CN117122549A - Anti-sugar composition containing hydrolyzed wheat protein and application thereof - Google Patents
Anti-sugar composition containing hydrolyzed wheat protein and application thereof Download PDFInfo
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- CN117122549A CN117122549A CN202311045648.6A CN202311045648A CN117122549A CN 117122549 A CN117122549 A CN 117122549A CN 202311045648 A CN202311045648 A CN 202311045648A CN 117122549 A CN117122549 A CN 117122549A
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- ACCCMOQWYVYDOT-UHFFFAOYSA-N hexane-1,1-diol Chemical compound CCCCCC(O)O ACCCMOQWYVYDOT-UHFFFAOYSA-N 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
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- DLRVVLDZNNYCBX-RTPHMHGBSA-N isomaltose Chemical group O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)C(O)O1 DLRVVLDZNNYCBX-RTPHMHGBSA-N 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
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- OEIJHBUUFURJLI-UHFFFAOYSA-N octane-1,8-diol Chemical compound OCCCCCCCCO OEIJHBUUFURJLI-UHFFFAOYSA-N 0.000 description 1
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- HBMJWWWQQXIZIP-UHFFFAOYSA-N silicon carbide Chemical compound [Si+]#[C-] HBMJWWWQQXIZIP-UHFFFAOYSA-N 0.000 description 1
- 229910010271 silicon carbide Inorganic materials 0.000 description 1
- 235000011888 snacks Nutrition 0.000 description 1
- FQENQNTWSFEDLI-UHFFFAOYSA-J sodium diphosphate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]P([O-])(=O)OP([O-])([O-])=O FQENQNTWSFEDLI-UHFFFAOYSA-J 0.000 description 1
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- 150000008163 sugars Chemical class 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 235000019818 tetrasodium diphosphate Nutrition 0.000 description 1
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- RZLVQBNCHSJZPX-UHFFFAOYSA-L zinc sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Zn+2].[O-]S([O-])(=O)=O RZLVQBNCHSJZPX-UHFFFAOYSA-L 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
- A61K8/20—Halogens; Compounds thereof
- A61K8/21—Fluorides; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/645—Proteins of vegetable origin; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/66—Enzymes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
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Abstract
The invention belongs to the technical field of oral care, and discloses an anti-sugar composition containing hydrolyzed wheat protein and application thereof. The invention relates to an anti-sugar composition containing hydrolyzed wheat protein, which comprises the following components in parts by mass: 0.001 to 0.1 part of glucose oxidase, 0.001 to 0.1 part of dextranase, 0.001 to 10 parts of hydrolyzed wheat protein, 0.01 to 0.5 part of arginine, 0.01 to 0.05 part of fluoride and 0.001 to 0.1 part of probiotics. The anti-sugar composition adopts scientific proportion and synergy, and the prepared oral care product can effectively decompose dental plaque, reduce dental calculus, inhibit bacteria generation, inhibit tooth demineralization and promote tooth remineralization, reduce toxic and side effects of fluoride and maintain oral health.
Description
Technical Field
The invention relates to the technical field of oral care, in particular to an anti-sugar composition containing hydrolyzed wheat protein and application thereof.
Background
With the improvement of living standard, the processing refinement degree of the sugar-containing food is also higher, and the intake of the sugar-containing food and the sugar-containing beverage is in an ascending trend, wherein the children are influenced the most. Epidemiological investigation of oral health showed that the caries rate of 12 years old children was 34.5%, which increased by 7.8% over ten years ago. The caries rate of children aged 5 is 70.9%, which is a surface that the oral health of children is not optimistic. Children are a high incidence group of caries, and caries occurrence is closely related to factors such as sugar intake and oral hygiene. In addition, some acidic foods and acidic beverages are also a major factor in the deterioration of the balance of the oral environment in children and in the erosion of enamel. Because of the lack of consciousness and necessity of oral care, good brushing habit cannot be actively developed, high sugar-content snacks such as candies, biscuits and the like are often taken at ordinary times, parents are not guided in time, and many children are subjected to oral diseases such as acid erosion, pain and the like prematurely, so that the study, life and general health of the children are directly affected.
Food residues in children's mouth contain more carbohydrates which become nutrients for oral bacteria. Common oral pathogenic bacteria in children's oral cavity mainly include streptococcus mutans, porphyromonas gingivalis, fusobacterium nucleatum, lactobacillus acidophilus and the like, and these oral bacteria ferment in the process of metabolizing sugar to produce a large amount of organic acids, and these organic acids can penetrate the enamel surface to cause mineral (hydroxyapatite) loss of teeth to thereby demineralize, so that the teeth are corroded by acid. Furthermore, as the bacteria in the oral cavity multiply to form a layer of adhered film-dental plaque on the surface of the teeth, acid generated by the bacteria can be adhered to the surface of the teeth for a long time, so that the hydroxyapatite is dissolved by the acid to generate hydrogen phosphate ions and calcium ions, and the hydrogen phosphate ions and the calcium ions are washed away by saliva to finally erode the teeth, so that the teeth are accelerated to be demineralized, and vicious circle is caused. Thus, the anti-glycaemic effect of the children's toothpaste is particularly important.
