JP2587680B2 - Method for producing fine powder of hyaluronic acid or sodium hyaluronate - Google Patents

Method for producing fine powder of hyaluronic acid or sodium hyaluronate

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Publication number
JP2587680B2
JP2587680B2 JP63116638A JP11663888A JP2587680B2 JP 2587680 B2 JP2587680 B2 JP 2587680B2 JP 63116638 A JP63116638 A JP 63116638A JP 11663888 A JP11663888 A JP 11663888A JP 2587680 B2 JP2587680 B2 JP 2587680B2
Authority
JP
Japan
Prior art keywords
hyaluronic acid
sodium hyaluronate
salt
mill
organic solvent
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.)
Expired - Fee Related
Application number
JP63116638A
Other languages
Japanese (ja)
Other versions
JPH01287103A (en
Inventor
徹 新藤
昌和 畠山
泰弘 黒川
純 平木
正弘 藤井
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
JNC Corp
Original Assignee
Chisso Corp
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Filing date
Publication date
Application filed by Chisso Corp filed Critical Chisso Corp
Priority to JP63116638A priority Critical patent/JP2587680B2/en
Priority to FR8906190A priority patent/FR2631344B1/en
Priority to GB8910811A priority patent/GB2218429B/en
Priority to DE19893915557 priority patent/DE3915557A1/en
Publication of JPH01287103A publication Critical patent/JPH01287103A/en
Application granted granted Critical
Publication of JP2587680B2 publication Critical patent/JP2587680B2/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/12—Powdering or granulating
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73—Polysaccharides
    • A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q1/00—Make-up preparations; Body powders; Preparations for removing make-up
    • A61Q1/12—Face or body powders for grooming, adorning or absorbing
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/41—Particular ingredients further characterized by their size
    • A61K2800/412—Microsized, i.e. having sizes between 0.1 and 100 microns
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
    • C08J2305/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Materials Engineering (AREA)
  • Biochemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Birds (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Cosmetics (AREA)

Description

【発明の詳細な説明】 （産業上の利用分野） 本発明は、平均粒径20μｍ以下の微粉末状ヒアルロン
酸もしくはヒアルロン酸ナトリウム（以下、ヒアルロン
酸（塩）という。）の製造法に関する。Description: TECHNICAL FIELD The present invention relates to a method for producing finely divided hyaluronic acid or sodium hyaluronate (hereinafter referred to as hyaluronic acid (salt)) having an average particle size of 20 μm or less.

ヒアルロン酸は、関節、硝子体、臍剤、軟骨、皮膚、
鳥類のとさか等の結合組織中にその構成成分として存在
し、組織の柔軟性、構造維持、細胞の代謝調節等に重要
な機能を果たしている。また該ヒアルロン酸は、高分子
物質であり、その溶液は強い粘弾性を持ち、保水作用を
有するところから、化粧品原料として広く使用されてい
る。Hyaluronic acid is used in joints, vitreous, umbilicus, cartilage, skin,
It is present as a component in connective tissues such as birds' crests and plays an important role in tissue flexibility, structure maintenance, cell metabolism regulation, and the like. Further, the hyaluronic acid is a polymer substance, and its solution has strong viscoelasticity and has a water retaining effect, and is therefore widely used as a raw material for cosmetics.

（従来の技術） 従来、ヒアルロン酸（塩）の製造は工業的にはにわと
りのとさか、牛の目の硝子体または臍帯等から抽出する
方法もしくはヒアルロン酸産生能を有する微生物を培地
に培養して製造する方法（醗酵法）により行われてい
る。(Prior art) Conventionally, the production of hyaluronic acid (salt) is industrially a method of extracting from chicken chick, vitreous body or umbilical cord of cattle, or culturing a microorganism having hyaluronic acid-producing ability in a medium. It is performed by a production method (fermentation method).

