Elepidogrammitis extracting solution as well as preparation method and application thereof
Through microjet technology assisted in enzymatic decomposition and antioxidant protection, the problems of low extraction efficiency of bird's horned bird's mint and easy to destroy active ingredients are solved, efficient extraction and stability are achieved, and the sustainable development of the bird's nest industry has been promoted.
Patent Information
- Application Number
- CN202411344549.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-25
- Publication Date
- 2025-01-10
AI Technical Summary
In the existing bird's nest processing technology, the extraction efficiency of bird's horns and bird's crushes is low, the energy consumption is high, and it is easy to destroy the active ingredients, resulting in waste of resources and unstable product quality.
Microjet technology assists enzymatic lysis, and a huge pressure difference is formed on the syringe horned sialic acid and other active ingredients through high-pressure and high-speed microjets, and an antioxidant and amino acid protector are added to ensure biological activity. and stability.
It significantly improves the extraction efficiency and purity of Yanjiao Yan crushed extract, retains biological activity, extends the shelf life, reduces equipment investment and operating costs, and improves the overall quality and market competitiveness of the product.
Smart Images
Figure BDA0005060586680000091 
Figure BDA0005060586680000101
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of bird's nest processing, and in particular to a bird's nest horn and bird's nest fragment extract, and a preparation method and application thereof. Background Art
[0002] Bird's nest mainly contains protein, carbohydrates, lipids, inorganic elements, vitamins and other ingredients. A study analyzed the amino acids contained in bird's nest and found that it contains 20 common amino acids such as aspartic acid and glutamic acid, and 8 essential amino acids. The content of carbohydrates in bird's nest is second only to protein, mainly sialic acid, mannose, glucosamine, galactosamine, etc. Lipids account for about 0.14-0.28% of bird's nest, and fatty acid components include stearic acid, palmitic acid, linolenic acid, linoleic acid, etc. Bird's nest has a high content of inorganic elements and contains trace elements necessary for humans. Studies have shown that the elements with higher content in bird's nest are four major elements Ca>Mg>Na>K, followed by five essential trace elements Fe>Zn>Mn>Cr>Cu. Bird's nest has always been considered a precious tonic because of its rich nutritional ingredients.
[0003] During the transportation and processing of bird's nest, about 15% of bird's nest will be formed into bird's nest corners and bird's nest fragments. The nutritional value of these bird's nest corners and bird's nest fragments is as rich as that of bird's nest strips, but their use is limited due to factors such as poor appearance or difficulty in soaking. They are often discarded or resold at low prices, which causes a great waste of resources. Therefore, exploring the reasonable disposal methods of bird's nest corners and bird's nest fragments is particularly important for improving the utilization rate of raw materials and even increasing the added value of enterprises.
[0004] At present, in the prior art, a series of extraction processes are used to extract liquid containing active ingredients from bird's nest horns and bird's nest pieces, and then the liquid is used in bird's nest products. Common extraction methods include decoction, acid-base extraction, and supercritical fluid extraction (SCFE). Decoction is a traditional Chinese medicine extraction method. The bird's nest horns are heated and boiled together with water to dissolve the active ingredients in the water, and then filtered to obtain the extract. This method is simple and easy to implement, but it has the disadvantages of low extraction efficiency, long extraction time, and high energy consumption. The acid-base extraction method uses acid-base solutions to react chemically with the active ingredients in bird's nest horns and bird's nest pieces to dissolve them in a solvent. By adjusting the pH value of the solution, the separation and purification of the target ingredients can be achieved, but this method may destroy some active ingredients, and requires strict control of pH, which is complicated to operate. Supercritical fluid extraction technology, especially supercritical CO2 extraction, has been widely used in the field of traditional Chinese medicine extraction. This technology controls pressure and temperature to give supercritical fluid selective dissolution ability, thereby efficiently extracting active ingredients from bird's nest and bird's nest fragments. This method has high extraction efficiency, no solvent residue, and is environmentally friendly, but it requires large equipment investment and high operating costs.