Fluoride inhibits bacterial acid production, inhibits enamel demineralization and promotes its remineralization, and has been widely used in the oral care industry. However, the safety of the oral liquid needs to be considered for long-term use so as to avoid complications such as fluoridontopathy. The toothpaste with the sugar-resistant effect on the market at present only depends on sodium fluoride to achieve the caries-preventing effect, the sugar-resistant path is single, no deep research is performed on the sugar-resistant components and the action mechanism of the toothpaste, and the removal of dental plaque biomembrane and removal of dental calculus are difficult to be achieved effectively. In addition, there is no children's toothpaste containing a remineralizing effective component for enamel restoration other than the fluorine-containing component toothpaste. Therefore, developing a novel children anti-sugar (anti-enamel erosion caused by glycolysis) oral care product can not only meet the requirements of wide consumers, but also have a wider market application prospect.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide an anti-sugar composition containing hydrolyzed wheat protein and application thereof.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
in a first aspect, the invention provides an anti-sugar composition containing hydrolyzed wheat protein, which comprises the following components in parts by mass: 0.001 to 10 parts of hydrolyzed wheat protein, 0.001 to 0.1 part of glucose oxidase, 0.001 to 0.1 part of dextranase, 0.01 to 0.5 part of arginine, 0.01 to 0.05 part of fluoride and 0.001 to 0.1 part of probiotics.
The anti-sugar composition adopts scientific proportion, and glucose oxidase can decompose glucose, reduce the generation of bacteria and inhibit the formation of dental plaque; the dextranase can effectively decompose dental plaque biomembrane and reduce dental calculus; the probiotics can inhibit the growth and reproduction of oral pathogenic bacteria, streptococcus mutans, porphyromonas gingivalis and fusobacterium nucleatum, and maintain the microecological balance of the oral cavity; the hydrolyzed wheat protein can be combined with calcium ions and adsorbed on the surface of tooth enamel to promote the tooth remineralization and repair process; arginine neutralizes acids produced by bacteria in the oral cavity, inhibits tooth demineralization and promotes tooth remineralization; the fluoride can enhance the hardness of teeth, promote tooth remineralization, and inhibit the growth of acid-producing bacteria; the components are synergistic, the stability of the prepared anti-sugar children toothpaste is obviously improved, and the activity of the bioactive components is high, so that the stability of various active substances in the shelf life of the product is ensured. Repeated use over a period of days is effective against sugar, including helping to inhibit oral bacteria, remove plaque biofilm, inhibit plaque formation, resist bacterial glycolysis acidogenesis, resist tooth demineralization due to glycolysis acidogenesis, promote tooth remineralization, and the like.
As a preferred embodiment of the anti-sugar composition of the present invention, the ratio of the addition amounts of both dextranase and glucose oxidase is 0.001:1 to 1:1. the preferable ratio is 0.01:1 to 1:1.
as a preferred embodiment of the anti-sugar composition of the present invention, the ratio of the addition amount of the dextranase or glucose oxidase to the hydrolyzed wheat protein or fluoride is 0.001:1 to 1:1. the preferred ratio is 0.01:1 to 1:1.
in a second aspect, the invention applies the anti-sugar composition in oral care.
In a third aspect, the present invention provides an anti-sugar oral care product comprising, in mass percent, from 0.001% to 5% of the anti-sugar composition. Preferably, the sugar-resistant composition comprises 0.01% -5%; more preferably, 0.01% to 3% of the anti-sugar composition is included.
As a preferred embodiment of the anti-glycation oral care product of the present invention, the anti-glycation oral care product is a glycation resistant child's dentifrice.
As a preferred embodiment of the anti-glycation oral care product of the present invention, the anti-glycation child toothpaste further comprises the following components in mass percent: 25 to 28 percent of humectant, 0.1 to 1 percent of natural antibacterial agent, 0.2 to 1 percent of thickening agent, 10 to 25 percent of soft abrasive, 0.01 to 20 percent of xylitol, 0.001 to 5 percent of calcium-containing compound, 0.01 to 2 percent of mild surfactant, 0.5 to 1 percent of essence and the balance of water.
As a preferred embodiment of the anti-sugar oral care product of the present invention, the natural antibacterial agent is at least one of benzyl alcohol, octylglycol, hexylene glycol, and p-hydroxyacetophenone.
As a preferred embodiment of the anti-sugar oral care product according to the invention, the calcium-containing compound is at least one of soluble calcium or sparingly soluble calcium.
As a preferred embodiment of the anti-sugar oral care product according to the present invention, the mild surfactant is at least one of sodium lauroyl sarcosinate, sodium lauroyl glutamate, cocoyl propyl betaine, sodium N-methyl cocoyl taurate, poloxamer, alkyl glycoside.
Compared with the prior art, the invention has the beneficial effects that:
the anti-sugar composition adopts scientific proportion, the components are synergistic, and the dextranase and the glucose oxidase decompose dental plaque biomembrane by dissolving extracellular polysaccharide of the dental plaque biomembrane, remove dental plaque and inhibit dental plaque formation. The anti-sugar composition can inhibit or kill harmful bacteria such as Escherichia coli, staphylococcus aureus, streptococcus mutans, porphyromonas gingivalis, fusobacterium nucleatum, etc., and has a bacteriostasis rate of 97.5% or more, thereby preventing or adjunctively treating oral diseases caused by oral pathogenic bacteria. Has effects of inhibiting pathogenic bacteria in oral cavity, resisting plaque acidogenesis, inhibiting plaque formation, removing dental plaque and dental calculus, resisting enamel erosion caused by glycolysis of oral bacteria, and promoting tooth remineralization. The prepared toothpaste can keep a formula with higher enzyme activity, and the enzyme activity is as high as 73-85% when the toothpaste is stored at 45 ℃ for 2 months.
