JP2021017460A - High-concentration dispersion of nanosized chitin - Google Patents
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- 229920002101 Chitin Polymers 0.000 title claims abstract description 199
- 239000006185 dispersion Substances 0.000 title claims abstract description 133
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- 238000003381 deacetylation reaction Methods 0.000 claims description 46
- 230000006196 deacetylation Effects 0.000 claims description 39
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 33
- 239000007864 aqueous solution Substances 0.000 description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 22
- 230000002378 acidificating effect Effects 0.000 description 21
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 15
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- 238000010438 heat treatment Methods 0.000 description 14
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- 239000002121 nanofiber Substances 0.000 description 10
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- 238000005406 washing Methods 0.000 description 8
- 239000002994 raw material Substances 0.000 description 7
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 6
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 6
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 5
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 5
- 230000005540 biological transmission Effects 0.000 description 5
- 239000002537 cosmetic Substances 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 235000013376 functional food Nutrition 0.000 description 5
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- 210000001724 microfibril Anatomy 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 229950006780 n-acetylglucosamine Drugs 0.000 description 5
- 229920001542 oligosaccharide Polymers 0.000 description 5
- 150000002482 oligosaccharides Chemical class 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 239000003513 alkali Substances 0.000 description 4
- 239000000306 component Substances 0.000 description 4
- 238000007865 diluting Methods 0.000 description 4
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- 238000004519 manufacturing process Methods 0.000 description 4
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- 239000007858 starting material Substances 0.000 description 4
- 241000238557 Decapoda Species 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- 238000007598 dipping method Methods 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229920003023 plastic Polymers 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- DLFVBJFMPXGRIB-UHFFFAOYSA-N Acetamide Chemical compound CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 2
- 229920001297 Chitin nanofibril Polymers 0.000 description 2
- 241000238424 Crustacea Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000007385 chemical modification Methods 0.000 description 2
- 239000013256 coordination polymer Substances 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 238000011033 desalting Methods 0.000 description 2
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- 239000000945 filler Substances 0.000 description 2
- 238000005227 gel permeation chromatography Methods 0.000 description 2
- MSWZFWKMSRAUBD-IVMDWMLBSA-N glucosamine group Chemical group OC1[C@H](N)[C@@H](O)[C@H](O)[C@H](O1)CO MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 238000011085 pressure filtration Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 229950003937 tolonium Drugs 0.000 description 2
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 2
- 229920002554 vinyl polymer Polymers 0.000 description 2
- 241000238366 Cephalopoda Species 0.000 description 1
- 102000012286 Chitinases Human genes 0.000 description 1
- 108010022172 Chitinases Proteins 0.000 description 1
- 241000254173 Coleoptera Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
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- 239000003480 eluent Substances 0.000 description 1
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- 235000012041 food component Nutrition 0.000 description 1
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- 238000002347 injection Methods 0.000 description 1
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- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
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- 235000005985 organic acids Nutrition 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- WZAPMUSQALINQD-UHFFFAOYSA-M potassium;ethenyl sulfate Chemical compound [K+].[O-]S(=O)(=O)OC=C WZAPMUSQALINQD-UHFFFAOYSA-M 0.000 description 1
- 235000013406 prebiotics Nutrition 0.000 description 1
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Abstract
Description
本発明は、ナノ化キチンの高濃度分散物に関し、詳しくは、均一な繊維幅を有するナノ化キチンを高濃度で含有する分散物に関する。 The present invention relates to a high concentration dispersion of nano-sized chitin, and more particularly to a dispersion containing a high concentration of nano-sized chitin having a uniform fiber width.
エビやカニ等の甲殻類の殻に豊富に含まれるキチンを解繊処理して得たキチンナノファイバーは、透明性や高い水膨潤性を示し、種々の分野における利用が検討されている。
キチンナノファイバーの製造方法としては、たとえば、結晶化度が９０％以下の精製β−キチンをｐＨが５以下の酸性液体に浸漬し、次いで浸漬されたβ−キチンを解繊処理する方法（特許文献１）、精製α−キチンを部分脱アセチル化し、ｐＨが５以下の酸性液体に浸漬した後、解繊処理する方法（特許文献２）等が提案されている。
特許文献１および２に記載されたキチンナノファイバーの製造方法においては、キチンナノフィブリルの表面に分布すると考えられるグルコサミン残基にプラス荷電を付与し、キチンナノフィブリルのミクロフィブリル間に荷電反発を生じさせて、ミクロフィブリルの解繊を容易にすることが提案されている。
Chitin nanofibers obtained by defibrating chitin, which is abundant in the shells of crustaceans such as shrimp and crab, show transparency and high water swelling properties, and their use in various fields is being studied.
As a method for producing chitin nanofibers, for example, a method of immersing purified β-chitin having a crystallinity of 90% or less in an acidic liquid having a pH of 5 or less and then defibrating the immersed β-chitin (Patent). Document 1), a method of partially deacetylating purified α-chitin, immersing it in an acidic liquid having a pH of 5 or less, and then defibrating it (Patent Document 2) has been proposed.
In the method for producing chitin nanofibers described in Patent Documents 1 and 2, a positive charge is applied to a glucosamine residue that is considered to be distributed on the surface of the chitin nanofibrils, and a charge repulsion is generated between the microfibrils of the chitin nanofibrils. It has been proposed to facilitate the defibration of microfibrils.
特許文献１、２に記載されたキチンナノファイバーの製造方法においては、精製β−キチンまたは精製α−キチンの部分脱アセチル化物の解繊処理は、プロペラミキサー、カッターミキサー、超音波ホモジナイザー、高圧ホモジナイザー、二軸混練機等の解繊、粉砕機器を用いて行われている。
また、キチンミクロフィブリルの解繊方法として、ウォータージェットの技術を応用して解繊する方法も提案されている（特許文献３）。
In the method for producing chitin nanofibers described in Patent Documents 1 and 2, the defibration treatment of a partially deacetylated product of purified β-chitin or purified α-chitin is performed by a propeller mixer, a cutter mixer, an ultrasonic homogenizer, or a high-pressure homogenizer. , It is carried out using defibrating and crushing equipment such as a twin-screw kneader.
Further, as a method for defibrating chitin microfibrils, a method for defibrating by applying a water jet technique has also been proposed (Patent Document 3).
特許文献１〜３に記載されるように、キチンナノファイバーは、一般的に、精製したキチンを機械的または物理的に解繊処理することにより得られており、かかる方法では、均一な繊維幅を有するキチンナノファイバーを簡便に得ることは困難であり、キチンナノファイバーの繊維幅を均一にするために解繊処理を繰り返す必要があった。
また、特許文献１〜３等に開示される製造方法では、キチンの水分散液を解繊処理に供するため、キチンナノファイバーは水に分散した状態で得られ、キチンナノファイバーを高濃度の分散物として得ることは困難であり、Ｎ−アセチルグルコサミンやオリゴ糖の原料として、また、医用材料、機能性食品、化粧品、飼料等に利用する上で、経済性に優れるとはいえなかった。
As described in Patent Documents 1 to 3, chitin nanofibers are generally obtained by mechanically or physically defibrating purified chitin, and in such a method, a uniform fiber width is obtained. It is difficult to easily obtain chitin nanofibers having the above, and it is necessary to repeat the defibration treatment in order to make the fiber width of the chitin nanofibers uniform.