該抽出法や醗酵法で得られたヒアルロン酸（塩）は、
限外過もしくは透析、溶剤沈澱、イオン交換樹脂処
理、分別沈澱、活性炭処理等を行つて精製したのち、ヒ
アルロン酸（塩）の水溶液に溶媒を加えてヒアルロン酸
（塩）の粉末状沈澱物を生成せしめ、これを乾燥して粉
末状ヒアルロン酸（塩）を製造している。Hyaluronic acid (salt) obtained by the extraction method or fermentation method,
After purification by ultrafiltration or dialysis, solvent precipitation, ion exchange resin treatment, fractional precipitation, activated carbon treatment, etc., the solvent is added to an aqueous solution of hyaluronic acid (salt) to remove the powdery precipitate of hyaluronic acid (salt). It is produced and dried to produce powdered hyaluronic acid (salt).

従来、化粧品原料として用いられるヒアルロン酸
（塩）は、水に溶解されてクリームもしくは乳液として
用いられることが多かつたが最近では、ヒアルロン酸
（塩）を微粉砕した微粉末状ヒアルロン酸（塩）をプレ
ス化粧品に少量添加すると、該プレス化粧品に微細なク
ラツクが発生するのを防止することができ、化粧したと
きのつつぱり感を改善することができる等の効果が出る
ため、粉体化粧品原料として微粉末の形で使用されるこ
とも多くなつてきた。Conventionally, hyaluronic acid (salt) used as a raw material for cosmetics has often been dissolved in water and used as a cream or emulsion, but recently, hyaluronic acid (salt) has been finely pulverized from finely divided hyaluronic acid (salt). ) Is added to press cosmetics in a small amount, so that fine cracks can be prevented from being generated in the press cosmetics, and the effect of improving the feeling of squeezing when applying makeup can be obtained. It is increasingly used in the form of fine powder as a raw material.

従来、該微粉末状ヒアルロン酸（塩）を製造するに
は、乾式粉砕法が行われていた。しかし、この乾式粉砕
法ではヒアルロン酸（塩）が天然物系高分子物質である
こと、ヒアルロン酸（塩）が吸湿性の強い物質であるこ
と等の理由で粉体化粧品として用いられる平均粒径20μ
ｍ以下の微粉末状ヒアルロン酸（塩）を製造することは
非常に困難であつた。Conventionally, dry pulverization has been performed to produce the fine powdered hyaluronic acid (salt). However, in this dry grinding method, the average particle size used for powder cosmetics is because hyaluronic acid (salt) is a natural polymer material and hyaluronic acid (salt) is a substance that has high hygroscopicity. 20μ
It has been very difficult to produce hyaluronic acid (salt) in the form of powder having a particle size of less than m.

（発明が解決しようとする課題） 本発明者らは平均粒径20μｍ以下の微粉末状ヒアルロ
ン酸（塩）の製造法について鋭意研究した。その結果、
ヒアルロン酸（塩）を溶解しない有機溶剤中に該ヒアル
ロン酸（塩）を懸濁させ、該懸濁液を湿式粉砕機を用い
て粉砕することにより、20μｍ以下の微粉末状ヒアルロ
ン酸（塩）が容易に得られることを見い出し、その知見
にもとづいて本発明を完成した。(Problems to be Solved by the Invention) The present inventors have intensively studied a method for producing fine powdery hyaluronic acid (salt) having an average particle size of 20 μm or less. as a result,
The hyaluronic acid (salt) is suspended in an organic solvent that does not dissolve the hyaluronic acid (salt), and the suspension is pulverized using a wet pulverizer. Have been found to be easily obtained, and the present invention has been completed based on the findings.

以上の記述から明らかなように、本発明の目的は、粉
体化粧品原料として用いられる20μｍ以下の微粉末状ヒ
アルロン酸（塩）の製造法を提供することである。As is apparent from the above description, an object of the present invention is to provide a method for producing fine powdery hyaluronic acid (salt) of 20 μm or less used as a powder cosmetic raw material.

（課題を解決するための手段） 本発明は下記の構成を有する。(Means for Solving the Problems) The present invention has the following configurations.

