KR20040022760A - Method for extracting hight purified hyaluronic acid from cocks comb - Google Patents
Method for extracting hight purified hyaluronic acid from cocks comb Download PDFInfo
- Publication number
- KR20040022760A KR20040022760A KR1020020053546A KR20020053546A KR20040022760A KR 20040022760 A KR20040022760 A KR 20040022760A KR 1020020053546 A KR1020020053546 A KR 1020020053546A KR 20020053546 A KR20020053546 A KR 20020053546A KR 20040022760 A KR20040022760 A KR 20040022760A
- Authority
- KR
- South Korea
- Prior art keywords
- hyaluronic acid
- solution
- salt
- extracting
- chloroform
- Prior art date
Links
Classifications
-
- 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
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Sustainable Development (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
Description
본 발명은 닭벼슬로부터 고순도 히아루론산의 추출방법에 관한 것으로, 구체적으로, 닭벼슬로부터 염과 클로로포름을 이용하여 고순도의 히아루론산을 추출하는 방법에 관한 것이다.The present invention relates to a method of extracting high purity hyaluronic acid from chicken rice, and more particularly, to a method of extracting high purity hyaluronic acid from a salt of chicken using salt and chloroform.
히아루론산은 생체 고분자의 일종으로 분자량이 5만 내지 1,300만 달톤에 이르는 무색 투명한 고점도의 선형 다당류이며, 반복 단위인 클루쿠론산과N-아세틸 글루코즈아민이 (1-3)과 (1-4)로 번갈아 결합되어 있다. 히아루론산은 피부, 눈의 초자체, 관절액, 탯줄 등에 널리 분포되어 있으며 스트렙토코커스 속 등의 박테리아에 의해 생성되기도 한다.Hyaluronic acid is a type of biopolymer, a colorless transparent high viscosity linear polysaccharide with a molecular weight of 50,000 to 13 million daltons, and the repeating units of glucuronic acid and N -acetyl glucosamine (1-3) and (1-4) Alternately combined. Hyaluronic acid is widely distributed in skin, eye vitreous body, joint fluid and umbilical cord, and is also produced by bacteria such as Streptococcus genus.
히아루론산은 보습 효과가 강하며 물리적 마찰에 대한 윤활 효과 및 세균 등의 침입에 대한 보호 효과가 있어 피부 노화방지용 고급 화장품 첨가제, (백내장, 녹내장, 각막 손상 등으로 인한)안과수술시 눈 조직의 보호제, 관절염 치료제, 외상 피복제, 외과 수술시 피복제 등과 같이 화장품 및 의료용으로 광범위하게 개발되고 있다.Hyaluronic acid has a strong moisturizing effect, lubricates against physical friction and protects against invasion of bacteria, etc. It is an advanced cosmetic additive to prevent skin aging, eye tissue protector during ophthalmic surgery (caused by cataracts, glaucoma, cornea damage, etc.), It is widely developed for cosmetics and medical applications, such as arthritis treatment, traumatic coating, and surgical coating.
미국특허 제2,585,546호(하디안(HADIAN))는 탯줄로부터 히아루론산을 추출하여 정제하는 방법을 설명한 것으로 정제과정이 매우 복잡하고 생산성이 낮아 효율적이지 못하다.U. S. Patent No. 2,585, 546 (HADIAN) describes a method of extracting and purifying hyaluronic acid from the umbilical cord and the purification process is very inefficient and low in productivity.
미국특허 제3,396,081호(빌렉(BILLEK))는 눈의 초자체, 탯줄 및 미생물로부터 히아루론산을 정제하는 방법을 제시하고 있는데, 여기서는 정제시 단백질 분해효소를 사용하여 단백질을 아미노산으로 분해시킨 후 이온 교환 수지를 사용하여 유리 아미노산 및 금속염을 제거함과 동시에 남아있는 단백질은 pH를 약 3 내지 4로 낮추어 침전에 의해 단백질을 제거하는방법을 사용하였다. 그러나, 이 방법의 단점은 pH를 산성으로 낮출 경우 고분자량의 히아루론산이 분해되는 것을 방지하기가 어려우므로 분자량 감소가 야기된다는 것이다.U.S. Patent No. 3,396,081 (BILLEK) discloses a method for purifying hyaluronic acid from the vitreous, umbilical cord and microorganisms of the eye, in which the protein is broken down into amino acids using proteolytic enzymes and then ion exchange resins are purified. In addition to the removal of free amino acids and metal salts, the remaining protein was used to remove the protein by precipitation by lowering the pH to about 3-4. However, a disadvantage of this method is that lowering the pH to acid makes it difficult to prevent the breakdown of high molecular weight hyaluronic acid, leading to a molecular weight reduction.
