KR101912418B1 - Antibiotic composition with Xylitol derivative and method for production of it - Google Patents

Abstract

The xylityl caprylate / citrate of the present invention exhibits excellent antibacterial activity. The novel synthesis method of xylityl caprylate / citrate used in the present invention has the advantage of overcoming the polymerization of a product, which is a problem of the conventional high temperature and acid catalyst synthesis method. An antibiotic composition includes xylityl caprylate / citrate and caprylyl glycol.

Description

TECHNICAL FIELD The present invention relates to an antimicrobial composition containing xylitol derivatives and an antimicrobial composition containing the xylitol derivative and a method for producing the same.

The present invention relates to a xylitylcaprylate / citrate composition having antimicrobial action and a process for its preparation.

Many of the cosmetic ingredients are emulsified products and contain nutrients of various bacteria such as animal and plant oil, fat and carbohydrate. As a result, microorganisms are easily contaminated during long-term storage and use, and preservatives are used to prevent them. Preservatives commonly used in cosmetics are parabens, phenoxyethanol, and chlorphenesin. These chemical preservatives cause allergic and irritation such as sensitive skin and contact dermatitis.

Meanwhile, recent studies in the UK have shown that parabens concentrations in breast cancer cells are significantly higher than those in normal controls, and that parabens can accumulate within the body's tissues. In addition, estrogen, a type of female hormone, has been suspected as a substance that can disturb the endocrine system due to its action, so it is necessary to research and develop natural antimicrobial substances that can replace parabens.

Also, there is a method of producing a composition for external application for skin which is not formulated with a conventional chemical preservative by using a diol having a recently known antimicrobial activity, a method of packing the microorganism in a container which inhibits the contamination of microorganisms in the manufacturing process, It is also possible to produce a skin external preparation which does not use a preservative or has a lower preservative content than usual. However, in the case of diol, skin irritation affects stability, solubilization, and stability of emulsified formulations. When a container that inhibits aseptic production or microbial inflow is used, the manufacturing process is complicated, Economic generality falls.

Therefore, it is necessary to develop a preservative having excellent repellency without using a chemical preservative.

Korean Patent Laid-Open No. 10-2015-0057589 (Aug. 25, 2015) "Antiseptic composition comprising xylitol fatty acid ester as an active ingredient" refers to a composition comprising xylitol fatty acid ester alone or an organic acid and its salt, a fatty acid and its ester, 1,2- , Or an aromatic compound, respectively, exhibit improved preservative activity against various microorganisms. Korean Patent Laid-Open No. 10-2015-0062394 (Jun. 20, 2015) " composition for skin external application having excellent buoyancy force " includes xylitylcaprylate having excellent buoyancy; And at least one substance selected from the group consisting of ethylhexyl glycerin, glyceryl caprylate, p-anisic acid, and citric mixed extract, thereby exhibiting a superior anti-buccal effect through a synergistic effect on the repellency of these substances have.

In the present invention, it is difficult to polymerize and purify the product due to the existing citric acid polymerization reaction. Thus, it is desired to synthesize xylitylcaprylate / citrate under low temperature synthesis conditions using an enzyme catalyst in the presence of a solvent.

The present invention also provides a preservative which enhances the antibacterial activity against fungi by mixing xylityl caprylate / citrate with caprylyl glycol.

The present invention provides an antimicrobial composition characterized by containing xylityl caprylate / citrate.

In the antimicrobial composition of the present invention, it is preferable to further include caprylyl glycol.

On the other hand, it is preferable that the caprylyl glycol is mixed at a weight ratio of 1: 1.0 to 2.0, preferably xylitylcaprylate / citrate: caprylyl glycol.

On the other hand, in the antimicrobial composition of the present invention, the composition preferably exhibits antibacterial activity against bacteria or fungi.

