JP2007008854A - Calcium chondroitin sulfate and method for producing the same - Google Patents
Calcium chondroitin sulfate and method for producing the same Download PDFInfo
- Publication number
- JP2007008854A JP2007008854A JP2005190959A JP2005190959A JP2007008854A JP 2007008854 A JP2007008854 A JP 2007008854A JP 2005190959 A JP2005190959 A JP 2005190959A JP 2005190959 A JP2005190959 A JP 2005190959A JP 2007008854 A JP2007008854 A JP 2007008854A
- Authority
- JP
- Japan
- Prior art keywords
- chondroitin sulfate
- calcium
- protein complex
- cartilage
- counter ion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 title claims abstract description 51
- 229920001287 Chondroitin sulfate Polymers 0.000 title claims abstract description 51
- 229940059329 chondroitin sulfate Drugs 0.000 title claims abstract description 51
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 title claims abstract description 39
- 239000011575 calcium Substances 0.000 title claims abstract description 39
- 229910052791 calcium Inorganic materials 0.000 title claims abstract description 39
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 12
- 210000000845 cartilage Anatomy 0.000 claims abstract description 55
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 38
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 38
- 238000000034 method Methods 0.000 claims abstract description 27
- 241001465754 Metazoa Species 0.000 claims abstract description 24
- 150000002500 ions Chemical class 0.000 claims abstract description 24
- 102000004190 Enzymes Human genes 0.000 claims abstract description 19
- 108090000790 Enzymes Proteins 0.000 claims abstract description 16
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims abstract description 13
- 239000011734 sodium Substances 0.000 claims abstract description 13
- 229910052708 sodium Inorganic materials 0.000 claims abstract description 13
- 239000002253 acid Substances 0.000 claims abstract description 11
- 238000000502 dialysis Methods 0.000 claims abstract description 11
- 239000012528 membrane Substances 0.000 claims abstract description 9
- 239000003929 acidic solution Substances 0.000 claims abstract description 8
- 229940043430 calcium compound Drugs 0.000 claims abstract description 7
- 150000001674 calcium compounds Chemical class 0.000 claims abstract description 7
- 230000001079 digestive effect Effects 0.000 claims abstract description 6
- 239000003014 ion exchange membrane Substances 0.000 claims abstract description 5
- 239000003456 ion exchange resin Substances 0.000 claims abstract description 5
- 229920003303 ion-exchange polymer Polymers 0.000 claims abstract description 5
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 5
- 238000000926 separation method Methods 0.000 claims abstract description 4
- 241000972773 Aulopiformes Species 0.000 claims description 8
- 235000019515 salmon Nutrition 0.000 claims description 8
- 241000283153 Cetacea Species 0.000 claims description 4
- 241000287828 Gallus gallus Species 0.000 claims description 4
- 235000013305 food Nutrition 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 3
- 239000002537 cosmetic Substances 0.000 claims description 2
- 229940079593 drug Drugs 0.000 claims description 2
- 239000002994 raw material Substances 0.000 abstract description 9
- 238000001035 drying Methods 0.000 abstract description 8
- KXKPYJOVDUMHGS-OSRGNVMNSA-N chondroitin sulfate Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](OS(O)(=O)=O)[C@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](C(O)=O)O1 KXKPYJOVDUMHGS-OSRGNVMNSA-N 0.000 abstract description 5
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 abstract description 4
- 238000000227 grinding Methods 0.000 abstract description 4
- 239000000243 solution Substances 0.000 abstract description 4
- 239000012530 fluid Substances 0.000 abstract description 2
- 229960005069 calcium Drugs 0.000 description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 229920002683 Glycosaminoglycan Polymers 0.000 description 9
- 238000007796 conventional method Methods 0.000 description 7
- 235000013372 meat Nutrition 0.000 description 7
- 238000005406 washing Methods 0.000 description 6
- 210000000988 bone and bone Anatomy 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 108091005804 Peptidases Proteins 0.000 description 4
- 238000010306 acid treatment Methods 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 238000010298 pulverizing process Methods 0.000 description 4
- 241000283690 Bos taurus Species 0.000 description 3
- 102000007474 Multiprotein Complexes Human genes 0.000 description 3
- 108010085220 Multiprotein Complexes Proteins 0.000 description 3
- 239000004365 Protease Substances 0.000 description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 235000013330 chicken meat Nutrition 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
- 108010043393 protease N Proteins 0.000 description 3
- 229910001415 sodium ion Inorganic materials 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 2