미국특허 제4,141,933호(발라즈(BALAZS))는 고도로 순수한 고분자량의 히아루론산 분획을 동물 조직으로부터 정제하는 방법을 기술하였다. 히아루론산을 함유한 동물 조직으로부터 클로로포름을 사용하여 pH 6 내지 7에서 단백질 및 미확인된 염증원인 물질을 제거하여 고순도의 히아루론산을 얻었다. 그러나 이 방법은 매우 복잡하고 생산수율이 낮고 생산 비용이 높을 뿐 아니라 염증원인 물질의 제거가 제한된 범위내에서만 가능한 것이 단점이다.US Pat. No. 4,141,933 (BALAZS) describes a method for purifying a highly pure high molecular weight hyaluronic acid fraction from animal tissue. Animal tissue containing hyaluronic acid was used to remove proteins and unidentified inflammatory agents at pH 6-7 using chloroform to obtain high purity hyaluronic acid. However, this method is very complicated, low production yield, high production cost, and only possible to remove the inflammatory material within a limited range.
즉 상기와 같이 추출에 의하여 얻어진 히아루론산에는 콘드로이틴 설페이트,글리칸 설페이트 등의 불순물이 혼입되어 있어 이들을 제거하기 위하여 복잡한 정제 공정이 필요할 뿐 아니라 비용이 많이 소모되어 추출에 의한 방법은 대량 생산에 부적합하다.That is, the hyaluronic acid obtained by the extraction as described above contains impurities such as chondroitin sulfate and glycan sulfate, so that a complicated purification process is required to remove them, as well as costly, and the extraction method is not suitable for mass production.
이에, 본 발명자들은 생체 조직인 닭 벼슬로부터 고순도의 히알우론산의 추출방법 개발하였으며, 상기 추출방법은 기존 방법에서의 문제점인 생산 수율이 낮고, 콘드로이틴 설페이트(Chondroitin sulfate)와 글루코사미노 글리칸 설페이트(Glycosamino glycan sulfate)등의 불순물이 혼입되어 있고, 제조 및 정제과정에서 비용이 많이 소요되는 단점을 개선할 수 있음을 알아내어 본 발명을 완성하였다.Therefore, the present inventors have developed a method of extracting high purity hyaluronic acid from chicken tissue, which is a living tissue, and the extraction method has a low production yield, which is a problem of the existing methods, and chondroitin sulfate and glucosamino glycan sulfate (Glycosamino glycan sulfate). The present invention has been completed by finding that impurities such as sulfate) can be mixed and the disadvantages of costly manufacturing and purification can be improved.
본 발명의 목적은 닭벼슬로부터 고순도의 히아루론산을 추출하는 방법을 제공하는 것이다.An object of the present invention is to provide a method for extracting high purity hyaluronic acid from chicken rice bran.
상기한 목적을 달성하기 위하여, 닭벼슬로부터 고순도의 히아루론산을 추출하는 방법을 제공한다.In order to achieve the above object, there is provided a method of extracting high purity hyaluronic acid from chicken rice.
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 닭벼슬로부터 고순도의 히아루론산의 추출방법을 포함한다.The present invention includes a method of extracting high purity hyaluronic acid from chicken rice bran.
구체적으로, 분쇄된 닭벼슬을 증류수와 클로로포름 혼합용매로 추출한 후 여과시켜 닭벼슬 용액을 얻는 단계(단계 1);Specifically, pulverized chicken rice extract with distilled water and chloroform mixed solvent and then filtered to obtain a chicken rice solution (step 1);
상기 닭벼슬 용액에 염 용액을 첨가한 후 교반하는 단계(단계 2);Adding a salt solution to the chicken rice solution and then stirring it (step 2);
상기 단계 2의 용액을 클로로포름에 첨가하고 교반하여 1차 상등액을 얻는 단계(단계 3);Adding the solution of step 2 to chloroform and stirring to obtain a primary supernatant (step 3);
상기 얻어진 1차 상등액을 원심분리하여 2차 상등액을 얻는 단계(단계 4); 및,Centrifuging the obtained primary supernatant to obtain a secondary supernatant (step 4); And,
상기 얻어진 2차 상등액에 에탄올을 첨가한 후 방치하여 침전된 히아루론산을 얻는 단계(단계 5)로 이루어진 추출방법을 포함한다.It includes an extraction method comprising the step (step 5) of obtaining the precipitated hyaluronic acid by adding ethanol to the secondary supernatant obtained and then left to stand.