(A) mixing and dissolving xylitol, caprylic acid, and citric acid in a solvent;

A step (b) of adding a lipase to the mixture of step (a) to perform a dehydration condensation reaction; and (b) a step of adding a lipase to the mixture of step (a).

On the other hand, the method of manufacturing a jailri butyl caprylate / citrate of the present invention, the solvent of step (a) is preferably t - any one of the preferably butanol (t -Butanol) or acetonitrile (Acetonitrile) .

On the other hand, in the method for producing xylitylcaprylate / citrate of the present invention, xylitol, caprylic acid and citric acid in step (a) are preferably mixed in a molar ratio of 3: 3: 1.

On the other hand, in the production method of the xylitylcaprylate / citrate of the present invention, the lipase of the step (b) is preferably added by 0.5 to 2% by weight of the total weight of the mixture prepared in the step (a) It is good.

On the other hand, in the production method of the xylitylcaprylate / citrate of the present invention, the step (b) is preferably carried out at 60 to 80 ° C.

The present invention shows that the xylityl caprylate / citrate composition exhibits excellent antimicrobial activity.

On the other hand, the process for preparing xylitylcaprylate / citrate of the present invention overcomes the polymerization of the product, which has been a problem of the existing high temperature, acid catalyst synthesis method.

1 shows FT-IR results of (a) xylitol, (b) caprylic acid, (c) citric acid and (d) xylitylcaprylate / citrate.
FIG. 2 shows the results of confirming the molecular weight of xylitylcaprylate / citrate according to the synthesis conditions by GPC. ((a) high-temperature acid catalyst synthesis conditions, and (b) low temperature enzyme catalyst synthesis conditions)

In the present invention, xylitol caprylate / citrate, which is obtained by the esterification reaction of xylitol, caprylic acid and citric acid, has been developed.

Xylitol, which is produced in the body as an intermediate in glucose, is mainly distributed in plants. It has a peculiar five-point structure and is mainly extracted from Finnish birch and is also called birch sugar. When the birch is cut into small pieces and heated in water, the polysaccharide, xylen, is decomposed and transformed into xylose. When the xylose is purified through various processes and then reduced, it becomes xylitol. Xylitol is not only birch, but also plum, strawberry, cauliflower and other natural ingredients such as vegetables and fruits.

Caprylic acid is a saturated fatty acid obtained from fat such as butter fat, palm oil, palm oil, fish oil, seaweed oil, etc. It is a natural flavoring agent which is colorless, has a slightly unpleasant smell, and has a sour taste. It has been reported that it has a strong antibacterial function according to the results of the study.

Citric acid is present in high concentration in citrus, lemon, lime, etc. and is one of weak organic acids. Citric acid is a natural preservative and is used to produce the taste or acidity of food or carbonated beverages. It is also an important intermediate of the citric acid cycle and is very important for the aerobic metabolism of all organisms.

In the present invention, xylitol caprylate / citrate is prepared by synthesizing xylitol, caprylic acid, and citric acid with lipase, and then xylitol caprylate / The preservative activity was confirmed. In addition, Capryllyl glycol was mixed with xylitylcaprylate / citrate for the antimicrobial activity against molds, which suggested the potential as a preservative for cosmetics.

Hereinafter, the present invention will be described in more detail in the following Examples and Experimental Examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[Example 1: Preparation of xylitylcaprylate / citrate of the present invention]

Xylitol, caprylic acid, and citric acid were dissolved in a solvent ( t- butanol, Acetonitrile) in an equivalent molar ratio of 3: 3: 1 respectively and mixed with an enzyme catalyst corresponding to 0.5, 1, , NOVOZYM435, Lipase acrylic resin from Candida antarctica ) was added and heat was applied to 70 ° C to dissolve and mix well. The mixed raw materials were dehydrated and condensed at 70 ℃ for up to 24 hours. After the completion of the reaction, the remaining catalyst was removed by filtration, and the resulting water and solvent were removed by distillation under reduced pressure (80 ° C, 5 torr or less). Decolorization and deodorization (80 DEG C, 1 torr or less) were carried out using activated carbon and diatomaceous earth to prepare xylitol caprylate / citrate.