- 239000004129 EU approved improving agent Substances 0.000 description 2
- 239000005909 Kieselgur Substances 0.000 description 2
- 239000004909 Moisturizer Substances 0.000 description 2
- 206010036790 Productive cough Diseases 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 241000282898 Sus scrofa Species 0.000 description 2
- 229940121363 anti-inflammatory agent Drugs 0.000 description 2
- 239000002260 anti-inflammatory agent Substances 0.000 description 2
- 208000003464 asthenopia Diseases 0.000 description 2
- VSGNNIFQASZAOI-UHFFFAOYSA-L calcium acetate Chemical compound [Ca+2].CC([O-])=O.CC([O-])=O VSGNNIFQASZAOI-UHFFFAOYSA-L 0.000 description 2
- 239000001639 calcium acetate Substances 0.000 description 2
- 235000011092 calcium acetate Nutrition 0.000 description 2
- 229960005147 calcium acetate Drugs 0.000 description 2
- 229910001424 calcium ion Inorganic materials 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 235000015872 dietary supplement Nutrition 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000002778 food additive Substances 0.000 description 2
- 235000013373 food additive Nutrition 0.000 description 2
- 239000000314 lubricant Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000001333 moisturizer Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000011085 pressure filtration Methods 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- 210000003802 sputum Anatomy 0.000 description 2
- 208000024794 sputum Diseases 0.000 description 2
- 108090000145 Bacillolysin Proteins 0.000 description 1
- 229920002567 Chondroitin Polymers 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 102000035092 Neutral proteases Human genes 0.000 description 1
- 108091005507 Neutral proteases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- -1 and among them Chemical compound 0.000 description 1
- 238000000889 atomisation Methods 0.000 description 1
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical compound [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 description 1
- 239000001527 calcium lactate Substances 0.000 description 1
- 229960002401 calcium lactate Drugs 0.000 description 1
- 235000011086 calcium lactate Nutrition 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- DLGJWSVWTWEWBJ-HGGSSLSASA-N chondroitin Chemical compound CC(O)=N[C@@H]1[C@H](O)O[C@H](CO)[C@H](O)[C@@H]1OC1[C@H](O)[C@H](O)C=C(C(O)=O)O1 DLGJWSVWTWEWBJ-HGGSSLSASA-N 0.000 description 1
- 229940006423 chondroitin sulfate sodium Drugs 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 235000020774 essential nutrients Nutrition 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 235000013611 frozen food Nutrition 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
Abstract
Description
本発明は、動物の軟骨を原料として得られるコンドロイチン硫酸カルシウムおよびその製造方法に関するものである。 The present invention relates to calcium chondroitin sulfate obtained from animal cartilage as a raw material and a method for producing the same.
コンドロイチン硫酸およびムコ多糖タンパク質複合体は、主に鮫、鯨、鮭、エイ、牛、豚、鶏などの動物軟骨を酵素またはアルカリ処理によって消化し、その消化液を乾燥することによって製造されている。
しかしながら、これら従来の方法で得られるムコ多糖タンパク質複合体は、コンドロイチン硫酸の対イオンがナトリウムであるコンドロイチン硫酸ナトリウムが主成分である。そのためこれらのムコ多糖タンパク質複合体を利用した食品などを摂取した場合、本来意図しない過剰のナトリウムを摂取することとなっていた。
ナトリウムの過剰摂取は様々な疾病の原因となることが指摘されており、健康維持のためにも必要量以上のナトリウムの摂取は好ましくないので、食品として利用されるコンドロイチン硫酸はナトリウム塩でなく、疾病の原因とならないカルシウム塩が求められていた。
Chondroitin sulfate and mucopolysaccharide protein complexes are mainly produced by digesting animal cartilage such as salmon, whales, salmon, rays, cows, pigs, chickens, etc. by enzymatic or alkaline treatment, and drying the digestive fluid .
However, the mucopolysaccharide protein complex obtained by these conventional methods is mainly composed of sodium chondroitin sulfate in which the counter ion of chondroitin sulfate is sodium. For this reason, when foods using these mucopolysaccharide protein complexes are ingested, excessive sodium that is not intended is to be ingested.
It has been pointed out that excessive intake of sodium causes various diseases, and it is not preferable to take more sodium than necessary to maintain health, so chondroitin sulfate used as food is not a sodium salt, There has been a need for calcium salts that do not cause disease.
しかしながら、コンドロイチン硫酸ナトリウムをコンドロイチン硫酸カルシウムに変える簡便な方法はこれまで開発されておらず、したがってこれまで産業上利用されていない。 However, a simple method for changing chondroitin sulfate sodium to chondroitin calcium sulfate has not been developed so far, and thus has not been used industrially.
一般にカルシウムの摂取は骨を丈夫にして健康維持に役立つと考えられており大変重要であるが、厚生労働省による統計調査などで日本人の必要栄養素としては常にその不足が指摘されている。 In general, calcium intake is very important because it is thought to be strong in bones and help maintain health, but statistical surveys by the Ministry of Health, Labor and Welfare have always pointed out the lack of essential nutrients for Japanese people.
本発明の課題は、動物の軟骨を原料として得られるコンドロイチン硫酸ナトリウムをコンドロイチン硫酸カルシウムの形で得る簡便な方法を提供することにある。 An object of the present invention is to provide a simple method for obtaining sodium chondroitin sulfate obtained from animal cartilage as a raw material in the form of calcium chondroitin sulfate.
