KR20180119936A - A manufacturing method of acetylation hyaluronic acid comprising activation process - Google Patents
A manufacturing method of acetylation hyaluronic acid comprising activation process Download PDFInfo
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Abstract
Description
본 발명은 반응속도 및 수율을 향상시키는 아세틸화 히알루론산의 제조 방법에 관한 것으로서, 구체적으로 히알루론산을 팽윤(swelling)시키는 단계 및 상기 팽윤된 히알루론산에 아세틸화제를 첨가하여 히알루론산을 아세틸화하는 단계를 포함하는 아세틸화 히알루론산의 제조방법에 관한 것이다.The present invention relates to a method for producing acetylated hyaluronic acid which improves the reaction rate and yield, and more particularly, to a method for producing hyaluronic acid by swelling hyaluronic acid and a method of acetylating hyaluronic acid by adding an acetylating agent to the swelled hyaluronic acid To a process for preparing acetylated hyaluronic acid.
히알루론산(Hyaluronic acid)은 분자량이 500,000 내지 13,000,000 Da에 이르는 무색의 고점도 다당류로서, 글루쿠론산과 N-아세틸 글루코오스아민이 (1-3)과 (1-4)로 결합된 반복단위를 가진다. 또한, 히알루론산은 보습효과를 가져 화장품 첨가제로 많이 사용되며, 히알루론산의 농도가 높을수록 보습력이 증가하게 된다. Hyaluronic acid is a colorless, highly viscous polysaccharide having a molecular weight of 500,000 to 13,000,000 Da. Glucuronic acid and N-acetylglucosamine have repeating units bonded with (1-3) and (1-4). In addition, hyaluronic acid is used as a cosmetic additive because of its moisturizing effect, and the higher the concentration of hyaluronic acid, the more the moisture is increased.
그러나 히알루론산은 친수성이 높은 물질로 이를 피부에 도포하였을 때, 소수성인 피부에 완벽히 부착되지 못하여 보습효과를 충분히 나타내지 못하는 단점이 있다. 위의 단점을 개선하기 위해 히알루론산에 아세틸화 반응을 가하여 친수/소수 정도를 조절함으로써, 보습효과가 개선된 아세틸화 히알루론산을 제조하는 연구가 진행되었다. 그러나 기존의 아세틸화 히알루론산 제조 기술은 반응이 일어나는 장소인 히알루론산의 하이드록시기(-OH)에 아세틸화제가 쉽게 접근하지 못하여 반응이 원활히 진행되지 않는 어려움이 있다. 이로 인해 히알루론산의 아세틸화 반응 속도 및 수율이 낮아 이를 개선하기 위한 공정 개발이 필요한 실정이다.However, hyaluronic acid is a highly hydrophilic substance, and when it is applied to the skin, it is not adhered perfectly to the hydrophobic skin and has a disadvantage that it can not sufficiently show the moisturizing effect. In order to improve the above disadvantages, researches have been conducted to prepare acetylated hyaluronic acid having improved moisturizing effect by controlling the degree of hydrophilicity / hydrophobicity by applying an acetylation reaction to hyaluronic acid. However, conventional acetylated hyaluronic acid production technology has a difficulty in that the reaction can not proceed smoothly because the acetylating agent can not easily approach the hydroxyl group (-OH) of hyaluronic acid, which is the place where the reaction takes place. As a result, the acetylation reaction rate and yield of hyaluronic acid are low, and a process for improving the process is required.
한편, 아세틸화 히알루론산을 저분자화하여 피부로의 흡수율을 높이려는 시도는 개시되어 있으나(한국공개특허 10-2015-0015209), 히알루론산에 활성화 공정을 가하여 팽윤(swelling)시키는 단계를 통해 히알루론산의 아세틸화도 및 수득률을 높이려는 연구는 아직 없는 실정이다. On the other hand, attempts have been made to increase the absorption rate of acetylated hyaluronic acid into the skin by lowering the molecular weight of the hyaluronic acid (Korean Patent Laid-open No. 10-2015-0015209), but the hyaluronic acid is swelled by an activation process, There has been no research to increase the degree of acetylation and the yield of the enzyme.
