CN117487782B

CN117487782B - Hydrolysis sponge and extraction and application thereof

Info

Publication number: CN117487782B
Application number: CN202311445464.9A
Authority: CN
Inventors: 吴宪忠
Original assignee: Guangzhou Xulang Biotechnology Co ltd
Current assignee: Guangzhou Xulang Biotechnology Co ltd
Priority date: 2023-09-19
Filing date: 2023-11-01
Publication date: 2024-11-29
Anticipated expiration: 2043-11-01
Also published as: CN117487782A

Abstract

The present invention provides a hydrolyzed sponge and its extraction and application. By using a specific composite hydrolase (composed of snail enzyme and papain in a mass ratio of 1 to 3:7 to 9) to extract the siliceous sponge, without using acid-base reagents, the hydrolyzed sponge can be completely and efficiently extracted without causing needle breakage, and the purity of the obtained hydrolyzed sponge can reach more than 99%. The product has an off-white appearance and is suitable for preparing skin care products or skin medicines.

Description

Hydrolysis sponge and extraction and application thereof

Technical Field

The invention belongs to the technical field of hydrolysis sponge. More particularly, to a hydrolysis sponge and extraction and application thereof.

Background

The hydrolyzed sponge is an extremely tiny needle-shaped bone extracted from siliceous sponge, can not be identified by naked eyes, and is in a needle shape with round ends or pointed ends under a microscope, so the hydrolyzed sponge is also called a sponge microneedle. The needle tip of the hydrolysis sponge can be pricked into skin, opens skin channels to reach basal layer, helps to activate skin microcirculation, relieves skin pore blockage, can accelerate skin metabolism and accelerate natural peeling of aged stratum corneum, achieves natural, safe and effective biophysical skin refreshing, and shortens the skin refreshing period from 28 days to 1 week. Therefore, the hydrolysis sponge can be used for preparing skin care products or skin medicines.

At present, the extraction method of the hydrolysis sponge mainly comprises acid hydrolysis, alkali bleaching and the like, for example, the sponge is sequentially crushed, acid treated, heated, cleaned and the like in the prior art, but the crushing operation of the method can lead to broken needles of partial hydrolysis sponge, further the skin care effect is reduced because the broken needles are not easy to prick into skin, the acid treatment can bring new impurities, and the impurities and the micro needles directly reach the basal layer together, so that unnecessary damage is brought to the skin, and for example, the sponge is sequentially subjected to alkali treatment, ultrasonic treatment, water washing, strong acid treatment and the like in the prior art, so that the strong acid and alkali treatment has operational danger, can bring new impurities and bring unnecessary damage to the skin.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide an extraction method of the hydrolysis sponge, which is used for extracting the siliceous sponge by using specific compound hydrolase, so that the hydrolysis sponge with low needle breakage rate and high purity is extracted on the premise of not using an acid-base reagent.

The first object of the present invention is to provide a complex hydrolase.

The second object of the invention is to provide the application of the complex hydrolase in extracting hydrolysis sponge.

The third object of the invention is to provide an extraction method of the hydrolysis sponge.

A fourth object of the present invention is to provide a hydrolyzed sponge extracted by the above method.

A fifth object of the present invention is to provide the use of the above-mentioned hydrolysis sponge for the preparation of skin care products or skin medicaments.

A sixth object of the present invention is to provide a skin care product.

A seventh object of the present invention is to provide a dermatological agent.

The above object of the present invention is achieved by the following technical scheme:

the invention provides a compound hydrolase, which consists of snailase and papain in a mass ratio of 1-3:7-9.