Further, in the production methods disclosed in Patent Documents 1 to 3 and the like, since the aqueous dispersion of chitin is subjected to the defibration treatment, the chitin nanofibers are obtained in a state of being dispersed in water, and the chitin nanofibers are dispersed in a high concentration. It is difficult to obtain as a product, and it cannot be said that it is excellent in economic efficiency when used as a raw material for N-acetylglucosamine and oligosaccharides, and for medical materials, functional foods, cosmetics, feeds and the like.
そこで、均一な繊維幅を有するナノ化キチンを高濃度で含む分散物であって、種々の分野における利用性に優れるナノ化キチンの分散物が望まれている。 Therefore, a dispersion containing nano-chitin having a uniform fiber width at a high concentration and having excellent usability in various fields is desired.
上記に鑑み、本発明は、均一な繊維幅を有するナノ化キチンを高濃度で含有する分散物を提供することを目的とする。 In view of the above, it is an object of the present invention to provide a dispersion containing a high concentration of nano-chitin having a uniform fiber width.
本発明者らは、上記課題を解決すべく鋭意検討した結果、高濃度のアルカリまたは酸により低温で処理し、希釈および中和することにより得られるキチンの析出物が、ナノ化されたキチンであることを見出し、さらに検討を行って、繊維幅が２０ｎｍ以下であるナノ化キチンを、１０重量％〜３０重量％の濃度で含有する分散物を得て、本発明を完成した。 As a result of diligent studies to solve the above problems, the present inventors have obtained nano-sized chitin, which is obtained by treating with a high concentration of alkali or acid at a low temperature, diluting and neutralizing the chitin precipitate. Further studies were carried out to obtain a dispersion containing nanochitin having a fiber width of 20 nm or less at a concentration of 10% by weight to 30% by weight, and the present invention was completed.
すなわち、本発明は以下に関する。
［１］繊維幅が２０ｎｍ以下であるナノ化キチンを１０重量％〜３０重量％の濃度で含有する、ナノ化キチンの分散物。
［２］ナノ化キチンの濃度が１５重量％〜２５重量％である、［１］に記載の分散物。
［３］ナノ化キチンの繊維幅が１０ｎｍ以下である、［１］または［２］に記載の分散物。
［４］ナノ化キチンの脱アセチル化度が８％以下である、［１］〜［３］のいずれかに記載の分散物。
［５］ウェットケーキ状の分散物である、［１］〜［４］のいずれかに記載の分散物。
［６］ナノ化キチンの脱アセチル化物の分散物。
［７］脱アセチル化度が１０％〜５０％である、［６］に記載の分散物。
［８］［６］または［７］に記載の分散物を含有する、ゲル。
That is, the present invention relates to the following.
[1] A dispersion of nano-sized chitin containing nano-sized chitin having a fiber width of 20 nm or less at a concentration of 10% by weight to 30% by weight.
[2] The dispersion according to [1], wherein the concentration of nano-sized chitin is 15% by weight to 25% by weight.
[3] The dispersion according to [1] or [2], wherein the fiber width of the nano-sized chitin is 10 nm or less.
[4] The dispersion according to any one of [1] to [3], wherein the degree of deacetylation of nano-sized chitin is 8% or less.
[5] The dispersion according to any one of [1] to [4], which is a wet cake-like dispersion.
[6] A dispersion of deacetylase of nano-sized chitin.
[7] The dispersion according to [6], wherein the degree of deacetylation is 10% to 50%.
[8] A gel containing the dispersion according to [6] or [7].
本発明により、均一な繊維幅を有するナノ化キチンを高濃度で含有する分散物を提供することができる。
本発明の分散物に含有されるナノ化キチンは、ナノ化キチンの形態を保ったまま脱アセチル化することができる。ナノ化キチンの脱アセチル化物の分散物は、液性を酸性とすることにより、または酸性水溶液に分散させることにより、低濃度でゲルを形成することができる。
上記分散物は、Ｎ−アセチルグルコサミンやオリゴ糖の原料、医用材料、プラスチック材料等として、また、機能性食品、化粧品、飼料等の成分として有用である。
INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a dispersion containing a high concentration of nano-sized chitin having a uniform fiber width.
The nano-sized chitin contained in the dispersion of the present invention can be deacetylated while maintaining the form of the nano-sized chitin. The dispersion of the deacetylase of nanonized chitin can form a gel at a low concentration by making the liquid acidic or by dispersing it in an acidic aqueous solution.
The above dispersion is useful as a raw material for N-acetylglucosamine and oligosaccharides, a medical material, a plastic material, and the like, and as a component for functional foods, cosmetics, feeds, and the like.
本発明は、均一な繊維幅を有するナノ化キチンを高濃度で含有する分散物（以下、本明細書にて「本発明の分散物」とも称する）を提供する。
本発明の分散物は、繊維幅が２０ｎｍ以下であるナノ化キチンを、１０重量％〜３０重量％の濃度で含有する。
The present invention provides a dispersion containing a high concentration of nano-sized chitin having a uniform fiber width (hereinafter, also referred to as “dispersion of the present invention” in the present specification).
The dispersion of the present invention contains nano-chitin having a fiber width of 20 nm or less at a concentration of 10% by weight to 30% by weight.
ここで、「ナノ化キチン」とは、繊維幅がナノメートルオーダー、すなわち１μｍ未満であるキチンをいう。
本発明の分散物に含有されるナノ化キチンの繊維幅は２０ｎｍ以下であり、好ましくは１０ｎｍ以下である。また、本発明の分散物に含有されるナノ化キチンの平均繊維幅は通常３ｎｍ〜１０ｎｍであり、好ましくは３ｎｍ〜５ｎｍである。
本発明の分散物に含有されるナノ化キチンの繊維幅は、本発明の分散物を水等で適宜希釈し、透過型電子顕微鏡にて観察した際の画像から計測される。
一方、本発明の分散物に含有されるナノ化キチンを透過型電子顕微鏡下にて観察したとき、長さ方向においてスパイラル状にねじれていることが認められ、繊維長を明確に計測することができない。しかし、後述するように、本発明の分散物を調製した際、分散物中に含有されるナノ化キチンの分子量は、出発原料としたキチンに比べて低下するものの、その程度は通常の物理的解繊方法により調製する場合と同程度であり、物理的解繊方法により調製されたキチンナノファイバーの繊維長とほぼ変わらないと推測される。
Here, "nano-sized chitin" refers to chitin having a fiber width on the order of nanometers, that is, less than 1 μm.
The fiber width of the nano-sized chitin contained in the dispersion of the present invention is 20 nm or less, preferably 10 nm or less. The average fiber width of the nano-sized chitin contained in the dispersion of the present invention is usually 3 nm to 10 nm, preferably 3 nm to 5 nm.
The fiber width of nano-sized chitin contained in the dispersion of the present invention is measured from an image obtained by appropriately diluting the dispersion of the present invention with water or the like and observing it with a transmission electron microscope.
On the other hand, when the nano-sized chitin contained in the dispersion of the present invention was observed under a transmission electron microscope, it was found that it was twisted in a spiral shape in the length direction, and the fiber length could be clearly measured. Can not. However, as will be described later, when the dispersion of the present invention is prepared, the molecular weight of nano-sized chitin contained in the dispersion is lower than that of chitin as a starting material, but the degree is usually physical. It is about the same as the case of preparing by the defibration method, and it is presumed that it is almost the same as the fiber length of the chitin nanofibers prepared by the physical defibration method.
