Toothpaste containing enzyme preparation and preparation method thereof

Abstract

The invention relates to the technical field of enzyme preparations, and particularly discloses toothpaste containing an enzyme preparation and a preparation method of the toothpaste, and the toothpaste comprises the following components in percentage by weight: 0.05%-0.2% of glucanase, 0.1%-0.3% of lysozyme, 0.02%-0.08% of lactoperoxidase, 3%-8% of nano-hydroxyapatite, 1%-3% of trehalose, 10%-20% of glycerol, 20%-40% of a friction agent, 1%-2% of a foaming agent and the balance of water. And the balance of deionized water. The glucanase, the lysozyme and the lactoperoxidase are respectively wrapped in the sodium alginate microspheres through a gradient pH embedding technology, the thickness of an embedding layer and the pH environment are controlled, external adverse factors are effectively isolated, and the storage stability and the use activity of the enzyme are remarkably improved.

Description

Technical Field

[0001] The invention belongs to the technical field of enzyme preparations, and particularly relates to a toothpaste containing an enzyme preparation and a preparation method thereof. Background Art

[0002] Traditional toothpaste mainly achieves cleaning and anti-caries functions through abrasives and fluoride, but mechanical friction has limited efficiency in removing dental plaque and is difficult to effectively inhibit the colonization of oral pathogens. Enzyme preparations are considered an important direction for improving oral care effectiveness because of their specific ability to decompose plaque biofilms and their antibacterial activity.

[0003] However, enzymes in toothpaste systems are easily affected by environmental factors and interactions with other ingredients, resulting in loss of activity or insufficient stability. At the same time, in the existing technology, the encapsulation and protection methods of enzyme preparations are single, making it difficult to achieve multi-enzyme synergy and long-term sustained release. Summary of the Invention

[0004] The object of the present invention is to provide a toothpaste containing an enzyme preparation and a preparation method thereof, so as to solve the problems raised in the above background technology.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] A toothpaste containing an enzyme preparation, comprising the following components in weight percentage: 0.05%-0.2% of glucanase, 0.1%-0.3% of lysozyme, 0.02%-0.08% of lactoperoxidase, 3%-8% of nanohydroxyapatite, 1%-3% of trehalose, 10%-20% of glycerol, 20%-40% of abrasive, 1%-2% of foaming agent, and the balance being deionized water;

[0007] The dextranase, lysozyme and lactoperoxidase are respectively encapsulated in the sodium alginate microspheres through gradient pH embedding, and the embedding layer thickness is 50-200 nm.

[0008] Preferably, the activity ratio of the glucanase to the lysozyme is 1:1.5-2.5, and the activity of the lactoperoxidase is maintained by a thiocyanate activation system.

[0009] Preferably, the nano-hydroxyapatite has a rod-shaped crystal structure with an aspect ratio of 10:1-20:1, and its surface is modified with polyaspartic acid.

[0010] Preferably, the friction agent is a mixture of modified hydrated silica with a particle size of D50 = 8-12 μm and calcium carbonate with a particle size of D50 = 3-5 μm in a ratio of 2:1-3:1.

[0011] A method for preparing a toothpaste containing an enzyme preparation comprises the following steps:

[0012] S1. Premix trehalose and glycerol at 35-40°C to form a protective matrix;

[0013] S2. Use a microfluidic device to sequentially embed three enzyme preparations in a pH gradient environment, and control the embedding temperature at 4-8°C;

[0014] S3, mixing the enzyme microsphere dispersion and the premixed matrix with low-speed stirring under the protection of inert gas;

[0015] S4. Add nano-hydroxyapatite and abrasive in stages, homogenizing at 2000-3000 rpm for 3-5 minutes after each addition;

[0016] S5. Finally, a foaming agent is added and degassing is performed under a vacuum degree of -0.08 to -0.1 MPa to obtain toothpaste.

[0017] Preferably, in step S2, the embedding pH of glucanase is 5.2-5.8, the embedding pH of lysozyme is 6.8-7.2, and the embedding pH of lactoperoxidase is 4.5-5.0.

[0018] Preferably, in step S4, the nano-hydroxyapatite is added in three times, each time with an interval of 10 minutes, and the addition temperature gradient is 25°C, 30°C, and 35°C.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] (1) Through the gradient pH encapsulation technology, dextranase, lysozyme and lactoperoxidase are respectively encapsulated in sodium alginate microspheres, and the thickness of the encapsulation layer and the pH environment are controlled to effectively isolate adverse external factors and significantly improve the storage stability and activity of the enzymes. Dextranase decomposes the dextran biofilm matrix, lysozyme directly lyses the bacterial cell wall, and lactoperoxidase generates antibacterial substances under the activation of thiocyanate. Multiple mechanisms work together to remove dental plaque.

