KR102273009B1 - Method for synthesis of cosmetic emulsifier derived from nature resource with immobilized enzyme - Google Patents

Abstract

The present invention uses immobilized lipases, a naturally derived emulsifier using diglycerin and capric acid, a natural emulsifier for cosmetics to obtain diglycerin caprate The present invention uses immobilized lipases rather than a synthesis method using a homogeneous catalyst, and thus the problem of excessive energy use, unreacted substances, and lowering of purity in high-temperature processes according to the homogeneous catalytic reaction. was solved, and through this, diglycerin caprate can be obtained with high purity.

Description

Synthesis method of natural emulsifier for cosmetics using immobilized enzyme {Method for synthesis of cosmetic emulsifier derived from nature resource with immobilized enzyme}

The present invention relates to a method for synthesizing a natural-derived emulsifier for cosmetics using an immobilized enzyme, and more specifically, using immobilized lipases, diglycerin and capric acid. It relates to a method for synthesizing a high-purity, naturally-derived emulsifier, diglycerin caprate using

Surfactants (surface activation agents or surfactants) have surface wetting, penetration, air bubbles, defoaming, emulsification, solubilization, dispersion, agglomeration and cleaning of the surface due to the ability to lower the surface tension (dyne/cm) through the activation of the interface between the phases. It is an essential raw material used in all detergents and cosmetics because of its action.

Recently, consumers prefer mildness products with excellent biodegradability and surfactants with excellent physical properties such as wettability, foaming power, cleaning power, and resistance to surface tension that conventional surfactants have while having little biostimulation. have. In addition, there is a demand for the development of multifunctional products that can exhibit a combination of flexibility, sterilization, whitening and anti-aging effects in addition to interfacial properties.

On the other hand, the level of manufacturing technology for surfactants derived from nature in Korea does not escape from the stage of imitation of advanced technologies such as the United States, Japan, and Germany. This is a difficult situation. Most companies have the technology for general-purpose products, but the technology for natural-derived functional high-value-added products is insignificant. In addition, due to the growing interest in human safety, there is an urgent need to develop an in-house technology such as an environmentally friendly biodegradable natural surfactant, but the technology development for this is still insufficient.

Korean Patent Laid-Open No. 10-1999-0009251 (199.02.05) describes an emulsified and dispersed type sensitive makeup cosmetic composition using only natural vegetable emulsifiers. Korean Patent Application Laid-Open No. 10-2014-0047443 (2014.04.22) describes a low-viscosity cosmetic composition using a natural emulsifier.

The present invention is to develop and provide a method for producing a high-purity natural emulsifier (Diglycerin Caprate) using immobilized lipases.

The present invention is a method for synthesizing diglycerin caprate, characterized in that by adding diglycerin and capric acid, using acetonitrile as a solvent, and reacting with immobilized lipase provides

In the synthesis method of diglycerin caprate of the present invention, the synthesis molar ratio of diglycerin and capric acid is preferably 2 to 3:1.

In the method for synthesizing diglycerin caprate of the present invention, the immobilized lipase is preferably added in an amount of 5 to 20% (w/w) based on the total amount of diglycerin and capric acid.

In the method for synthesizing diglycerin caprate of the present invention, the solvent acetonitrile is preferably added by 50 to 1,800% (w/w) based on the total amount of diglycerin and capric acid. .

In the method for synthesizing diglycerin caprate of the present invention, the reaction is preferably performed for 2 to 24 hours.

The present invention uses immobilized lipases rather than a synthesis method using a homogeneous catalyst, thereby solving the problem of excessive energy use and unreacted substances and lowering purity of the high-temperature process according to the homogeneous catalytic reaction. Glycerin caprate can be obtained in high purity.

1 is a result of comparing the mono-ester content (% (w / w)) according to the change in the molar ratio of diglycerin (Diglycerin) and capric acid (Capric acid).
Figure 2 shows the reaction results according to the change in the content of immobilized lipase.
Figure 3 shows the reaction results according to the change in the solvent acetonitrile (Acetonitrile) content.
4 is a result of comparing the degree of reaction according to the reaction time.