本発明の分散物に含有されるナノ化キチンは、カニ、エビ等の甲殻類に存在するα−キチン、イカの中骨やハオリムシに存在するβ−キチンのいずれのキチンから得られるものでもよい。 The nano-chitin contained in the dispersion of the present invention may be obtained from any of α-chitin present in crustaceans such as crabs and shrimp, and β-chitin present in squid bones and haori beetles. ..
本発明の分散物は、通常１０重量％〜３０重量％の濃度で、好ましくは１５重量％〜２５重量％の濃度で、上記したナノ化キチンを含有する。
ここで、本発明の分散物に含有されるナノ化キチンの濃度は、本発明の分散物中におけるキチン含有量を、常圧加熱乾燥法により固形分量として定量し、得られた定量値と、乾燥前の本発明の分散物の重量とから算出される。
The dispersion of the present invention usually contains the above-mentioned nanonized chitin at a concentration of 10% by weight to 30% by weight, preferably 15% by weight to 25% by weight.
Here, the concentration of the nano-sized chitin contained in the dispersion of the present invention is the quantitative value obtained by quantifying the chitin content in the dispersion of the present invention as a solid content by a normal pressure heating and drying method. It is calculated from the weight of the dispersion of the present invention before drying.
本発明の分散物に含有されるナノ化キチンの脱アセチル化度は、好ましくは８％以下であり、より好ましくは５％以下である。また、原料として用いるキチンにおける脱アセチル化度を考慮すると、本発明の分散物に含有されるナノ化キチンの脱アセチル化度は、通常１％〜５％程度である。
ナノ化キチンの脱アセチル化度は、トルイジンブルーを指示薬とした１／４００Ｎポリビニル硫酸カリウム水溶液によるコロイド滴定により測定することができる。
The degree of deacetylation of the nano-chitin contained in the dispersion of the present invention is preferably 8% or less, more preferably 5% or less. Further, considering the degree of deacetylation of chitin used as a raw material, the degree of deacetylation of nano-sized chitin contained in the dispersion of the present invention is usually about 1% to 5%.
The degree of deacetylation of nano-sized chitin can be measured by colloidal titration with a 1 / 400N aqueous potassium polyvinyl sulfate solution using toluidine blue as an indicator.
本発明の分散物においては、上記したように、微細かつ均一な繊維幅を有するナノ化キチンが、水等の溶媒に１０重量％〜３０重量％の高濃度で分散されており、ウェットケーキ状の外観を呈する。 In the dispersion of the present invention, as described above, nano-sized chitin having a fine and uniform fiber width is dispersed in a solvent such as water at a high concentration of 10% by weight to 30% by weight, and is in the form of a wet cake. It presents the appearance of.
本発明の分散物は、たとえば、キチンを４２メッシュの篩を通過する程度に粉砕し、前記キチン粉末を高濃度のアルカリ水溶液に浸漬し、室温以下の温度で静置した後、氷を加えて希釈し、高濃度の酸で中和してキチンを析出させ、前記キチンの分散物を水で洗浄して脱塩し、必要に応じて脱水することにより、調製することができる。
また、本発明の分散物は、粉砕したキチン粉末を、高濃度の酸の水溶液に浸漬処理し、高濃度のアルカリで中和することによっても調製することができるが、目的とするナノ化キチンの低分子化を考慮すると、アルカリ水溶液に浸漬処理し、酸で中和することが好ましい。
In the dispersion of the present invention, for example, chitin is pulverized to the extent that it passes through a 42-mesh sieve, the chitin powder is immersed in a high-concentration alkaline aqueous solution, allowed to stand at a temperature below room temperature, and then ice is added. It can be prepared by diluting and neutralizing with a high concentration acid to precipitate chitin, washing the dispersion of chitin with water, desalting it, and dehydrating it if necessary.
The dispersion of the present invention can also be prepared by immersing the crushed chitin powder in an aqueous solution of a high concentration acid and neutralizing it with a high concentration alkali, but the desired nano-sized chitin In consideration of the reduction in molecular weight, it is preferable to immerse the product in an alkaline aqueous solution and neutralize it with an acid.
キチンの粉砕は、通常の粉砕機にて行うことができ、たとえば、カッターミル粉砕機、ハンマーミル粉砕機、転動ボールミル粉砕機、乾式気流粉砕機、対向気流乾式粉砕機等の一般的な乾式粉砕機を用いて行うことができる。 The chitin can be crushed by a normal crusher, for example, a general dry type such as a cutter mill crusher, a hammer mill crusher, a rolling ball mill crusher, a dry air flow crusher, and an oncoming air flow dry crusher. It can be done using a crusher.
高濃度のアルカリ水溶液としては、４０重量％〜４８重量％程度の水酸化ナトリウム水溶液、水酸化カリウム水溶液等を用いることができる。高濃度のアルカリ水溶液は、キチン粉末が十分に浸漬される程度の量を用いる。
キチン粉末の高濃度のアルカリ水溶液での浸漬処理は、好ましくは室温（２５℃）以下、より好ましくは２０℃以下、さらに好ましくは１５℃以下の温度で、１０時間〜２４時間行うことが好ましく、１５時間〜２０時間行うことがより好ましい。
As the high-concentration alkaline aqueous solution, a sodium hydroxide aqueous solution, a potassium hydroxide aqueous solution, or the like of about 40% by weight to 48% by weight can be used. The high-concentration alkaline aqueous solution should be used in such an amount that the chitin powder is sufficiently immersed.
The dipping treatment of the chitin powder in a high-concentration alkaline aqueous solution is preferably carried out at a temperature of room temperature (25 ° C.) or lower, more preferably 20 ° C. or lower, still more preferably 15 ° C. or lower, for 10 to 24 hours. More preferably, it is carried out for 15 to 20 hours.
上記浸漬処理の後、氷を加えるとキチンは粘調な状態で溶け出すので、キチン浸漬液が透明になるまで攪拌する。前記の氷の添加による希釈処理は、２５℃以下、好ましくは０℃以下、より好ましくは−１０℃以下にて、キチン粉末の浸漬物が均一な溶液となるまで（アルカリ濃度が１０（ｗ／ｖ）％程度となるまで）行う。
上記希釈処理により得たキチン溶液の中和は、３０重量％〜４０重量％程度の酸、たとえば濃塩酸等を添加して、２５℃以下、好ましくは０℃以下にて、前記溶液のｐＨが７〜８．５程度となるように行う。
上記のアルカリ水溶液での浸漬処理を好ましくは２５℃以下、より好ましくは２０℃以下の低温で、希釈および中和を、２５℃以下、好ましくは０℃以下の低温で行うことにより、キチンの脱アセチル化を抑制することができる。
After the above dipping treatment, when ice is added, chitin melts in a viscous state, so the mixture is stirred until the chitin dipping solution becomes transparent. The dilution treatment by adding ice is carried out at 25 ° C. or lower, preferably 0 ° C. or lower, more preferably -10 ° C. or lower, until the soaked chitin powder becomes a uniform solution (alkali concentration: 10 (w / w /). v) Perform until it reaches about%.
To neutralize the chitin solution obtained by the above dilution treatment, an acid of about 30% by weight to 40% by weight, for example, concentrated hydrochloric acid is added, and the pH of the solution is 25 ° C. or lower, preferably 0 ° C. or lower. Do so so that it is about 7 to 8.5.
Desorption of chitin by diluting and neutralizing with the above alkaline aqueous solution at a low temperature of preferably 25 ° C. or lower, more preferably 20 ° C. or lower, and at a low temperature of 25 ° C. or lower, preferably 0 ° C. or lower. Acetylation can be suppressed.