단계 1은 분쇄된 닭벼슬을 증류수와 클로로포름 혼합용매로 추출한 후 여과시켜 닭벼슬 용액을 얻는 것으로, 증류수와 클로로포름의 혼합비로 가장 바람직한 비율은 20:1(v/v)이다. 상기 닭벼슬에 상기 혼합용매를 첨가한 후 3∼5일 동안 방치시키고, 이때 1일에 1번 정도 교반시키는 것이 바람직하다. 이후 닭벼슬 용액을 얻은 후 여과과정을 거치는데, 바람직하게는 나일론 메쉬(mesh)를 이용하여 여과시킨다.Step 1 is to extract the pulverized chicken rice with distilled water and chloroform mixed solvent and then filtered to obtain a chicken rice solution, the most preferred ratio is 20: 1 (v / v) in the mixing ratio of distilled water and chloroform. After the mixed solvent is added to the chicken rice, it is left for 3 to 5 days, at which time it is preferable to stir about once a day. After the chicken rice solution to obtain a filtration process, it is preferably filtered using a nylon mesh (mesh).
단계 2는 상기 닭벼슬 용액에 염(NaCl, Sodium Chloride) 용액을 첨가한 후 교반하는 것으로, 상기 염 용액을 첨가하여 용액 내의 이온세기를 증가시켜 닭벼슬염 추출물을 얻는다. 상기 염 용액의 농도는 염의 최종 농도가 0.3∼4 M이 되도록 하며, 염 용액을 첨가한 후 20 분∼1 시간 동안 교반한다.Step 2 is to add a salt (NaCl, Sodium Chloride) solution to the chicken rice solution and then stirred. The salt solution is added to increase the ionic strength in the solution to obtain a chicken rice salt extract. The concentration of the salt solution is such that the final concentration of the salt is 0.3 to 4 M, and the salt solution is added and stirred for 20 minutes to 1 hour.
단계 3은 상기 단계 2의 용액을 클로로포름에 첨가하고 교반하여 1차 상등액을 얻는 것으로, 상기 클로로포름의 양은 상기 닭벼슬 염 추출물과 같은 부피비가 바람직하다. 상기 교반은 3∼4 시간동안 수행하여 상분리를 시키며, 이때 1차 상등액을 얻는다.In step 3, the solution of step 2 is added to chloroform and stirred to obtain a primary supernatant. The amount of chloroform is preferably in the same volume ratio as that of the chicken rice salt extract. The stirring is performed for 3 to 4 hours to perform phase separation, whereby a primary supernatant is obtained.
단계 4는 상기 얻어진 1차 상등액을 원심분리하여 2차 상등액을 얻는 것으로, 원심분리기는 본 분야에 사용되는 통상적인 원심분리기를 사용하며, 이 때 원심분리기의 속도는 10,000∼20,000 rpm이며, 바람직하게는 12,000 rpm이며, 10 분∼50 분 동안 수행하여 2차 상등액을 얻는다.Step 4 is to obtain a secondary supernatant by centrifuging the obtained primary supernatant, the centrifuge uses a conventional centrifuge used in the art, wherein the speed of the centrifuge is 10,000 to 20,000 rpm, preferably Is 12,000 rpm, and is carried out for 10 to 50 minutes to obtain a secondary supernatant.
단계 5는 상기 얻어진 2차 상등액에 에탄올을 첨가한 후 방치하여 침전된 히아루론산을 얻는 것으로, 첨가되는 에탄올의 양은 2차 상등액에 대하여 부피비로 2∼4 배를 첨가한다. 에탄올을 첨가한 후 2∼10℃에서 20∼30 시간동안 방치시켜 침전시키며, 바람직하게는 4℃에서 24 시간동안 방치시켜 침전시킨다. 침전물을 수거한 후 동결건조하여 분말로 이루어진 히아루론산을 얻는다.Step 5 is obtained by adding ethanol to the secondary supernatant obtained to obtain a precipitated hyaluronic acid, the amount of ethanol added is added 2 to 4 times by volume relative to the secondary supernatant. After adding ethanol, it is allowed to stand for 20 to 30 hours at 2 to 10 ° C. to precipitate, preferably for 24 hours at 4 ° C. to precipitate. The precipitate is collected and then lyophilized to obtain a powder of hyaluronic acid.