本発明者は上記課題に鑑みて鋭意検討した結果、動物軟骨のカルシウムを利用する方法、すなわち動物軟骨を粉砕し、酸で処理することによって軟骨中に一部存在する硬骨を溶解させ、生成したカルシウムイオンと軟骨中のコンドロイチン硫酸に結合していたナトリウムイオンを、交換させ遊離したナトリウムイオンと余分な酸を透析または洗浄によって除去し、得られた軟骨を酵素などで消化し、これをフィルタープレスなどでろ過し清澄化処理後、乾燥固化または乾燥粉末化させることによって目的とするコンドロイチン硫酸タンパク質複合体を得る方法を開発した。また、カルシウム源としてカルシウム化合物を利用する方法をも開発した。
本発明の方法によれば、特別な装置を必要とせず、さらにカルシウム源としての食品添加物なども必要とせずに、非常に簡便な方法で目的とする主成分がコンドロイチン硫酸カルシウムであるムコ多糖タンパク質複合体を製造することができる。
As a result of intensive studies in view of the above problems, the present inventor has produced a method using calcium of animal cartilage, that is, pulverizing animal cartilage and treating it with acid to dissolve bone bones partially present in cartilage. Calcium ions and sodium ions bound to chondroitin sulfate in cartilage are exchanged to remove free sodium ions and excess acid by dialysis or washing, and the resulting cartilage is digested with enzymes, etc., and this is filtered. The method of obtaining the target chondroitin sulfate protein complex was developed by filtering and clarifying after drying, etc., and making it dry-solidify or dry-powder. We have also developed a method using calcium compounds as a calcium source.
According to the method of the present invention, a mucopolysaccharide whose target main component is calcium chondroitin sulfate is a very simple method without requiring a special device and without requiring a food additive as a calcium source. Protein complexes can be produced.
すなわち、本発明は下記の１〜６の、対イオンがカルシウムであるコンドロイチン硫酸タンパク質複合体の製造方法およびその方法で得られるコンドロイチン硫酸タンパク質複合体に関する。
１．動物軟骨を粉砕し、酸処理したのち、酵素で処理し、得られた消化液を乾燥し粉末化することを特徴とする対イオンがカルシウムであるコンドロイチン硫酸タンパク質複合体の製造方法。
２．動物軟骨から得られる対イオンがナトリウムであるコンドロイチン硫酸タンパク質複合体を酸性溶液下でカルシウム化合物と反応させイオン交換樹脂、イオン交換膜、透析膜または限外ろ過膜等の物理的または化学的分離方法によって、遊離したナトリウムと余剰のカルシウムを除去し、得られた消化液を乾燥し粉末化することを特徴とする対イオンがカルシウムである精製されたコンドロイチン硫酸タンパク質複合体の製造方法。
３．動物軟骨が、鮫、鯨、鮭、エイ、牛、豚及び鶏軟骨から選択されるものである前記１または２に記載のコンドロイチン硫酸タンパク質複合体の製造方法。
４．前記１に記載の方法で得られる対イオンがカルシウムであるコンドロイチン硫酸タンパク質複合体。
５．前記２に記載の方法で得られる対イオンがカルシウムである精製されたコンドロイチン硫酸タンパク質複合体。
６．前記４または５に記載のコンドロイチン硫酸タンパク質複合体を含んでなる食品、医薬品、医薬部外品または化粧品。
That is, this invention relates to the manufacturing method of the chondroitin sulfate protein complex whose counter ion is calcium of the following 1-6, and the chondroitin sulfate protein complex obtained by the method.
1. A method for producing a chondroitin sulfate protein complex in which the counter ion is calcium, characterized in that animal cartilage is pulverized, acid-treated, then treated with an enzyme, and the resulting digestive juice is dried and powdered.
2. Physical or chemical separation method such as ion exchange resin, ion exchange membrane, dialysis membrane or ultrafiltration membrane by reacting chondroitin sulfate protein complex with sodium counterion obtained from animal cartilage with calcium compound in acidic solution A method for producing a purified chondroitin sulfate protein complex in which the counter ion is calcium, which comprises removing free sodium and excess calcium and drying and pulverizing the resulting digested liquid.
3. 3. The method for producing a chondroitin sulfate protein complex according to 1 or 2 above, wherein the animal cartilage is selected from salmon, whale, salmon, ray, cow, pig and chicken cartilage.
4). 2. A chondroitin sulfate protein complex in which the counter ion obtained by the method described in 1 is calcium.
5. 3. A purified chondroitin sulfate protein complex in which the counter ion obtained by the method described in 2 is calcium.
6). A food, pharmaceutical, quasi drug or cosmetic comprising the chondroitin sulfate protein complex according to 4 or 5 above.
本発明は原料の動物軟骨中に存在するコンドロイチン硫酸ナトリウムを固体のままでそのナトリウムを動物軟骨由来のカルシウムに交換する方法、または従来の方法で得られたコンドロイチン硫酸ナトリウムを水溶液中においてカルシウム化合物由来のカルシウムに交換する方法を提供したものであり、本発明によれば動物軟骨からきわめて簡便な方法で、主成分がコンドロイチン硫酸カルシウムであるムコ多糖タンパク質複合体を得ることができる。 The present invention is a method for exchanging sodium chondroitin sulfate present in animal cartilage as a raw material with calcium derived from animal cartilage while remaining solid, or sodium chondroitin sulfate obtained by a conventional method derived from a calcium compound in an aqueous solution. According to the present invention, a mucopolysaccharide protein complex whose main component is calcium chondroitin sulfate can be obtained from animal cartilage by a very simple method.
本発明の第一方法において原料として使用される動物軟骨は、特に限定されないが鮫、鯨、鮭、エイ、牛、豚、鶏などの動物軟骨が挙げられる。これらの中でも好ましいのは、鮫、鮭、エイの軟骨である。 The animal cartilage used as a raw material in the first method of the present invention is not particularly limited, and examples thereof include animal cartilage such as salmon, whale, salmon, ray, cow, pig and chicken. Of these, sputum, sputum and ray cartilage are preferred.