水による洗浄は、１回あたり、析出したキチンの分散物に対し２５０重量倍〜５００重量倍の水により行い、かかる洗浄操作を通常１０回〜２０回繰り返す。かかる洗浄操作により、析出したキチンの分散物の脱塩を十分に行うことができる。本発明において、水による洗浄は、析出したキチンの分散物の塩分濃度が、通常０．０１重量％以下となるまで行う。
なお、脱塩の程度は、デジタル塩分濃度計を用いて塩分濃度を測定することにより、確認することができる。
Washing with water is carried out with 250 to 500 times by weight of water with respect to the precipitated dispersion of chitin, and such a washing operation is usually repeated 10 to 20 times. By such a washing operation, the precipitated dispersion of chitin can be sufficiently desalted. In the present invention, washing with water is carried out until the salt concentration of the precipitated chitin dispersion is usually 0.01% by weight or less.
The degree of desalting can be confirmed by measuring the salinity using a digital salinity meter.
析出したキチンの分散物の脱水は、通常の脱水方法により適宜行うことができるが、たとえば、遠心濾過、加圧濾過等により、好適に行うことができる。 Dehydration of the precipitated chitin dispersion can be appropriately carried out by a usual dehydration method, but can be preferably carried out by, for example, centrifugal filtration, pressure filtration or the like.
本発明の分散物に含有されるナノ化キチンは、キチナーゼ等の酵素による分解性が良好であり、脱アセチル化等の化学修飾も容易である、という特性を有する。
また、本発明の分散物は、繊維幅が微小かつ均一であるナノ化キチンを、従来にはない高濃度で含有するため、Ｎ−アセチルグルコサミンやオリゴ糖の原料、医用材料、プラスチック材料のフィラー等として、また、機能性食品、化粧品、飼料等の成分として、効率よく利用することができる。
The nano-sized chitin contained in the dispersion of the present invention has the characteristics that it has good degradability by an enzyme such as chitinase and is easily chemically modified such as deacetylation.
In addition, since the dispersion of the present invention contains nano-sized chitin having a fine and uniform fiber width at an unprecedented high concentration, it is a filler for N-acetylglucosamine and oligosaccharide raw materials, medical materials, and plastic materials. It can be efficiently used as a component of functional foods, cosmetics, feeds and the like.
さらに本発明は、本発明の分散物に含有されるナノ化キチンの化学修飾の一態様として、該ナノ化キチンの脱アセチル化物の分散物（以下、本明細書にて「本発明のナノ化キチンの脱アセチル化物の分散物」とも称する）を提供する。
本発明において、ナノ化キチンの脱アセチル化物の脱アセチル化度は、１０％〜５０％であることが好ましく、１５％〜４０％であることがより好ましい。
Further, the present invention, as one aspect of chemical modification of the nano-sized chitin contained in the dispersion of the present invention, is a dispersion of a deacetylase of the nano-sized chitin (hereinafter, "nano-sized of the present invention" in the present specification. Also referred to as "dispersion of deacetylase of chitin").
In the present invention, the degree of deacetylation of the deacetylase of nano-chitin is preferably 10% to 50%, more preferably 15% to 40%.
本発明のナノ化キチンの脱アセチル化物の分散物は、酸を少量加えて、該分散物の液性を酸性とするか、または酸性水溶液に分散させると、ナノ化キチンの脱アセチル化物の分散性が向上し、ナノ化キチンの脱アセチル化物の濃度が１重量％〜２重量％の低濃度にて均一なゲルを形成し得る。ナノ化キチンの脱アセチル化度が１８％を超えると、形成されるゲルの透明性が高まり、ナノ化キチンの脱アセチル化度が４０％程度になると、ほぼ透明なチキソトロピー性を呈する粘稠なゲルが得られる。
本発明のナノ化キチンの脱アセチル化物の分散物の分散性を向上させ、またはゲルを形成させるために添加される酸としては、該分散物の液性を酸性とすることができれば特に限定されないが、ナノ化キチンの脱アセチル化物の低分子化を抑制する観点からは弱酸であることが好ましく、有機酸であることがより好ましい。また、食品成分として利用するためには、可食性の酸であることが好ましい。可食性の有機酸としては、乳酸、クエン酸等が挙げられる。
また、本発明のナノ化キチンの脱アセチル化物の分散物に添加される酸の量は少量でよく、該分散物における酸の濃度が０．５重量％〜１重量％程度となる量でよい。
本発明のナノ化キチンの脱アセチル化物を酸性水溶液に分散させる場合も、酸性水溶液としては上記した酸の水溶液を用いることができ、その濃度は、０．５重量％〜１重量％程度の低濃度でよい。
The dispersion of the deacetylated product of nano-sized chitin of the present invention can be dispersed by adding a small amount of acid to make the liquid property of the dispersion acidic or by dispersing it in an acidic aqueous solution. The properties are improved, and a uniform gel can be formed at a low concentration of 1% by weight to 2% by weight of the deacetylated product of nano-chitin. When the degree of deacetylation of nano-sized chitin exceeds 18%, the transparency of the formed gel increases, and when the degree of deacetylation of nano-sized chitin reaches about 40%, it is viscous and exhibits almost transparent thixotropy. A gel is obtained.
The acid added to improve the dispersibility of the deacetylase dispersion of the nano-chitin of the present invention or to form a gel is not particularly limited as long as the liquid property of the dispersion can be made acidic. However, from the viewpoint of suppressing the reduction of molecular weight of the deacetylase of nano-chitin, a weak acid is preferable, and an organic acid is more preferable. Further, in order to use it as a food component, it is preferably an edible acid. Examples of edible organic acids include lactic acid and citric acid.
Further, the amount of acid added to the dispersion of the deacetylase of nano-sized chitin of the present invention may be small, and the concentration of acid in the dispersion may be about 0.5% by weight to 1% by weight. ..
When the deacetylase of nano-sized chitin of the present invention is dispersed in an acidic aqueous solution, the above-mentioned acid aqueous solution can be used as the acidic aqueous solution, and the concentration thereof is as low as about 0.5% by weight to 1% by weight. The concentration may be sufficient.
本発明のナノ化キチンの脱アセチル化物の分散物は、上記した本発明のナノ化キチンの分散物を、アルカリ水溶液中で加温または加熱し、水洗することにより、得ることができる。
アルカリ水溶液としては、水酸化ナトリウム、水酸化カリウム等の水溶液が好ましく用いられる。アルカリ水溶液の濃度は、キチンの結晶構造が膨潤しない範囲で目的とする脱アセチル化度に応じて適宜設定され、通常１０（ｗ／ｖ）％〜５５（ｗ／ｖ）％、好ましくは１０（ｗ／ｖ）％〜４０（ｗ／ｖ）％、より好ましくは３０（ｗ／ｖ）％〜３６（ｗ／ｖ）％の濃度のアルカリ水溶液が用いられる。
アルカリ水溶液中での加温または加熱温度は、目的とする脱アセチル化度に応じて適宜設定され、通常４５℃〜８０℃であり、好ましくは４５℃〜６０℃である。
アルカリ水溶液中での加温または加熱時間も、目的とする脱アセチル化度に応じて適宜設定され、通常５時間〜７２時間であり、好ましくは５時間〜１７時間であり、より好ましくは５時間〜６時間である。
アルカリ水溶液の濃度、加温または加熱温度、および加温または加熱時間を調整することにより、ナノ化キチンの脱アセチル化度を調整することができる。たとえば、３０（ｗ／ｖ）％〜３６（ｗ／ｖ）％の水酸化ナトリウム水溶液中、６０℃で５時間脱アセチル化反応を行った場合、脱アセチル化度は１５％〜１６％であり、８０℃で５時間脱アセチル化反応を行った場合、脱アセチル化度は１８％〜１９％であり、４５℃で７２時間脱アセチル化反応を行った場合、脱アセチル化度は２５％〜２６％である。
The dispersion of the deacetylase of the nano-sized chitin of the present invention can be obtained by heating or heating the above-mentioned dispersion of the nano-sized chitin of the present invention in an alkaline aqueous solution and washing with water.