[0005] In view of this, the present invention is proposed. Summary of the invention
[0006] The purpose of the present invention is to provide an extract of bird's horn and bird's nest pieces, and a preparation method and application thereof. The method adopts microfluidization technology to assist enzymatic hydrolysis, so as to efficiently dissolve and release the effective ingredients such as protein, sialic acid, etc. in the bird's horn and bird's nest pieces tissue, thereby improving the extraction efficiency and retaining the biological activity of the extracted substances to the maximum extent.
[0007] The first aspect of the present invention provides a method for preparing a bird's nest and bird's nest extract, comprising the following steps:
[0008] S1. Soaking bird's nest horns and bird's nest pieces in water to obtain expanded bird's nest horns and bird's nest pieces;
[0009] S2, immersing the bird's nest and bird's nest pieces soaked in step S1 in a reducing agent aqueous solution and / or water for ultrasonic cleaning several times to obtain cleaned bird's nest and bird's nest pieces;
[0010] S3, crushing the bird's nest pieces cleaned in step S2;
[0011] S4, adding protease to the bird's nest pieces crushed in step S3 for enzymolysis, and using microfluidization technology to assist enzymolysis;
[0012] S5, heating the enzymatic solution obtained after the treatment in step S4 to inactivate the protease;
[0013] S6. Centrifuge, filter and separate the enzymatic hydrolysate after inactivation in step S5 to obtain bird's nest and bird's nest extract.
[0014] Preferably, in step S1, the mass ratio of bird's nest and bird's nest pieces to water is 1:(10-30), and the mixture is soaked at room temperature for 2-6 hours; more preferably, the mass ratio of bird's nest and bird's nest pieces to water is 1:20, and the mixture is soaked at room temperature for 3-4 hours.
[0015] Preferably, in step S2, the reducing agent includes: one or more of vitamin C, tea polyphenols, glutathione and gallic acid; more preferably, the reducing agent includes: at least two of vitamin C, tea polyphenols, glutathione and gallic acid, for example: a combination of vitamin C and tea polyphenols, glutathione and gallic acid, vitamin C and tea polyphenols and glutathione, vitamin C and tea polyphenols and glutathione and gallic acid, etc.
[0016] The contents of nitrite, nitrate and heavy metal in the bird's nest and bird's nest pieces after soaking are measured. When the contents of nitrite and nitrate exceed the preset values, the bird's nest is soaked in a reducing agent aqueous solution, filtered and then soaked several times, and finally ultrasonically cleaned several times in pure water to obtain the cleaned bird's nest and bird's nest pieces; when the contents of nitrite and nitrate do not exceed the preset values, ultrasonic cleaning is directly performed several times in pure water. The bird's nest and bird's nest pieces are cleaned with a reducing agent. The reducing agent can react with nitrite and reduce it to non-toxic nitrogen or other nitrogen compounds, thereby effectively degrading the nitrite content in the bird's nest, improving the safety of bird's nest consumption, reducing the health risks caused by excessive intake of nitrite, and improving the taste and color of the bird's nest and bird's nest pieces to a certain extent.
[0017] Preferably, in step S2, the conditions for ultrasonic cleaning are: ultrasonic power 100-200 W, ultrasonic frequency 25-35 kHz, and ultrasonic time of 5-10 min each time.
[0018] Preferably, step S3 includes:
[0019] S31, wet-grinding the cleaned bird's nest horns and bird's nest pieces into millimeter-sized particles;
[0020] S32, subjecting the ground bird's nest horns and bird's nest pieces to ultrasonic crushing.
[0021] Preferably, in step S4, the pressure range of the microjet is 20 MPa-220 MPa, and the temperature range is 40° C.-60° C.; more preferably, the pressure of the microjet is 150 MPa.
[0022] Preferably, in step S4, the protease comprises: one or more of trypsin, cathepsin, papain, alkaline protease and subtilisin.