Drawings
FIG. 1 is a statistical graph of the anti-tartar effect of a sugary child toothpaste;
FIG. 2 is a graphical representation of an in vitro inhibition plaque glycolysis evaluation experiment.
Detailed Description
For a better description of the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to the following specific examples. It will be appreciated by persons skilled in the art that the specific embodiments described herein are for purposes of illustration only and are not intended to be limiting.
The test methods used in the examples are conventional methods unless otherwise specified; the materials, reagents and the like used, unless otherwise specified, are all commercially available.
The anti-sugar composition of the present invention in various embodiments, is added in an amount of 0.001% to 5%, preferably 0.01% to 5%, more preferably 0.01% to 3%, depending on the form of the oral care product.
The raw materials of the anti-sugar children toothpaste comprise humectant, natural antibacterial agent, thickener, soft abrasive, xylitol, calcium-containing compound, anti-sugar composition, mild surfactant, essence and deionized water; the content ranges of the raw materials are as follows by weight percent: 25 to 28 percent of humectant, 0.1 to 1 percent of natural antibacterial agent, 0.2 to 1 percent of thickening agent, 10 to 25 percent of soft abrasive, 0.01 to 20 percent of xylitol, 0.001 to 5 percent of calcium-containing compound, 0.001 to 5 percent of sugar-resistant composition, 0.01 to 2 percent of mild surfactant, 0.5 to 1 percent of essence and the balance of water (preferably deionized water).
The humectant is at least one of sorbitol, glycerol, polyethylene glycol and propylene glycol.
The natural antibacterial agent is at least one of benzyl alcohol, octanediol, hexanediol and p-hydroxyacetophenone. The natural antibacterial agent belongs to food-grade components, is safe and free of toxic and side effects, has good antibacterial and bacteriostatic properties, and can prolong the shelf life of foods.
The thickener is at least one of xanthan gum, cellulose gum and carbomer.
The soft abrasive is silicon dioxide.
The calcium-containing compound is at least one of soluble calcium or slightly soluble calcium. The soluble calcium or slightly soluble calcium is at least one of calcium lactate, calcium glycerophosphate and calcium gluconate. The child resistant toothpaste also adds xylitol, which is a natural plant sweetener that is not metabolized by bacteria in the oral cavity and therefore does not produce acid to attack the teeth. The combination of xylitol and calcium-containing compound can inhibit bacterial metabolism and promote tooth remineralization, thereby resisting bacterial erosion and strengthening teeth.
The mild surfactant is at least one of sodium lauroyl sarcosinate, sodium lauroyl glutamate, cocoyl propyl betaine, N-methyl cocoyl taurate sodium, poloxamer and alkyl glycoside. The mild surfactant can clean teeth without wound cavity mucous membrane, is safe and non-irritating, can be used independently, and can also be used by compounding the mild surfactant with sodium laurylsulfate.
The anti-sugar composition comprises, by weight, 0.001-0.1 part of glucose oxidase, 0.001-0.1 part of dextran, 0.001-10 parts of hydrolyzed wheat protein, 0.01-0.5 part of arginine, 0.01-0.05 part of fluoride and 0.001-0.1 part of probiotics, wherein a plurality of active ingredients in the anti-sugar composition can play a synergistic effect in a toothpaste product, inhibit the growth of cariogenic bacteria and other oral pathogenic bacteria, inhibit the formation of dental plaque, resist enamel acid erosion and promote tooth remineralization. The ratio of the addition amount of the dextranase to the glucose oxidase in the anti-sugar composition is 0.001:1 to 1:1, preferably in a ratio of 0.01:1 to 1:1. the addition ratio of the dextranase or the glucose oxidase to the hydrolyzed wheat protein or the fluoride is 0.001:1 to 1:1, preferably in a ratio of 0.01:1 to 1:1.
in the following examples and comparative examples, the performance test method was:
(1) High temperature stability investigation of physicochemical index of toothpaste formula
The method for detecting the pH of the toothpaste refers to the national standard GB 8372 2017 toothpaste.
The method for investigating the stability of the toothpaste by referring to national standard GB 8372 2017 toothpaste is slightly modified: taking 2 sample toothpastes, wherein one sample is stored at normal temperature, the other sample is placed in a constant temperature incubator at 45+/-1 ℃, taken out at different time intervals, and compared with the normal temperature, the stability is checked: comparing paste extrusion with a sample stored at normal temperature, judging whether the appearance color and smell are normal, judging whether abnormal phenomena such as coarsening and water diversion exist, then cutting a pipe to check whether abnormal phenomena such as grain forming, coarsening, water diversion and shelling exist in the paste, and if not, judging that the stability is normal.
The toothpaste consistency method refers to the national standard GB 8372 2001 toothpaste consistency test method.
(2) Evaluation method for tooth scale removal effect of toothpaste
1) Inoculating streptococcus mutans to 8mL of BHI broth, and standing and culturing overnight at 37 ℃ in an aerobic state for 20-24 h;
2) Taking a 24-hole plate, adding 200 microliters of bacterial liquid into each hole, adding 1800 microliters of BHI containing 5% of sucrose, and carrying out aerobic culture at 37 ℃ for 20 hours;
3) The culture broth was discarded with a 5mL pipette, and 2mL of 0.1mol/L (ph=6.0) PBS was added along the pore wall for washing once;
4) Adding 2ml of a toothpaste solution (10 times diluted) with the pre-heat preservation temperature of 37 ℃ and preserving the temperature for 5 minutes in a water bath kettle or a constant temperature incubator with the temperature of 37 ℃;
5) Washing 3 times with 2mL PBS, and washing off residual toothpaste solution;
6) Oven drying at 60deg.C for 30min;
7) 2mL of 0.1% crystal violet staining solution is added into each hole for 10min;
8) Discarding the dyeing liquid, continuously cleaning for 3 times by using 2mL PBS, and drying for 10-15 min at 60 ℃;
9) 2mL of acetone was added: ethanol = 2:8 eluate, absorbance values measured at 590 nm;
tartar removal = [ (blank absorbance-enzyme preparation treatment solution absorbance)/blank absorbance ] ×100%.