상기와 같은 배경 하에, 본 발명자들은 히알루론산의 아세틸화 반응 전 활성화 공정을 통해 히알루론산을 팽윤(swelling)시킴으로써 아세틸화제의 접근을 용이하게 하여 아세틸화 반응 속도 및 수율을 현저히 개선할 수 있음을 규명함으로써, 본 발명을 완성하였다. Under the above background, the present inventors have found that the swelling of hyaluronic acid through the activation process before the acetylation reaction of hyaluronic acid facilitates the access of the acetylating agent, thereby remarkably improving the acetylation reaction rate and yield Thereby completing the present invention.
본 발명의 목적은 히알루론산을 팽윤(swelling)시키는 단계 및 히알루론산을 아세틸화하는 단계를 포함하는 아세틸화 히알루론산의 제조방법을 제공하는 것이다. It is an object of the present invention to provide a method for producing acetylated hyaluronic acid comprising swelling hyaluronic acid and acetylating hyaluronic acid.
상기 목적을 달성하기 위한 양태로서, 본 발명은 히알루론산을 팽윤(swelling)시키는 단계 및 상기 팽윤된 히알루론산에 아세틸화제를 첨가하여 히알루론산을 아세틸화하는 단계를 포함하는, 아세틸화 히알루론산의 제조방법을 제공한다. In order to achieve the above object, the present invention provides a method for producing a hyaluronic acid, comprising swelling hyaluronic acid and acetylating hyaluronic acid by adding an acetylating agent to the swelled hyaluronic acid, ≪ / RTI >
이하, 본 발명의 제조방법을 구체적으로 설명한다.Hereinafter, the production method of the present invention will be described in detail.
본 발명은 아세틸화 히알루론산을 제조하는데 있어서, 히알루론산의 아세틸화 반응 전에 히알루론산을 활성화시키는 활성화 공정을 포함하는 것을 특징으로 한다. The present invention is characterized by comprising an activation step of activating hyaluronic acid before the acetylation reaction of hyaluronic acid in producing acetylated hyaluronic acid.
상기 활성화 공정은 히알루론산을 팽윤(swelling) 시키는 단계를 포함한다. The activation process comprises swelling hyaluronic acid.
기존의 아세틸화 히알루론산의 제조 기술은 아세틸화 반응이 일어나는 장소인 히알루론산의 하이드록시기(-OH)에 아세틸화제가 쉽게 접근하지 못하여 반응속도가 느리고, 아세틸화 히알루론산의 수율이 낮은 문제점이 있었다. Conventional production technology of acetylated hyaluronic acid has a problem that the acetylation agent is not easily accessible to the hydroxyl group (-OH) of hyaluronic acid, which is the place where the acetylation reaction takes place, so that the reaction rate is slow and the yield of acetylated hyaluronic acid is low there was.
이에 본 발명에서는 히알루론산을 아세틸화 하기 전에 히알루론산을 팽윤시켜 아세틸화제가 히알루론산의 하이드록시기에 쉽게 접근하게 하도록 하였다. Therefore, in the present invention, hyaluronic acid is swelled before acetylation of hyaluronic acid so that the acetylating agent easily accesses the hydroxyl group of hyaluronic acid.
본 발명의 용어 "히알루론산"은 β-D-N-아세틸글루코사민(N-Acetylglucosamine)과 β-D-글루쿠론산(Glucuronic Acid)이 교대로 결합된 직쇄상의 고분자로, 본 발명에서는 히알루론산 자체, 이의 염 또는 이들의 조합을 모두 포함하는 의미로 사용될 수 있다. 상기 히알루론산의 분자량은 100,000 내지 5,000,000 Da일 수 있으나 이에 제한되지 않는다. The term "hyaluronic acid" of the present invention is a linear polymer in which β-DN-acetylglucosamine and β-D-glucuronic acid are alternately bonded. In the present invention, hyaluronic acid itself, A salt thereof, or a combination thereof. The molecular weight of the hyaluronic acid may be 100,000 to 5,000,000 Da, but is not limited thereto.