본 발명은 대부분 히알우론산과 단백질이 이온결합을 형성하고 있는 점을 착안하여 용액내의 이온세기를 화학적으로 변화시켜 히알우론산과 단백질의 결합을 유리 시켜 제거하는 방법을 도입함으로써 방법적인 면이나 생산 수율적인 면을 비교할 때 단순하고 간단한 방법으로 높은 히알우론산의 수율을 얻을 수 있으며, 더욱이 종래의 활액(synovial fluid)에 의해 얻어진 히아루론산 제조 및 정제과정에서 사용된 비용의 10배 이상 절감되어 미생물에서 추출한 히알우론산과 비슷한 생산비로 고순도의 히알우론산을 정제하여 얻을 수 있다.The present invention focuses on the fact that most of the hyaluronic acid and protein form an ionic bond, thereby chemically changing the ionic strength in the solution, thereby introducing a method of releasing and removing the binding of the hyaluronic acid and the protein, thereby improving the method and production yield. In comparison, a high yield of hyaluronic acid can be obtained by a simple and simple method. Furthermore, the production cost of hyaluronic acid obtained by the conventional synovial fluid is reduced by more than 10 times, and the production cost similar to that of hyaluronic acid extracted from microorganisms. It can be obtained by purifying hyaluronic acid of high purity.
또한, 염의 농도를 조절하여 용액내의 이온세기를 조절해 줌으로써 히아루론산의 수율과 단백질 및 헥산 제거 수준이 현저히 높일 수 있었다. 헥산 농도는 모든 조건에서 두 번의 클로로포름 처리과정을 통해 확인되지 않는 수준으로 감소함을 나타내었다.In addition, by adjusting the salt concentration to control the ionic strength in the solution, the yield of hyaluronic acid and the protein and hexane removal level could be significantly increased. Hexane concentrations were reduced to levels not seen through two chloroform treatments under all conditions.
이하 본 발명을 실시예에 의하여 더욱 상세히 설명한다.Hereinafter, the present invention will be described in more detail with reference to Examples.
단 하기 실시예는 본 발명을 예시하는 것으로 본 발명의 내용이 실시예에 의해 한정되는 것은 아니다.However, the following examples are illustrative of the present invention, and the content of the present invention is not limited by the examples.
<실시예 1> 닭벼슬로부터 히아루론산 추출Example 1 Extraction of Hyaluronic Acid from Chicken Crest
노계(생후 6개월 이상)의 계관 100g(비 건조 중량)을 잘게 조각 내어 99%에탄올로 1일 수세하여 혈액 및 오염물질을 제거 한 후 증류수로 4∼5회 수세 한 후 분쇄기로 2차 분쇄하였다.100 g (non-dry weight) of the broth of the old system (more than 6 months old) was finely crushed, washed with 99% ethanol for 1 day to remove blood and contaminants, washed 4 to 5 times with distilled water, and then pulverized with a grinder. .
(단계 1) 분쇄된 닭 벼슬을 유리조에 넣고 증류수와 클로로포름을 20:1 부피비로 섞은 혼합용매 1 ℓ를 첨가하고 1일을 1회 단위로 교반하여 총 3∼4회에 걸쳐 닭 벼슬로부터 닭벼슬 용액을 추출한 후 나일론 mesh를 이용하여 여과하였다.(Step 1) Put the ground chicken rice bran in a glass bath, add 1 L of a mixed solvent mixed with distilled water and chloroform in a 20: 1 volume ratio, and stir once a day for three to four times. The solution was extracted and filtered using a nylon mesh.
(단계 2) 상기 닭벼슬 용액을 수거하여 염의 최종 농도가 0.3∼1 몰농도 수준이 되도록 염을 첨가하여 용액 내의 이온세기를 증가시킨 후 30분간 교반하였다.(Step 2) The chicken rice bran solution was collected and salt was added to increase the final concentration of the salt to a level of 0.3 to 1 molar concentration, and then stirred for 30 minutes after increasing the ionic strength in the solution.