Gas chromatography was performed to compare compositions according to% enzyme added.

The gas chromatographic results of Example 1 Solvent Catalyst (%) mono (%) di (%) tri (%) polymer (%) ¹⁾ Acetonitrile 0.5 36.6 40.5 10.48 0 One 39.4 41.3 5.88 0 2 45.44 35.06 5.43 0 t- butanol 0.5 34.1 42.5 7.26 0 One 40.3 39.3 6.2 0 2 46.27 43.82 6.99 0

1) Polymer means that polymerization of tetra or more occurs.

As shown in Table 1, it was confirmed that the xylitol caprylate / citrate prepared using NOVOZYM435 overcomes the polymerization problem of the prior art.

[Comparative Example 1: Manufacture using p-TSA instead of the enzyme catalyst NOVOZYM435 of the present invention]

Instead of the enzyme catalyst NOVOZYM435 of the present invention, p-TSA (para toluene sulfoic acid) was added in an amount of 2 wt%.

[Experimental Example 1: Comparison of composition according to enzyme catalyst and solvent]

In this Experimental Example, gas chromatography was performed to compare the composition of Comparative Example 1 with that of the enzyme catalyst added in an amount of 2 wt% in Example 1.

Gas Chromatographic Results of Xylitylcaprylate / Citrate on Catalysts Catalyst Solvent mono (%) di (%) tri (%) polymer (%) ¹⁾ p-TSA Acetonitrile 37.94 27.27 4.53 15.09 t- butanol 48.38 31.40 4.75 15.47 NOVOZYM
435 Acetonitrile 45.44 35.06 5.43 0 t- butanol 46.27 43.82 6.99 0

1) Polymer means that polymerization of tetra or more occurs.

In Table 2, it can be seen that when the enzyme catalyst NOVOZYM435 of the present invention is used, the polymerization does not proceed.

[Experimental Example 2: Confirmation of molecular structure of xylitylcaprylate / citrate using FT-IR]

In this Experimental Example, the molecular structure of xylitylcaprylate / citrate containing 2 wt% of NOVOZYM435 of Example 1 was confirmed using FT-IR (Fourier transform infrared spectroscopy). IR spectra were obtained by irradiating the specimens while changing the wavelength of light in the infrared region using a Fourier transform infrared spectrometer and measuring the absorption transmission and reflectance. OH functional groups of xylitol was confirmed in the wavelength range 3000-3600cm ^-1, C = O bond of the ester compound was determined in the wavelength range 1735-1750cm ^-1.

The FT-IR analysis spectrum of xylitylcaprylate / citrate is shown in Fig. 1 in comparison with the corresponding raw material. The broad and strong peaks at 3200-3600 cm ^-1 in xylitol spectrum (a) are due to the stretching of -OH bonds. In the caprylic acid (b) and citric acid (c) spectra, the -OH and C═O groups a strong peak can be found in each 2500-3300cm ^-1, 1700-1725cm ^-1. In the xylitylcaprylate / citrate (d) spectrum, peaks corresponding to alcohol stretching 3200 ^- 3600 cm ^-1 and Ester C = O 1735 ^- 1750 cm ^-1 were identified.

[Experimental Example 3: Confirmation of molecular weight of xylitylcaprylate / citrate using GPC]

In this example, the molecular weight of the xylitylcaprylate / citrate containing 2 wt% of NOVOZYM435 of Example 1 was confirmed by gel permeation chromatography (GPC). Tosoh EcoSEC HLC-8320 GPC was used, and 2x TSKgel SupermultiporeHZ-M + TSKgel SuperHZ-2500 (4.6x150mm) column was used. Data were processed using EcoSEC software.