본 발명에서, 상기 히알루론산의 염은 히알루론산 나트륨, 히알루론산 칼륨, 히알루론산 칼슘, 히알루론산 마그네슘, 히알루론산 아연, 히알루론산 코발트 등의 무기염과, 히알루론산 테트라부틸암모늄 등의 유기염이 모두 포함될 수 있다. In the present invention, the salt of hyaluronic acid is preferably an inorganic salt such as sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronate, and organic salts such as tetrabutylammonium hyaluronate .
본 발명에서, 히알루론산은 히알루론산 자체 또는 이의 염을 단독으로, 또는 히알루론산 자체 또는 이의 염을 2종 이상 조합하여 사용할 수 있다. 상기 히알루론산 또는 이의 염은 미생물로부터 분리되거나, 합성, 또는 구입한 것일 수 있으나 이에 제한되지 않는다. 일 예로 상기 히알루론산은 스트렙토코커스(Streptococcus) 속 미생물(Streptocossus equi, Streptococcus zooepidemicus)에서 분리, 정제된 것일 수 있다.In the present invention, hyaluronic acid may be used alone or in combination with hyaluronic acid itself or a salt thereof, or hyaluronic acid itself or a combination of two or more thereof. The hyaluronic acid or a salt thereof may be isolated from a microorganism, synthesized, or purchased, but is not limited thereto. For example, the hyaluronic acid may be one isolated from a Streptococcus sp. Strain (Streptococcus zooepidemicus).
본 발명에서, 히알루론산의 팽윤(swelling)은 히알루론산의 사슬이 풀어져서 아세틸화제의 접근을 용이하게 하는 것일 수 있다.In the present invention, the swelling of hyaluronic acid may be such that the chain of hyaluronic acid is loosened to facilitate access of the acetylating agent.
본 발명의 히알루론산을 팽윤시키는 단계는 히알루론산에 촉매 흡착 공정 또는 공융 용매를 첨가하는 것일 수 있고, 상기 촉매 흡착 공정은 히알루론산에 산촉매를 첨가하는 것일 수 있다. The step of swelling the hyaluronic acid of the present invention may be a step of adding a catalyst adsorption process or a eutectic solvent to hyaluronic acid, and the step of adsorbing the catalyst may include adding an acid catalyst to hyaluronic acid.
구체적으로 상기 산촉매는 황산을 첨가하는 것일 수 있고, 상기 황산은 진한 황산으로 96% 이상, 보다 구체적으로 98% 황산일 수 있다. Specifically, the acid catalyst may be sulfuric acid, and the sulfuric acid may be 96% or more, more specifically 98% sulfuric acid as concentrated sulfuric acid.
본 발명에서 히알루론산을 팽윤시키는 단계는 히알루론산에 산촉매 또는 공융 용매를 첨가하여 교반하는 단계로서, 아세틸화 반응 전에 일어나는 것을 특징으로 한다. In the present invention, the step of swelling hyaluronic acid is a step of adding an acid catalyst or a eutectic solvent to hyaluronic acid and stirring, which is characterized in that it occurs before the acetylation reaction.
본 발명에서 촉매 흡착 공정에 사용되는 산촉매의 양은 히알루론산 대비 5 내지 30 중량% 포함될 수 있다. 5 중량% 미만의 경우 흡착되는 촉매의 양이 적어 용매에 대한 용해도 증진을 기대할 수 없으며, 30 중량% 초과이면 히알루론산의 분해가 상당히 진행되는 단점이 있을 수 있다.In the present invention, the amount of the acid catalyst used in the catalyst adsorption process may be 5 to 30 wt% relative to hyaluronic acid. If the amount is less than 5% by weight, the amount of the adsorbed catalyst is small, and the solubility in the solvent is not expected to increase. If the amount is more than 30% by weight, the degradation of hyaluronic acid may be considerably accelerated.