原料となる動物軟骨を分離回収する方法は、特に限定されないが、軟骨が含まれる肉部分からあらかじめ内臓や硬骨を取り除き、加熱して肉を熱変成させた後、肉を排除して軟骨を分離回収する方法等が挙げられる。例えば特開2002-315499号公報に記載のように、加熱処理後の軟骨が含まれる肉部分を、貫通細孔を有する容器壁と容器壁に向かって水を噴射するノズルを備えた回転軸を有する回転ドラムを用いて、回転ドラムの回転力、肉同士あるいは肉と回転容器との摩擦力、及びノズルから噴射される水圧力により、肉を細片にして貫通細孔を通して容器外に排除し軟骨を容器内に残留させて回収することができる。 The method for separating and recovering animal cartilage as a raw material is not particularly limited, but after removing the internal organs and bones from the meat part containing cartilage and heating to thermally transform the meat, the meat is removed and the cartilage is separated. The method of collection | recovery etc. are mentioned. For example, as described in Japanese Patent Application Laid-Open No. 2002-315499, a meat portion containing cartilage after heat treatment is provided with a rotating shaft having a container wall having penetrating pores and a nozzle for injecting water toward the container wall. Using the rotating drum, the rotating force of the rotating drum, the frictional force between the meats or between the meat and the rotating container, and the water pressure sprayed from the nozzle remove the meat out of the container through the through-holes. The cartilage can remain in the container and be collected.
動物軟骨は、有効に利用するためには細かく粉砕して使用する。
細かく粉砕した動物軟骨は酸性溶液に浸漬して放置し、その後洗浄または透析によって酸を除去する。使用する酸としては、酢酸（0.2〜５質量％、好ましくは約１質量％水溶液、希硫酸（0.05〜３質量％、好ましくは約0.2質量％），希塩酸（0.1〜５質量％、好ましくは約0.5質量％）、クエン酸（0.2〜５質量％、好ましくは約１質量％水溶液）が挙げられるが、中でも酢酸が好ましい。この酸処理は、例えば、軟骨の等倍量〜２倍量の酸性溶液中で５〜１５℃の温度で撹拌下で１８〜２４時間浸漬して行うことが好ましい。
Animal cartilage is used after being finely crushed for effective use.
The finely pulverized animal cartilage is left immersed in an acidic solution, and then the acid is removed by washing or dialysis. Examples of the acid to be used include acetic acid (0.2 to 5% by mass, preferably about 1% by mass aqueous solution, dilute sulfuric acid (0.05 to 3% by mass, preferably about 0.2% by mass), dilute hydrochloric acid (0.1 to 5% by mass, preferably about 0.5 mass%) and citric acid (0.2 to 5 mass%, preferably about 1 mass% aqueous solution), among which acetic acid is preferred, and this acid treatment is, for example, an acidic amount of 1 to 2 times the amount of cartilage. It is preferable to immerse in a solution at a temperature of 5 to 15 ° C. with stirring for 18 to 24 hours.
酸の除去は洗浄または透析によって、軟骨のｐＨが６〜７になるまで行う。軟骨のｐＨは、軟骨をすり潰したものにｐＨ試験紙当てること等によって測定する。洗浄及び透析の方法は特に制限されないが、例えば洗浄は、軟骨容量の２倍程度の容器中、５〜１５℃の新鮮な水道水を常に下部から加えて上部からオーバーフローさせながら行う。透析は、例えば透析用セルロースチューブ（分画分子量約12,000〜14,000）に酸処理した軟骨を入れ、両端を結束して５〜１０倍量の５〜１５℃の水中に浸漬して適宜水を換えながら行う。多量の軟骨を処理する工業生産スケールの場合には作業が簡便である洗浄処理が好ましい。洗浄及び透析処理後の軟骨は、遠心分離機等を用いて脱水する。 Acid removal is performed by washing or dialysis until the cartilage pH is 6-7. The pH of the cartilage is measured by applying a pH test paper to the ground cartilage. The method of washing and dialysis is not particularly limited. For example, washing is performed in a container having about twice the capacity of cartilage while always adding fresh tap water of 5 to 15 ° C. from the bottom and overflowing from the top. For dialysis, for example, put acid-treated cartilage into a cellulose tube for dialysis (fraction molecular weight of about 12,000-14,000), bind both ends and immerse in 5-10 times the amount of water at 5-15 ° C to change the water appropriately. While doing. In the case of an industrial production scale in which a large amount of cartilage is processed, a cleaning process is preferable because the operation is simple. The cartilage after washing and dialysis is dehydrated using a centrifuge or the like.
次に、酸処理を施した軟骨を酵素等を用いて消化させる。
本発明に使用する酵素は、食品添加物用酵素製剤として市販されているタンパク質分解酵素であればいずれも使用可能であるが、分解速度や処理条件の簡便性から中性プロテアーゼが好ましい。例えば、プロテアーゼ（天野エンザイム（株）製；プロテアーゼＮ「アマノ」Ｇ）が挙げられる。酵素による処理条件（酵素の使用量、時間、温度）は、使用する酵素の力価に適した条件を選択することができる。例えば、本発明において酵素としてプロテアーゼ（天野エンザイム（株）製；プロテアーゼＮ「アマノ」Ｇ）を使用した場合、酵素処理は、酸処理後の軟骨に0.1〜0.5質量％の酵素を添加し、５０〜６０℃で２〜４時間撹拌し、目視で軟骨が完全に消化するまで行う。
Next, the acid-treated cartilage is digested using an enzyme or the like.