The present invention is to prepare a naturally-derived emulsifier diglycerin caprate by esterification using immobilized lipases, and lowers the production of side reactants than a method using a general homogeneous catalyst and to prepare diglycerin caprate of high purity.

Accordingly, the present invention is diglycerin caprate, characterized in that by adding diglycerin and capric acid, using acetonitrile as a solvent, and reacting with immobilized lipase. A synthesis method is provided.

Enzymatic reactions have high substrate specificity compared to reactions with normal chemical catalysts. In addition, it has advantages such as being able to react under mild conditions such as room temperature and normal pressure. Enzymes are originally water-soluble proteins, and when performing an enzymatic reaction, the enzyme is generally dissolved in a reaction solution. However, in this general method, it is not easy to recover the used enzyme from the reaction solution after completion of the reaction, and the enzyme is disposable. This is particularly a big problem when the enzyme cannot be obtained cheaply or when the operation for producing the enzyme is complicated.

For effective use of the enzyme, the enzyme must be easily recovered from the reaction solution by any method. An enzyme processed for this purpose is an immobilized enzyme. Initially, the enzyme was called an insoluble enzyme because it was used as water-insoluble, but for the above purpose, it is not necessarily necessary to make the enzyme water-insoluble, so it is collectively called an immobilized enzyme. The immobilized enzyme usually made insoluble in water can be easily recovered from the reaction solution by a method such as decantation or centrifugation.

On the other hand, by filling the column with the immobilized enzyme and flowing the reaction solution, it becomes possible to continuously conduct the enzyme reaction. Specifically, there are three major methods for immobilizing an enzyme protein: a carrier binding method, a crosslinking method, and a blanket method. The carrier binding method is a method of binding an enzyme to an insoluble carrier by any method, but the binding mode is roughly divided into an ionic binding method, a physical adsorption method, and a covalent bonding method. The crosslinking method is a method to increase the molecular weight by covalent bonding between enzyme molecules to make them insoluble. The entrapment method includes a grid method in which an enzyme is immobilized in a matrix such as a gel and a microcapsule method in which the enzyme is encapsulated in microcapsules.

By other methods, the enzyme is covalently bound to an enteric polymer (a polymer that is soluble in a neutral or alkaline region, insoluble in an acidic region), and is water-soluble during the reaction and the pH is adjusted to an acidic region after the reaction is completed. A method of recovering by insolubilization has also been reported. In the present invention, the enzyme is filled in the column by the immobilized enzyme method, but the present invention is not limited thereto, and other methods described above or methods well known in the art may be used.

The method of synthesizing diglycerin caprate of the present invention is not a synthesis method using a general homogeneous catalyst, but uses immobilized lipases to solve the problem of excessive energy use and the high-temperature process according to the homogeneous catalytic reaction. The problem of unreacted substances and lowering of purity was solved. More specifically, 'CalB-IM' sold by Xenofocus was used as the immobilized lipase. In addition, substrate reaction molar ratio, immobilized enzyme content, solvent amount, and reaction time were optimized.

In the synthesis method of diglycerin caprate of the present invention, the synthesis molar ratio of diglycerin and capric acid is preferably 2 to 3:1, and the immobilized lipase is , preferably 5 to 20% (w/w) is added based on the total amount of diglycerin and capric acid, and the solvent acetonitrile is preferably 50 to 1,800% (w/w) based on the total amount of diglycerin and capric acid ( w/w) is preferably added, preferably reacting for 2 to 24 hours. Under the above conditions, diglycerin caprate of high purity and high yield can be prepared.