(2) Evaluation method for inhibiting in-vitro dental plaque glycolysis
Principle of: by culturing dental plaque, after treatment with the sample, the pH difference between the sample group and the control group was observed to see whether the sample group could inhibit the growth of dental plaque.
And (3) preparation of a reagent: (1) plaque growth medium: 6g BBL trypticase soyase, 20g sucrose, were dissolved in 160.8g deionized water, sterilized at 121℃for 20min and cooled. 13.2g of fresh saliva was added before use. (2) Glycolytic medium: 6g BBL pancreatic casein soytone, 10g sucrose are dissolved in 184g deionized water, sterilized at 121 ℃ for 20min and cooled; (3) experimental grouping: sample treatment liquid: toothpaste samples were formulated into toothpaste: water = 1:3 toothpaste slurry; negative control: physiological saline; positive control: glycolytic mediators.
The operation steps are as follows: (1) plaque formation: 3 to 5 persons were collected, mixed (about 100 mL), sucrose was added to the saliva to a concentration of 0.1%, and 5mL of the saliva was added to a test tube. The cleaned polished glass rod was inserted and immersed overnight at 37 ℃. (2) Plaque growth: 13.2g of fresh saliva was collected and added to the growth medium and mixed well. 5mL of the medium is taken in a new test tube, the glass rod treated in the step (1) is transferred into the test tube, and the glass rod is soaked for more than 6 hours at 37 ℃. The glass rod was then transferred to a fresh tube containing 5mL of fresh saliva and soaked overnight at 37 ℃. (3) 6-8 mL of the treatment solution is added into a test tube. Immersing the glass rod treated in the step (2) in the treatment liquid once per second, and treating for 60S. The glass rod was then washed by immersing it in deionized water in a test tube for 2 washes at 10S each. (4) 5mL of glycolytic medium was added to a fresh tube, and the glass rod after washing in (3) was immersed in the tube and treated at 37℃for 6 hours. (5) The pH was measured and recorded.
Efficacy evaluation: efficacy% compared to positive control = [1- (pH mean of positive control-pH mean of experimental group)/(pH mean of positive control-pH mean of negative control) ] ×100%
The above values of greater than or equal to 50% indicate efficacy, i.e., inhibition of plaque glycolysis in vitro.
(3) Method for detecting enzyme activity stability of biological enzyme in toothpaste
Principle of: dextranase is capable of hydrolysing alpha-1, 6 glycosidic linkages in dextran. The reaction product is isomaltooligosaccharide. The activity of the dextranase was determined by measuring how much reducing sugar was formed. Reducing sugars were determined using the llanes method.
And (3) preparation of a reagent: substrate solution: dextran (Sigma, NO. 31390) from Leuconostoc was dissolved in water in a proportion of 0.5g dry matter (105 ℃,4 h) and diluted to 50ml. (as-prepared); reagent A:8.25g of potassium ferricyanide and 10.6g of anhydrous sodium carbonate, the volume is fixed to 1L by water, and the mixture is placed in shade for 2 to 3 days. (lifetime 3 months); reagent B:25g potassium iodide +50g zinc sulphate heptahydrate +250g sodium chloride, and water to 1L (3 months of life); reagent C:5ml acetic acid was diluted to 100ml with water;
measurement procedure: mixing 10ml of substrate solution and 4ml of 0.1M acetic acid buffer in a test tube (1), incubating the mixture at 37 ℃ for 10min, then adding 1ml of enzyme solution in the test tube (1) and mixing, and further incubating at 37 ℃ for 30min; another test tube (2) was prepared in advance and 5ml of reagent A+3ml of water were added for use. Taking 2ml of reaction liquid to the test tube (2) after the incubation of the test tube (1) is completed, placing the test tube (2) in a boiling water bath for 5min, taking out the test tube (2), cooling, continuously adding 5ml of reagent B+3ml of reagent C, uniformly mixing, titrating by using 0.01mol/L sodium thiosulfate, changing the solution into milky white and keeping the color of 30s unchanged as an end point, and recording the titration volume.
And (3) calculating: one unit of dextranase activity is defined as: under the above conditions, an enzyme amount having the same sugar as a micromole of sodium thiosulfate reducing ability can be produced in one minute.
Unit/ml= (B-T) ×0.01×1000× (1/30) × (15/2) ×f×n
B, blank drop quantification (Unit: ml)
T: titration amount of enzyme sample (unit: ml)
F: coefficient of 0.01mol/L sodium thiosulfate
N: dilution factor of enzyme
(4) Detection method for inhibiting pathogenic bacteria in oral cavity by toothpaste
The performance of the children's anti-glycaemic toothpaste in inhibiting oral pathogenic bacteria is evaluated by referring to the detection method of GB 15979-2002 appendix C4.