（１） ヒアルロン酸もしくはヒアルロン酸ナトリウム
を溶解しない有機溶剤中に、ヒアルロン酸もしくはヒア
ルロン酸ナトリウムを懸濁させたのち、該懸濁液を湿式
粉砕機で粉砕し、有機溶剤を除去したのち、粉砕された
ヒアルロン酸もしくはヒアルロン酸ナトリウムを乾燥す
ることにより、微粉末状のヒアルロン酸もしくはヒアル
ロン酸ナトリウムを製造する方法。(1) After suspending hyaluronic acid or sodium hyaluronate in an organic solvent that does not dissolve hyaluronic acid or sodium hyaluronate, the suspension is pulverized with a wet pulverizer to remove the organic solvent, and then pulverized. A method for producing hyaluronic acid or sodium hyaluronate in the form of fine powder by drying hyaluronic acid or sodium hyaluronate.

（２）有機溶剤として、エチルアルコール、メチルアル
コール、アセトン、ｎ−プロパノール、イソプロパノー
ル、アセトニトリルもしくはこれらの２種以上の混合物
を用いる前記第１項記載の方法。(2) The method according to the above (1), wherein ethyl alcohol, methyl alcohol, acetone, n-propanol, isopropanol, acetonitrile or a mixture of two or more thereof is used as the organic solvent.

（３）湿式粉砕機として、媒体攪拌式粉砕機、湿式高速
回転ミル式粉砕機もしくはこれらを連結したものである
前記第１項記載の方法。(3) The method according to the above (1), wherein the wet pulverizer is a medium stirring pulverizer, a wet high-speed rotary mill pulverizer, or a combination thereof.

（４）微粉末状のヒアルロン酸もしくはヒアルロン酸ナ
トリウムの平均粒径が20μｍ以下である前記第１項記載
の方法。(4) The method according to the above (1), wherein the average particle size of the fine powdery hyaluronic acid or sodium hyaluronate is 20 μm or less.

（５）媒体攪拌式粉砕機として、塔式粉砕機、攪拌槽型
粉砕機、流通管型粉砕機もしくはアニユラー型粉砕機を
用いる前記第３項記載の方法。(5) The method according to the above (3), wherein as the medium stirring type pulverizer, a tower type pulverizer, a stirring tank type pulverizer, a flow tube type pulverizer or an Anyurer type pulverizer is used.

（６）湿式高速回転ミル式粉砕機として、コロイドミル
式粉砕機を用いる前記項３項記載の方法。(6) The method according to item 3, wherein a colloid mill is used as the wet high-speed rotary mill.