More specifically, a monoester content of 84.4% (w/w) could be obtained at a 3:1 reaction molar ratio of diglycerin and capric acid, and 3:1 was derived as the best reaction molar ratio. In addition, based on the total amount of diglycerin and capric acid, it showed a diglycerin caprate conversion rate of 99.5% (w/w) or more at an immobilized enzyme addition amount of 20% (w/w). In addition, the use of an acetonitrile solvent of 200% (w/w) based on the total amount of diglycerin and capric acid was most suitable for the reaction. In addition, as the reaction time, it was confirmed that the optimal reaction time was to react for 8 hours. As a result of synthesizing diglycerin caprate by combining the above optimal reaction conditions, it was possible to produce high-purity diglycerin caprate of 85.9% (w/w) or more of mono ester, and 75.4% (w/w) of high-purity diglycerin caprate. It was possible to obtain diglycerin caprate in yield.

Hereinafter, the configuration of the present invention will be described in detail through the following examples. However, the scope of the present invention is not limited only to the following examples, and includes modifications of technical ideas equivalent thereto.

[ Example 1: Synthesis of natural emulsifier of the present invention]

1. Experimental Materials

In order to synthesize diglycerin caprate, which is a naturally derived emulsifier of the present invention, diglycerin was purchased from KCI, and capric acid was used as Emery product. In addition, the immobilized enzyme was used by purchasing the product name CalB-IM from Xenofocus, and other reagents were purchased from reagent-grade and used.

2. Analysis method

Before proceeding with the GC analysis, all analysis was performed after inducing silylation. As analysis conditions, an Agilent DB-5 (25m×0.2mm×0.11㎛, 19091J-002) was used for the column, and the initial temperature of the oven was maintained at 150°C for 5 minutes, and then at a rate of 10°C/min to 280°C. The temperature was raised and held at 280° C. for 10 minutes. Thereafter, the content purity of the compound was obtained using the area percentage method.

3. Explore optimal production conditions

3-1. Reaction for synthesizing natural emulsifier of the present invention molar ratio optimization

For esterification with diglycerin caprate, CalB-IM was selected among immobilized lipases to derive a molar ratio with a high monoester content according to a change in the molar ratio. The experiment was performed as follows. did.

The synthesis molar ratio of diglycerin and capric acid was set to 1:1 mol, 2:1 mol, 3:1 mol molar ratio. The content of the immobilized lipase CalB-IM was 20% (w/w) based on the total amount of diglycerin and capric acid, and acetonitrile was used as the solvent.

As a specific experimental method, diglycerin and capric acid were added in a molar ratio of 1:1 mol, 2:1 mol, and 3:1 mol, respectively, to a 500ml round-bottom flask, and then immobilized lipase (CalB-IM) 20% (w / w) was added based on the total amount of diglycerin and capric acid, and 200% (w / w) of acetonitrile was added based on the total amount of diglycerin and capric acid, and the stopper was closed. It was closed and reacted for 24 hours at 60°C with stirring. After completion of the reaction, the immobilized lipase (CalB-IM) and the solvent were removed by filtration, and the ester content was comparatively analyzed through GC (gas chromatography) analysis (Table 1, FIG. 1). 1 is a result of comparing the mono-ester content (% (w/w)) according to the change in the molar ratio of diglycerin (Diglycerin) and capric acid (Capric acid).

Monoester content (% (w/w)) according to the change in the molar ratio of diglycerin and capric acid no Molar ratio of diglycerin and capric acid Ester content (% (w/w)) remark Mono Di Tri One 1:1 60.7 37.0 2.3 2 2:1 78.2 21.1 0.7 3 3:1 84.4 15.6 -

As a result of the experiment, as an optimal reaction condition for the enzymatic ester reaction of diglycerin caprate, a molar ratio of 3:1 was the most excellent with a monoester content of 84.4% (w/w).

3-2. Optimization of the amount of immobilized lipase (CalB-IM) added

Based on the results of the above molar ratio optimization experiment, a molar ratio of diglycerin and capric acid was selected as 3:1, and the reaction was performed with different amounts of immobilized lipase (CalB-IM) added. The amount added was 1% (w/w), 3% (w/w), 5% (w/w), 10% (w/w), 20% (w/w), and acetonite was used as a solvent. Reel (Acetonitrile) was used.