As the alkaline aqueous solution, an aqueous solution of sodium hydroxide, potassium hydroxide or the like is preferably used. The concentration of the alkaline aqueous solution is appropriately set according to the desired degree of deacetylation within a range in which the crystal structure of chitin does not swell, and is usually 10 (w / v)% to 55 (w / v)%, preferably 10 ( An alkaline aqueous solution having a concentration of w / v)% to 40 (w / v)%, more preferably 30 (w / v)% to 36 (w / v)% is used.
The heating or heating temperature in the alkaline aqueous solution is appropriately set according to the desired degree of deacetylation, and is usually 45 ° C to 80 ° C, preferably 45 ° C to 60 ° C.
The heating or heating time in the alkaline aqueous solution is also appropriately set according to the desired degree of deacetylation, and is usually 5 hours to 72 hours, preferably 5 hours to 17 hours, and more preferably 5 hours. ~ 6 hours.
The degree of deacetylation of nano-chitin can be adjusted by adjusting the concentration of the alkaline aqueous solution, the heating or heating temperature, and the heating or heating time. For example, when the deacetylation reaction is carried out at 60 ° C. for 5 hours in a 30 (w / v)% to 36 (w / v)% sodium hydroxide aqueous solution, the degree of deacetylation is 15% to 16%. When the deacetylation reaction was carried out at 80 ° C. for 5 hours, the deacetylation degree was 18% to 19%, and when the deacetylation reaction was carried out at 45 ° C. for 72 hours, the deacetylation degree was 25% to 19%. It is 26%.
アルカリ水溶液中で加温または加熱した後、酸による中和処理を行ってもよいが、上述したように、本発明のナノ化キチンの脱アセチル化物が、低濃度の酸によりゲルを形成することから、ナノ化キチンの脱アセチル化物の分散物からアルカリを除去するには、該分散物に対し、２５０重量倍〜５００重量倍程度の水により、１０回〜２０回程度繰り返し洗浄することが好ましい。
また、本発明のナノ化キチンの脱アセチル化物の分散物の液性が弱酸性になると、膨潤、ゲル化して、ろ過による回収が不可能となることがあるので、水による洗浄は、ろ過した際のろ液のｐＨが８．５〜９を示す程度で終了することが好ましい。
After heating or heating in an alkaline aqueous solution, neutralization treatment with an acid may be performed, but as described above, the deacetylated product of the nano-chitin of the present invention forms a gel with a low concentration of acid. Therefore, in order to remove the alkali from the dispersion of the deacetylated nano-chitin, it is preferable to repeatedly wash the dispersion with water of about 250 to 500 times by weight about 10 to 20 times. ..
Further, if the liquidity of the dispersion of the deacetylase of nano-sized chitin of the present invention becomes weakly acidic, it may swell and gel, and recovery by filtration may not be possible. Therefore, washing with water was performed by filtration. It is preferable to finish the process when the pH of the filtrate shows 8.5-9.
上述したように、本発明のナノ化キチンの脱アセチル化物の分散物は、該分散物の液性を酸性とするか、または酸性水溶液に分散させると、ナノ化キチンの脱アセチル化物の分散性が向上し、低濃度で均一なゲルを形成することができ、脱アセチル化度を調整することによって、透明性の高いゲルや、チキソトロピー性を示すゲルを得ることができる。
本発明のナノ化キチンの脱アセチル化物の分散物は、さらに化学修飾に供することができる。
As described above, the dispersion of the deacetylated product of the nano-sized chitin of the present invention has the dispersibility of the deacetylated product of the nano-sized chitin when the liquid property of the dispersion is made acidic or dispersed in an acidic aqueous solution. It is possible to form a uniform gel at a low concentration, and by adjusting the degree of deacetylation, a highly transparent gel or a gel exhibiting thixotropy can be obtained.
The dispersion of the deacetylase of nanonized chitin of the present invention can be further subjected to chemical modification.
以下、本発明について、実施例により詳細に説明する。 Hereinafter, the present invention will be described in detail with reference to Examples.
［実施例１］ナノ化キチンを高濃度で含有する分散物（ウェットケーキ状ナノ化キチン）
下記の通り、ナノ化キチンを高濃度で含有する分散物を調製した。
カニ殻由来のα−キチン粉末（４２メッシュ篩過品、脱アセチル化度＝１．０％〜２．０％（「キチンＬ−ＰＣ」、甲陽ケミカル株式会社製）に、１０重量倍の４８重量％の水酸化ナトリウム水溶液を加え、キチン粉末に十分に水酸化ナトリウム水溶液を浸透させた後、２０℃で一昼夜静置した。
次いで、水酸化ナトリウムの濃度が１０（ｗ／ｖ）％程度になるまで、砕いた氷を添加し、−１０℃以下にて均一な液状になるまで攪拌した。
次に、氷を加えながら濃塩酸を添加して、溶液のｐＨが８．０〜８．５程度になるまで中和し、キチンを析出させた。
析出したキチンの分散物を、該分散物に対し大量の水で洗浄を繰り返し行い、デジタル塩分濃度計（「ＥＳ−４２１」、株式会社アタゴ製）で測定される塩分が０．０１重量％以下になるまで脱塩を行った。
脱塩したキチンの分散物を、加圧ろ過（フィルタープレス）（薮田機械株式会社製）により脱水し、ウェットケーキ状の分散物を得た。
[Example 1] Dispersion containing nano-sized chitin at a high concentration (wet cake-like nano-sized chitin)
As described below, a dispersion containing a high concentration of nano-sized chitin was prepared.
Α-chitin powder derived from crab shell (42 mesh sieved product, degree of deacetylation = 1.0% to 2.0% ("chitin L-PC", manufactured by Koyo Chemical Co., Ltd.), 10 times by weight 48 A wt% sodium hydroxide aqueous solution was added, and the chitin powder was sufficiently impregnated with the sodium hydroxide aqueous solution, and then allowed to stand at 20 ° C. for 24 hours.
Then, crushed ice was added until the concentration of sodium hydroxide reached about 10 (w / v)%, and the mixture was stirred at −10 ° C. or lower until a uniform liquid was obtained.
Next, concentrated hydrochloric acid was added while adding ice to neutralize the solution until the pH became about 8.0 to 8.5, and chitin was precipitated.
The precipitated dispersion of chitin is repeatedly washed with a large amount of water, and the salt content measured by a digital salinity meter (“ES-421”, manufactured by Atago Co., Ltd.) is 0.01% by weight or less. Desalination was performed until
The desalted chitin dispersion was dehydrated by pressure filtration (filter press) (manufactured by Yabuta Kikai Co., Ltd.) to obtain a wet cake-like dispersion.