[0021] (2) After surface modification with polyaspartic acid, nanohydroxyapatite is evenly dispersed in a rod-shaped crystal structure, which promotes the remineralization of tooth enamel and reduces the risk of abrasive wear on teeth. Nanohydroxyapatite and compound abrasives are added in stages, combined with temperature gradient control and high-speed homogenization process to improve the uniformity of ingredients and paste stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a process flow chart of the present invention. DETAILED DESCRIPTION

[0023] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0024] Example 1:

[0025] See also Figure 1 As shown, a toothpaste containing an enzyme preparation comprises the following components in percentage by weight:

[0026] Glucanase 0.1%, lysozyme 0.2%, lactoperoxidase 0.05%, nanohydroxyapatite 5%, trehalose 2%, glycerol 15%, friction agent 30%, foaming agent 1.5%, and the balance is deionized water.

[0027] A method for preparing a toothpaste containing an enzyme preparation comprises the following steps:

[0028] S1. Mix trehalose and glycerol in a 38°C constant temperature water bath and stir at 300 rpm for 20 minutes to form a transparent mucus;

[0029] S2. Use a microfluidic device (flow rate 0.5 mL / min) to embed three enzymes separately:

[0030] Dextranase pH = 5.5, embedding layer thickness 80 nm;

[0031] Lysozyme pH = 7.0, embedding layer thickness 120 nm;

[0032] Lactoperoxidase: pH = 4.8, embedding layer thickness 60 nm;

[0033] The particle size distribution of microspheres after embedding is 200-400nm;

[0034] S3. Under nitrogen protection, the enzyme microsphere dispersion and the matrix were mixed at a low speed of 500 rpm for 30 minutes, and nanohydroxyapatite was added in three stages:

[0035] First time: 25℃, addition amount 2%;

[0036] Second time: 30℃, adding amount 1.5%;

[0037] The third time: 35℃, adding amount 1.5%;

[0038] After each addition, homogenize at 2500 rpm for 4 minutes;

[0039] S4. After adding the foaming agent, degassing was performed at a vacuum degree of -0.09 MPa for 15 minutes to obtain toothpaste.

[0040] Example 2:

[0041] A toothpaste containing an enzyme preparation comprises the following components in percentage by weight:

[0042] Glucanase 0.15%, lysozyme 0.25%, lactoperoxidase 0.06%, nanohydroxyapatite 6%, trehalose 2.5%, glycerol 18%, friction agent 35%, foaming agent 1.8%, and the balance is deionized water.

[0043] The preparation method comprises the following steps:

[0044] S1. Mix trehalose and glycerol in a 36°C constant temperature water bath and stir at 400 rpm for 25 minutes to form a transparent mucus;

[0045] S2. Use a microfluidic device (flow rate 0.6 mL / min) to embed three enzymes separately:

[0046] Dextranase: pH = 5.6, embedding layer thickness 100 nm;

[0047] Lysozyme: pH = 7.1, embedding layer thickness 150 nm;

[0048] Lactoperoxidase: pH = 4.9, embedding layer thickness 70 nm;

[0049] The particle size distribution of microspheres after embedding is 220-450nm;

[0050] S3. Under argon protection, the enzyme microsphere dispersion and the matrix were mixed at a low speed of 600 rpm for 40 minutes;

[0051] Nanohydroxyapatite is added in three stages:

[0052] First time: 28℃, addition amount 2.5%;

[0053] Second time: 32℃, addition amount 2.0%;

[0054] The third time: 37°C, adding 1.5%;

[0055] After each addition, homogenize at 2800 rpm for 4.5 minutes;

[0056] S4. After adding the foaming agent, degassing was performed at a vacuum degree of -0.085 MPa for 20 minutes to obtain toothpaste.

[0057] Example 3:

[0058] A toothpaste containing an enzyme preparation comprises the following components in percentage by weight:

[0059] Dextranase 0.08%, lysozyme 0.18%, lactoperoxidase 0.04%, nanohydroxyapatite 4%, trehalose 1.8%, glycerol 12%, friction agent 25%, foaming agent 1.2%, and the balance is deionized water.