本発明の製造法で用いられるヒアルロン酸（塩）は、
牛の目の硝子体、臍帯、にわとりのとさか等の生体から
抽出法により得たもの、ヒアルロン酸産生能を有する微
生物、たとえばストレプトコツカス・ピオゲネス、スト
レプトコツカス・エクイ、ストレプトコツカス・エクイ
シミリス、ストレプトコツカス、デイスガラクテイエ、
ストレプトコツカス・ズーエピデミカス等を、たとえば
培養液１当りブドウ糖20〜30g、酵母エキス5g、リン
酸１カルシウム3g、リン酸２カリウム2g、チオ硫酸ナト
リウム0.1g、硫酸マグネシウム７水塩0.1g、亜硫酸ナト
リウム0.02g、塩化コバルト0.01g、塩化マンガン0.01
g、消泡剤5gを含むPH6.0〜8.5、好ましくは7.0の培養液
中で培養温度25℃〜40℃、好ましくは30℃〜35℃で１〜
３日間振とう培養もしくは通気培養し、ヒアルロン酸
（塩）を該培養液中に生成蓄積させ、該培養液を遠心分
離もしくは過して菌体を除去したのち、液を限外
過もしくは透析することによつて低分子量物質を除去
し、低分子量物質を除去した液に水溶性有機溶剤を添
加してヒアルロン酸ナトリウムを沈澱させ、ついで該沈
澱物を臭化セチルトリメチルアンモニウム等による分画
沈澱、イオン交換クロマトグラフイー処理、活性炭処
理、（脱色処理）濃縮工程を経て濃縮された精製ヒアル
ロン酸（塩）水溶液を得たのち、該水溶液をスプレー乾
燥、凍結乾燥などの方法により粉末状ヒアルロン酸
（塩）としたものもしくは上述の濃縮された精製ヒアル
ロン酸（塩）水溶液にヒアルロン酸（塩）を溶解しない
水溶性の有機溶剤（たとえばエチルアルコール、メチル
アルコール、アセトン、イソプロパノールなど）を添加
してヒアルロン酸（塩）を沈澱させ、該沈澱を分離し、
さらに該水溶性有機溶剤で該沈澱を洗浄したのち、該沈
澱を乾燥して得られたものなどをあげることができる。Hyaluronic acid (salt) used in the production method of the present invention is
Vitreous of cattle eyes, umbilical cord, those obtained by extraction from living organisms such as chicken comb, microorganisms having hyaluronic acid-producing ability, such as Streptococcus pyogenes, Streptococcus equi, Streptococcus equisimilis, Streptococcus, Dysgalacteier,
Streptococcus zooepidemicus, etc., for example, 20-30 g of glucose, 5 g of yeast extract, 3 g of monocalcium phosphate, 2 g of dipotassium phosphate, 0.1 g of sodium thiosulfate, 0.1 g of magnesium sulphate heptahydrate 0.1 g, sodium sulfite 0.02 g, cobalt chloride 0.01 g, manganese chloride 0.01
g, containing a defoamer 5 g PH6.0-8.5, preferably 7.0 in the culture medium 25 ℃ ~ 40 ℃, preferably 30 ℃ ~ 35 ℃ 1 ~
After shaking culture or aeration culture for 3 days, hyaluronic acid (salt) is produced and accumulated in the culture solution, and the culture solution is centrifuged or passed to remove bacterial cells, and then the solution is subjected to ultrafiltration or dialysis. As a result, the low-molecular-weight substance is removed, and a water-soluble organic solvent is added to the liquid from which the low-molecular-weight substance has been removed to precipitate sodium hyaluronate, and the precipitate is fractionated and precipitated with cetyltrimethylammonium bromide or the like. After obtaining a purified aqueous solution of hyaluronic acid (salt) concentrated through an ion exchange chromatography treatment, an activated carbon treatment, and a (decolorizing treatment) concentration step, the aqueous solution is subjected to powdery hyaluronic acid (spray drying, freeze-drying, etc.). Salt) or a water-soluble organic solvent that does not dissolve hyaluronic acid (salt) in the concentrated aqueous solution of purified hyaluronic acid (salt) described above (for example, ethyl acetate). Call, methyl alcohol, acetone, isopropanol, etc.) was added to precipitate the hyaluronic acid (salt), separating the 該沈 lees,
Further, those obtained by washing the precipitate with the water-soluble organic solvent and then drying the precipitate can be exemplified.

本発明の製造法で用いるヒアルロン酸（塩）を溶解し
ない有機溶剤としては、該ヒアルロン酸（塩）を溶解し
ない有機溶剤であれば特に制限はないが、エチルアルコ
ール、メチルアルコール、アセトン、ｎ−プロパノー
ル、イソプロパノール、アセトニトリルもしくはこれら
の２種以上の混合物が好ましい。ヒアルロン酸（塩）を
溶解しない有機溶剤に該ヒアルロン酸（塩）を懸濁させ
るには、乾燥した粉末状ヒアルロン酸（塩）を該有機溶
剤に添加してもよいが、上述の水溶性有機溶剤で洗浄を
行つたヒアルロン酸（塩）の沈澱物を引きつづき、水溶
性有機溶剤を添加して懸濁させた懸濁液をそのまま用い
ることもできる。該有機溶剤中に懸濁されるヒアルロン
酸（塩）の量は、用いる粉砕機の種類によつて異なる
が、懸濁液１当り20〜400g程度が好ましい。該ヒアル
ロン酸（塩）の懸濁液中の量が多すぎると粉砕しても平
均粒径が20μｍ以下にならない場合があり、また該量が
少なすぎると、粉砕機本体の摺動部分の磨耗が大きくな
り、該磨耗物が微粉砕されたヒアルロン酸（塩）中に異
物として混入する原因となるので注意する必要がある。
このため、該有機溶剤中のヒアルロン酸（塩）の量は粉
砕テストを繰り返してその結果から決めることが好まし
い。The organic solvent that does not dissolve hyaluronic acid (salt) used in the production method of the present invention is not particularly limited as long as it is an organic solvent that does not dissolve the hyaluronic acid (salt), but ethyl alcohol, methyl alcohol, acetone, n- Preferred are propanol, isopropanol, acetonitrile or a mixture of two or more thereof. In order to suspend the hyaluronic acid (salt) in an organic solvent that does not dissolve the hyaluronic acid (salt), dried powdery hyaluronic acid (salt) may be added to the organic solvent. The precipitate of hyaluronic acid (salt) washed with a solvent can be subsequently used, and a suspension obtained by adding a water-soluble organic solvent to the suspension can be used as it is. The amount of hyaluronic acid (salt) suspended in the organic solvent varies depending on the type of crusher used, but is preferably about 20 to 400 g per suspension. If the amount of the hyaluronic acid (salt) in the suspension is too large, the average particle size may not be reduced to 20 μm or less even when the material is pulverized, and if the amount is too small, the sliding portion of the pulverizer body is worn. It is necessary to pay attention to this because the abrasion becomes a foreign substance in the finely pulverized hyaluronic acid (salt).
For this reason, it is preferable that the amount of hyaluronic acid (salt) in the organic solvent is determined based on the results of repeated pulverization tests.