上記で得られたウェットケーキ状の分散物について、透過型電子顕微鏡（ＴＥＭ）による観察を行った。
すなわち、上記で得られた分散物を、精製水にてキチン濃度が２．５重量％程度となるように希釈し、ＴＥＭ観察用試料を作製して、「ＪＥＭ−２１００」（日本電子株式会社製）にて５０，０００倍の倍率で観察した。ＴＥＭによる観察画像を図１に示した。
The wet cake-like dispersion obtained above was observed with a transmission electron microscope (TEM).
That is, the dispersion obtained above is diluted with purified water so that the chitin concentration is about 2.5% by weight to prepare a TEM observation sample, and "JEM-2100" (JEOL Ltd.) It was observed at a magnification of 50,000 times. The observation image by TEM is shown in FIG.
図１に示されるように、得られた分散物は、キチンがエレメンタリーミクロフィブリル単位にまで解繊されたナノ化キチンの分散物であることが確認された。
図１に示すナノ化キチンの観察画像から、ナノ化キチンの繊維幅を計測したところ、１０ｎｍ以下であり、繊維長は正確には計測できなかった。また、繊維幅の計測値から、算出された平均繊維幅は３ｎｍ〜５ｎｍであった。
一方、観察されるナノ化キチンは、長さ方向においてスパイラル状にねじれており、上記した通り、繊維長を正確に計測することはできなかった。
As shown in FIG. 1, it was confirmed that the obtained dispersion was a dispersion of nano-sized chitin in which chitin was defibrated to elementary microfibril units.
When the fiber width of the nano-sized chitin was measured from the observation image of the nano-sized chitin shown in FIG. 1, it was 10 nm or less, and the fiber length could not be measured accurately. The average fiber width calculated from the measured value of the fiber width was 3 nm to 5 nm.
On the other hand, the observed nano-chitin was twisted in a spiral shape in the length direction, and as described above, the fiber length could not be measured accurately.
［試験例１］実施例１のナノ化キチンの分散物におけるナノ化キチン含有濃度およびナノ化キチンの脱アセチル化度の測定
実施例１のナノ化キチンの分散物について、下記の通りナノ化キチン含有濃度およびナノ化キチンの脱アセチル化度の測定を行った。
[Test Example 1] Measurement of nano-chitin content concentration and deacetylation degree of nano-chitin in the nano-chitin dispersion of Example 1 The nano-chitin dispersion of Example 1 is as follows. The content concentration and the degree of deacetylation of nano-chitin were measured.
（１）ナノ化キチン含有濃度の測定
実施例１のナノ化キチンの分散物について、常圧加熱乾燥法により固形分量を求め、乾燥前の分散物の重量とから、ナノ化キチンの含有濃度を算出した。
(1) Measurement of Nanonized Chitin Concentration The solid content of the nanonized chitin dispersion of Example 1 was determined by the atmospheric heating and drying method, and the nanonized chitin content was determined from the weight of the dispersion before drying. Calculated.
（２）脱アセチル化度の測定
Ｎ，Ｎ−ジメチルアセトアミド１３８０ｇを、あらかじめスターラーを入れておいた２Ｌのガラス製ビーカーに量り取り、スターラーにて撹拌し、そこに塩化リチウム１２０ｇを加えて溶解させて、キチン溶解液を調製した。
実施例１のナノ化キチンの分散物を常圧にて加熱乾燥し、得られた固形分２．５ｇを量り取り、上記キチン溶解液を加えて全量を５００ｇとした。一晩撹拌して溶解させ、得られたナノ化キチン溶液１．０ｇを量り取り、脱イオン水を加えて５０ｍＬとし、０．１（ｗ／ｖ）％トルイジンブルーを３滴加えて混合し、１／４００Ｎポリビニル硫酸カリウム水溶液（ＰＶＳＫ）にて滴定した（液色が青色から赤紫色に変化し、赤紫色を３秒以上保持した時を終点とした）。
ナノ化キチン中のグルコサミン単位およびアセチルグルコサミン単位のモル質量をそれぞれ１６１および２０３とし、滴定値から脱アセチル化度を算出した。
(2) Measurement of degree of deacetylation Weigh 1380 g of N, N-dimethylacetamide into a 2 L glass beaker containing a stirrer in advance, stir with a stirrer, and add 120 g of lithium chloride to dissolve it. To prepare a chitin solution.
The dispersion of nano-sized chitin of Example 1 was heated and dried at normal pressure, the obtained solid content of 2.5 g was weighed, and the above-mentioned chitin solution was added to bring the total amount to 500 g. Stir overnight to dissolve, weigh 1.0 g of the resulting nanonized chitin solution, add deionized water to make 50 mL, add 3 drops of 0.1 (w / v)% toluidine blue and mix. It was titrated with a 1/400 N polyvinyl sulfate aqueous solution (PVSK) (the end point was when the liquid color changed from blue to magenta and the magenta was retained for 3 seconds or longer).
The molar masses of the glucosamine unit and the acetylglucosamine unit in the nano-sized chitin were set to 161 and 203, respectively, and the degree of deacetylation was calculated from the titration value.
上記（１）、（２）の測定結果を、表１に示した。 The measurement results of (1) and (2) above are shown in Table 1.
表１に示されるように、実施例１のナノ化キチンの分散物は、ナノ化キチンを２０重量％という高濃度で含有するものであった。
また、表１に示されるように、実施例１のナノ化キチンの分散物中のナノ化キチンの脱アセチル化度は５％であり、出発原料として用いたキチン粉末に比べて、脱アセチル化があまり進行していないことが認められた。
As shown in Table 1, the dispersion of nano-sized chitin of Example 1 contained nano-sized chitin at a high concentration of 20% by weight.
Further, as shown in Table 1, the degree of deacetylation of the nano-sized chitin in the dispersion of the nano-sized chitin of Example 1 was 5%, which was deacetylated as compared with the chitin powder used as a starting material. Was found to be less advanced.
［実施例２］ナノ化キチンの脱アセチル化物の分散物
実施例１のナノ化キチンの分散物を希釈し、その９．０ｇ（ナノ化キチン含有量＝７重量％）を水に分散させて全量３０ｍＬとし、３０ｍＬの４８重量％水酸化ナトリウムを加えて攪拌し、６０℃に加温して、５時間脱アセチル化反応を行わせた。反応時の水酸化ナトリウム濃度は、約３６（ｗ／ｖ）％であった。
反応後、ナノ化キチンの脱アセチル化物の分散物に対し大量の水で、洗浄液（ろ液）のｐＨが８．５〜９．０となるまで、繰り返し洗浄した。
得られた分散物中のナノ化キチンの脱アセチル化物について、上記試験例１における脱アセチル化度の測定と同様の方法で脱アセチル化度を測定したところ、１５．７％〜１６．２％であった。
得られた脱アセチル化物の分散物に酢酸を数滴添加して酸性とすると、低濃度（１．０重量％〜２．０重量％）のナノ化キチン濃度においてゲル化することが認められた。得られた脱アセチル化物のゲルの外観を図２に示した。
図２に示されるように、実施例２のナノ化キチンの脱アセチル化物の分散物を酸性とした場合、該分散物は、均一で粘稠なゲル状を示した。
[Example 2] Dispersion of deacetylated nano-chitin The dispersion of nano-chitin of Example 1 was diluted, and 9.0 g (nano-chitin content = 7% by weight) thereof was dispersed in water. The total volume was 30 mL, 30 mL of 48 wt% sodium hydroxide was added, the mixture was stirred, and the mixture was heated to 60 ° C. to carry out a deacetylation reaction for 5 hours. The sodium hydroxide concentration during the reaction was about 36 (w / v)%.