[0023] Preferably, the amount of protease added is 4‰ to 10‰ of the dry matter weight of the bird's nest horns and chopped bird's nest, the enzymolysis conditions are 40-60°C, the enzymolysis time is 1-2h, and the solid-liquid ratio is 2:100-5:100 (g / ml); more preferably, the amount of protease added is 6‰ of the dry matter weight of the bird's nest horns and chopped bird's nest, the enzymolysis conditions are 55°C, the enzymolysis time is 1.5h, and the solid-liquid ratio is 3:100 (g / ml).
[0024] Preferably, in step S5, the conditions for heating to inactivate the protease include: heating temperature 80-90° C., heating time 20-50 min.
[0025] Preferably, an antioxidant may be added to the enzymatic hydrolysis solution of step S6, and the antioxidant includes: one or more of catalase, glutathione reductase, glucose-6-phosphate dehydrogenase, vitamin E and vitamin C; more preferably, the antioxidant includes: at least two of catalase, glutathione reductase, glucose-6-phosphate dehydrogenase, vitamin E and vitamin C, for example: a combination of glutathione reductase and vitamin E, glucose-6-phosphate dehydrogenase and vitamin C, catalase, etc., and vitamin E and vitamin C.
[0026] The addition of antioxidants can significantly enhance its antioxidant properties, effectively remove or neutralize free radicals generated during storage, processing and use, thereby protecting the active ingredients in the bird's nest and bird's nest extract from being destroyed by oxidation; it can significantly extend the shelf life of the bird's nest and bird's nest extract, and reduce the risk of quality degradation and deterioration due to oxidation, which is of great significance for maintaining product stability and market competitiveness; antioxidants help maintain the stability and activity of nutrients in the bird's nest and bird's nest extract, such as protein, carbohydrates, amino acids and various trace elements; adding antioxidants can keep the bird's nest and bird's nest extract bright color and pleasant aroma for a longer period of time, thereby improving the user experience.
[0027] Preferably, in step S6, an amino acid protecting agent may be added, including one or more of tert-butyloxycarbonyl (di-tert-butyl dicarbonate), benzyloxycarbonyl (benzyloxycarbonyl chloride), and fluorenylmethoxycarbonyl (9-fluorenylmethoxycarbonyl chloride); more preferably, the amino acid protecting agent includes at least two of tert-butyloxycarbonyl (di-tert-butyl dicarbonate), benzyloxycarbonyl (benzyloxycarbonyl chloride), and fluorenylmethoxycarbonyl (9-fluorenylmethoxycarbonyl chloride), for example, a combination of tert-butyloxycarbonyl and benzyloxycarbonyl, benzyloxycarbonyl and fluorenylmethoxycarbonyl, tert-butyloxycarbonyl, benzyloxycarbonyl, and fluorenylmethoxycarbonyl.
[0028] In addition, the amino acid protecting agent also includes other specific protecting groups: according to the characteristics of the amino acid side chain group, other specific protecting groups can also be selected for protection. For example, for cysteine containing a thiol group, a trityl (Trt) protecting group can be used for protection; for histidine containing an imidazole group, a p-toluenesulfonyl (Tos) group can be used for protection, etc.
[0029] Since the acid-base environment may change during the proteolysis process, thereby destroying the amino acids or causing condensation reactions, affecting their biological activity and nutritional value, the addition of amino acid protective agents can stabilize the amino acids, prevent them from being destroyed or undergoing condensation reactions, maintain their biological activity, and ensure the nutritional value of the product. In addition, the addition of amino acid protective agents can improve the stability of the product, help extend the shelf life of the product, and reduce the loss of nutritional value of the product during storage and transportation. In addition, the addition of amino acid protective agents can prevent the amino acids from being destroyed when the bird's nest and bird's nest extract is compounded with other products due to the different chemical properties of the compounded products, thereby protecting their integrity.
[0030] A second aspect of the present invention provides a bird's horn and bird's nest shreds extract, which is obtained by the bird's horn and bird's nest shreds extract preparation method described above.
[0031] Preferably, the extract is combined with a carrier such as chitosan-nano iron to form a stable inclusion complex, thereby further improving the stability of the product and reducing degradation and inactivation during storage and transportation.