該ヒアルロン酸（塩）懸濁液の粉砕は１回粉砕機にか
けるだでもよいが、数回たとえば３〜５回循環して粉砕
機にかける方が均一に微粉砕されたヒアルロン酸（塩）
が得られるので好ましい。Although the hyaluronic acid (salt) suspension may be pulverized once with a pulverizer, it is more preferable to circulate the mixture with the pulverizer several times, for example, three to five times, and to pulverize the hyaluronic acid (salt) uniformly finely.
Is preferred.

また、該懸濁液を粉砕するときの該懸濁液の温度は常
温でよいが、低温の方が微粉砕処理が円滑に行われるの
で好ましい。The temperature of the suspension at the time of pulverizing the suspension may be room temperature, but a lower temperature is preferred because the fine pulverization process is performed smoothly.

本発明に用いる湿式粉砕機は湿式粉砕機であればどん
な型式のものでもよいが、好ましくは媒体攪拌式粉砕
機、湿式高速回転ミル式粉砕機がよい。The wet pulverizer used in the present invention may be of any type as long as it is a wet pulverizer, but a medium stirring pulverizer and a wet high-speed rotary mill pulverizer are preferred.

微粉砕処理後、水溶性有機溶剤は過もしくは遠心分
離によつて除去する。得られたケーキを減圧乾燥するこ
とにより微粉末状のヒアルロン酸（塩）が得られる。After pulverization, the water-soluble organic solvent is removed by excess or centrifugation. By drying the obtained cake under reduced pressure, hyaluronic acid (salt) in the form of fine powder is obtained.

（実施例） 以下、実施例、比較例を用いて本発明を具体的に説明
するが、本発明はこれら実施例に限定されるものではな
い。(Examples) Hereinafter, the present invention will be specifically described with reference to Examples and Comparative Examples, but the present invention is not limited to these Examples.

実施例１ 醗酵法で製造したヒアルロン酸ナトリウムの乾燥粉末
（平均粒径:150μｍ）400gを2000mlのエチルアルコール
中に加え、よく攪拌して均一な膨潤したスラリーとす
る。このスラリーを媒体攪拌式粉砕機（商品名：コボー
ルミル、神鋼フアウドラー製、機種:MS−18）を用い
て、下記条件で粉砕した。Example 1 400 g of a dry powder (average particle size: 150 μm) of sodium hyaluronate produced by a fermentation method was added to 2000 ml of ethyl alcohol, and stirred well to form a uniformly swollen slurry. This slurry was pulverized under the following conditions by using a medium stirring type pulverizer (trade name: Koball Mill, manufactured by Shinko Faudler, model: MS-18).