[0060] The preparation method comprises the following steps:

[0061] S1. Mix trehalose and glycerol in a 40°C constant temperature water bath and stir at 250 rpm for 15 minutes to form a transparent mucus;

[0062] S2. Use a microfluidic device (flow rate 0.4 mL / min) to embed three enzymes separately:

[0063] Dextranase: pH = 5.3, embedding layer thickness 60 nm;

[0064] Lysozyme: pH = 6.9, embedding layer thickness 100 nm;

[0065] Lactoperoxidase: pH = 4.7, embedding layer thickness 50 nm;

[0066] The particle size distribution of microspheres after embedding is 180-380nm;

[0067] S3. Under nitrogen protection, the enzyme microsphere dispersion and the matrix were mixed at a low speed of 400 rpm for 25 minutes;

[0068] Nanohydroxyapatite is added in three stages:

[0069] First time: 24℃, addition amount 1.5%;

[0070] Second time: 29℃, addition amount 1.5%;

[0071] The third time: 34°C, adding amount 1.0%;

[0072] After each addition, homogenize at 2200 rpm for 3.5 minutes;

[0073] S4. After adding the foaming agent, degassing was performed at a vacuum degree of -0.095 MPa for 18 minutes to obtain toothpaste.

[0074] Application experiment example:

[0075] An experimental group and a control group were set up. The experimental group was the enzyme-containing toothpaste prepared in Example 1 (containing glucanase, lysozyme, and lactoperoxidase), and the control group was a commercially available ordinary toothpaste (without enzyme preparation, and the main ingredients were sodium fluoride and calcium carbonate);

[0076] Hydroxyapatite sheets (simulating tooth enamel surfaces) were pre-coated with salivary protein and then inoculated with bacterial solution for 48 hours to obtain artificial dental plaque models. The artificial dental plaque models were randomly divided into an experimental group and a control group (n = 10 per group). The model surfaces were brushed with toothpaste using an electric toothbrush (pressure 200 g, frequency 200 times / min) for 2 minutes. After rinsing, the residual plaque area was quantified using crystal violet staining (analyzed with ImageJ software) and the clearance rate was calculated:

[0077] Clearance rate (%) = (1-plaque area before treatment / plaque area after treatment) × 100;

[0078] The experimental data are as follows:

[0079] Group Average clearance rate (%) Standard deviation (%) Experimental group 82.4 3.1 control group 58.7 2.5

[0080] From the above, we can see that the clearance rate of the experimental group was significantly higher than that of the control group (p<0.05).

[0081] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A toothpaste containing an enzyme preparation, characterized in that Contains the following components in weight percentage: Glucanase 0.05%-0.2%, lysozyme 0.1%-0.3%, lactoperoxidase 0.02%-0.08%, nanohydroxyapatite 3%-8%, trehalose 1%-3%, glycerol 10%-20%, abrasive 20%-40%, foaming agent 1%-2%, and the balance is deionized water; The dextranase, lysozyme and lactoperoxidase are respectively encapsulated in the sodium alginate microspheres through gradient pH embedding, and the embedding layer thickness is 50-200 nm.

2. The toothpaste containing an enzyme preparation according to claim 1, characterized in that: The activity ratio of the glucanase to the lysozyme is 1:1.5-2.5, and the activity of the lactoperoxidase is maintained by a thiocyanate activation system.

3. The toothpaste containing an enzyme preparation according to claim 1, characterized in that: The nano-hydroxyapatite has a rod-shaped crystal structure, an aspect ratio of 10:1-20:1, and a surface modified with polyaspartic acid.

4. The toothpaste containing an enzyme preparation according to claim 1, characterized in that: The friction agent is a mixture of modified hydrated silica with a particle size of D50 = 8-12 μm and calcium carbonate with a particle size of D50 = 3-5 μm in a ratio of 2:1-3:

1.

5. A method for preparing toothpaste containing enzyme preparation, characterized in that: The following steps are involved: S1. Premix trehalose and glycerol at 35-40°C to form a protective matrix; S2. Use a microfluidic device to sequentially embed three enzyme preparations in a pH gradient environment, and control the embedding temperature at 4-8°C; S3, mixing the enzyme microsphere dispersion and the premixed matrix with low-speed stirring under the protection of inert gas; S4. Add nano-hydroxyapatite and abrasive in stages, homogenizing at 2000-3000 rpm for 3-5 minutes after each addition; S5. Finally, a foaming agent is added and degassing is performed under a vacuum degree of -0.08 to -0.1 MPa to obtain toothpaste.

6. The method for preparing a toothpaste containing an enzyme preparation according to claim 5, characterized in that: In the step S2, the embedding pH of glucanase is 5.2-5.8, the embedding pH of lysozyme is 6.8-7.2, and the embedding pH of lactoperoxidase is 4.5-5.

0.

7. The method for preparing a toothpaste containing an enzyme preparation according to claim 5, characterized in that: In step S4, the nano-hydroxyapatite is added in three times, each time with an interval of 10 minutes, and the addition temperature gradient is 25°C, 30°C, and 35°C.

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