GPC results of xylitylcaprylate / citrate are shown in Fig. Spectrum (a) shows that polymerized by the polymerization of citric acid after synthesizing ester by applying acid catalyst at the existing high temperature condition, and spectrum (b) shows that the polymerization problem is solved after synthesizing with enzyme catalyst at low temperature.

As a result of the analysis through the above experimental examples, the structural formula of the synthesized xylitylcaprylate / citrate compound is shown in the following Chemical Formula 1.

[Experimental Example 4: Measurement of antimicrobial activity using Disc diffusion assay]

In this experimental example, the antimicrobial activity of the xylitylcaprylate / citrate (XCC) synthesized material containing 2 wt% of NOVOZYM435 of Example 1 was measured by the disc diffusion assay on the test strains. Antimicrobial activity was compared using phenoxyethanol which is used as a cosmetic preservative. The cultured strains were used for this experiment after controlling with 10 ⁶ CFU / ml of bacteria and 10 ⁵ CFU / ml of fungi. Each sample was prepared at a concentration of 5%, absorbed by 0.02 ml into the disc, and dried to volatilize the solvent. Antimicrobial activity of each sample was measured by measuring the size of the clear zone (mm) produced around the disc after the absorbed paper disc was incubated on a plate-coated plate. Disc diffusion assays were performed for one species of S. aureus , two E. coli, P. aeruginosa , five fungi ( C. albicans ), and five fungi ( A. niger ) Are shown in Table 3.

Antimicrobial activity measurement using Disc diffusion assay Strains Zone diameter (mm) S. aureus
(10 mg / disc) P. aeruginosa
(10 mg / disc) E. coli
(10 mg / disc) C. albicans
(10 mg / disc) A. niger
(10 mg / disc) Xylitol - - - - - Citric acid 10 13 11 - - XCC 11 12 11 10 - Phenoxy
ethanol (control group) 11 10 10 11 10

The antimicrobial activity of xylitol caprylate / citrate of the present invention against one gram-positive strain, two gram-negative strains and fungi was confirmed according to the results of the experiment.

[Example 2: Combination of xylitylcaprylate / citrate and caprylyl glycol]

XCCG was prepared by combining xylityl caprylate / citrate with 2 wt% of NOVOZYM435 of Example 1 and carpyryl glycol (Carprylyl glycol) at a weight ratio of 1: 1.5.

[Experimental Example 5: Antibacterial activity test of XCCG by measurement of minimum inhibitory concentration (MIC)] [

In this experiment, Agar dilution method was used to measure the minimum inhibitory concentration (MIC) of xylitol caprylate / citrate, which showed antimicrobial activity through disc diffusion assay. Each strain was adjusted to 10 ⁶ to 10 ⁷ CFU / ml and used in this experiment. Serial dilution method was used to determine the amount of xylityl caprylate / citrate to which 2% by weight of NOVOZYM435 of Example 1 was added as a control group and phenoxyethanol as a control group, microorganisms of XCC and Caprylyl glycol combined (XCCG) The minimum inhibitory concentrations were compared. The samples were used after diluting the concentration with TSB and PDB using a 2-fold dilution method. (Sample concentration: 8%, 4%, 2%, 1%, 0.5%, 0.25%, 0.125%). After culturing for 2 days, diluted samples were plated on the medium and the proliferation of the bacteria was confirmed after 2 days. When cultured and visually observed, the concentration at which each microorganism was not proliferated was determined as the MIC concentration.

Minimum inhibitory concentration measurement result (w / v%) Strains Citric acid XCC XCCG Phenoxyethanol
(Control group) S. aureus One 0.5 0.5 0.5 P. aeruginosa 0.25 One 0.25 0.5 E. coli One One 0.25 0.5 C. albicans - One 0.5 One A. niger - - One One

As shown in Table 4 above, the minimum inhibitory concentration was 0.5% when the control group, phenoxyethanol, was applied to Gram-positive S. aureus and Gram-negative bacteria such as E. coli and P. aeruginosa , and the concentration of xylitol caprylate / citrate The minimum inhibitory concentrations were 1% and 0.5%, similar to phenoxyethanol.

The minimum inhibitory concentration of fungal, C. albicans , was 1% for phenoxyethanol and XCC, similar to phenoxyethanol, but not for citric acid alone. XCCG showed antimicrobial activity against A. niger at 1% and antimicrobial activity against E. coli and P. aeruginosa at 0.25% ppm.

The combination of xylitol caprylate / citrate and caprylyl glycol synthesized in this study showed an increase in the antimicrobial effect against five fungi.

[Experimental Example 6: Challenge test of XCCG]

In this Experimental Example, a challenge test was performed to measure the repelling force in cosmetics based on the results of the antibacterial activity of XCCG combined with xylitol caprylate / citrate and caprylyl glycol in Experimental Example 5, and CTFA (The Cosmetic, Toiletry, and Fragrance Association) standards. Based on the results of antibacterial activity, lotion containing XCCG was prepared. To compare the flotation power, a conventional lotion containing 1% of phenoxyethanol and a lotion without preservative were prepared, respectively, and the ingredients thereof are shown in Table 5 below .

The content of O / W type emulsion (a: XCC + lotion, b: phenoxyethanol + lotion, c: Preservative free lotion) INCI Name Content (% (w / w)) a b c Water qs 100 qs 100 qs 100 Glycerin 3.0 3.0 3.0 Xanthan gum 0.2 0.2 0.2 Isostearyl isostearate 2.00 2.00 2.00 Cetearyl alcohol 0.5 0.5 0.5 pentylene glycol 3.0 3.0 3.0 Polyglyceryl-isostearate / Dimer dilinoleate copolymer 2.0 2.0 2.0 Polyglyceryl-10 stearate 1.5 1.5 1.5 XCCG 1.0 - - Phenoxyethanol - 1.0 -

The test strains S. aureus, P, aeruginosa, E. coli, C. albicans and A. niger, which showed antibacterial activity on the lotion, were inoculated into each lotion and stored in a 25 ℃ incubator. The survival rate was calculated and the buoyancy was measured.

Tables 6 to 10 below show the measured number of bacteria. In each table, T ₀ is the number of microorganisms immediately after inoculation and T ₂ , T ₇ , T ₁₄ , T ₂₁ , and T ₂₈ are the number of bacteria measured on days 2, 7, 14, 21, to be. Control is phenoxyethanol and PFL is a presercative free lotion with no preservative added.

The measured value of S. aureus in 1% XCCG (CFU / g) Time T ₀ T ₂ T ₇ T ₁₄ T ₂₁ T ₂₈ Results
(CFU / g) 1.65x10 ⁶ ND ND ND ND ND C (CFU / g) 1.65x10 ⁶ 1.0x10 ⁴ ND ND ND ND PFL 1.65x10 ⁶ 7.1 x 10 ⁶ 9.05x10 ⁶ 1.03x10 ⁷ 2.75 x 10 ⁷ 3.5x10 ⁷

The measured value of E. coli (CFU / g) in 1% XCCG Time T ₀ T ₂ T ₇ T ₁₄ T ₂₁ T ₂₈ Results
(CFU / g) 6.55x10 ⁵ 9.0x10 ³ ND ND ND ND C (CFU / g) 6.55x10 ⁵ 5.53x10 ⁴ ND ND ND ND PFL 6.55x10 ⁵ 8.09x10 ⁵ 2.0x10 ⁶ 4.54x10 ⁶ 6.9x10 ⁶ 7.0 x 10 ⁶

The measured value of P. aeruginosa (CFU / g) in 1% XCCG Time T ₀ T ₂ T ₇ T ₁₄ T ₂₁ T ₂₈ Results
(CFU / g) 1.66x10 ⁶ 3.35x10 ⁴ ND ND ND ND C (CFU / g) 1.66x10 ⁶ 6.5x10 ⁵ ND ND ND ND PFL 1.66x10 ⁶ 4.65x10 ⁶ 8.7x10 ⁶ 9.98x10 ⁶ 3.8 x 10 ⁷ 7.36x10 ⁷

Measured value of C. albicans in 1% XCCG (CFU / g) Time T ₀ T ₂ T ₇ T ₁₄ T ₂₁ T ₂₈ Results
(CFU / g) 7.7x10 ⁵ 2.3x10 ⁴ ND ND ND ND C (CFU / g) 7.7x10 ⁵ 1.2x10 ⁴ ND ND ND ND PFL 7.7x10 ⁵ 9.36x10 ⁵ 1.23x10 ⁶ 2.54x10 ⁶ 5.2x10 ⁶ 7.0 x 10 ⁶

The measured value (CFU / g) of A. niger in 1% XCCG Time T ₀ T ₂ T ₇ T ₁₄ T ₂₁ T ₂₈ Results
(CFU / g) 1.28x10 ⁵ 3.65x10 ³ ND ND ND ND C (CFU / g) 1.28x10 ⁵ 5.95x10 ³ 1.2x10 ² ND ND ND PFL 1.28x10 ⁵ 2.4 x 10 ⁶ 7.0 x 10 ⁶ 2.39x10 ⁷ 3.32x10 ⁷ 2.07x10 ⁸

As shown in Tables 6 to 10, the number of bacteria was increased after 4 weeks in the lotion containing no preservative. When the combination of phenoxyethanol alone and xylitol caprylate / citrate and caprylyl glycol was used, Day of the day.

The xylitylcaprylate / citrate of the present invention complements the disadvantages of the existing high temperature, acid catalyzed synthesis method and overcomes the product polymerization by using an enzyme catalyst in the presence of a solvent. In addition, the antimicrobial activity of xylitylcaprylate / citrate against gram-positive bacteria and fungi was confirmed.

On the other hand, in order to enhance the antimicrobial activity against the mold of xylitylcaprylate / citrate, it was combined with caprylyl glycol, and it was confirmed that the activity was equivalent to that of phenoxyethanol used as a conventional preservative.

Thus, it was confirmed that the composition comprising xylitylcaprylate / citrate and icaprylilyl glycol of the present invention could be used as a preservative substitute.

Claims (9)

(Xylityl caprylate / citrate) and caprylyl glycol (represented by the following formula (1)),
Antibacterial activity against Staphylococcus aureus , Escherichia coli , Pseudomonas aeruginosa , Candida albicans , and Aspergillus niger . Antibacterial activity against Staphylococcus aureus , Escherichia coli , Pseudomonas aeruginosa , Candida albicans and Aspergillus niger . Wherein the antimicrobial composition has an activity of inhibiting the activity of the antibiotic.
[Chemical Formula 1]

delete The method according to claim 1,
The caprylyl glycol may be, for example,
Xylityl caprylate / citrate: caprylyl glycol is mixed at a weight ratio of 1: 1.0 to 2.0.
delete (A) mixing and dissolving xylitol, caprylic acid and citric acid in a solvent;
(B) a step (b) of adding a lipase to the mixture of step (a) to perform a dehydration condensation reaction.
6. The method of claim 5,
The solvent of step (a)
t - butanol (t -Butanol) or a method of producing a jailri butyl caprylate / citrate, characterized in that at least one of acetonitrile (Acetonitrile).
6. The method of claim 5,
Wherein the xylitol, caprylic acid, and citric acid in step (a) are mixed at a molar ratio of 3: 3: 1.
6. The method of claim 5,
Wherein the lipase of step (b) is added in an amount of 0.5 to 2% by weight based on the total weight of the mixture prepared in step (a).
6. The method of claim 5,
Wherein step (b) is carried out at 60 to 80 占 폚.

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