[0032] The third aspect of the present invention provides an application of an extract of bird's horns and bird's nest fragments, wherein the extract of bird's horns and bird's nest fragments is applied in bird's nest products.
[0033] Specifically, the bird's nest products include but are not limited to: health products, foods, beauty products and beverages with whitening, anti-oxidation, anti-fatigue, anti-aging, anti-virus and immunity-enhancing effects.
[0034] The present invention has the following beneficial effects:
[0035] The present invention adopts microfluidization technology to assist enzymolysis, and utilizes high-pressure and high-speed microjets to form a huge pressure difference between the inside and outside of the swallow's horn and swallow's shreds tissue, so as to efficiently dissolve and release the effective ingredients in the swallow's horn and swallow's shreds tissue, such as protein, sialic acid, etc., thereby improving the extraction efficiency; the microfluidization technology can be carried out under mild operating conditions, avoiding the destruction of active ingredients such as protein by high temperature or strong acid and alkali conditions, and effectively retaining the nutrients and biological activity in the swallow's horn and swallow's shreds; the protein structure of the swallow's horn and swallow's shreds liquid after microjet treatment becomes looser, which is easy to be recognized and cut into small molecule peptides by proteases, which can not only improve the efficiency of the enzymolysis reaction, but also shorten the time of the enzymolysis reaction; by optimizing the conditions of microjet treatment, the molecular weight distribution of the enzymolysis product can be accurately controlled, so as to obtain small molecule peptides with specific biological activity; the microjet-assisted proteolysis technology can significantly improve the purity and activity of the swallow's horn and swallow's shreds extract, and by removing impurities and ineffective components, retaining and enriching the effective components, thereby improving the overall quality of the product; the microjet-assisted proteolysis technology has low equipment investment and low operating cost, which is helpful to promote the sustainable development and transformation and upgrading of the swallow's horn and swallow's shreds industry. DETAILED DESCRIPTION
[0036] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.
[0037] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form also includes the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.
[0038] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0039] Example 1
[0040] This embodiment provides a method for preparing a bird's nest and bird's nest extract, comprising the following steps:
[0041] (1) Soak bird's nest shreds and bird's nest shreds in water to obtain expanded bird's nest shreds, with the mass ratio of bird's nest shreds and bird's nest shreds to water being 1:20, and soak at room temperature for 3 hours.
[0042] (2) Soaking the soaked bird's nest and bird's nest pieces in a mixed aqueous solution of vitamin C and tea polyphenols and in water for ultrasonic cleaning three times each to obtain cleaned bird's nest and bird's nest pieces; the ultrasonic cleaning conditions are: ultrasonic power 150 W, ultrasonic frequency 30 kHz, and ultrasonic time 5 min each time.
[0043] (3) Place the cleaned bird's nest and bird's nest pieces in a grinding device with a rotation speed of 600 r / min for wet grinding for 10 minutes, and grind them into a particle size of about 0.25 mm; place the ground bird's nest and bird's nest pieces in an ultrasonic device with an ultrasonic intensity of 150 W, an ultrasonic frequency of 20 KHz, and an ultrasonic time of 30 minutes.
[0044] (4) Papain and alkaline protease were added in a ratio of 1:1 to the crushed bird's nest and bird's nest pieces for enzymatic hydrolysis. The total amount of protease added was 6‰ of the dry matter weight of the bird's nest and bird's nest pieces. The enzymatic hydrolysis temperature was 55°C, the enzymatic hydrolysis time was 1.5 h, the solid-liquid ratio was 3:100 (g / ml), and microfluidization technology was used to assist enzymatic hydrolysis at a microfluidization pressure of 20 MPa.
[0045] (5) The enzymatic hydrolysate obtained after the treatment is heated to inactivate the protease; the inactivation temperature is 85° C. and the time is 30 min.
[0046] (6) The inactivated enzymatic hydrolysate was centrifuged at 4500 r / min for 8 min, and then passed through 40-mesh and 200-mesh vibrating sieves in turn to obtain bird's nest horn and bird's nest shreds extract.