상기 황산 촉매를 이용한 활성화 공정은 아세트산에 분산된 히알루론산에 황산 촉매를 첨가함으로써 히알루론산의 하이드록시기(-OH)를 술폰산기(-OSO3H)로 치환하여, 용매에 대한 히알루론산의 용해도를 높이고 히알루론산 분자간 수소결합을 끊어 아세틸화제의 접근을 용이하게 함으로써 아세틸화 히알루론산의 수율 및 반응속도가 향상된다.In the activation process using the sulfuric acid catalyst, the hydroxyl group (-OH) of hyaluronic acid is substituted with a sulfonic acid group (-OSO 3 H) by adding a sulfuric acid catalyst to hyaluronic acid dispersed in acetic acid, and the solubility of hyaluronic acid And the hyaluronic acid intermolecular hydrogen bond is broken to facilitate the access of the acetylating agent, thereby improving the yield and the reaction rate of the acetylated hyaluronic acid.
본 발명의 용어 “공융 용매”는 2개 이상 용매의 혼합물을 만들기 위해 제1용매에 추가로 첨가되는 용매를 의미한다. 구체적으로 제1용매는 아세트산일 수 있고, 공융 용매는 하이드로카본(hydrocarbon)류를 제외한 물, 메탄올, 에탄올을 포함한 알코올류, 또는 테트라하이드로퓨란(THF)을 포함하는 비양성자성(aprotic) 용매일 수 있고, 보다 구체적으로 물일 수 있으나 이에 제한되지 않는다.The term " eutectic solvent " of the present invention means a solvent that is added to the first solvent to make a mixture of two or more solvents. Specifically, the first solvent may be acetic acid, and the eutectic solvent may be water, methanol, alcohol including alcohols or tetrahydrofuran (THF) for aprotic days, such as hydrocarbons, And, more specifically, but not exclusively, water.
본 발명에서 공융 용매 첨가 공정에 사용되는 용매의 양은 히알루론산 대비 5 내지 30 중량% 포함될 수 있다. 5 중량% 미만의 경우 용매의 양이 적어 히알루론산의 사슬이 풀어지는 효과를 기대하기 어려우며, 30 중량% 초과이면 물에 의해 많은 양의 아세트산무수물이 분해되어 아세트산무수물의 양이 추가로 첨가되어야 하는 단점이 있을 수 있다. In the present invention, the amount of the solvent used in the eutectic solvent addition step may be 5 to 30 wt% based on hyaluronic acid. If the amount is less than 5% by weight, the effect of loosening the chain of hyaluronic acid is not expected to be expected, and if it is more than 30% by weight, a large amount of acetic anhydride is decomposed by water and an additional amount of acetic anhydride is added There may be disadvantages.
상기 공융 용매 첨가를 이용한 활성화 공정은 히알루론산에 용매를 첨가함으로써 히알루론산 사슬을 풀어지게 하여, 용매 및 아세틸화제의 확산을 용이하게 함으로써 아세틸화 히알루론산의 수율 및 반응속도가 향상된다.In the activation process using the eutectic solvent addition, a solvent is added to the hyaluronic acid so that the hyaluronic acid chain is released, and the solvent and the acetylating agent are easily diffused, thereby improving the yield and the reaction rate of the acetylated hyaluronic acid.