(5) Method for detecting inhibition glycolysis effect of toothpaste
(1) And (3) preparation of a reagent:
plaque growth medium: 6g BBL trypticase soyase, 20g sucrose, were dissolved in 160.8g deionized water, sterilized at 121℃for 20min and cooled. 13.2g of fresh saliva was added before use. Glycolytic medium: 6g BBL pancreatic casein soytone, 10g sucrose are dissolved in 184g deionized water, sterilized at 121 ℃ for 20min and cooled; preparing a treatment fluid: toothpaste samples were formulated into toothpaste: water = 1:3 toothpaste slurry, other bacteriostatic substances were formulated as a solution of appropriate concentration.
(2) Polishing glass rod
New glass rods were polished 25mm from one end using silicon carbide sandpaper nos. 240, 320, 400 and 600 in sequence on a lathe. After the preliminary sanding, the samples were polished again using No. 600 sandpaper prior to each experiment. The experiments are set up to a negative control group and a positive control group, and each experiment needs up to 1-2 experiment groups, and each group of experiments needs 4 glass rods.
(3) Experimental procedure
The first day: the glass rod is cleaned by dilute hydrochloric acid in an ultrasonic way, washed, dried and polished by No. 600 sand paper. And washing the glass rod with deionized water and drying. 3 to 5 persons were collected, mixed (about 100 mL), sucrose was added to the saliva to a concentration of 0.1%, and 5mL of the saliva was added to each of 16 test tubes. The glass rod was inserted and immersed overnight at 37 ℃. Preparing a bacterial plaque growth medium, and sterilizing at high temperature for the next day.
The following day: 13.2g of fresh saliva was collected and added to the growth medium and mixed well. 5mL of the medium is taken in a new test tube, the glass rod of the previous day is transferred into a new 16 test tubes, and the glass rod is soaked for more than 6 hours at 37 ℃. Then transferred to 16 tubes each containing 5mL of fresh saliva and soaked overnight at 37 ℃. Preparing glycolytic medium, and sterilizing at high temperature for the third day.
Third day: the prepared treatment solutions are respectively placed into 4 test tubes, so that the treatment solutions can fully cover bacterial plaque (about 6-8 mL) on a glass rod. 10mL of deionized water was pipetted into 32 tubes, respectively. The glass rod was taken out of the incubator at 37℃and immersed once every S in the treatment liquid, and treated for 60S. The glass rod was then washed by immersing it in deionized water in a test tube for 2 washes of 10S each. 5mL of glycolytic medium is sucked into 16 test tubes, and the washed glass rod is soaked into the glycolytic test tubes for 6 hours at 37 ℃. The pH of the experimental and control groups was determined and recorded. The glass rod is placed into a beaker, added with 1M HCl, soaked, washed and recovered.
(4) Efficacy evaluation
Efficacy% compared to positive control = [1- (pH mean of positive control-pH mean of experimental group)/(pH mean of positive control-pH mean of negative control) ] ×100%
The above values greater than or equal to 50% indicate effectiveness.
(6) Evaluation method of in-vitro acid corrosion resistance model
Principle of: the tooth paste soaking treatment is carried out on bovine enamel anthropomorphic teeth, and the glycolysis solution is used for pickling teeth, so that whether the hardness change of the anterior and posterior bovine tooth samples is obviously different from that of a control is observed to evaluate the pickling resistance.
And (3) preparation of a reagent: (1) artificial saliva: 200g/L MUCIN TYPE II, 0.38g/L NaCl, 0.183g/LCaCl 2 、0.528g/L KH 2 PO 4 1.114g/L KCl. Firstly, stirring MUCIN with hot water at 80 ℃ until the MUCIN is completely dissolved, then sequentially adding other components, stirring and dissolving, fixing the volume, and finally, regulating the pH to 7.0 with 1mol/L KOH solution; (2) glycolytic solution: 6g of trypticase soytone broth, 20g of sucrose, were sterilized in 160.8g of deionized water at 121℃for 20min and cooled. 3-4 people's fresh saliva was collected and 13.2g fresh saliva was added before use. The culture medium added with saliva is placed in a 37 ℃ incubator for culturing for 18-24 hours, and the pH value is measured to be about 4.00. Finally sterilizing at high temperature and high pressure for 30min at 121 ℃ and cooling for standby; (3) sample treatment liquid: toothpaste samples were formulated into toothpaste: water = 1: 3.
The operation steps are as follows:
1) Screening hardness value is (230-360) kg/mm 2 Is a dental enamel sample;
2) The screened enamel specimens were subjected to extracorporeal circulation treatment as shown in table 1:
TABLE 1 cycle treatment
Note that: the processing of sequence numbers 1 to 3 is performed 6 times and then sequence number 4 is performed.
3) The hardness value of the enamel sample after the extracorporeal circulation treatment is tested and recorded as HV Before acid etching Soaking in glycolytic solution for 30min, measuring hardness, and recording as HV After acid etching 。
4) Remineralizing treatment: the enamel sample after acid etching is subjected to extracorporeal circulation treatment again, the operation is the same as 2), and then the hardness value is tested and recorded as HV Remineralisation 。
Efficacy evaluation:
1) Evaluation of acid etching resistance: calculation of the change in microhardness values before and after acid etching Δhv1=hv Before acid etching -HV After acid etching Independent sample t-test was used. If the Δhv value of the sample group is less than the Δhv value of the control group and the two groups of Δhv values are statistically significant (P < 0.05), the sample group is considered to be resistant to enamel erosion by glycolytic acidogenesis in the oral cavity.