(단계 3) 상기 단계 2의 용액을 동일한 용량의 클로로포름에 100 ㎖/min속도로 첨가하고, 첨가가 완료된 후 4시간 교반한 후 상 분리된 용액의 1차 상등액을 수거하였다.(Step 3) The solution of step 2 was added to the same volume of chloroform at a rate of 100 ml / min, and after the addition was completed, the mixture was stirred for 4 hours, and the primary supernatant of the phase-separated solution was collected.
(단계 4) 상기 1차 상등액을 원심분리기를 이용하여 12000 rpm에서 20분동안 원심분리 한 후 2차 상등액을 수거한다.(Step 4) After centrifuging the primary supernatant at 12000 rpm for 20 minutes using a centrifuge, the secondary supernatant is collected.
(단계 5) 상기 2차 상등액에 3배수 에탄올을 50 ㎖/min로 첨가한 후 4℃에서 24 시간 동안 방치하여 침전시키고, 침전물을 수거한 후 동결 건조 하여 분말의 히알우론산(5 ㎎/g)을 얻었다.(Step 5) To the secondary supernatant, 3-fold ethanol was added at 50 ml / min, and then allowed to stand at 4 ° C. for 24 hours to precipitate. The precipitate was collected and freeze-dried to obtain hyaluronic acid (5 mg / g) as a powder. Got it.
<실험예 1> 염의 첨가 농도에 따른 히아루론산의 농도Experimental Example 1 Concentration of Hyaluronic Acid According to Concentration of Salt
본 발명의 염의 첨가 농도에 따른 히아루론산의 농도를 측정하기 위하여,상기 실시예 1의 단계 2에서 염 용액의 농도를 변화시켜 이때 얻어진 히아루론산의 농도를 측정하였다. 측정장비는 흡광기(Backman, USA)를 이용하여 측정하였다. 결과는 하기표 1에 나타내었다.In order to measure the concentration of hyaluronic acid according to the concentration of the salt of the present invention, the concentration of the hyaluronic acid obtained by changing the concentration of the salt solution in step 2 of Example 1 was measured. The measuring equipment was measured using an absorber (Backman, USA). The results are shown in Table 1 below.
상기표 1에서 보는 바와 같이, 염의 농도가 1.5 M에서 가장 높은 광학 밀도(0.208)를 나타내었으며, 0.3 M 및 4 M에서 각각 0.195 및 0.173의 광학밀도를 나타내었다. 이는 염의 농도가 1.0 M에서 히아루론산의 농도가 가장 높음을 알 수 있다.As shown in Table 1 , the salt concentration showed the highest optical density (0.208) at 1.5 M, the optical density of 0.195 and 0.173 at 0.3 M and 4 M, respectively. It can be seen that the highest concentration of hyaluronic acid at the salt concentration of 1.0 M.
<실험예 2> 히아루론산의 독성 실험Experimental Example 2 Toxicity Test of Hyaluronic Acid
본 발명의 추출방법에 의해 얻어진 히아루론산의 독성 실험은 세포의 MG63을 이용하여 MTT assay법으로 실시하였다.(Culture of animal cells, R. Ian Freshney, Wiley-Liss) 결과는표 2에 나타내었다.Toxicity experiments of hyaluronic acid obtained by the extraction method of the present invention was carried out by MTT assay using the cell MG63. (Culture of animal cells, R. Ian Freshney, Wiley-Liss) The results are shown in Table 2 .
상기표 2에서 보는 바와 같이, 히아루론산의 농도별로 검토한 MG63이 점점 증가하여 이 결과로 부터 세포독성이 발현되지 않아 인체 내 주입 물질로 사용 가능함을 나타내었다.As shown in Table 2 , the MG63 examined for each concentration of hyaluronic acid is gradually increased, indicating that the cytotoxicity is not expressed from this result, so that it can be used as an injection material in the human body.
<실험예 3> 히아루론산의 순도 측정Experimental Example 3 Measurement of Purity of Hyaluronic Acid
본 발명의 추출방법에 얻어진 히아루론산의 순도를 측정하기 위하여, 히아루론산 분석, 단백질 함량 측정 및 헥산을 측정하였다. 구체적으로, 히아론산 분석은 변형된 카바졸 방법( Nelly Blumankrantz-abose-hansen, Analytical Biochemistry 1973)을 사용하였고, 단백질 함량은 공지의 방법인 Bio-Rad BCA microassay법을 이용하였고, 헥산은 260 nm의 파장에서 흡광도로 측정하였다.In order to measure the purity of hyaluronic acid obtained in the extraction method of the present invention, hyaluronic acid analysis, protein content measurement and hexane were measured. Specifically, the hyaluronic acid analysis was performed using a modified carbazole method (Nelly Blumankrantz-abose-hansen, Analytical Biochemistry 1973), the protein content was a known method Bio-Rad BCA microassay method, hexane was 260 nm Absorbance was measured at the wavelength.