본 발명의 구체적인 실시예에 따르면 아세틸화 반응에 앞서 히알루론산에 황산 또는 물을 첨가하여 팽윤시킨 경우, 아세틸화 반응이 더욱 원활하게 일어나고 반응 속도가 빨라짐을 확인하였다(도 2 및 도 4). 특히 아세틸화 반응 전 히알루론산을 팽윤시키는 활성화 공정이 없는 경우, 아세틸화 반응이 거의 일어나지 않거나 24시간이 지나도 반응이 일어나지 않았으나, 본 발명의 방법에 의해 10분 만에 높은 수율로 아세틸화 히알루론산을 제조할 수 있음을 확인하였다.According to a specific example of the present invention, it was confirmed that when the hyaluronic acid was swelled by adding sulfuric acid or water to the hyaluronic acid prior to the acetylation reaction, the acetylation reaction occurred more smoothly and the reaction rate became faster (FIGS. 2 and 4). In particular, in the absence of the activation step of swelling the hyaluronic acid before the acetylation reaction, the acetylation reaction hardly occurred or the reaction did not occur even after 24 hours, but the acetylated hyaluronic acid was obtained at a high yield in 10 minutes by the method of the present invention .
본 발명은 상기 팽윤된 히알루론산에 아세틸화제를 첨가하여 히알루론산을 아세틸화하는 단계를 포함한다.The present invention includes a step of acetylating hyaluronic acid by adding an acetylating agent to the swelled hyaluronic acid.
본 발명의 아세틸화 히알루론산 제조방법은 아세틸화 히알루론산의 수율 및 반응속도를 향상시킴으로써 대량 생산에 용이하고 제조 경쟁력을 확보하는 효과가 있다. The method for producing acetylated hyaluronic acid of the present invention has an effect of improving the yield and the reaction rate of acetylated hyaluronic acid, thereby facilitating mass production and securing manufacturing competitiveness.
도 1은 히알루론산이 활성화 공정을 거쳐 팽윤되는 것을 나타낸 것으로, 활성화 공정 이전 바닥에 침전된 것 및 활성화 공정 이후 용매에 팽윤되어 섞인 것을 나타낸 것이다.
도 2는 실시예 1 및 비교예 1의 히알루론산의 아세틸화 반응 결과 (a)색 변화 및 (b)성상을 나타낸 것이다.
도 3은 실시예 1 및 비교예 1의 히알루론산의 아세틸화 반응 경과에 따른 용액의 투명도 변화를 나타낸 것이다.Figure 1 shows that hyaluronic acid is swollen through the activation process, indicating that the hyaluronic acid has precipitated on the bottom prior to the activation process and that it is swollen and mixed with the solvent after the activation process.
Fig. 2 shows (a) color change and (b) characteristics of the acetylation reaction of hyaluronic acid in Example 1 and Comparative Example 1.
FIG. 3 shows the transparency of the solution according to the progress of the acetylation reaction of hyaluronic acid in Example 1 and Comparative Example 1. FIG.
이하, 실시예를 통하여 본 발명을 보다 상세히 설명하고자 한다. 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.
비교예Comparative Example 1: One: 아세틸화Acetylation 히알루론산의 제조 Production of hyaluronic acid
히알루론산 하이드록시기(-OH)를 아세틸화 반응을 통해 아세틸기로 치환하여 아세틸화 히알루론산을 제조하였다. 구체적으로, 20 ml 아세트산(acetic acid)에 2g의 히알루론산나트륨(sodium hyaluronate)을 첨가하고, 이어서 80 ml의 아세트산무수물(acetic anhydride)을 넣고 촉매인 진한 황산(98%)을 2ml를 추가로 용액에 천천히 투입하여 아세틸화 반응을 진행하였다. 반응 종결 후 물에 천천히 첨가하여 2번 수세 후 여과하여 아세틸화 히알루론산을 수득하였다. Acylated hyaluronic acid was prepared by replacing the hyaluronic acid hydroxyl group (-OH) with an acetyl group through an acetylation reaction. Specifically, 2 g of sodium hyaluronate was added to 20 ml of acetic acid, then 80 ml of acetic anhydride was added, and 2 ml of concentrated sulfuric acid (98%) as a catalyst was further added to the solution To proceed the acetylation reaction. After completion of the reaction, the reaction mixture was slowly added to water, washed twice with water and filtered to obtain acetylated hyaluronic acid.