Any enzyme can be used as the enzyme used in the present invention as long as it is a proteolytic enzyme commercially available as an enzyme preparation for food additives, but a neutral protease is preferable from the standpoint of degradation rate and simple processing conditions. For example, protease (Amano Enzyme Co., Ltd .; Protease N “Amano” G) can be mentioned. The conditions suitable for the titer of the enzyme to be used can be selected as the conditions for the treatment with the enzyme (amount of enzyme used, time, temperature). For example, when a protease (manufactured by Amano Enzyme Co., Ltd .; protease N “Amano” G) is used as an enzyme in the present invention, the enzyme treatment is performed by adding 0.1 to 0.5% by mass of enzyme to the cartilage after acid treatment, Stir at ~ 60 ° C for 2-4 hours, until the cartilage is completely digested visually.
酵素処理によって得られた軟骨消化液は、珪藻土等のろ過助剤を加えてフィルタープレス等によりろ過し、清澄化処理をしたろ液を得る。
得られたろ液を加熱蒸発、減圧乾燥、噴霧乾燥等の方法により、乾燥固化あるいは粉末化させることにより本発明目的物の対イオンがカルシウムであるコンドロイチン硫酸タンパク質複合体を得ることができる。
The cartilage digestive liquid obtained by the enzyme treatment is filtered with a filter press or the like after adding a filter aid such as diatomaceous earth to obtain a clarified filtrate.
A chondroitin sulfate protein complex in which the counter ion of the object of the present invention is calcium can be obtained by drying, solidifying or pulverizing the obtained filtrate by a method such as heat evaporation, drying under reduced pressure, or spray drying.
得られた対イオンがカルシウムであるコンドロイチン硫酸タンパク質複合体は、例えば、特開2000-273102号公報記載の方法により、分画分子量30,000〜50,000程度の膜で限外ろ過を行うことによって、雑多な低分子やペプチドを除去でき、精製することができる。 The obtained chondroitin sulfate protein complex whose counter ion is calcium is miscellaneous by performing ultrafiltration with a membrane having a molecular weight cut off of about 30,000 to 50,000, for example, by the method described in JP-A-2000-273102. Small molecules and peptides can be removed and purified.
本発明によって得られる、対イオンがカルシウムであるコンドロイチン硫酸タンパク質複合体のカルシウム源は、軟骨中に一部存在する硬骨由来のカルシウムであり、動物軟骨を粉砕し、酸処理を施すことによって硬骨が酸性溶液中に溶解したものである。粉砕した軟骨は、酸処理中のカルシウムイオンが存在する酸性溶液中において、イオン交換樹脂やイオン交換膜のような挙動を示し、軟骨中のコンドロイチン硫酸に結合しているナトリウムイオンを溶液中のカルシウムイオンと交換する。 The calcium source of the chondroitin sulfate protein complex obtained by the present invention in which the counter ion is calcium is calcium derived from the bone that is partly present in the cartilage, and the bone is obtained by crushing the animal cartilage and applying acid treatment. It is dissolved in an acidic solution. The pulverized cartilage behaves like an ion exchange resin or an ion exchange membrane in an acidic solution in which calcium ions are present during acid treatment, and sodium ions bound to chondroitin sulfate in cartilage are converted into calcium in the solution. Exchange with ions.
本発明の第二の方法によれば、従来の方法により得られるコンドロイチン硫酸タンパク質複合体の対イオンの交換を簡便に行うことができる。
従来の方法により得られる対イオンがナトリウムである動物軟骨由来のコンドロイチン硫酸タンパク質複合体を、酸性溶液中でカルシウム化合物と反応させてイオン交換樹脂、イオン交換膜、透析膜または限外ろ過膜等の物理的または化学的分離方法によって、遊離したナトリウムと余剰のカルシウムを除去した後、乾燥粉末化することによって対イオンがカルシウムである精製されたコンドロイチン硫酸タンパク質複合体が得られる。
According to the second method of the present invention, the exchange of the counter ion of the chondroitin sulfate protein complex obtained by the conventional method can be easily performed.
A chondroitin sulfate protein complex derived from animal cartilage whose counter ion obtained by a conventional method is sodium is reacted with a calcium compound in an acidic solution to produce an ion exchange resin, an ion exchange membrane, a dialysis membrane, or an ultrafiltration membrane. After removing free sodium and excess calcium by a physical or chemical separation method, a purified chondroitin sulfate protein complex in which the counter ion is calcium is obtained by dry pulverization.
本発明で従来の方法により得られるコンドロイチン硫酸タンパク質複合体を原料として使用する場合においても、酸性溶液としては、軟骨を原料とする方法と同様のものが使用され、乾燥粉末化についても軟骨を原料とする方法と同様に行う。
本発明において使用されるカルシウム化合物としては、酢酸カルシウム、乳酸カルシウム等が挙げられ、中でも酢酸カルシウムが好ましい。
Even when the chondroitin sulfate protein complex obtained by the conventional method in the present invention is used as a raw material, the acidic solution is the same as the method using cartilage as a raw material, and cartilage is used as a raw material for dry powdering. This is done in the same way as
Examples of the calcium compound used in the present invention include calcium acetate and calcium lactate, and among them, calcium acetate is preferable.