After the reaction, the dispersion of the deacetylase of nano-chitin was repeatedly washed with a large amount of water until the pH of the washing solution (filter solution) became 8.5-9.0.
The degree of deacetylase of nano-chitin in the obtained dispersion was measured by the same method as the measurement of the degree of deacetylation in Test Example 1 above, and found to be 15.7% to 16.2%. Met.
When several drops of acetic acid were added to the obtained dispersion of deacetylase to make it acidic, gelation was observed at a low concentration of nano-chitin (1.0% by weight to 2.0% by weight). .. The appearance of the obtained deacetylase gel is shown in FIG.
As shown in FIG. 2, when the dispersion of the deacetylase of nano-sized chitin of Example 2 was made acidic, the dispersion showed a uniform and viscous gel state.
［実施例３］ナノ化キチンの脱アセチル化物の分散物
脱アセチル化反応を８０℃で５時間行わせた他は、実施例２と同様にして、ナノ化キチンの脱アセチル化物の分散物を調製した。
得られた分散物中のナノ化キチンの脱アセチル化物について、上記試験例１における脱アセチル化度の測定と同様の方法で脱アセチル化度を測定したところ、１８．８％であった。上記の脱アセチル化条件にて、さほど脱アセチル化が進まないのは、ナノ化キチンの結晶構造内部は膨潤することなく、繊維表面あるいは非晶化している部分のアセタミドが脱アセチル化されるためであると考えられる。
実施例３のナノ化キチンの脱アセチル化物の分散物に酢酸を数滴添加して酸性とすると、１．６重量％のナノ化キチン濃度で透明性の高いゲルを形成した（図３）。
[Example 3] Dispersion of deacetylase of nano-sized chitin A dispersion of deacetylase of nano-sized chitin was prepared in the same manner as in Example 2 except that the deacetylation reaction was carried out at 80 ° C. for 5 hours. Prepared.
Regarding the deacetylase of nano-sized chitin in the obtained dispersion, the deacetylation degree was measured by the same method as the measurement of the deacetylation degree in Test Example 1 above, and it was 18.8%. Under the above deacetylation conditions, deacetylation does not proceed so much because the inside of the crystal structure of nano-chitin does not swell and the acetamide on the fiber surface or in the amorphous part is deacetylated. Is considered to be.
When a few drops of acetic acid were added to the dispersion of the deacetylase of nano-sized chitin of Example 3 to make it acidic, a highly transparent gel was formed at a concentration of 1.6% by weight of nano-sized chitin (Fig. 3).
［実施例４］ナノ化キチンの脱アセチル化物の分散物
脱アセチル化反応を、５２．７（ｗ／ｖ）％の水酸化ナトリウム水溶液中にて６０℃で５時間行わせた他は、実施例２と同様にして、ナノ化キチンの脱アセチル化物の分散物を調製した。
得られた分散物中のナノ化キチンの脱アセチル化物について、上記試験例１における脱アセチル化度の測定と同様の方法で脱アセチル化度を測定したところ、４０％であった。
実施例４のナノ化キチンの脱アセチル化物の分散物に酢酸を数滴添加して酸性とすると、１．２重量％のナノ化キチン濃度でほぼ透明なゲルを形成した（図４）。
実施例４のナノ化キチンの脱アセチル化物により形成されるゲルは、図４に示されるように、チキソトロピー性を示し、ゲルが充填されたボトルを倒置させても、ゲルが落下しない程度の強い粘稠性を示した。
また、実施例４のナノ化キチンの脱アセチル化物により形成されるゲルについて、実施例１のナノ化キチンの分散物の場合と同様にＴＥＭによる観察を行った。その結果を図５に示した。
図５に示されるように、ＴＥＭ観察画像において、ナノ化キチンの均一な分散が認められたものの、軸方向に短い結晶が多く観察された。
[Example 4] Dispersion of deacetylase of nano-chitin The deacetylation reaction was carried out in a 52.7 (w / v)% aqueous sodium hydroxide solution at 60 ° C. for 5 hours. A dispersion of deacetylases of nanochitin was prepared in the same manner as in Example 2.
Regarding the deacetylase of nano-sized chitin in the obtained dispersion, the deacetylation degree was measured by the same method as the measurement of the deacetylation degree in Test Example 1 above, and it was found to be 40%.
When a few drops of acetic acid were added to the dispersion of the deacetylase of nano-sized chitin of Example 4 to make it acidic, an almost transparent gel was formed at a concentration of 1.2% by weight of nano-sized chitin (FIG. 4).
As shown in FIG. 4, the gel formed by the deacetylase of nano-sized chitin of Example 4 exhibits thixotropy and is strong enough that the gel does not fall even when the bottle filled with the gel is inverted. It showed viscousness.
In addition, the gel formed by the deacetylase of nano-sized chitin in Example 4 was observed by TEM in the same manner as in the case of the dispersion of nano-sized chitin in Example 1. The result is shown in FIG.
As shown in FIG. 5, in the TEM observation image, although uniform dispersion of nano-sized chitin was observed, many crystals short in the axial direction were observed.
［実施例５］ナノ化キチンの脱アセチル化物の分散物
脱アセチル化反応を、３０（ｗ／ｖ）％の水酸化ナトリウム水溶液中にて４５℃で７２時間行わせた他は、実施例２と同様にして、ナノ化キチンの脱アセチル化物の分散物を調製した。
得られた分散物中のナノ化キチンの脱アセチル化物について、上記試験例１における脱アセチル化度の測定と同様の方法で脱アセチル化度を測定したところ、２６．４％であった。
実施例５のナノ化キチンの脱アセチル化物の分散物に酢酸を数滴添加して酸性とすると、１．８重量％のナノ化キチン濃度でゲルを形成した。
[Example 5] Dispersion of deacetylase of nano-sized chitin Example 2 except that the deacetylase reaction was carried out in a 30 (w / v)% aqueous sodium hydroxide solution at 45 ° C. for 72 hours. A dispersion of deacetylases of nano-chitin was prepared in the same manner as above.
Regarding the deacetylase of nano-sized chitin in the obtained dispersion, the deacetylation degree was measured by the same method as the measurement of the deacetylation degree in Test Example 1 above, and it was 26.4%.
When several drops of acetic acid were added to the dispersion of the deacetylase of nano-sized chitin of Example 5 to make it acidic, a gel was formed at a nano-sized chitin concentration of 1.8% by weight.
［試験例２］ナノ化キチンおよびナノ化キチンの脱アセチル化物の分子量の測定
実施例１のナノ化キチンの分散物および実施例５のナノ化キチンの脱アセチル化物の分散物に含有されるナノ化キチンおよびナノ化キチンの脱アセチル化物の分子量を、以下の通り測定した。
実施例１のナノ化キチンの分散物および実施例５のナノ化キチンの脱アセチル化物の分散物を、４８重量％水酸化ナトリウム水溶液にそれぞれ浸漬し、８５℃で１７時間脱アセチル化反応を行い、中和後、脱アセチル化物（脱アセチル化度≧８５％）を回収し、０．５重量％酢酸水溶液に溶解して、下記の条件下にてゲル浸透クロマトグラフィー（ＧＰＣ）分析を行った。
なお、比較のため、出発原料として用いたキチン（「キチンＬ−ＰＣ」、甲陽ケミカル株式会社製）および精製キチン（「キチンＴＣ−Ｌ」、甲陽ケミカル株式会社製）についても、同様に処理して分子量を測定した。
＜分子量測定条件＞
（ｉ）カラム：ＴＳＫゲルＧ６０００ＰＷＸＬ−ＣＰおよびＴＳＫゲルＧ３０００ＰＷＸＬ−ＣＰ（東ソー株式会社）
（ｉｉ）溶離液：０．２５Ｍ酢酸−０．２５Ｍ酢酸ナトリウム緩衝液
（ｉｉｉ）流速：０．５ｍＬ／ｍｉｎ
（ｉｖ）試料注入量：２００μＬ
（ｖ）オーブン温度：４０℃
（ｖｉ）検出器：ＲＩ（示差屈折）検出器
（ｖｉｉ）標準試料：プルラン
（ｖｉｉｉ）分析時間：６０ｍｉｎ
測定結果を表２に示した。
[Test Example 2] Measurement of molecular weight of nano-sized chitin and deacetylated product of nano-sized chitin Nano contained in the dispersion of nano-sized chitin of Example 1 and the dispersion of deacetylated product of nano-sized chitin of Example 5. The molecular weights of deacetylated chitins and nano-chitins were measured as follows.