실시예Example 1: 활성화 공정을 포함하는 1: containing the activation process 아세틸화Acetylation 히알루론산의 제조 Production of hyaluronic acid
활성화 공정을 통해 히알루론산을 팽윤(swelling)시킨 후 아세틸화 반응을 진행하여 아세틸화 히알루론산을 제조하였다. After the hyaluronic acid was swelled through the activation process, the acetylation reaction proceeded to produce acetylated hyaluronic acid.
1-1. 촉매흡착 단계를 포함하는 아세틸화 히알루론산의 제조1-1. Preparation of acetylated hyaluronic acid including catalyst adsorption step
20 ml 아세트산에 2g의 히알루론산나트륨 및 0.2 ml의 진한 황산(98%)을 첨가하고 교반하여 활성화 공정을 진행하였다. 활성화 공정이 완료된 후 반응 용액에 80 ml의 아세트산무수물을 넣고 1.8 ml 진한 황산(98%)을 추가로 용액에 천천히 첨가하여 아세틸화 반응을 진행하였다. 반응 종결 후 물에 천천히 첨가하여 2번 수세 후 침전물을 여과한 뒤 진공 건조하여 흰색 파우더 형태의 아세틸화 히알루론산을 수득하였다.2 g of sodium hyaluronate and 0.2 ml of concentrated sulfuric acid (98%) were added to 20 ml of acetic acid and stirred to conduct the activation process. After the activation process was completed, 80 ml of acetic anhydride was added to the reaction solution, and 1.8 ml of concentrated sulfuric acid (98%) was slowly added to the solution to conduct the acetylation reaction. After completion of the reaction, the reaction mixture was slowly added to water and the mixture was washed twice. The precipitate was filtered and vacuum-dried to obtain acetylated hyaluronic acid in the form of a white powder.
1-2. 공융 용매 첨가 단계를 포함하는 아세틸화 히알루론산의 제조1-2. Preparation of acetylated hyaluronic acid including eutectic solvent addition step
20 ml 아세트산에 2g의 히알루론산나트륨 및 1g의 공융 용매를 첨가하고 교반하여 활성화 공정을 진행하였다. 활성화 공정이 완료된 후 반응 용액에 80 ml의 아세트산무수물을 넣고 2 ml 진한 황산(98%)을 추가로 용액에 천천히 첨가하여 아세틸화 반응을 진행하였다. 반응 종결 후 물에 천천히 첨가하여 2번 수세 후 침전물을 여과한 뒤 진공 건조하여 흰색 파우더 형태의 아세틸화 히알루론산을 수득하였다.2 g of sodium hyaluronate and 1 g of eutectic solvent were added to 20 ml of acetic acid and stirred to carry out the activation process. After the activation process was completed, 80 ml of acetic anhydride was added to the reaction solution, and 2 ml of concentrated sulfuric acid (98%) was slowly added to the solution to conduct the acetylation reaction. After completion of the reaction, the reaction mixture was slowly added to water and the mixture was washed twice. The precipitate was filtered and vacuum-dried to obtain acetylated hyaluronic acid in the form of a white powder.
히알루론산이 활성화 공정을 거치면서 어떠한 형태의 변화가 일어나는지 육안으로 관찰하였다. 활성화 공정 이전에는 히알루론산이 용매인 아세트산에 침전된 형태로 존재하다가 활성화 공정을 거친 후 침전물이 사라지고 용매 내에서 팽윤(swelling)되는 것을 확인할 수 있었다(도 1).Hyaluronic acid was visually observed as it underwent the activation process. Before the activation process, it was confirmed that hyaluronic acid was present in a form precipitated in acetic acid as a solvent. After the activation process, the precipitate disappeared and swelled in the solvent (FIG. 1).