As a specific experimental method, a molar ratio of diglycerin and capric acid was added to a 500 ml round-bottom flask so that the molar ratio of diglycerin and capric acid was 3:1, and then, acetonitrile, a solvent, was added to diglycerin and capric acid. 200% (w/w) based on the total amount of acid was added, added, stirred and dissolved, and immobilized lipase (CalB-IM) was added to 1% (w/w), respectively, based on the total amount of diglycerin and capric acid, 3 % (w/w), 5% (w/w), 10% (w/w), and 20% (w/w) were added. Thereafter, the flask was capped and reacted at 60° C. for 24 hours while stirring. After completion of the reaction, the immobilized lipase (CalB-IM) and the solvent were removed by filtration, and then, the content and conversion of unreacted capric acid were comparatively analyzed through gas chromatography (GC) analysis (Table 2, FIG. 2). Figure 2 shows the reaction results according to the change in the content of immobilized lipase (CalB-IM).

Reaction results according to changes in immobilized lipase (CalB-IM) content no CalB-IM content Capric acid conversion rate remark (%(w/w)) (%(w/w)) (%(w/w)) One One 22.34 77.6 2 3 12.7 87.3 3 5 7.7 92.3 4 10 0.6 99.4 5 20 0.4 99.6

As a result of the experiment, as the optimal reaction conditions for the enzymatic ester of diglycerin caprate, the immobilized enzyme (CalB-IM) was 99.5% when applied at 20% (w/w) based on the total amount of diglycerin and capric acid. (w/w) or higher conversion was the most excellent.

3-3. Solvent optimization

Based on the optimal molar ratio and enzyme addition amount selected through the above experiment, the degree of reaction according to the content of the solvent (Acetonitrile) was compared. Specific solvent content is 0% (w/w), 25% (w/w), 25% (w/w), 50% (w/w), 100%, respectively, based on the total amount of diglycerin and capric acid (w/w), 200% (w/w), and 1,800% (w/w) were added and reacted.

As a specific experimental method, the molar ratio of diglycerin and capric acid was added to a 500 ml round-bottom flask so that the molar ratio was 3:1, and then acetonitrile as a solvent was added to the total amount of diglycerin and capric acid. 0% (w / w) ~ 1800% (w / w) applied, respectively, as a standard, stirred and melted, and the immobilized enzyme (CalB-IM) was 20% (w / w) based on the total amount of diglycerin and capric acid ) was added. Thereafter, the flask was capped and reacted at 60° C. for 24 hours while stirring. After completion of the reaction, the reaction was filtered to remove the immobilized lipase (CalB-IM) and the solvent, and then the unreacted capric acid content and the ester content were comparatively analyzed through GC (gas chromatography) analysis (Table 3, FIG. 3). Figure 3 shows the reaction results according to the change in the solvent acetonitrile (Acetonitrile) content.

Reaction results according to the change in the solvent acetonitrile content no Solvent content (% (w/w)) Capric acid Ester content (% (w/w)) remark % (w/w) Mono Di Tri One 0 20.9 50.8 28.3 - 2 25 11.6 62.2 26.2 - 3 50 10.2 77.5 12.3 - 4 100 10.1 80.8 9.1 - 5 200 2.7 86.2 11.1 - 6 1,800 4.2 89.2 6.6 -

As a result of the experiment, as the optimal reaction conditions for the enzymatic ester of diglycerin caprate, the optimal solvent ratio was 200% (w/w) based on the total amount of diglycerin and capric acid.

3-4. Reaction time optimization

Based on the molar ratio, catalyst content, and solvent content selected through the above experiment, the degree of reaction according to the lapse of reaction time was confirmed. The reaction time was carried out for 2 to 24 hours, and samples were taken according to each time to check the degree of reaction.