The dispersion of nano-sized chitin of Example 1 and the dispersion of deacetylase of nano-sized chitin of Example 5 were each immersed in a 48 wt% sodium hydroxide aqueous solution, and a deacetylation reaction was carried out at 85 ° C. for 17 hours. After neutralization, the deacetylase (deacetylation degree ≥ 85%) was recovered, dissolved in a 0.5 wt% acetic acid aqueous solution, and subjected to gel permeation chromatography (GPC) analysis under the following conditions. ..
For comparison, chitin (“chitin L-PC”, manufactured by Koyo Chemical Co., Ltd.) and purified chitin (“chitin TC-L”, manufactured by Koyo Chemical Co., Ltd.) used as starting materials were also treated in the same manner. The molecular weight was measured.
<Molecular weight measurement conditions>
(I) Column: TSK gel G6000PWXL-CP and TSK gel G3000PWXL-CP (Tosoh Corporation)
(Ii) Eluent: 0.25M acetic acid-0.25M sodium acetate buffer (iii) Flow rate: 0.5 mL / min
(Iv) Sample injection volume: 200 μL
(V) Oven temperature: 40 ° C.
(Vi) Detector: RI (Differential Refractometer) Detector
(Vii) Standard sample: Pullulan (vii) Analysis time: 60 min
The measurement results are shown in Table 2.
上述した通り、図１に示すＴＥＭ画像からは、実施例１の分散物中のナノ化キチンについて、十分な長さを有する繊維の存在を確認することができなかったが、表２に示す分子量の測定結果からは、該ナノ化キチンにおいて平均分子量の顕著な減少（低分子量化）は認められず、繊維の切断はさほど生じていないことが示唆された。
また、実施例５の分散物中に存在するナノ化キチンの脱アセチル化物においても、平均分子量の顕著な減少（低分子量化）は認められず、ナノ化キチンの形態を保ったまま脱アセチル化されたことが示唆された。
As described above, from the TEM image shown in FIG. 1, the presence of fibers having a sufficient length could not be confirmed for the nano-chitin in the dispersion of Example 1, but the molecular weights shown in Table 2 were not confirmed. From the measurement results of, no significant decrease in average molecular weight (reduced molecular weight) was observed in the nano-sized chitin, suggesting that fiber cutting did not occur so much.
Further, even in the deacetylase of nano-sized chitin present in the dispersion of Example 5, no significant decrease in average molecular weight (reduction in molecular weight) was observed, and deacetylation was performed while maintaining the form of nano-sized chitin. It was suggested that it was done.
上述したように、実施例１の分散物は、１０ｎｍ以下の均一な繊維幅を有し、ほぼエレメンタリーミクロフィブリル単位にまで解繊されたナノ化キチンが、２０重量％とこれまでにない高濃度で分散された分散物であった。
かかる分散物は、均一なナノ化キチンを高濃度で含有するため、実施例２〜５に示す通り、これを原料として、あるいはこれを出発物質として、ナノ化キチンの形態を保ったまま脱アセチル化することができ、脱アセチル化されたナノ化キチンは、その分散物の液性を酸性とし、または酸性水溶液に分散させると、低濃度においてゲルを形成することができる。ナノ化キチンの脱アセチル化度の程度により、透明性の高いゲルや、粘度が高くチキソトロピー性を示すゲルを得ることができる。
また、ナノ化キチンの脱アセチル化物からさらに誘導体化を検討することもできる。
本発明の分散物は、Ｎ−アセチルグルコサミンやオリゴ糖の原料としても有用である。
さらに、本発明の分散物は、医用材料、樹脂中のフィラー、紙質向上剤等として有用であり、また、機能性食品（特にプレバイオティクスとして）、化粧品、飼料等の成分として利用することもできる。
As described above, the dispersion of Example 1 has a uniform fiber width of 10 nm or less, and nano-sized chitin defibrated to almost elementary microfibril units is 20% by weight, which is unprecedentedly high. It was a dispersion dispersed by concentration.
Since such a dispersion contains a uniform nano-chitin at a high concentration, as shown in Examples 2 to 5, deacetylation using this as a raw material or using this as a starting material while maintaining the form of nano-chitin. The deacetylated nano-chitin can form a gel at a low concentration when the liquid property of the dispersion is made acidic or dispersed in an acidic aqueous solution. Depending on the degree of deacetylation of nano-sized chitin, a gel having high transparency or a gel having high viscosity and exhibiting thixotropy can be obtained.
Further derivatization from the deacetylase of nano-chitin can also be investigated.
The dispersion of the present invention is also useful as a raw material for N-acetylglucosamine and oligosaccharides.
Further, the dispersion of the present invention is useful as a medical material, a filler in a resin, a paper quality improver, etc., and can also be used as a component of functional foods (particularly as prebiotics), cosmetics, feeds, etc. it can.
以上、詳述したように、本発明により、均一な繊維幅を有するナノ化キチンを高濃度で含有するナノ化キチンの分散物を提供することができる。
本発明の分散物に含有されるナノ化キチンは、ナノ化キチンの形態を保ったまま脱アセチル化することができる。ナノ化キチンの脱アセチル化物の分散物は、液性を酸性とすることにより、または酸性水溶液に分散させることにより、低濃度でゲルを形成することができる。
本発明の分散物は、Ｎ−アセチルグルコサミンやオリゴ糖の原料、医用材料、プラスチック材料等として、また、機能性食品、化粧品、飼料等の成分として好適に利用され得る。
As described in detail above, according to the present invention, it is possible to provide a dispersion of nano-chitin containing a high concentration of nano-chitin having a uniform fiber width.
The nano-sized chitin contained in the dispersion of the present invention can be deacetylated while maintaining the form of the nano-sized chitin. The dispersion of the deacetylase of nanonized chitin can form a gel at a low concentration by making the liquid acidic or by dispersing it in an acidic aqueous solution.
The dispersion of the present invention can be suitably used as a raw material for N-acetylglucosamine and oligosaccharides, a medical material, a plastic material and the like, and as a component of functional foods, cosmetics, feeds and the like.
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| CN113024688A (en) * | 2021-05-24 | 2021-06-25 | 贝尔纳丝新材料(江苏)有限公司 | Preparation method of water phase and organic phase solvent stable dispersion nano chitin |
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| CN113024688B (en) * | 2021-05-24 | 2021-09-17 | 贝尔纳丝新材料(江苏)有限公司 | Preparation method of water phase and organic phase solvent stable dispersion nano chitin |
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