2) Evaluation of remineralization promoting ability: calculation of hardness value change before and after remineralization Δhv2=hv Remineralisation -HV After acid etching By using an independent sample t-test, if the Δhv value of the sample group is greater than that of the control group, and the two groups of Δhv values are statistically significant (P < 0.05), the sample group is considered to be able to promote remineralization of enamel eroded by glycolytic acidogenesis in the oral cavity.
The composition of the sugarresistant child toothpaste of examples 1-10 and comparative examples 1-11 is shown in Table 2.
The preparation methods of the anti-sugar child toothpaste of examples 1 to 10 and comparative examples 1 to 11 are:
adding sodium pyrophosphate, xylitol, calcium lactate and sodium fluoride into deionized water, stirring to dissolve completely, adding sorbitol, glycerol, polyethylene glycol-8 and benzyl alcohol, and stirring for 5min to obtain liquid phase. Adding silicon dioxide, xanthan gum and stirring well into a container, adding powder into the liquid phase, and stirring for 20min. Finally adding essence, cocamidopropyl betaine, lactobacillus salivarius, glucose oxidase, arginine and hydrolyzed wheat protein, stirring for 15min, and degassing to obtain the final product.
Table 2 anti-sugar child toothpaste ingredients
Table 2 subsequent sugar-resistant children toothpaste component
The results of examination of the physical and chemical index stability of the anti-glycal child toothpastes of examples 1 to 5 are shown in Table 3:
table 3 physical and chemical index stability investigation of sugar-resistant child toothpaste
As shown in Table 3, the sugar-resistant child toothpastes of examples 1 to 5 were accelerated aged at 45℃for 3 months, and then tested for various indexes related to the products. The detection results show that the anti-sugar children toothpaste products of examples 1-5 have good overall stability, normal physical and chemical indexes, good compatibility of formula components, no water-oil separation phenomenon and excellent quality.
The anti-sugar child toothpaste of examples 1 to 5 has the effect of removing dental calculus as shown in fig. 1, and the anti-sugar child toothpaste of examples 1 to 5 has the effect of removing dental calculus obviously, wherein the effect of removing dental calculus is derived from the decomposition effect of biological enzymes in the composition on dental plaque biological membranes in cooperation with other components, and the effect of removing dental calculus is obviously improved along with the increase of the concentration of the biological enzymes.
The results of the detection of the stability of the biological enzyme activity of the anti-glycal children toothpastes of examples 1 to 5 are shown in Table 4:
TABLE 4 enzymatic Activity of anti-sugar child toothpaste
The average enzyme activity of the anti-sugar children toothpastes of examples 1 to 5 is about 75% at high temperature for two months, and the residual activity is increased as the enzyme concentration is increased, which is up to 83%, so that the biological enzyme can keep higher enzyme activity and play an active role in the formula system of the anti-sugar toothpastes.
The oral pathogenic bacteria inhibitory effects of the anti-diabetic child teeth 1 of examples 1 to 5 and comparative examples 1 to 11 are shown in Table 5:
TABLE 5 antibacterial Rate against oral pathogenic bacteria of sugar-resistant child toothpaste
| Antibacterial rate | Streptococcus mutans% | Porphyromonas odontovorans% | Fusobacterium nucleatum% |
| Example 1 | ＞99.8 | ＞99.8 | ＞97.5 |
| Example 2 | ＞99.8 | ＞99.8 | ＞97.5 |
| Example 3 | ＞99.8 | ＞99.8 | ＞97.5 |
| Example 4 | ＞99.8 | ＞99.8 | ＞97.5 |
| Example 5 | ＞99.8 | ＞99.8 | ＞97.5 |
| Comparative example 1 | 66 | 67 | 58 |
| Comparative example 2 | 60 | 59 | 65 |
| Comparative example 3 | 76 | 82 | 75 |
| Comparative example 4 | 31 | 35 | 28 |
| Comparative example 5 | 26 | 29 | 31 |
| Comparative example 6 | 77 | 86 | 89 |
| Comparative example 7 | 83 | 90 | 88 |
| Comparative example 8 | 92 | 87 | 66 |
| Comparative example 9 | 72 | 76 | 78 |
| Comparative example 10 | 91 | 92 | 86 |
| Comparative example 11 | 89 | 90 | 92 |
The anti-glycaemic child toothpaste of examples 1 to 5 is effective in inhibiting the growth of oral pathogenic bacteria.
The inhibition of glycolysis by the anti-glycation childhood toothpaste of example 5 and the toothpaste without the anti-glycation composition is shown in figure 2, and plaque growth is inhibited and proliferation is prevented by the anti-glycation childhood toothpaste of example 5. Plaque still grew vigorously compared to toothpaste without the anti-sugar composition, showing turbidity of the solution and acidity of the solution.
The evaluation results of the dental plaque-inhibiting acid generating ability of the anti-sugar child toothpastes of examples 1 to 5 and comparative examples 1 to 11 are shown in Table 6:
table 6 evaluation of the plaque inhibition acid generating ability of the anti-glycal child toothpaste
As shown in table 6, in the test for evaluating the plaque acid production inhibition ability, the anti-sugar compositions in the anti-sugar child toothpastes of examples 1 to 5 had a synergistic effect, and were capable of synergistically inhibiting plaque acid production, and the plaque acid production inhibition effect was also remarkably improved when the concentration was increased. Meanwhile, the individual components of the anti-glycemic child toothpastes of comparative examples 1 to 5 were less effective in inhibiting plaque acidogenesis than the individual components, much less synergistic inhibition in multiple components. Biological enzymes such as dextranase, glucose oxidase and the like have the effects of decomposing dental plaque biological films and reducing bacterial adhesion, can inhibit bacterial proliferation to a certain extent, and experimental data show that hydrolyzed wheat protein is taken as a wheat polypeptide and also has the effect of inhibiting bacterial proliferation. Sodium fluoride and arginine have weak bacteriostasis to bacteria, so that the acid production capacity of dental plaque is weak. It is understood that the anti-sugar composition in the anti-sugar child toothpaste of examples 1 to 5 has an anti-sugar effect of inhibiting plaque acid production. In addition, as can be seen by comparing each example with comparative examples 6-11, the various compositions act synergistically, which can significantly promote inhibition of plaque acidogenesis and have a synergistic effect.