このようにして調製したコンドロイチン硫酸タンパク質複合体は、主成分であるコンドロイチン硫酸の対イオンが常法によって得られるナトリウムではなくカルシウムであることに特徴がある。 The chondroitin sulfate protein complex thus prepared is characterized in that the counter ion of chondroitin sulfate, which is the main component, is calcium instead of sodium obtained by a conventional method.
本発明により得られる対イオンがカルシウムであるコンドロイチン硫酸およびこれを含むムコ多糖タンパク複合体は、例えば高齢者や病弱者、体力が衰えた者などに対してムコ多糖、タンパク質およびカルシウムの栄養補助剤としての機能を有し、また従来のコンドロイチン硫酸と同様に、抗炎症剤、保湿剤、関節潤滑剤、眼精疲労緩和剤、皮膚の代謝改善剤などきわめて多様な用途に活用可能である。 The chondroitin sulfate whose counter ion obtained by the present invention is calcium and the mucopolysaccharide protein complex containing the same are, for example, nutritional supplements of mucopolysaccharide, protein and calcium for elderly people, sick people, those with weak physical strength, etc. In addition, like conventional chondroitin sulfate, it can be used for a wide variety of applications such as anti-inflammatory agents, moisturizers, joint lubricants, eye strain relief agents, and skin metabolism improving agents.
以下、実施例を挙げて説明するが、本発明は以下の記載に限定されるものではない。 Hereinafter, although an example is given and explained, the present invention is not limited to the following statements.
実施例１：エイ軟骨由来コンドロイチン硫酸の製造
反応槽に軟骨粉砕物２００ｋｇと水２００ｋｇを入れ、これに９０％酢酸８ｋｇを入れ、撹拌した。このときの水温は８℃であった。２４時間後、遠心分離によって液を除き、さらに水２００ｋｇを入れて２時間撹拌したのち遠心分離によって液を除いた。この操作を３回繰り返した。次に水を毎分１リットル程度の割合で注入しながら１晩放置した。翌朝、遠心分離によって水分を除き、さらに水２００ｋｇを入れて２時間撹拌したのち遠心分離によって水分を除いた。この操作を３回繰り返した。次に水を毎分１リットル程度の割合で注入しながら１晩放置した。この間、随時水分のｐＨを測定し、６〜７以下であれば次操作に移ってもよい。翌朝、遠心分離によって液を除き、斜軸ニーダー（２００Ｌ容量）に酸処理した軟骨とプロテアーゼ（天野エンザイム（株）製；プロテアーゼＮ「アマノ」Ｇ）２００ｇ入れて、５８℃で２時間撹拌した後、９２℃に加熱し１５分間撹拌して酵素失活処理を行った。これにろ過助剤として珪藻土＃１００（昭和化学工業（株）製）５ｋｇを加えて、均質化後、フィルタープレスで加圧ろ過を行った。なお、使用した加圧ろ過装置は、薮田機械（株）製，FR66-4型，ろ過面積：2.8ｍ2，ろ過容積：３６Ｌである。次に、得られた抽出液を９０℃に加熱し１０分間加熱殺菌後、ディスク型スプレードライヤー（（株）坂本技研製，DA220-10S）にて噴霧乾燥させコンドロイチン硫酸タンパク質複合体の白色粉末14.5ｋｇを得た。噴霧乾燥条件は、微粒化方式：φ９０ｍｍ回転ピン型ディスク、12,000ｒｐｍ、入口熱風温度：１８５℃、出口排風温度：９０℃、乾燥室内圧：0.1ｋｐａで行った。
Example 1 Production of Ray Cartilage-derived Chondroitin Sulfate 200 kg of cartilage pulverized product and 200 kg of water were placed in a reaction tank, and 8 kg of 90% acetic acid was added thereto and stirred. The water temperature at this time was 8 ° C. After 24 hours, the liquid was removed by centrifugation, 200 kg of water was further added and stirred for 2 hours, and then the liquid was removed by centrifugation. This operation was repeated three times. Next, the water was allowed to stand overnight while being poured at a rate of about 1 liter per minute. The next morning, the water was removed by centrifugation, 200 kg of water was further added and stirred for 2 hours, and then the water was removed by centrifugation. This operation was repeated three times. Next, the water was allowed to stand overnight while being poured at a rate of about 1 liter per minute. During this time, the pH of the moisture is measured at any time, and if it is 6-7 or less, the next operation may be started. The next morning, the liquid was removed by centrifugation, and acid-treated cartilage and protease (Amano Enzyme Co., Ltd .; Protease N “Amano” G) (200 g) were placed in a slanted kneader (200 L capacity) and stirred at 58 ° C. for 2 hours. The enzyme was inactivated by heating to 92 ° C. and stirring for 15 minutes. To this was added 5 kg of diatomaceous earth # 100 (manufactured by Showa Chemical Industry Co., Ltd.) as a filter aid, and after homogenization, pressure filtration was performed with a filter press. In addition, the used pressure filtration apparatus is Iwata Kikai Co., Ltd., FR66-4 type, filtration area: 2.8 m 2 , and filtration volume: 36 L. Next, the obtained extract was heated to 90 ° C., sterilized by heating for 10 minutes, and then spray-dried with a disk-type spray dryer (manufactured by Sakamoto Giken Co., Ltd., DA220-10S) to obtain a white powder 14.5 of chondroitin sulfate protein complex. kg was obtained. The spray drying conditions were as follows: atomization method: φ90 mm rotating pin type disk, 12,000 rpm, inlet hot air temperature: 185 ° C., outlet exhaust air temperature: 90 ° C., drying chamber pressure: 0.1 kpa.