According to the invention, the properties of the siliceous sponge are subjected to targeted research, and the fact that the siliceous sponge is subjected to enzymolysis by adopting the compound hydrolase with specific types and proportions is found, so that the hydrolytic sponge can be completely and efficiently extracted, breakage of needles is not easy to occur, the purity of the obtained hydrolytic sponge can reach more than 99%, the siliceous sponge is suitable for preparing skin care products or skin medicaments, the appearance of the obtained hydrolytic sponge is white, foreign body sensation cannot be generated when the obtained hydrolytic sponge is added into the skin care products or skin medicaments, and the hydrolytic sponge is more easily accepted by consumers. In addition, the invention uses the specific compound hydrolase to extract the hydrolysis sponge, replaces the traditional acid-base treatment method, does not need to use acid-base reagents, avoids acid-base residues in the hydrolysis sponge, and effectively reduces the risks of damaging skin and polluting the environment. Therefore, the application of the compound hydrolase in extracting hydrolysis sponge is within the protection scope of the invention.

Preferably, the mass ratio of the snailase to the papain is 2:8.

Preferably, the hydrolysis sponge is extracted from a siliceous sponge. The siliceous sponge, also called siliceous sponge, refers to a sponge with siliceous spicules or spongy filaments and siliceous spicules in bones.

The invention also provides an extraction method of the hydrolysis sponge, which is obtained by carrying out enzymolysis on the siliceous sponge by using the compound hydrolase, and then carrying out solid-liquid separation and drying.

Preferably, the siliceous sponge is further added to water before the enzymolysis, and heated at 70-100 ℃ for 20-40 min (most preferably, heated at 90 ℃ for 30 min). At this point the siliceous sponge will loosen.

Further preferably, the dosage ratio of the siliceous sponge to water is 3-8 g/100-360 mL, most preferably 5 g/200 mL.

Further preferably, before the siliceous sponge is added to the water, the siliceous sponge is further subjected to pretreatment, such as washing, filtering, and centrifuging in order to remove macroscopic impurities such as sand in the siliceous sponge.

More preferably, the washing is with water.

More preferably, the dosage ratio of the siliceous sponge to the water is 3-8 g/100-360 mL, most preferably 5 g/200 mL.

More preferably, the cleaning is 2 to 4 times.

More preferably, the centrifugation is at 1000 to 3000rpm for 1 to 3min, most preferably at 2000rpm for 2min.

Further preferably, after the heating, filtration is also performed.

Preferably, the mass ratio of the composite hydrolase to the siliceous sponge is 1:20-30, and most preferably 1:25.

Preferably, the enzymatic hydrolysis is performed in an aqueous environment.

Further preferably, the dosage ratio of the siliceous sponge to water is 1-3 g:25-150 mL, most preferably 1g:45mL.

Preferably, the temperature of the enzymolysis is 30-50 ℃, and most preferably 40 ℃. At this temperature, the complex hydrolase can exert an enzymatic hydrolysis better.

Preferably, the enzymolysis time is 8-16 h, and most preferably 12h. At the moment, the composite hydrolase can better play the enzymolysis effect, the enzymolysis rate is low when the time is too short, the time cost is wasted when the time is too long, and the enzymolysis rate can not be further improved.

Preferably, the enzymolysis is carried out while stirring, so that the composite hydrolase is fully contacted with the siliceous sponge, and the cellulose, sugar and other impurities are more efficiently digested and decomposed, so that the impurities are dissolved in water.

Further preferably, the stirring speed is 50 to 150rpm, most preferably 100rpm.

Preferably, the enzyme is inactivated after the enzymolysis, for example, the enzyme is heated at 110-130 ℃ for 0.4-0.6 h, and most preferably, the enzyme is heated at 120 ℃ for 0.5h.

Preferably, the means for solid-liquid separation include, but are not limited to, filtration, centrifugation, and the like. The operation of solid-liquid separation can effectively remove the compound hydrolase and avoid the residue in the product.

Further preferably, the centrifugation is at 1000 to 3000rpm for 1 to 3min, most preferably at 2000rpm for 2min.

Preferably, after the solid-liquid separation, cleaning and adsorption are also performed to remove the tiny impurities.

Further preferably, the washing is with water.

More preferably, the dosage ratio of the siliceous sponge to water is 1-10 g/30-500 mL, most preferably 5 g/200 mL.