The evaluation results of the acid etching resistance and remineralization promotion ability of the sugary child toothpastes of examples 1 to 5 and comparative examples 1 to 11 are shown in tables 7 and 8:
TABLE 7 evaluation of acid resistance of a child anti-Glycerol toothpaste
Table 8 evaluation of remineralization promoting ability of children's anti-sugar toothpaste
As can be seen from tables 7 and 8, the anti-acid etching ability and enamel remineralization promoting ability of the anti-sugar child toothpastes of examples 1 to 5 were significantly different from those of the negative control group. By comparison of the single components of comparative examples 1-5, the single components of probiotics, glucanase and dextranase do not resist dental erosion and promote enamel remineralization, but fluoride, hydrolyzed wheat protein and arginine are both capable of resisting dental erosion and promoting enamel remineralization, but the acid erosion resistance and the tooth remineralization promotion ability of hydrolyzed wheat protein are both significantly better than those of sodium fluoride and arginine; in addition, the anti-glycal child toothpastes of comparative examples 1 to 5 were significantly lower in effect than the anti-acid etching ability and remineralization promoting ability of the multi-component anti-glycation composition. Meanwhile, it can be seen from comparison with comparative examples 6 to 11 that the anti-sugar child toothpastes lacking any one single anti-sugar component are inferior in both acid etching resistance and remineralization resistance to the anti-sugar child toothpastes entirely containing the anti-sugar component (examples 1 to 5). As can be seen from Table 7, the anti-glycaemic child toothpastes of examples 1 to 5 have an increased resistance to acid etch as the composition and concentration of the glycaemic active ingredient increases. Meanwhile, as can be seen from table 8, the anti-glycal children toothpastes of examples 1 to 5 have increased their ability to promote remineralization of enamel with increasing composition and concentration of the anti-glycal active ingredient, but the hardness of enamel is relatively stable after increasing to a certain level, and the remineralization effect is not obvious (examples 3 to 5). Therefore, glucose oxidase, dextranase, fluoride, hydrolyzed wheat protein, arginine and probiotics in the anti-sugar composition play a synergistic role and jointly play an anti-sugar effect.
In summary, the present invention provides an anti-glycal composition comprising glucose oxidase, dextranase, hydrolyzed wheat protein, sodium fluoride, probiotics and arginine, and an anti-glycal child toothpaste comprising the same, having anti-glycal efficacy, in particular, multi-path anti-glycal efficacy in inhibiting plaque acidogenesis, resisting enamel erosion, promoting enamel remineralization. Can resist sugar from multiple paths, protect children teeth, inhibit dental plaque formation, remove dental calculus, resist tooth demineralization caused by bacterial glycolysis and acid production due to sugar in food, promote tooth remineralization, and maintain oral health.
The glucose oxidase in the sugar-resistant composition of the present invention is an aerobic dehydrogenase obtained by fermentation with Aspergillus niger or the like. Has no toxic and side effects on human body, and has the functions of removing glucose, deoxidizing, sterilizing and the like. The glucose oxidase has the advantages of catalytic specificity, high activity, high catalytic efficiency and the like, and can decompose and gradually decompose glucan in food residues adhered to teeth, thereby inhibiting dental plaque formation. In addition, glucose oxidase has good adhesiveness on teeth and can continuously act. The hydrogen peroxide generated in the process of decomposing glucose by glucose oxidase can be gradually released, so that excessive hydrogen peroxide can not be generated at one time to stimulate other tissues of an oral cavity, dental stains and dental plaque can be safely and effectively removed, dental enamel is not damaged, gum is not stimulated, and teeth can be efficiently whitened in a short time. The dextranase in the anti-sugar composition is one of glucanases, and can cut off alpha-1, 6-glycosidic bond to degrade glucan. The biological membrane of dental plaque is mainly composed of extracellular polysaccharide, and the main component of the biological membrane is glucan. The research shows that the dextranase can inhibit the initial formation of oral pathogenic bacteria biomembrane, reduce the adhesion of bacteria, change the structure of biomembrane and destroy the cross-linking between bacterial colonies. The dextranase can effectively decompose dental plaque biomembrane, thereby inhibiting the formation of dental plaque. The invention solves the problem that the stability and activity of biological enzyme in toothpaste are easy to change.