상기표 3에서 보는 바와 같이, 단백질 함량이 0.08% 미만이고, 260 ㎚의 파장에서 측정한 헥산의 흡광도는 0.00±0.01로 나타났다. 이는 본 발명의 닭벼슬로부터 추출된 히아루론산의 순도가 우수함을 나타내고 있음을 알 수 있다.As shown in Table 3 , the protein content is less than 0.08%, the absorbance of hexane measured at a wavelength of 260 nm was 0.00 ± 0.01. It can be seen that the purity of hyaluronic acid extracted from the chicken shackle of the present invention is excellent.
상술한 바와 같이, 본 발명의 닭벼슬로부터 염과 클로로포름을 이용하여 고순도의 히아루론산을 추출 방법은 기존의 미생물을 이용한 히아루론산 추출방법과 달리하여 생체 조직 내에 히아루론산과 단백질 사이의 이온 결합을 형성하고 있는 점을 착안하여 용액 상에서 첨가하는 염의 이온세기를 변화시켜 생산 수율을 높이고 추출과정을 단순화하여 고순도의 히아루론산을 추출할 수 있다.As described above, the method of extracting high purity hyaluronic acid from salt of chicken and salt of chloroform of the present invention is different from the conventional method of extracting hyaluronic acid using microorganisms to form an ionic bond between hyaluronic acid and protein in biological tissues. The hyaluronic acid of high purity can be extracted by changing the ionic strength of the salt added in solution, increasing the production yield and simplifying the extraction process.
한편, 히아루론산은 생체조직에서 추출할 경우 제조 원가가 매우 높다는 단점이 있었는데 현재 폐기물로 처리되고 있는 닭벼슬을 폐기물의 재활용 차원에서 고부가가치 물질을 추출하여 활용시 원가 절감뿐만 아니라 미개발 자원의 효율적 이용 측면에서 이용가치를 높일 수 있으며, 기존의 생체재료에서 추출한 히알우론산의 낮은 면역성 및 불순물의 혼합 등의 문제점을 해결하여 고순도의 히알우론산을 얻을 수 있다. 또한, 닭 벼슬에서 고순도, 히아루론산을 추출하여 사용시 미생물에서 추출한 히알우론산보다 물리적 안정성이 높고 면역 반응에서 안정성이 우수한 관절염 치료제, 안과용 보충제와 같은 의용 재료 또는 특이한 보습력이 있는 화장품 원료로서 응용 가능하다.On the other hand, hyaluronic acid has a disadvantage in that the manufacturing cost is very high when extracted from biological tissues. In addition, cost reduction and efficient use of undeveloped resources are made by extracting high value-added materials from chicken rice which is currently treated as waste in order to recycle waste. In order to increase the value of use, it is possible to obtain high purity hyaluronic acid by solving problems such as low immunity and mixing of impurities of hyaluronic acid extracted from existing biomaterials. In addition, high purity, hyaluronic acid extracted from the chicken shackles when using the physical stability than the hyaluronic acid extracted from microorganisms and excellent in stability in the immune response can be applied as a medical material such as arthritis treatments, ophthalmic supplements or cosmetic ingredients with a specific moisturizing power.