활성화 공정 적용에 따른 색 변화 및 성상 비교Comparison of color change and property according to activation process application
활성화 공정의 적용을 통한 히알루론산의 팽윤(swelling) 효과가 히알루론산의 아세틸화 반응에 미치는 영향을 확인하였다. 히알루론산의 아세틸화 반응이 진행됨에 따라 소수성이 증가, 용매인 아세트산에 대한 용해도가 증가하여 용액이 투명해질 것이므로, 비교예 1과 실시예 1의 색 변화 및 분리·정제 후 수득한 성상을 비교하여 하기의 표 1에 기재하였다.The effect of swelling of hyaluronic acid on the acetylation reaction of hyaluronic acid was confirmed by the application of the activation process. As the acetylation reaction of hyaluronic acid proceeds, the hydrophobicity increases and the solubility to acetic acid, which is a solvent, increases, so that the solution becomes transparent. Therefore, the color change of Comparative Example 1 and Example 1 are compared with those obtained after separation and purification Are shown in Table 1 below.
도 2는 히알루론산의 아세틸화 반응 결과 용액의 색 변화 및 성상을 관찰한 것이다. 활성화 공정, 즉 팽윤단계를 거치지 않은 비교예 1은 아세틸화 반응 후에도 아세틸화 반응이 잘 일어나지 않아 색 변화가 없는 반면, 활성화 공정을 거친 실시예 1의 경우, 아세틸화 반응이 진행되면서 반응 용액이 투명해지는 것을 관찰할 수 있었다(도 2-a). FIG. 2 shows the color change and properties of the solution as a result of the acetylation reaction of hyaluronic acid. In Comparative Example 1, which did not undergo the activation process, that is, the swelling step, the acetylation reaction did not occur well and the color did not change after the acetylation reaction. On the other hand, in the case of Example 1 in which the activation process was performed, (Fig. 2-a).
활성화 공정을 거친 실시예 1에서는 흰색 파우더(powder) 형태의 아세틸화 히알루론산을 얻을 수 있었지만, 활성화 공정을 거치지 않은 비교예 1의 경우 히알루론산이 물에 용해되어 겔(gel)형태의 아세틸화 히알루론산을 수득하였다(도 2-b).Acetylated hyaluronic acid in the form of a white powder was obtained in Example 1 after the activation process, but in Comparative Example 1 in which the activation process was not performed, hyaluronic acid was dissolved in water to form an acetylated hyaluronic acid (Fig. 2-b).
이와 같은 결과는, 아세틸화 반응 전 히알루론산을 팽윤시켜 활성화 공정을 거친 경우 아세틸화 반응이 더욱 원활하게 일어남을 뒷받침한다. These results support that the acetylation reaction occurs more smoothly when the hyaluronic acid is swelled before the acetylation reaction and then activated.
활성화 공정 적용에 따른 수득률 및 표면장력 비교Comparison of yield rate and surface tension according to activation process
활성화 공정의 적용에 따른 아세틸화 히알루론산의 수득률 및 표면장력을 비교하였다. The yield and surface tension of acetylated hyaluronic acid according to the application of the activation process were compared.
상기 결과를 통해 활성화 공정을 거친 실시예 1의 아세틸화 히알루론산은 화장품에 일반적으로 사용되는 아세틸화 히알루론산(표면장력 55.0 mN/m, J.Cosmetic.Sci, 50, 171-184, 1999 참조)과 비교할 때, 표면장력 등의 성질이 유사한 것으로 확인되었다. 즉, 아세틸화 반응 전 활성화 공정은 반응 결과로 수득하는 아세틸화 히알루론산의 성질에 영향을 미치지 않음을 뒷받침한다.The acetylated hyaluronic acid of Example 1 which had been subjected to the activation process through the above results was found to be acetylated hyaluronic acid (surface tension 55.0 mN / m, see J. Cosmetic. Sci, 50, 171-184, 1999) , It was confirmed that the properties such as surface tension were similar. That is, the activation process before the acetylation reaction does not affect the properties of the acetylated hyaluronic acid obtained as a result of the reaction.