As a specific experimental method, a molar ratio of diglycerin and capric acid was added to a 500 ml round-bottom flask so that the molar ratio was 3:1, and then acetonitrile as a solvent was added to the total amount of diglycerin and capric acid. 200% (w/w) was added as a standard, stirred and melted, and 20% (w/w) of the immobilized enzyme (CalB-IM) was added based on the total amount of diglycerin and capric acid. The flask was capped and reacted at 60° C. for 24 hours while stirring, and 10 ml of samples were taken and filtered using a pipette according to the reaction times of 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, and 24 hours. . Then, after removing the solvent, the degree of reaction according to the reaction time was checked through GC (gas chromatography) analysis, and the optimal reaction time was derived (Table 4, FIG. 4). 4 is a result of comparing the degree of reaction according to the reaction time.

Comparison of reaction degree according to reaction time no reaction time
(hr) Capric acid Ester content (% (w/w)) remark % (w/w) Mono Di Tri One 2 6.5 84.8 8.7 - 2 4 5.2 85.4 9.4 - 3 6 4.5 85.3 10.2 - 4 8 3.7 86.1 10.2 - 5 10 3.3 85.9 10.8 - 6 24 2.8 86.1 11.1 -

As a result of the experiment, it was confirmed that the optimal reaction time was to react for 8 hours as the optimal reaction conditions for the enzymatic ester of diglycerin caprate.

3-5 synthesis and purification

The experiment was conducted by reflecting the optimal conditions derived from the experimental results of 3-1 to 3-4. Specifically, 498 g (3.0 mol) of diglycerin, 172.2 g (1.0 mol) of capric acid, 1,340.4 g (200% (w/w)) of acetonitrile in a 3L round bottom flask , immobilized enzyme (CalB-IM) 134.04g (20% (w/w)), molecular sieve (Molecularseive) 13.404g (2% (w/w)) were added, the temperature was raised to 60° C., and stirred to 8 reacted for hours.

After the reaction, after cooling to room temperature, ethanol (200 g) was added and dissolved, followed by filtration to separate the immobilized enzyme. The filtrate was concentrated under reduced pressure at 65° C./10 torr/1 hr to remove the solvent, and the concentrate was transferred to a separatory funnel, ethyl acetate (300 g), and water (100 g) were added, followed by vigorous shaking and stagnant separation.

The completely separated supernatant (Ethyl acetate layer) layer was separated and concentrated under reduced pressure under the conditions of 65° C./5.0 torr/1 hr to remove the solvent. Deodorization was carried out while purging with nitrogen (N2gas) under the conditions of 80°C/5torr/1hr, and after decolorization using acid clay, high-purity diglycerin caprate was synthesized by filtration. Purity analysis of the synthesis product (Diglycerin caprate) was analyzed using gas chromatography (GC).

As a result of the experiment, it was confirmed that the diglycerin caprate of the present invention contained 85.9% (w/w) of high-purity monoester. In addition, the synthetic yield was calculated by measuring the weight to the first decimal place, and as a result, a high yield of 75.4% (w/w) was confirmed.

* Theoretical production amount = 3 moles of diglycerol (Mw. 166g/mol, 166g x 3mol) + 1 mole of capric acid (Mw 172.2, 172.2g) = 1 mole of diglycerol caprate (Mw. 320.2), theoretical production amount: 320.2g

Claims (5)

Diglycerin and capric acid are added, acetonitrile is used as a solvent, and an esterification reaction is induced with immobilized lipase,
The synthetic molar ratio of diglycerin and capric acid is 2 to 3:1,
The immobilized lipase is added by 5-20% (w/w) based on the total amount of diglycerin and capric acid,
The acetonitrile is added by 50 to 1,800% (w/w) based on the total amount of diglycerin and capric acid,
The reaction is a method of synthesizing diglycerin monocaprate, characterized in that it is carried out for 2 to 24 hours. delete delete delete delete

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