メデイア ガラス メヂイア径 １〜1.5mm ローター周速 13m/sec フイード速度 500ml/min スラリー温度 （入口:7℃、出口:7℃） 粉砕はスラリーを３回粉砕機に循環させて微粉砕を行
つた。微粉砕されたスラリーは、吸引過によつてエチ
ルアルコールと分離し、減圧乾燥を行つた。得られた乾
燥微粉砕品の量は388gであつた。粒度を光透視法で測定
した結果、平均粒径は3.0μｍであつた。Media glass Media diameter 1 to 1.5 mm Rotor peripheral speed 13 m / sec Feed speed 500 ml / min Slurry temperature (inlet: 7 ° C, outlet: 7 ° C) In the pulverization, the slurry was circulated through a pulverizer three times to perform fine pulverization. The finely pulverized slurry was separated from ethyl alcohol by suction and dried under reduced pressure. The amount of the obtained dry finely pulverized product was 388 g. As a result of measuring the particle size by a light transmission method, the average particle size was 3.0 μm.

実施例２ 醗酵法で製造したヒアルロン酸ナトリウムの精製水溶
液300（ヒアルロン酸ナトリウム900g含有）に、0.2モ
ル／になるように食塩を溶解せしめる。この溶液を厳
しく攪拌しつつ、エタノール900を徐々に添加してヒ
アルロン酸ナトリウムを析出させる。攪拌を防止し、
過法で母液を分離したのち、得られた沈澱物をエタノー
ル900を添加して、再度攪拌を行い洗浄を行う。洗浄
終了後、過法で母液を分離する。Example 2 Salt is dissolved in a purified aqueous solution of sodium hyaluronate 300 (containing 900 g of sodium hyaluronate) produced by fermentation at a concentration of 0.2 mol /. While stirring this solution vigorously, ethanol 900 is gradually added to precipitate sodium hyaluronate. Prevent agitation,
After separating the mother liquor by an excess method, the obtained precipitate is added with ethanol 900, and stirred again for washing. After the washing is completed, the mother liquor is separated by an excessive method.

得られたヒアルロン酸ナトリウム沈澱物にエタノール
30を攪拌しつつ添加し、均一なスラリーとする。この
スラリーを湿式高速回転ミル（商品名：ボツクボルトホ
モジナイザー、中央機工製、機種:100型）を用いて、下
記条件で粉砕した。Ethanol was added to the obtained sodium hyaluronate precipitate.
Add 30 with stirring to make a uniform slurry. This slurry was pulverized using a wet high-speed rotation mill (trade name: Bolt bolt homogenizer, manufactured by Chuo Kiko, model: 100 type) under the following conditions.

クリアランス 0.07〜0.08mm フイード速度 10/min 微粉砕は循環して行い、10分間行つた。微粉砕された
スラリーは、吸引過によつてエチルアルコールと分離
し、減圧乾燥を行つた。得られた乾燥微粉砕品の量は72
2gであつた。粒度を光透過法で測定した結果、平均粒径
は8.5μｍであつた。Clearance 0.07-0.08mm Feed rate 10 / min The pulverization was performed by circulation and was performed for 10 minutes. The finely pulverized slurry was separated from ethyl alcohol by suction and dried under reduced pressure. The amount of the dried finely pulverized product obtained is 72
It was 2g. As a result of measuring the particle size by a light transmission method, the average particle size was 8.5 μm.

実施例３ 醗酵法で製造したヒアルロン酸ナトリウムの乾燥粉末
（平均粒径:150μｍ）5gを300mlのエチルアルコール中
に加え、よく攪拌して均一な膨潤したスラリーとする。
このスラリーを媒体攪拌式粉砕機（商品名:DYNO−MILL
シンマルエンタープライゼス製、機種:KDL型）を用い
て、下記条件で粉砕した。Example 3 5 g of a dry powder (average particle size: 150 μm) of sodium hyaluronate produced by a fermentation method was added to 300 ml of ethyl alcohol, and the mixture was stirred well to form a uniform swollen slurry.
This slurry is crushed with a medium agitation type (trade name: DYNO-MILL)
(Shinmaru Enterprises, model: KDL type) under the following conditions.