[0047] The present embodiment also provides a bird's nest and bird's nest chopped extract, which is obtained by the bird's nest and bird's nest chopped extract preparation method described above.
[0048] Example 2
[0049] This embodiment is basically the same as Embodiment 1, except that the microjet pressure in this embodiment is 50 MPa.
[0050] Example 3
[0051] This embodiment is basically the same as Embodiment 1, except that the microjet pressure in this embodiment is 80 MPa.
[0052] Example 4
[0053] This embodiment is basically the same as Embodiment 1, except that the microjet pressure in this embodiment is 120 MPa.
[0054] Example 5
[0055] This embodiment is basically the same as Embodiment 1, except that the microjet pressure in this embodiment is 150 MPa.
[0056] Example 6
[0057] This embodiment is basically the same as Embodiment 1, except that the microjet pressure in this embodiment is 180 MPa.
[0058] Example 7
[0059] This embodiment is basically the same as Embodiment 1, except that the microjet pressure in this embodiment is 200 MPa.
[0060] Example 8
[0061] This embodiment is basically the same as Embodiment 1, except that the microjet pressure in this embodiment is 220 MPa.
[0062] Comparative Example
[0063] This comparative example is basically the same as Example 1, except that in this comparative example, only protease is used for enzymolysis, and microfluidization technology is not used to assist enzymolysis.
[0064] Test Case
[0065] According to the existing measurement standards and methods, the bird's nest and bird's nest extracts obtained in Examples 1-8 and the comparative example were measured. The protein content of the extracts was tested according to GB5009.5-2016, and the sialic acid content of the extracts was tested according to GB / T30636-2014. The results are shown in Table 1.
[0066] Table 1
[0067] project Protein (g / 100g) Sialic acid (g / kg) Comparative Example 0.45 0.78 Example 1 0.86 1.09 Example 2 0.98 1.21 Example 3 1.16 1.33 Example 4 1.37 1.56 Example 5 1.56 1.68 Example 6 1.51 1.62 Example 7 1.47 1.55 Example 8 1.32 1.49
[0068] As can be seen from Table 1, the protein and sialic acid content of Examples 1-8 using microfluidics to assist enzymatic extraction of protein and sialic acid content has a significant improvement compared to the comparative example without using microfluidics. In addition, as the microfluidic pressure increases from 20Mpa to 220Mpa, protein and sialic acid show a trend of first increasing and then decreasing. At 150Mpa, the protein and sialic acid content are the highest. This is because appropriate pressure can promote the contact of protease with bird's nest and bird's nest fragments, accelerate the enzymatic reaction, and when the pressure is too high, the enzyme molecules will be overactive, resulting in an excessively fast enzymatic reaction rate, and may even cause nonspecific enzymolysis, reducing the purity and yield of the target product. Excessive pressure will also destroy the spatial structure of the enzyme molecules, resulting in enzyme inactivation, thereby reducing the efficiency of enzymatic hydrolysis.
[0069] The total colony count of the extract was tested according to GB4789.2-2016. At the same time, to ensure that the extract can be used directly, the nitrite, heavy metals, microorganisms and other indicators of the extract were tested with reference to the enterprise standard QB / BZXXD 0001-2021. The results are shown in Table 2.
[0070] Table 2 Safety index content data table of extract
[0071]
[0072]
[0073] It can be seen from several indicators in Table 2 that the safety of the bird's nest and bird's nest extract of the present invention meets the standards, and can be directly used as an extract stock solution for the preparation of health products, foods, beauty products, beverages and other products with whitening, anti-oxidation, anti-fatigue, anti-aging, anti-virus and immunity-enhancing effects.