활성화 공정 적용에 따른 Depending on the activation process application 아세틸화Acetylation 반응 속도 비교 Comparison of reaction rate
히알루론산의 아세틸화 반응 시작 후 반응 용액이 투명해 지는 시간을 측정함으로써, 활성화 공정 적용에 따른 아세틸화 반응 속도를 비교하였다.The rate of acetylation reaction according to the application of the activation process was compared by measuring the time that the reaction solution became transparent after starting the acetylation reaction of hyaluronic acid.
실시예 1의 경우 아세틸화 반응 시작 후 10분이 지나면서부터 용액이 투명해지는 것을 육안으로 확인할 수 있었다. 반면 비교예 1에서는 반응 시작 후 24시간이 지나도 용액이 투명해지지 않는 것을 확인함으로써, 반응속도가 매우 느리고 거의 반응이 진행되지 않음을 알 수 있었다(도 3).In Example 1, it was visually confirmed that the solution became transparent from 10 minutes after the start of the acetylation reaction. On the other hand, in Comparative Example 1, it was confirmed that the solution did not become transparent even after 24 hours from the start of the reaction, so that the reaction rate was very slow and the reaction hardly proceeded (FIG. 3).
이와 같은 결과는 활성화 공정을 거친 히알루론산은 아세틸화 반응 속도가 빨라진다는 것을 뒷받침하는 것이다. These results support the fact that hyaluronic acid after the activation process accelerates the acetylation reaction rate.
Claims (9)
상기 팽윤된 히알루론산에 아세틸화제를 첨가하여 히알루론산을 아세틸화하는 단계를 포함하는, 아세틸화 히알루론산의 제조방법.
Swelling hyaluronic acid;
And adding acetylating agent to the swelled hyaluronic acid to acetylate hyaluronic acid.
The method of claim 1, wherein the swelling enhances the solubility of hyaluronic acid and facilitates access of the acetylating agent to hyaluronic acid.
The method according to claim 1, wherein the swelling step comprises adding an acid catalyst or a eutectic solvent to hyaluronic acid.
4. The method according to claim 3, wherein the acid catalyst is sulfuric acid.
4. The method according to claim 3, wherein the eutectic solvent is at least one solvent selected from the group consisting of water, methanol, ethanol, and an aprotic solvent.
5. The method according to claim 4, wherein the sulfuric acid is obtained by substituting a hydroxyl group (-OH) of a hyaluronic acid with a sulfonic acid group (-OSO 3 H) to increase the solubility of hyaluronic acid and breaking the hydrogen bond between hyaluronic acid molecules, Lt; / RTI >
6. The method of claim 5, wherein the eutectic solvent is used in combination with a first solvent to increase the solubility of hyaluronic acid in the first solvent to break the hyaluronic acid intermolecular hydrogen bond to facilitate access of the acetylating agent.
The manufacturing method according to claim 3, wherein the swelling step comprises adding an acid catalyst or a eutectic solvent to hyaluronic acid and stirring.
The process according to claim 1, wherein the acetylating agent is acetic anhydride.
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| JPH069707A (en) * | 1992-04-21 | 1994-01-18 | Shiseido Co Ltd | Highly acetylated hyaluronic acid and production thereof |
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| JP2004010622A (en) * | 2002-06-03 | 2004-01-15 | National Institute Of Advanced Industrial & Technology | Method for acylating polysaccharide substance |
| KR20160067887A (en) * | 2013-10-08 | 2016-06-14 | 큐피가부시키가이샤 | Carboxymethyl-group-containing modified hyaluronic acid and/or salt thereof and/or production method for carboxymethyl-group-containing modified hyaluronic acid and/or salt thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110423291A (en) * | 2019-09-06 | 2019-11-08 | 焦点生物医药有限公司 | A kind of instant Sodium Hyaluronate preparation method |
| CN110423291B (en) * | 2019-09-06 | 2020-06-16 | 焦点生物医药有限公司 | Preparation method of instant sodium hyaluronate |
| CN113150184A (en) * | 2021-04-14 | 2021-07-23 | 浙江理工大学 | Method for preparing acetylated sodium hyaluronate |
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| KR102371689B1 (en) | 2022-03-07 |
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