グラインデイングエレメント ガラスビーズ グラインデイングエレメント径 0.5〜0.75mmφ アジテーターデイスク周速 10m/sec フイード速度 50ml/min スラリー温度 20℃ 粉砕は、スラリーをマグネチツクスターラーで攪拌し
ながら、粉砕機にペリスタルテツクポンプでフイードさ
せ、微粉砕を行つた。Grinding element Glass beads Grinder element diameter 0.5 ~ 0.75mmφ Agitator disk peripheral speed 10m / sec Feed speed 50ml / min Slurry temperature 20 ° C Grinding is performed by stirring the slurry with a magnetic stirrer and feeding it to a crusher using a peristaltic pump. And pulverized.

微粉砕されたスラリーは、吸引過によつてエチルア
ルコールと分離し、減圧乾燥を行つた。得られた乾燥微
粉砕品の量は4.82gであつた。粒度を光透過法で測定し
た結果、平均粒径は4.1μｍであつた。The finely pulverized slurry was separated from ethyl alcohol by suction and dried under reduced pressure. The amount of the obtained dried finely pulverized product was 4.82 g. As a result of measuring the particle size by a light transmission method, the average particle size was 4.1 μm.

比較例１ 実施例１で使用したと同じヒアルロン酸ナトリウムの
乾燥粉末（平均粒径:150μｍ）20gを容積950mlのアルミ
ナ製ボールミルポツトにアルミナボールとともに入れ、
100rpmで４時間乾式粉砕した。得られた粉砕品の粒度を
光透過法で測定した結果、平均粒径は40μｍであつた。Comparative Example 1 20 g of the same dry powder of sodium hyaluronate (average particle size: 150 μm) used in Example 1 was put together with alumina balls into a 950 ml alumina ball mill pot.
Dry milling was performed at 100 rpm for 4 hours. As a result of measuring the particle size of the obtained pulverized product by a light transmission method, the average particle size was 40 μm.

（発明の効果） 本発明を用いることにより、平均粒径20μｍ以下の微
粉末状ヒアルロン酸（塩）を容易に製造することができ
る。(Effect of the Invention) By using the present invention, fine powdery hyaluronic acid (salt) having an average particle diameter of 20 µm or less can be easily produced.

Claims (6)

(57)【特許請求の範囲】(57) [Claims] 【請求項１】ヒアルロン酸もしくはヒアルロン酸ナトリ
ウムを溶解しない有機溶剤中に、ヒアルロン酸もしくは
ヒアルロン酸ナトリウムを懸濁させたのち、該懸濁液を
湿式粉砕機で粉砕し、有機溶剤を除去したのち、粉砕さ
れたヒアルロン酸もしくはヒアルロン酸ナトリウムを乾
燥することにより、微粉末状のヒアルロン酸もしくはヒ
アルロン酸ナトリウムを製造する方法。(1) After suspending hyaluronic acid or sodium hyaluronate in an organic solvent which does not dissolve hyaluronic acid or sodium hyaluronate, the suspension is pulverized with a wet pulverizer to remove the organic solvent. A method of producing finely divided hyaluronic acid or sodium hyaluronate by drying the ground hyaluronic acid or sodium hyaluronate. 【請求項２】有機溶剤として、エチルアルコール、メチ
ルアルコール、アセトン、ｎ−プロパノール、イソプロ
パノール、アセトニトリルもしくはこれらの２種以上の
混合物を用いる請求項１記載の方法。2. The method according to claim 1, wherein ethyl alcohol, methyl alcohol, acetone, n-propanol, isopropanol, acetonitrile or a mixture of two or more thereof is used as the organic solvent. 【請求項３】湿式粉砕機として、媒体攪拌式粉砕機、湿
式高速回転ミル式粉砕機もしくはこれらを連結したもの
である請求項１記載の方法。3. The method according to claim 1, wherein the wet mill is a medium stirring mill, a wet high-speed rotary mill mill, or a combination thereof. 【請求項４】微粉末状のヒアルロン酸もしくはヒアルロ
ン酸ナトリウムの平均粒径が20μｍ以下である請求項１
記載の方法。4. The fine powdery hyaluronic acid or sodium hyaluronate has an average particle size of 20 μm or less.
The described method. 【請求項５】媒体攪拌式粉砕機として、塔式粉砕機、攪
拌槽型粉砕機、流通管型粉砕機もしくはアニユラー型粉
砕機を用いる請求項３記載の方法。5. The method according to claim 3, wherein a tower type pulverizer, a stirring tank type pulverizer, a flow tube type pulverizer or an Anyurer type pulverizer is used as the medium stirring type pulverizer. 【請求項６】湿式高速回転ミル式粉砕機として、コロイ
ドミル式粉砕機を用いる請求項３記載の方法。6. The method according to claim 3, wherein a colloid mill is used as the wet high-speed rotary mill.
JP63116638A 1988-05-13 1988-05-13 Method for producing fine powder of hyaluronic acid or sodium hyaluronate Expired - Fee Related JP2587680B2 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP63116638A JP2587680B2 (en) 1988-05-13 1988-05-13 Method for producing fine powder of hyaluronic acid or sodium hyaluronate
FR8906190A FR2631344B1 (en) 1988-05-13 1989-05-11 PROCESS FOR THE PREPARATION OF FINE HYALURONIC ACID POWDER OR SODIUM HYALURONATE AND FINE POWDER THUS OBTAINED
GB8910811A GB2218429B (en) 1988-05-13 1989-05-11 Finely powdered hyalluronic acid or sodium hyaluronate and a process for producing it
DE19893915557 DE3915557A1 (en) 1988-05-13 1989-05-12 FINE POWDER OF HYALURONIC ACID OR SODIUM HYALURONATE AND METHOD OF PREPARING THEREOF