[0074] In summary, the present invention adopts microfluidization technology to assist enzymolysis, and utilizes high-pressure and high-speed microjets to form a huge pressure difference between the inside and outside of the bird's nest and bird's nest tissue, so as to efficiently dissolve and release the effective ingredients in the bird's nest and bird's nest tissue, such as protein, sialic acid, etc., thereby improving the extraction efficiency; the microjet technology can be carried out under mild operating conditions, avoiding the destruction of active ingredients such as protein under conditions such as high temperature or strong acid and strong alkali, and effectively retaining the nutrients and biological activity in the bird's nest and bird's nest; the bird's nest and bird's nest liquid after microjet treatment has a looser protein structure, which is easy to be recognized and cut into Small molecule peptides can not only improve the efficiency of enzymatic hydrolysis reactions, but also shorten the time of enzymatic hydrolysis reactions; by optimizing the conditions of microfluidization treatment, the molecular weight distribution of enzymatic hydrolysis products can be precisely controlled to obtain small molecule peptides with specific biological activity; microfluidic-assisted proteolysis technology can significantly improve the purity and activity of bird's nest and bird's nest extracts, and by removing impurities and ineffective ingredients, retaining and enriching effective ingredients, the overall quality of the product is improved; microfluidic-assisted proteolysis technology requires small equipment investment and low operating costs, which will help promote the sustainable development and transformation and upgrading of the bird's nest and bird's nest industry.
[0075] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.
Claims
1. A method for preparing an extract of bird's nest horns and broken bird's nests, characterized in that: The steps include: S1. Soaking bird's nest horns and bird's nest pieces in water to obtain expanded bird's nest horns and bird's nest pieces; S2, immersing the bird's nest and bird's nest pieces soaked in step S1 in a reducing agent aqueous solution and / or water for ultrasonic cleaning several times to obtain cleaned bird's nest and bird's nest pieces; S3, crushing the bird's nest pieces cleaned in step S2; S4, adding protease to the bird's nest pieces crushed in step S3 for enzymolysis, and using microfluidization technology to assist enzymolysis; S5, heating the enzymatic solution obtained after the treatment in step S4 to inactivate the protease; S6. Centrifuge, filter and separate the enzymatic hydrolysate after inactivation in step S5 to obtain bird's nest and bird's nest extract.
2. The method for preparing the bird's nest horn and bird's nest extract according to claim 1, characterized in that: In step S1, the mass ratio of bird's nest and bird's nest pieces to water is 1:(10-30), and the mixture is soaked at room temperature for 2-6 hours.
3. The method for preparing the bird's nest horn and bird's nest extract according to claim 1, characterized in that: In step S2, the reducing agent includes: one or more of vitamin C, tea polyphenols, glutathione and gallic acid.
4. The method for preparing the bird's nest horn and bird's nest extract according to claim 1, characterized in that: In step S2, the ultrasonic cleaning conditions are: ultrasonic power 100-200W, ultrasonic frequency 25-35kHz, and ultrasonic time of 5-10min each time.
5. The method for preparing the bird's nest horn and bird's nest extract according to claim 1, characterized in that: Step S3 includes: S31, wet-grinding the cleaned bird's nest horns and bird's nest pieces into millimeter-sized particles; S32, subjecting the ground bird's nest horns and bird's nest pieces to ultrasonic crushing.
6. The method for preparing the bird's nest horn and bird's nest extract according to claim 1, characterized in that: In step S4, the pressure range of the micro jet is 20 MPa-220 MPa, and the temperature range is 40° C.-60° C.
7. The method for preparing the extract of bird's nest horns and broken bird's nests according to claim 1, characterized in that: In step S4, the protease includes one or more of trypsin, cathepsin, papain, alkaline protease and subtilisin.
8. The method for preparing the bird's nest horn and bird's nest extract according to claim 1, characterized in that: In step S5, the conditions for heating to inactivate the protease include: heating temperature 80-90° C., heating time 20-50 min.
9. An extract of bird's nest horns and bird's nest pieces, characterized in that: The bird's horn and bird's nest crumbs extract is obtained by the method for preparing the bird's horn and bird's nest crumbs extract according to any one of claims 1 to 8.
10. An application of bird's nest horns and bird's nest pieces extract, characterized in that: The bird's nest horns and bird's nest fragments extract as claimed in claim 9 is applied in bird's nest products.