製造したコンドロイチン硫酸タンパク質複合体のミネラル成分を（財）日本冷凍食品検査協会に依頼して実施して、表１の結果を得た。なお、表１中Ｌｏｔ．１は常法によって、Ｌｏｔ．２は本法によって製造したものである。
表１の結果より、本発明の方法によって得られたコンドロイチン硫酸タンパク質複合体は、コンドロイチン硫酸カルシウムを主成分とするものであることが確認された。 From the results in Table 1, it was confirmed that the chondroitin sulfate protein complex obtained by the method of the present invention was mainly composed of calcium chondroitin sulfate.
本発明のコンドロイチン硫酸およびこれを含むムコ多糖タンパク複合体は、例えば高齢者や病弱者、体力が衰えた者などに対してムコ多糖、タンパク質およびカルシウムの栄養補助剤としての機能を有し、また従来のコンドロイチン硫酸と同様に、抗炎症剤、保湿剤、関節潤滑剤、眼精疲労緩和剤、皮膚の代謝改善剤などきわめて多様な用途に活用可能なものである。
The chondroitin sulfate of the present invention and a mucopolysaccharide protein complex containing the chondroitin sulfate have a function as a nutritional supplement for mucopolysaccharides, proteins and calcium, for example, for the elderly, the sick, and those with weak physical strength. Like conventional chondroitin sulfate, it can be used for a wide variety of applications such as anti-inflammatory agents, moisturizers, joint lubricants, eye strain relief agents, and skin metabolism improving agents.
Claims (6)
A food, medicine, quasi-drug or cosmetic comprising the chondroitin sulfate protein complex according to claim 4 or 5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005190959A JP4955944B2 (en) | 2005-06-30 | 2005-06-30 | Chondroitin calcium sulfate and method for producing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005190959A JP4955944B2 (en) | 2005-06-30 | 2005-06-30 | Chondroitin calcium sulfate and method for producing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2007008854A true JP2007008854A (en) | 2007-01-18 |
| JP4955944B2 JP4955944B2 (en) | 2012-06-20 |
Family
ID=37747809
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2005190959A Active JP4955944B2 (en) | 2005-06-30 | 2005-06-30 | Chondroitin calcium sulfate and method for producing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4955944B2 (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011136159A1 (en) * | 2010-04-28 | 2011-11-03 | 日本薬品株式会社 | Composition for the treatment of aesthenopia |
| CN103172764A (en) * | 2013-04-01 | 2013-06-26 | 管桂义 | Method for producing chondroitin by taking duck tracheas as raw material |
| CN103243079A (en) * | 2013-05-16 | 2013-08-14 | 青岛农业大学 | Extracting method of duck chondroitin sulfate by compound enzymolysis |
| JP5355682B2 (en) * | 2009-03-19 | 2013-11-27 | 丸共バイオフーズ株式会社 | Antigen-specific T cell proliferation promoter |
| JP2016029150A (en) * | 2014-07-16 | 2016-03-03 | 地方独立行政法人青森県産業技術センター | High water-retentive proteoglycans, cosmetics and method for production of high water-retentive proteoglycans |
| CN105622779A (en) * | 2016-01-20 | 2016-06-01 | 定陶县地元生化制品有限公司 | Method for preparing clear chondroitin sulfate enzymatic hydrolysate |
| JP2017125164A (en) * | 2016-01-15 | 2017-07-20 | 地方独立行政法人青森県産業技術センター | High water holding modified proteoglycan |
| CN111467480A (en) * | 2020-04-26 | 2020-07-31 | 山东朱氏药业集团有限公司 | Glucosamine chondroitin sulfate calcium tablet and preparation method thereof |
| CN114230685A (en) * | 2021-12-21 | 2022-03-25 | 美泰科技（青岛）股份有限公司 | Preparation process of chondroitin sulfate chelated calcium with anti-osteoporosis function |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001064687A (en) * | 1999-08-27 | 2001-03-13 | Tomita Pharmaceutical Co Ltd | Solution for contact lens care |
| JP2001247453A (en) * | 2000-03-09 | 2001-09-11 | Marin Pharm:Kk | Film for soft capsule and soft capsule formulation |
| WO2003062279A1 (en) * | 2002-01-23 | 2003-07-31 | Institute Of Nutraceutical Research Pty Ltd | Nutraceuticals for the treatment, protection and restoration of connective tissues |
-
2005
- 2005-06-30 JP JP2005190959A patent/JP4955944B2/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001064687A (en) * | 1999-08-27 | 2001-03-13 | Tomita Pharmaceutical Co Ltd | Solution for contact lens care |
| JP2001247453A (en) * | 2000-03-09 | 2001-09-11 | Marin Pharm:Kk | Film for soft capsule and soft capsule formulation |
| WO2003062279A1 (en) * | 2002-01-23 | 2003-07-31 | Institute Of Nutraceutical Research Pty Ltd | Nutraceuticals for the treatment, protection and restoration of connective tissues |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5355682B2 (en) * | 2009-03-19 | 2013-11-27 | 丸共バイオフーズ株式会社 | Antigen-specific T cell proliferation promoter |
| JP5865242B2 (en) * | 2010-04-28 | 2016-02-17 | 日本薬品株式会社 | Composition for treating eye strain |
| US8663716B2 (en) | 2010-04-28 | 2014-03-04 | Nihon Pharmaceutical Co., Ltd. | Composition for treatment of aesthenopia |
| WO2011136159A1 (en) * | 2010-04-28 | 2011-11-03 | 日本薬品株式会社 | Composition for the treatment of aesthenopia |
| CN103172764A (en) * | 2013-04-01 | 2013-06-26 | 管桂义 | Method for producing chondroitin by taking duck tracheas as raw material |