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JPH01287103A JPH01287103A (en) 1989-11-17
JP2587680B2 true JP2587680B2 (en) 1997-03-05

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DE (1) DE3915557A1 (en)
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GB (1) GB2218429B (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100990301B1 (en) * 2007-09-28 2010-10-26 가부시키가이샤 시세이도 Swellable crosslinked hyaluronic acid powder and its manufacturing method
KR101285626B1 (en) * 2004-05-27 2013-07-15 노보자임스 바이오파마 디케이 에이/에스 A dried and agglomerated hyaluronic acid product

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH02215707A (en) * 1989-02-15 1990-08-28 Chisso Corp Skin cosmetic
JPH0630605B2 (en) * 1990-10-23 1994-04-27 チッソ株式会社 Method for producing sodium hyaluronate aqueous solution
JPH08842B2 (en) * 1991-05-22 1996-01-10 信越化学工業株式会社 Purification method of polysaccharides
US7956180B2 (en) 2004-05-27 2011-06-07 Novozymes A/S Dried and agglomerated hyaluronic acid product
JP4932166B2 (en) * 2005-02-28 2012-05-16 焼津水産化学工業株式会社 Method for producing particulate chitosan
WO2019073363A1 (en) * 2017-10-12 2019-04-18 Solyplus Berlin Gmbh Processing method for biopolymers using solvent combinations
CN109851822B (en) * 2018-12-07 2022-02-11 山东众山生物科技有限公司 Preparation method of instant sodium hyaluronate
CN112981613B (en) * 2021-03-01 2022-05-31 浙江景嘉医疗科技有限公司 Preparation method of fibrous sodium hyaluronate

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4404346A (en) * 1980-08-05 1983-09-13 Rohm And Haas Company Production of powdered resin and the powdered resin so produced
JPS62289197A (en) * 1986-06-06 1987-12-16 Kyowa Hakko Kogyo Co Ltd Production of sodium hyaluronate powder

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101285626B1 (en) * 2004-05-27 2013-07-15 노보자임스 바이오파마 디케이 에이/에스 A dried and agglomerated hyaluronic acid product
KR100990301B1 (en) * 2007-09-28 2010-10-26 가부시키가이샤 시세이도 Swellable crosslinked hyaluronic acid powder and its manufacturing method

Also Published As

Publication number Publication date
JPH01287103A (en) 1989-11-17
DE3915557C2 (en) 1991-10-24
DE3915557A1 (en) 1989-11-23
FR2631344A1 (en) 1989-11-17
FR2631344B1 (en) 1993-12-31
GB8910811D0 (en) 1989-06-28
GB2218429B (en) 1991-06-26
GB2218429A (en) 1989-11-15

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