Oral probiotics are included in the anti-sugar composition of the present invention. Probiotics are a general term for microorganisms beneficial to hosts, and include active strains and inactivated strains, and can help human bodies to adjust or maintain microecological balance, prevent diseases and promote body health. The probiotics are at least one of Bifidobacterium adolescentis, lactobacillus bifidus, lactobacillus crispatus, bifidobacterium longum, bifidobacterium breve, lactobacillus bulgaricus, lactobacillus gasseri, lactobacillus helveticus, lactobacillus johnsonii, lactobacillus plantarum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus salivarius and Streptococcus thermophilus. Aiming at the defect that probiotics cannot be stored at normal temperature, the invention utilizes the inactivated freeze-dried powder of the probiotics to replace living bacteria. After entering the oral cavity along with toothpaste, probiotics occupy binding points on oral mucosa by adhesion, thereby reducing the colonization of oral pathogenic bacteria in the oral environment, inhibiting the growth and reproduction of pathogenic bacteria, and reducing the number of harmful bacteria such as cariogenic bacteria, periodontal pathogenic bacteria, oral candida and the like. The probiotics can also have self aggregation and copolymerization with harmful bacteria to form a biological film, and the biological film is wrapped and then is taken out of the oral cavity, so that the growth of pathogenic bacteria is inhibited, and a barrier for preventing the colonization and infection of the pathogenic bacteria is formed.
The hydrolysis-resistant wheat protein in the sugar-resistant composition takes wheat protein as a raw material, and small molecular polypeptide substances obtained by hydrolysis by adopting methods of chemical hydrolysis, microbial fermentation, proteolysis and the like have the characteristics of good water solubility, stable dispersion, easy absorption, strong biological activity and the like. The main component of the hydrolyzed wheat protein is wheat peptide, and the wheat peptide can chelate mineral elements to promote the absorption and utilization of minerals by animals. The hydrolyzed wheat protein can chelate calcium ions, adhere to the surface of hydroxyapatite (main component of enamel), increase the hardness of teeth, promote remineralization of teeth and repair acid etched teeth. In addition, the hydrolyzed wheat protein and fluoride are combined, so that the synergistic effect is achieved, the remineralization of fluorine can be further enhanced, and the toxic and side effects of the fluoride are reduced.
Arginine in the anti-sugar composition of the present invention plays an important role in the human body, and as a constituent of salivary mucin, it can adhere to the tooth surface along with the semi-permeable membrane action of salivary mucin, thereby regulating the balance of oral flora and dynamic balance between ions. Meanwhile, the oxygenic acid can buffer and regulate the pH value of the sleeping liquid by participating in the metabolism of the hydrogen-producing neutrop, and is easy to adsorb on the tooth surface and dentin tubules due to the better solubility of the oxygenic acid, so that calcium and phosphorus ions are promoted to be deposited on the tooth surface and in the dentin tubules, and tooth remineralization is promoted. In addition, arginine has the ability to promote the deposition of hydroxyapatite and to close dentinal tubules, thus achieving the effect of anti-sensitivity.
The product of the invention adopts a scientific use method, can be used repeatedly within a period of multiple days to effectively resist sugar, and comprises the functions of inhibiting oral bacteria, removing dental plaque biomembrane, inhibiting dental plaque formation, resisting bacterial glycolysis and acid production, resisting tooth demineralization caused by glycolysis and acid production, promoting tooth remineralization and the like. The method comprises the following steps: the anti-sugar component is used for decomposing glucose, reducing the metabolism source of oral bacteria, removing dental calculus polysaccharide, reducing soft calculus, reducing dental plaque, inhibiting the growth of acid-producing bacteria, removing dental plaque biomembrane, neutralizing acid produced by bacteria, further enhancing the hardness of teeth, promoting the remineralization of teeth, and inhibiting the demineralization of teeth to achieve the effect of inhibiting glycolysis acid etching of teeth.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted equally without departing from the spirit and scope of the technical solution of the present invention.
Claims (10)
1. An anti-sugar composition containing hydrolyzed wheat protein, which is characterized by comprising the following components in parts by mass: 0.001 to 10 parts of hydrolyzed wheat protein, 0.001 to 0.1 part of glucose oxidase, 0.001 to 0.1 part of dextranase, 0.01 to 0.5 part of arginine, 0.01 to 0.05 part of fluoride and 0.001 to 0.1 part of probiotics.
2. The anti-sugar composition of claim 1, wherein the ratio of the added amounts of both dextranase and glucose oxidase is 0.001:1 to 1:1.
3. the anti-sugar composition of claim 1, wherein the ratio of the added amounts of the dextranase or glucose oxidase to the hydrolyzed wheat protein or fluoride is 0.001:1 to 1:1.
4. use of an anti-sugar composition according to any one of claims 1 to 3 in oral care.
5. An anti-sugar oral care product comprising, in mass percent, from 0.001% to 5% of the anti-sugar composition of any one of claims 1 to 3.
6. The anti-glycation oral care product of claim 5, wherein the anti-glycation oral care product is a glycation resistant child's toothpaste.
7. The anti-glycation oral care product of claim 5 or 6, wherein the glycation resistant child toothpaste further comprises the following components in mass percent: 25 to 28 percent of humectant, 0.1 to 1 percent of natural antibacterial agent, 0.2 to 1 percent of thickening agent, 10 to 25 percent of soft abrasive, 0.01 to 20 percent of xylitol, 0.001 to 5 percent of calcium-containing compound, 0.01 to 2 percent of mild surfactant, 0.5 to 1 percent of essence and the balance of water.
8. The anti-sugar oral care product of claim 7, wherein the natural antibacterial agent is at least one of benzyl alcohol, octylglycol, hexylene glycol, p-hydroxyacetophenone.
9. The anti-sugar oral care product of claim 7, wherein the calcium-containing compound is at least one of soluble calcium or sparingly soluble calcium.
10. The anti-sugar oral care product of claim 7, wherein the mild surfactant is at least one of sodium lauroyl sarcosinate, sodium lauroyl glutamate, cocoyl propyl betaine, sodium N-methyl cocoyl taurate, poloxamer, alkyl glycoside.
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