| CN103243079A (en) * | 2013-05-16 | 2013-08-14 | 青岛农业大学 | Extracting method of duck chondroitin sulfate by compound enzymolysis |
| CN103243079B (en) * | 2013-05-16 | 2014-06-11 | 青岛农业大学 | Extracting method of duck chondroitin sulfate by compound enzymolysis |
| JP2016029150A (en) * | 2014-07-16 | 2016-03-03 | 地方独立行政法人青森県産業技術センター | High water-retentive proteoglycans, cosmetics and method for production of high water-retentive proteoglycans |
| JP2017125164A (en) * | 2016-01-15 | 2017-07-20 | 地方独立行政法人青森県産業技術センター | High water holding modified proteoglycan |
| CN105622779A (en) * | 2016-01-20 | 2016-06-01 | 定陶县地元生化制品有限公司 | Method for preparing clear chondroitin sulfate enzymatic hydrolysate |
| CN111467480A (en) * | 2020-04-26 | 2020-07-31 | 山东朱氏药业集团有限公司 | Glucosamine chondroitin sulfate calcium tablet and preparation method thereof |
| CN114230685A (en) * | 2021-12-21 | 2022-03-25 | 美泰科技（青岛）股份有限公司 | Preparation process of chondroitin sulfate chelated calcium with anti-osteoporosis function |
| CN114230685B (en) * | 2021-12-21 | 2022-12-20 | 美泰科技（青岛）股份有限公司 | Preparation process of chondroitin sulfate chelated calcium with anti-osteoporosis function |
Also Published As
| Publication number | Publication date |
|---|---|
| JP4955944B2 (en) | 2012-06-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4955944B2 (en) | Chondroitin calcium sulfate and method for producing the same | |
| CN101061827B (en) | Industry method of producing fish collagen peptide from fish skin and bone by an enzyme method | |
| JP5232831B2 (en) | Oil seed meal fractionation and processing | |
| JPH0310659A (en) | Iron-rich hemoferrum and production thereof | |
| CN101724677B (en) | Method for extracting collagen polypeptide and hydroxyapatite in fish scales by cooking hot extrusion | |
| WO2016173222A1 (en) | Hypoallergenic, odor-reduced fish protein oligopeptide, industrialized preparation method for same, and applications thereof | |
| CN102533914A (en) | High-purity fishy smell and foreign odor-free fish collagen protein peptide and preparation method thereof | |
| JPH06505982A (en) | Method for producing collagen, collagen produced by this method, and uses of collagen | |
| CN101892278A (en) | Method for extracting micromolecular collagen oligopeptide from fish | |
| JP2008206470A (en) | Method for producing collagen peptide of fish scale origin | |
| CN107365824A (en) | A kind of preparation method for hydrolyzing Isin glue collagen Gly-His-Lys | |
| CN102732592A (en) | Method for preparing freshwater fish bone gelatin by enzyme process | |
| EA006435B1 (en) | Method for producing a nutrition additive, an additive and its use | |
| CN108048514A (en) | The method of fresh bone extraction chondroitin sulfate, collagen peptide and albumen fatty powder | |
| RU2487152C2 (en) | Method of producing gelatine | |
| JPS5837001A (en) | Production of hyaluronic acid | |
| JPH03168064A (en) | Making of powdered flavoring foodstuff using blood of domestic animal | |
| JP2003299497A (en) | Mucopolysaccharide and method for producing the same | |
| CN103952456A (en) | Extraction method of embryonated egg phosvitin phosphopeptides | |
| JP2003245055A (en) | Skin-improving food composition and skin-improving method | |
| RU2287959C2 (en) | Method for producing of natural structurizers from fish wastes | |
| JP7423803B2 (en) | Process method for efficiently producing carnosine-rich compounds | |
| RU2390252C1 (en) | Method of producing protein additive from animal-origin raw materials | |
| RU2562581C1 (en) | Method of producing biologically active agent from sea cucumber, having general tonic and immunomodulating properties | |
| JP3155746B1 (en) | Method for producing type II collagen |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| RD04 | Notification of resignation of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7424 Effective date: 20070705 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20071217 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20110204 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110331 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20110909 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20111206 |
|
| A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20111222 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20120210 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20120217 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20120306 |
|
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20120316 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 4955944 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20150323 Year of fee payment: 3 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |