaccharide in which D-glucosamine is β (1 → 4) linked. Also, the usual commercially available chitin is 1
0 to 20% is deacetylated, while chitosan is biomass that is 15 to 25% N-acetylated. These are for industrial use, chemical use, reagent use,
It may be one for food. All of these are used as the chitin-based material of the present invention in the raw material of the present invention.

Glucosamine and chitosan oligomers are produced by hydrolysis of chitin-based materials, but their degree of deacetylation is 45 to 5 due to alkali decomposition or acid decomposition.
When it reaches 5%, chitin becomes water-soluble and decomposition proceeds in water as a uniform reaction. On the other hand, chitin is uniformly dissolved in concentrated hydrofluoric acid from the initial stage of the reaction, and hydrolysis and hydrofluoric acid decomposition using hydrofluoric acid as a catalyst proceed in a competitive manner for a relatively short time. A chitosan oligomer is produced at. However, if a large amount of glucosamine hydrofluoride is produced in the early stage of the reaction, it will be susceptible to air oxidation, and the reaction liquid will change from dark brown to black, so it is good to pay attention to air blocking and light shielding.

In the method of the present invention, when the chitin-based material is uniformly dissolved in concentrated hydrogen fluoride, the insoluble portion is removed by filtration, centrifugation, adsorption or the like, and a solute solid substance is obtained from the solution portion. In practice, concentration is carried out through vacuum distillation, spray-drying, freeze-drying, neutralization filtration, neutralization centrifugation to recover hydrofluoric acid. The glucosamine or / and chitosan oligomer composition thus obtained is in principle in the hydrofluoride form. The glucosamine or (and) chitosan oligomer composition thus obtained is then treated with the desired inorganic acid salt (eg, hydrochloride) or organic acid salt (eg, acetate, lactate, ascorbate, benzoate). ) Etc. are often put to practical use after being changed by the double decomposition method.

Here, the amount of hydrogen fluoride used in the present invention is a range determined by proper use of flaky chitin. First, the concentration of hydrofluoric acid is 75 to 95% HF, as will be described later, and this concentration suppresses the production of fluorine-containing lower sugars, which causes gelation of dissolved chitin-based materials and the occurrence of side reactions. It is necessary to avoid a decrease in the yield of the target product. Furthermore, the reaction temperature and the reaction time can be determined in consideration of the degree of polymerization and the yield of the chitosan oligomer.

Next, the general method of the present invention will be described with reference to specific examples. However, the reaction equipment, the filter material, and the like used in this experimental example are all made of polypropylene, polytetrafluoroethylene, or hydrofluoric acid-resistant metal. 80 in a stirrer flask (3 l)
% Hydrofluoric acid (1000 g) and cooled to 5 ° C. or lower, and then flaked chitin (prepared from king crab shell) (500 g) was added, and the mixture was placed under a nitrogen atmosphere at 0-5.
The reaction is allowed to stir at 2.5 ° C. for 2.5 hours. A dark brown, viscous slurry is obtained which is rapidly filtered at low temperature to remove insolubles. Hydrofluoric acid is recovered from the filtrate under reduced pressure by a spray drying method or the like to obtain a residue of about 700 g. 28% ammonia water and ice water are added to this residue and stirred to obtain about 1000 g of a mixed solution having a pH of about 4. The mixture is filtered and the filtrate is concentrated under reduced pressure to obtain about 300 g of a mixture of glucosamine and a hydrofluoric acid salt of a chitosan oligomer. When this composition is poured into a saturated aqueous solution of sodium chloride and heated and cooled once, a brown precipitate of a mixture of glucosamine and chitosan oligomer hydrochloride is obtained. About 330 g is obtained. These hydrochloride mixtures were dissolved in a hydrochloric acid / methanol mixed solution, the solution was deep-chilled, and the precipitated crystals were separated and precipitated, and the yield of each of them was measured. Glucosamine hydrochloride was about 55%,
It was found to be a mixture of about 20% chitobiose hydrochloride, about 16% chitotriose hydrochloride, about 7% chitotetraose hydrochloride.

The present invention relates to a method for producing a glucosamine or / and chitosan oligomer composition, and the form of the present product may be solid (powder, agglomerate) or liquid (solution, dispersion slurry). However, the chemical composition thereof may be a simple salt or a mixed salt of glucosamine or / and a chitosan oligomer, or a mixed salt containing a partially acetylated product thereof, that is, a composition. Among the inorganic acid salts, the most easily obtained composition is hydrofluoride, but this composition usually has other safe anions (for example, hydrochloride) in view of the corrosiveness and toxicity of fluoride ion. It is common to convert it to a new one for easier use. Of course, even such anion-converted product is included in the glucosamine or (and) chitosan oligomer composition of the present invention.

The glucosamine or (and) chitosan oligomer composition of the present invention generally has a large physiological activity on living organisms (especially) animals, for example, functional dietary fiber, feed additive, feed binder, food additive, antibacterial agent. Widely used in agents, preservatives, wound healing agents, arthritis remedies, health food ingredients, cosmetic ingredients, wastewater treatment agents, soil conditioners, natural agrochemical ingredients, other applications, and cationic flocculants, paints, dyes. ,
Widely used in industrial applications such as the textile industry and paper industry.

The inventors of the present invention have conducted a number of experiments on the above method to clarify the superiority of the present invention. Further, the method of the present invention is not limited to the examples shown below. It should be understood that the present invention should not be construed as being limited to the above, and the embodiment can be arbitrarily changed and implemented without departing from the spirit and the spirit of the present invention.

[0016]

Example 1 A flask (3 l) equipped with a stirrer was charged with 1000 g of 80% hydrofluoric acid and cooled to 0 ° C., and then 500 g of flaky chitin (flaky powder prepared from the shell of red king crab) was added thereto. 0 ° C in a nitrogen atmosphere
The mixture was cooled to −10 ° C. under stirring at 2 ° C. for 5 hours, the reaction solution was rapidly filtered under reduced pressure, the filtrate was concentrated under reduced pressure to 950 to 1000 g, the residue was ice-cooled, and the pH was adjusted with 28% ammonia water. Neutralize to about 3.0 and cool with ice. Precipitated crystals are filtered off, and the filtrate is concentrated by a flash evaporator until the volume is reduced to about half. A blackish brown residue is obtained. Then, the residue is poured into 600 ml of saturated saline and boiled for 1 hour while stirring, concentrated again under reduced pressure and cooled, and the residue obtained by separating the filtrate is dehydrated and dried. This solid is extracted with methanol,
When the methanol is distilled off from the extract, a blackish brown powder is about 45
0 g was obtained. When this product was analyzed by liquid chromatography, it was found to contain about 40% glucosamine hydrochloride and about 20% chitosan oligomer hydrochloride. This chitosan oligomer salt is mostly chitobiose hydrochloride, chitotriose hydrochloride, and chitotetraose hydrochloride, and it is found that a small amount of chitopentaose hydrochloride, chitohexaose hydrochloride, and chitoheptaose hydrochloride is contained. ing.

[0017]

Example 2 In the method of Example 1, 1200 g of 70% hydrofluoric acid was used instead of 80% hydrofluoric acid, and 400 g of flaky chitin was reacted at 25-30 ° C. for 5 hours. It turned black. Reaction mixture 5
After stirring for 3 hours on a water bath at 0 ° C., all chitin flakes were not dissolved and about half the amount was suspended in the liquid. A 75% hydrofluoric acid concentration significantly reduced the amount of this unreacted suspension. The amount of this suspension was adjusted to a concentration of hydrofluoric acid of 7
No significant reduction was observed unless it was 6% or more, and particularly 77-80% or more.

[0018]

[Example 3] When 96-100% hydrofluoric acid was used instead of 80% hydrofluoric acid in the method of Example 1, the time required to dissolve 400 g of chitin at 10 ° C was shortened within 1 hour. However, the reaction mixture becomes a completely black solution and the chitin flakes disappear. In this case, even if the reaction mixture was diluted with water, no precipitate was formed at all, and it is considered that glucosamine hydrofluoride was mostly formed. However, when this solution was distilled off under reduced pressure, a large amount of 2-acetamide-α-D-glucopyranosylfluorad was produced as a by-product, which was mixed with glucosamine hydrofluoride to obtain a considerable amount of a tar-like substance. The concentration of hydrofluoric acid had to be 95% or less.

[0019]

Example 4 300 g of 86% hydrofluoric acid was placed in a flask (1 l) equipped with a stirrer and cooled to -5 ° C, and 100 g of chitin (flakes powder prepared from shells of spiny lobster or sea crayfish) was added thereto. A viscous liquid with vigorous stirring is obtained. Then, this viscous liquid was poured into 2 liters of crushed ice to form a slurry, and hydrofluoric acid was distilled by vacuum distillation as quickly as possible along with humidification at 50 ° C.
0 to 50 g was collected, concentrated ammonia water was added to the residue, and p
H was neutralized to 3.5 and the deposited precipitate was filtered off. The precipitate filtered off here is the recovered chitin and is the raw material for the next experiment. Since the chitosan oligomer is dissolved in the filtrate, concentrated calcium chloride solution is added to the filtrate and filtered, and the filtrate is concentrated under reduced pressure to obtain about 30 chitosan oligomer hydrochloride.
get g. A liquid chromatographic analysis of this hydrochloride revealed that most of it was chitotriose hydrochloride, and that it contained some amounts of chitobiose hydrochloride, chitotetraose hydrochloride and chitopentaose hydrochloride.

[0020]

EXAMPLE 5 250 g of 90% hydrofluoric acid was placed in a flask (1 l) equipped with a stirrer, cooled to -10 ° C, and then chitin (flake powder prepared from the bran shell of Abrasemi) 9
Add 0 g and stir vigorously to obtain a yellowish brown viscous slurry. After mixing this with crushed ice, 28% ammonia water is added to neutralize and adjust the pH to about 4.0. Then, a concentrated calcium chloride solution is added and mixed, the reaction mixture is filtered, the precipitate on the filter paper is washed with a small amount of cold water, and then the whole filtrate is freeze-dried to give a composition containing 96% glucosamine hydrochloride and 4% chitobiose hydrochloride. The thing is obtained. This product was used as a feed additive after it was further defluorinated by adding calcium chloride.

[0021]

Example 6 1000 g of 78% hydrofluoric acid was placed in a flask (3 l) equipped with a stirrer and kept at 0 ° C. to form flaky chitin powder (flake powder prepared from the shell of king crab) 5
00 g was added and the mixture was stirred at 0 to 3 ° C for 5 hours to form a slurry. This slurry is filtered at a low temperature to separate insoluble chitin, and this is sent to the next charging material. The filtrate is concentrated under reduced pressure to collect hydrogen fluoride as much as possible, and the resulting residue is put into 1200 g of a 20% calcium chloride aqueous solution and vigorously stirred. The formed calcium fluoride precipitate is filtered off, the filtrate is extracted with methanol and the extract is concentrated to obtain a composition crystal of glucosamine hydrochloride. After fractionating glucosamine hydrochloride from this composition, the chitosan oligomer hydrochloride was analyzed by liquid chromatography to find that as an oligomer, about 20% chitobiose hydrochloride, about 16% chitotriose hydrochloride, about 8% chitotetraose hydrochloride. And a composition containing a small amount of chitin oligomer.

[0022]

Example 7 Chitin fine powder (flaky powder prepared from snow crab shell) 100 in a flask (2 l) equipped with a stirrer.
g, 300 g of 85% hydrofluoric acid was added thereto while maintaining at 5 ° C., and the mixture was reacted at 5 ° C. for 2 hours to form a brown slurry. Then, the mixture was kept on a water bath at 10 ° C for 9 hours, and then 500 ml of ice-cooled concentrated ammonia water was added and stirred to adjust the pH
Neutralize to 3.5 and chill with ice. Initially, a large amount of ammonium acid fluoride crystals are deposited, so this is filtered off and recrystallized to be recovered as ammonium acid fluoride for glass corrosion. The black filtrate is decolorized with activated carbon as much as possible, and then concentrated under reduced pressure on a water bath at 50 to 60 ° C. Then, 100 ml of concentrated hydrochloric acid was added to the distillation residue, the mixture was heated and stirred, and then concentrated under reduced pressure again to obtain a light yellowish brown crystalline residue. Therefore, when this was recrystallized from an aqueous alcohol solution, (a) glucosamine hydrochloride was obtained. A pale brown mixture of (b) chitobiose hydrochloride, (c) chitotriose hydrochloride is obtained. This mixture was analyzed by liquid chromatography.
It was found that (a) contained 82%, (b) contained 11%, and (c) contained 6%. Glucosamine and chitosan oligomer compositions as a mixture were dissolved in water and the solution was added to fish feeds, poultry feeds, livestock feeds, etc. in an amount of about 0.05 to 1.0%. Contributed to growth.

[0023]

Example 8 A residue of the glucosamine and chitosan oligomer composition obtained in Example 6 was concentrated under reduced pressure to 200 m.
1 of water, to which a concentrated aqueous solution of sodium lactate or sodium ascorbate is added until a precipitate no longer forms, is filtered and the filtrate is heated under reduced pressure on a water bath. By this method, a glucosamine / chitosan oligomer composition is obtained in the form of a lactate or ascorbate in the form of a light yellow to light brown powder. This composition is 0.05 to 1.0% as an additive that promotes the healthy growth of domestic animals.
It is used by being mixed with feed.

[0024]

Example 9 A nutrient solution for animal feed containing a chitosan oligomer composition and a feed additive using the same are shown below.
Glycine 150 g (2 mol), L-glutamic acid 147
g (1 mol) and 500 ml of water are kept at 100 ° C. for 100 hours to prepare a mixed amino acid solution. This mixed amino acid solution was found to be an excellent solvent for chitin oligomers and chitosan oligomers. The purified product obtained by recrystallizing glucosamine hydrochloride and chitosan oligomer hydrochloride, which are the products in Example 7, from methanol is dissolved in the above-mentioned mixed amino acid solution to be 1%. This solution was a colorless to pale yellow seasoning solution and nutrient solution with a strong sweetness similar to vinegar kelp, and showed great palatability with aquatic animals and insects, and thus was useful as a feed additive. Further, as a component of the mixed amino acid solution of this example, 2 mol of glycine was replaced with 1 mol of glycine and 1 mol of α-alanine, and under the other conditions, the mixed amino acid salt solution obtained in the same manner as above was used as a solvent and glucosamine was added thereto. A solution in which 2% of hydrochloride was dissolved was suitable as an additive for a compounded feed composed mainly of soybean meal.

[0025]

Example 10 When a chitin-based material is infiltrated into concentrated hydrofluoric acid containing 75 to 95% by weight of hydrogen fluoride, the crystalline region of chitin swells from the infiltrated portion and gradually begins to dissolve. This shows that protons from hydrogen fluoride or water are added to the proton-accepting portion of the swollen portion of chitin, and then the dissolution begins, and a HF-H ₂ O mixture suitable for this is 75-95% HF, especially HF is preferably 7
It was 8 to 92%. The state of adsorption of this proton seems to be quite different from that in the case of anhydrous hydrofluoric acid, and it seems to be slightly different from the reaction conditions for anhydrous hydrogen fluoride such as polyethylene terephthalate and polyurethane previously announced by the present inventors. There was (Japanese Patent Laid-Open No. 11-586
No. 6).

In other words, however, the chitin-based material is much more soluble in a solvent containing hydrogen fluoride than a synthetic product such as polyester, and when the solution is added to water or an alkaline aqueous solution, an oligomer having a low polymerization degree is formed. It is easy to generate. This method was first used by the present inventors for the purification of chitin, and was particularly applied for decolorization of chitin. After that, the present study was advanced, and 75 to 95%, particularly preferably 78 to 92% of hydrofluoric acid was added to glucosamine. Or (and) it became clear that it was suitable for the production of a chitosan oligomer composition, and the present invention was completed.

Many chitosan oligomers obtained by the method of the present invention have relatively low molecular weights, and in particular, glucosamine, chitobiose, chitotriose, chitotetraose, chitopentaose, chitohexaose, chitoheptaose, etc. are obtained in many cases. ing.

[0028]

INDUSTRIAL APPLICABILITY The present invention relates to a method for producing a glucosamine or / and chitosan oligomer composition, which comprises adding chitin to 100 to 2000 parts by weight of concentrated hydrofluoric acid containing 75 to 95% by weight of hydrogen fluoride. Glucosamine or (and) chitosan oligomer composition having a wide range of industrial applications by dehydrofluorinating a reaction mixture obtained by dispersing and dissolving 100 parts by weight of a base material at −20 to + 20 ° C. for hydrolysis. To provide a convenient way to get The glucosamine or (and) chitosan oligomer composition obtained by the method of the present invention does not have a special bitterness or pungency, and has only a slight saltiness or sourness in an almost neutral aqueous solution. It is useful as a feed additive and the like.

Continued front page    (72) Inventor Nao Kitano             6-2 Minamisikunigaoka-cho, Sakai City, Osaka Prefecture (72) Inventor Fumio Tanimoto             Neutral Sales Road Nishiiri 3chome, Kamigyo-ku, Kyoto-shi, Kyoto             471 Muromachi Sky Heights 112 (72) Inventor Hirohisa Kikuyama             Funakoko going down Ayakoji, Shinmachi-dori, Shimogyo-ku, Kyoto Prefecture, Kyoto Prefecture             Town 381-7 (72) Inventor Masahide Waki             56-134 Otani, Kumatori-cho, Sennan-gun, Osaka (72) Inventor Shinji Hashiguchi             3-21-5 Nishitoriishi, Takaishi-shi, Osaka Verde             Kamo 20 F-term (reference) 4C057 CC03 DD01

Claims (3)

[Claims] 1. Hydrolysis by dispersing and dissolving 100 parts by weight of chitin-based material in 100 to 2000 parts by weight of concentrated hydrofluoric acid containing 75 to 95% by weight of hydrogen fluoride at -20 to + 20 ° C. A method for producing a glucosamine or / and chitosan oligomer composition, which comprises dehydrofluorinating a reaction mixture carried out. 2. The glucosamine or the glucosamine according to claim 1, wherein the chitin-based material is a single or a mixture obtained, purified or recovered from crustaceans, insects, molluscs or microorganisms. (And) A method for producing a chitosan oligomer composition. 3. After dehydrofluoric acid removes insoluble matter from the reaction mixture, concentration is carried out by a vacuum distillation method, a spray drying method, a freeze drying method, a neutralization filtration method, a neutralization centrifugation method. A method for producing the glucosamine or / and chitosan oligomer composition according to claim 1.