More preferably, the cleaning is performed 1 to 3 times.

Further preferably, the adsorbent used for the adsorption is one or more of carbon powder, agarose and resin.

Further preferably, the adsorption time is 25 to 35min, and most preferably 30min.

Preferably, the drying is performed at 110-130 ℃ for 20-40 min, and most preferably at 120 ℃ for 30min.

Preferably, after said drying, further post-treatments, such as sieving, are performed.

Further preferably, a 80-120 mesh screen is used for sieving to obtain the hydrolysis sponge with different length particle diameters.

The method can not only completely and efficiently extract the hydrolysis sponge and is not easy to cause broken needle, but also can ensure that the purity of the obtained hydrolysis sponge reaches more than 99 percent, and the appearance of the product is white, and is suitable for preparing skin care products or skin medicaments, so the hydrolysis sponge extracted by the method, the application of the hydrolysis sponge in preparing the skin care products or skin medicaments, and the skin care products and skin medicaments containing the hydrolysis sponge are all within the protection scope of the invention.

The invention has the following beneficial effects:

The invention adopts the specific compound hydrolase to extract the siliceous sponge, can extract the hydrolytic sponge completely and efficiently without using acid-base reagents, is not easy to cause broken needles, can ensure that the purity of the obtained hydrolytic sponge reaches more than 99 percent, and the appearance of the product is white, thereby being suitable for preparing skin care products or skin medicaments.

Drawings

FIG. 1 is a macroscopic photograph of the siliceous sponge used in example 1.

FIG. 2 is a macroscopic photograph of the hydrolyzed sponge extracted in example 1.

FIG. 3 is a photograph of the hydrolyzed sponge extracted in example 1 under a microscope.

FIG. 4 is a macroscopic photograph of the hydrolyzed sponge extracted in example 2.

FIG. 5 is a photograph of the hydrolyzed sponge extracted in example 2 under a microscope.

FIG. 6 is a macroscopic photograph of the hydrolyzed sponge extracted in example 3.

FIG. 7 is a photograph of the hydrolyzed sponge extracted in example 3 under a microscope.

FIG. 8 is a macroscopic photograph of the siliceous sponge used in example 4.

FIG. 9 is a macroscopic photograph of the hydrolyzed sponge extracted in example 4.

FIG. 10 is a photograph of the hydrolyzed sponge extracted in example 4 under a microscope.

FIG. 11 is a macroscopic photograph of the hydrolyzed sponge extracted in comparative example 1.

FIG. 12 is a photograph of the hydrolyzed sponge extracted in comparative example 1 under a microscope.

FIG. 13 is a macroscopic photograph of the hydrolyzed sponge extracted in comparative example 2.

FIG. 14 is a photograph of the hydrolyzed sponge extracted in comparative example 2 under a microscope.

FIG. 15 is a macroscopic photograph of the hydrolyzed sponge extracted in comparative example 3.

FIG. 16 is a photograph of the hydrolyzed sponge extracted in comparative example 3 under a microscope.

FIG. 17 is a macroscopic photograph of the hydrolyzed sponge extracted in comparative example 4.

FIG. 18 is a photograph of the hydrolyzed sponge extracted in comparative example 4 under a microscope.

Detailed Description

The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.

Reagents and materials used in the following examples are commercially available unless otherwise specified.

Example 1 extraction method of hydrolysis sponge

S1, cleaning 12kg of siliceous sponge (produced from Jiangsu, and shown in a picture of FIG. 1) with 480L of water for 3 times, filtering, centrifuging filtrate at 2000rpm for 2min, adding the precipitate obtained by centrifugation into 480L of water, heating at 90 ℃ for 30min, and filtering again;

S2, placing the precipitate obtained by filtering the S1 into an extraction tank, adding 540L of water, adding 0.48kg of compound hydrolase (the mass ratio of snailase to papain is 2:8), carrying out enzymolysis for 12 hours at 40 ℃ (stirring at 100rpm while carrying out enzymolysis), and then heating for 0.5 hour at 120 ℃;

s3, taking the product obtained in the step S2 out of an extraction tank, filtering, centrifuging at 2000rpm for 2min, washing the precipitate obtained by centrifugation with 480L of water for 3 times, and adsorbing with agarose for 30min;

s4, drying the product obtained in the step S3 at 120 ℃ for 30min, and then passing through a 100-mesh screen to obtain the hydrolysis sponge.

The hydrolyzed sponge extracted in this example is shown in fig. 2, has an appearance of white-like, is observed under a microscope, and as shown in fig. 3, is a needle-like body with two pointed ends, the diameter is 10-20 micrometers, the length is 200-400 micrometers, the purity is >99%, the needle breakage rate is <1%, and the recovery rate is up to 33%.

Example 2 extraction method of hydrolysis sponge

S1, washing 12kg of siliceous sponge (produced from Jiangsu) with 400L of water for 4 times, filtering, centrifuging filtrate at 1000rpm for 3min, adding the precipitate obtained by centrifugation into 400L of water, heating at 70 ℃ for 40min, and filtering again;

s2, placing the precipitate obtained by filtering the S1 into an extraction tank, adding 300L of water, adding 0.6kg of compound hydrolase (the mass ratio of snailase to papain is 1:9), performing enzymolysis for 8 hours at 30 ℃ (stirring at 50rpm during enzymolysis), and then heating for 0.6 hours at 110 ℃;

S3, taking the product obtained in the step S2 out of an extraction tank, filtering, centrifuging at 1000rpm for 3min, washing the precipitate obtained by centrifugation with 360L of water for 2 times, and adsorbing with carbon powder for 25min;

s4, drying the product obtained in the step S3 at 110 ℃ for 40min, and then passing through a 80-mesh screen to obtain the hydrolysis sponge.

The hydrolyzed sponge extracted in this example is shown in fig. 4, has an appearance of white-like, is observed under a microscope, and as shown in fig. 5, is a needle-like body with two pointed ends, the diameter is 10-20 micrometers, the length is 200-400 micrometers, the purity is >99%, the needle breakage rate is <3%, and the recovery rate is up to 26%.

Example 3 extraction method of hydrolysis sponge

S1, washing 12kg of siliceous sponge (produced from Jiangsu) with 540L of water for 2 times, filtering, centrifuging filtrate at 3000rpm for 1min, adding the obtained precipitate into 540L of water, heating at 100 ℃ for 20min, and filtering again;

S2, placing the precipitate obtained by filtering the S1 into an extraction tank, adding 600L of water, adding 0.4kg of compound hydrolase (the mass ratio of snailase to papain is 3:7), performing enzymolysis for 16h at 50 ℃ (stirring at 150rpm during enzymolysis), and then heating for 0.4h at 130 ℃;

S3, taking the product obtained in the step S2 out of an extraction tank, filtering, centrifuging at 3000rpm for 1min, washing the precipitate obtained by centrifugation with 600L of water for 1 time, and adsorbing with resin for 35min;

s4, drying the product obtained in the step S3 at 130 ℃ for 20min, and then passing through a 120-mesh screen to obtain the hydrolysis sponge.

The hydrolyzed sponge extracted in this example is shown in fig. 6, has an appearance of white-like, and is observed under a microscope, and as shown in fig. 7, the hydrolyzed sponge is a needle-like body with two pointed ends, the diameter is 10-20 micrometers, the length is 100-300 micrometers, the purity is >99%, the needle breakage rate is <2%, and the recovery rate is up to 25%.

Example 4 extraction method of hydrolysis sponge

The difference is that the siliceous sponge produced in Jiangsu was replaced with the siliceous sponge produced in Henan (photograph is shown in FIG. 8) as in example 1.

The hydrolyzed sponge extracted in this example is shown in fig. 9, has an appearance of white-like, and is observed under a microscope, and as shown in fig. 10, the hydrolyzed sponge is a needle-like body with two round ends, the diameter is 10-20 micrometers, the length is 100-400 micrometers, the purity is >99%, the needle breakage rate is <5%, and the recovery rate is up to 30%.

Comparative example 1A method for extracting a hydrolysis sponge

The difference is that the complex hydrolase is replaced with equal quality helicase as in example 1.

The hydrolyzed sponge extracted in this comparative example was shown in fig. 11, and was gray in appearance, and as a result of observation under a microscope, as shown in fig. 12, it was found that it contained needle-like bodies with pointed ends at both ends, the diameter was 10 to 20 μm, the length was 200 to 400 μm, the purity was >91%, the needle breakage rate was <10%, and the recovery rate was 16%.

Comparative example 2 extraction method of hydrolysis sponge

The difference from example 1 is that the complex hydrolase is replaced by papain of equal quality.

The hydrolyzed sponge extracted in this comparative example was shown in fig. 13, and was gray in appearance, and as a result, as shown in fig. 14, it was found that it contained needle-like bodies with pointed ends at both ends, the diameter was 10 to 20 μm, the length was 200 to 400 μm, the purity was >72%, the needle breakage rate was <14%, and the recovery rate was 13%.

Comparative example 3 extraction method of hydrolysis sponge

The difference is that the snailase is replaced with pectase of equal mass as in example 1.

The hydrolyzed sponge extracted in this comparative example was shown in FIG. 15, and was grayish yellow in appearance, and as a result, as shown in FIG. 16, it was found that it contained needle-like bodies with pointed ends at both ends, the diameter was 10 to 20 microns, the length was 200 to 400 microns, the purity was >53%, the needle breakage rate was <19%, and the recovery rate was 10%.

Comparative example 4A method for extracting a hydrolysis sponge

The difference is that papain is replaced with bromelain of equal quality as in example 1.

The hydrolyzed sponge extracted in this comparative example was shown in FIG. 17 as having a gray yellow appearance, and as a result of observation under a microscope, as shown in FIG. 18, it was found that it contained needle-like bodies with pointed ends at both ends, the diameter was 10 to 20 μm, the length was 200 to 400 μm, the purity was >46%, the needle breakage rate was <21%, and the recovery rate was 8%.

The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.

Claims

1. The extraction method of the hydrolysis sponge is characterized by comprising the following steps of:

s1, adding a siliceous sponge into water, heating for 20-40 min at 70-100 ℃, and filtering, wherein the dosage ratio of the siliceous sponge to the water is 3-8 g/100-360 mL;

S2, carrying out enzymolysis on the siliceous sponge for 8-16 hours at the temperature of 30-50 ℃ by utilizing compound hydrolase, and then carrying out solid-liquid separation and drying, wherein the compound hydrolase consists of snailase and papain with the mass ratio of 1-3:7-9, the mass ratio of the compound hydrolase to the siliceous sponge is 1:20-30, the enzymolysis is carried out in an aqueous environment, and the dosage ratio of the siliceous sponge to water is 1-3 g:25-150 mL.

2. The extraction method according to claim 1, wherein the heating at S1 is at 90 ℃ for 30min.

3. The extraction method according to claim 1, wherein the ratio of the amount of the siliceous sponge to the amount of water used in S1 is 5 g/200 mL.

4. The extraction method according to claim 1, wherein the mass ratio of the snailase to the papain of S2 is 2:8.

5. The extraction method according to claim 1, wherein the mass ratio of the complex hydrolase to the siliceous sponge of S2 is 1:25.

6. The extraction method according to claim 1, wherein the ratio of the amount of the siliceous sponge to the amount of water used in S2 is 1g:45mL.

7. The extraction method according to claim 1, wherein the temperature of the enzymolysis of S2 is 40 ℃.

8. The extraction method according to claim 1, wherein the time for the enzymolysis of S2 is 12 hours.