Claims (4)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020020053546A KR20040022760A (en) | 2002-09-05 | 2002-09-05 | Method for extracting hight purified hyaluronic acid from cocks comb |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020020053546A KR20040022760A (en) | 2002-09-05 | 2002-09-05 | Method for extracting hight purified hyaluronic acid from cocks comb |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20040022760A true KR20040022760A (en) | 2004-03-18 |
Family
ID=37326482
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020020053546A KR20040022760A (en) | 2002-09-05 | 2002-09-05 | Method for extracting hight purified hyaluronic acid from cocks comb |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR20040022760A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102627702A (en) * | 2012-04-24 | 2012-08-08 | 哈尔滨工业大学 | Extraction method of squid hyaluronic acid |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4713448A (en) * | 1985-03-12 | 1987-12-15 | Biomatrix, Inc. | Chemically modified hyaluronic acid preparation and method of recovery thereof from animal tissues |
| KR19980080116A (en) * | 1997-03-27 | 1998-11-25 | 히라따 다다시 | Purification Method of Sodium Hyaluronate |
| US20020120132A1 (en) * | 2000-12-15 | 2002-08-29 | Al Prescott | Method for purifying high molecular weight hyaluronic acid |
| KR20020071256A (en) * | 2001-03-05 | 2002-09-12 | (주)신스바이오 | A manufacturing method of hyaluronic acid |
| KR20020071772A (en) * | 2001-03-06 | 2002-09-13 | (주)신스바이오 | Manufacturing method of hyaluronic acid to have sterilization activity |
-
2002
- 2002-09-05 KR KR1020020053546A patent/KR20040022760A/en not_active Application Discontinuation
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4713448A (en) * | 1985-03-12 | 1987-12-15 | Biomatrix, Inc. | Chemically modified hyaluronic acid preparation and method of recovery thereof from animal tissues |
| KR19980080116A (en) * | 1997-03-27 | 1998-11-25 | 히라따 다다시 | Purification Method of Sodium Hyaluronate |
| US20020120132A1 (en) * | 2000-12-15 | 2002-08-29 | Al Prescott | Method for purifying high molecular weight hyaluronic acid |
| KR20020071256A (en) * | 2001-03-05 | 2002-09-12 | (주)신스바이오 | A manufacturing method of hyaluronic acid |
| KR20020071772A (en) * | 2001-03-06 | 2002-09-13 | (주)신스바이오 | Manufacturing method of hyaluronic acid to have sterilization activity |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102627702A (en) * | 2012-04-24 | 2012-08-08 | 哈尔滨工业大学 | Extraction method of squid hyaluronic acid |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| ES2811101T3 (en) | Procedure for the Preparation and Purification of Hyaluronic Acid Sodium Salt | |
| AU756864B2 (en) | Sulfated saccharides | |
| ES2329731T3 (en) | DERIVATIVES OF ARIAL HIALURONIC ACID / SULFONA VINYL RENTAL. | |
| WO2004080388A2 (en) | Preparation of hyaluronic acid from eggshell membrane | |
| US20220304355A1 (en) | Identification and selection of a plant starting material of a plant chondroitin sulfate and hyaluronic acid, and transformation of such plant starting material to obtain ingredients for use in foods, supplements, medical devices or drugs | |
| KR20000064523A (en) | Oligosulfate Hyaluronic Acid | |
| NZ585699A (en) | Mixed butyric-formic esters of acid polysaccharides, and their preparation and use as skin cosmetics | |
| KR101132114B1 (en) | Method of molecular weight control of hyaluronic acid | |
| CN116003577A (en) | Recombinant collagen for skin soothing and repairing and application thereof | |
| US9598505B2 (en) | Silyl-derivatives of polysaccharides | |
| IL229432A (en) | Shark-like chondroitin sulphate and process for the preparation thereof | |
| KR20040022760A (en) | Method for extracting hight purified hyaluronic acid from cocks comb | |
| Hiramatsu et al. | Acid mucopolysaccharides in porcine gingiva | |
| KR100577075B1 (en) | Method for purifying hyaluronic acid using calcium salt and phosphate salt, or calcium phosphate salt | |
| EP4162062A1 (en) | Process for extracting a hyaluronic acid from a fungus, a hyaluronic acid of plant origin and use thereof | |
| JP3433989B2 (en) | Cosmetic raw material and method for producing the same | |
| KR100236766B1 (en) | Purification method for hyaluronic acid and its salts | |
| JPH0782132A (en) | Active oxygen suppressing agent | |
| JP3762234B2 (en) | Sulfated polysaccharide cross-linked with polycationic substance and process for producing the same | |
| CN110964100B (en) | Method for extracting high-purity phycocyanin and co-producing polysaccharide from spirulina | |
| JPH06219954A (en) | Hyaluronidase inhibitor | |
| CA2009730A1 (en) | Cosmetic formulations and isolation and purification of fractions of sodium hyaluronate used in cosmetic formulations | |
| CN116687959A (en) | Chondroitin sulfate biological polyamine complex, and preparation method and application thereof | |
| JPH06279255A (en) | Hyaluronidase inhibitor | |
| CZ202497A3 (en) | A portable device for measuring the impedance spectrum of steels and a measuring method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E601 | Decision to refuse application |