CN113995744B - Composition and antibacterial application thereof - Google Patents

Composition and antibacterial application thereof Download PDF

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Publication number
CN113995744B
CN113995744B CN202111195294.4A CN202111195294A CN113995744B CN 113995744 B CN113995744 B CN 113995744B CN 202111195294 A CN202111195294 A CN 202111195294A CN 113995744 B CN113995744 B CN 113995744B
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acid
composition
glycine
percent
escherichia coli
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CN113995744A (en
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朱绍和
陈丽
宋坤坪
田雪晨
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Wenzhou Lien Biotechnology Co.,Ltd.
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Wenzhou Kean University
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Priority to CN202111195294.4A priority Critical patent/CN113995744B/en
Priority to LU102887A priority patent/LU102887B1/en
Publication of CN113995744A publication Critical patent/CN113995744A/en
Priority to PCT/CN2022/124947 priority patent/WO2023061416A1/en
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/02—Saturated carboxylic acids or thio analogues thereof; Derivatives thereof
    • A01N37/04—Saturated carboxylic acids or thio analogues thereof; Derivatives thereof polybasic
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/06—Unsaturated carboxylic acids or thio analogues thereof; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
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    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
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    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/194—Carboxylic acids, e.g. valproic acid having two or more carboxyl groups, e.g. succinic, maleic or phthalic acid
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid, pantothenic acid
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid, pantothenic acid
    • A61K31/198—Alpha-aminoacids, e.g. alanine, edetic acids [EDTA]
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • 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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
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    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
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    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
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Abstract

The inhibition of organic acids inhibits the next generation of bacteria by entering the bacterial cells in a free diffusion manner. The composition of the present invention can control the number of bacteria to a degree harmless to the human body. The invention relates to a composition and its antibacterial application, used for human body and environment application, can reduce the reproduction trend of its existing microorganism, the composition comprises: 1.2% malic acid, 0.3% hippuric acid, 0.1% fumaric acid, 0.1% glycine and water. Wherein the minimum inhibitory concentration of the composition for inhibiting escherichia coli and staphylococcus aureus is 2.5 times lower than that of the composition for inhibiting escherichia coli and staphylococcus aureus by using 1.2% of malic acid, 0.3% of hippuric acid, 0.1% of fumaric acid or 0.1% of glycine alone.

Description

Composition and antibacterial application thereof
Technical Field
The invention relates to the technical field of medicines, in particular to a composition and antibacterial application thereof.
Background
The pangolin scales are used in the Xianfang Huoming drink in the 'woman good prescription' written by Song Dynasty Chen Ming Shuang, and the whole formula has the effects of clearing away heat and toxic materials, eliminating swelling and dissipating hard mass, promoting blood circulation to arrest pain and the like, and has the symptoms of local inflammation and pain. Pangolin scales recorded in Ben Cao gang mu of Ming Dynasty Li Shizhen records that it can eliminate phlegm, malaria, chills and fever, wind-arthralgia, rigidity and pain, dredge meridians, relieve pain and swelling, expel pus and blood, dredge orifices and kill parasites. In the recipe of the recipe, it is mentioned that "diarrhea with grief", pangolin scales and clam powder are stir-baked and ground into powder. As can be seen from the above ancient medical records, pangolin scales are used to treat diseases that modern medicine believes to be caused by bacterial infections. Guo Yi et al reported in the reports of Hunan institute of traditional Chinese medicine (academy of academic), reported that the water decoction of squama Manis has antibacterial effect on 11 kinds of bacteria such as Escherichia coli, wherein the main component of the water decoction is squama Manis.
However, as pangolins are in an extremely endangered state in modern society, 8 pangolins are listed in red record of endangered species in the world natural protection alliance (IUCN) 2014, and are listed in CITES appendix I level of protected animals in the Washington convention in 2016, so that the global trade is prohibited. China is the contracting country of International trade Convention (CITES) on endangered wild animal and plant species. Meanwhile, in 2020, pangolin scales are excluded from Chinese pharmacopoeia and used as medicine. Therefore, the ingredients or ingredient compositions which have the effect of inhibiting the flora in the squama of the pangolin are researched, and the artificial synthesis is carried out to be used as a substitute, so that the endangered animal pangolin can be protected while the relevant treatment scheme for replacing the ingredients of the pangolin is not influenced.
In the prior art, an organic acid is used in various fields as an organic acid additive, and among them, citric acid is used as the organic acid which is used most frequently for preservation and flavor addition. Chinese patent with publication number CN108670897B relates to a high-efficiency bacteriostatic wet tissue composition and a preparation method thereof. The raw materials for preparing the efficient bacteriostatic wet tissue composition comprise, by weight, 1-6 parts of ethanol, 0.5-3 parts of butanediol, 0.2-0.6 part of solubilizer, 0.01-0.02 part of menthol, 0.1-0.12 part of essence, 0.5-1.0 part of tea extract, 0.5-1.0 part of olive leaf extract, nano-silver, 0.5-1.0 part of purslane extract, 0.1-0.2 part of citric acid, 0.05-0.15 part of sodium citrate and 0.01-0.05 part of phenoxyethanol. The composition comprises organic acid citric acid as antibacterial component, wherein phenoxyethanol is used for assisting in dissolving citric acid. However, the existing organic acids are various, and the bacteriostatic effect of citric acid cannot reach the highest bacteriostatic efficiency.
Chinese patent with publication number CN108904374A relates to an oral cavity cleanser, which comprises the following components in parts by weight: 3-8 parts of tea extract, 1-3 parts of persimmon astringent, 1-2 parts of fructose, 2-4 parts of citric acid and 1-3 parts of spice.
Generally speaking, the pH of the mouth wash or other oral cleaning agent is in the range of 6.5-7.5, which is biased toward neutral, so that the use of the mouth wash does not destroy the proteins in the oral cavity and provides excellent bactericidal action in a neutral environment. The glycine buffer consisting of glycine is capable of maintaining the liquid in the range of 2.2-10.6. Pseudomonas aeruginosa and Serratia marcescens are pathogens of opportunistic infections causing immune impairment, and are mostly present at wounds. The oral cavity cleanser prevents pseudomonas aeruginosa and serratia marcescens from entering respiratory tract to cause harm to patients by the function of inhibiting the reproduction of the pseudomonas aeruginosa and the serratia marcescens in the oral cavity.
In the prior art, various weakly acidic bacteriostatic preservative effects are proved, but the application of organic acids with the most extensive and the highest inhibitory effects on human diseases such as escherichia coli and staphylococcus aureus is not found.
According to the experiment, the metabolite composition for inhibiting bacteria in the squama manitis is explored through the component research of the squama manitis with good antibacterial effect, so that the antibacterial effect of replacing the squama manitis with multi-combination weak acid is realized.
Furthermore, on the one hand, due to the differences in understanding to those skilled in the art; on the other hand, since the applicant has studied a great deal of literature and patents when making the present invention, but the disclosure is not limited thereto and the details and contents thereof are not listed in detail, it is by no means the present invention has these prior art features, but the present invention has all the features of the prior art, and the applicant reserves the right to increase the related prior art in the background.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a composition. Is used for human body or environment in which human body is located, and can reduce the propagation tendency of microorganism existing in the environment. The composition comprises: one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine and citric acid.
Preferably, the ingredients in the composition comprise: (1) malic acid; (2) hippuric acid; (3) optionally at least one of the following weakly acidic components: 1) Glycine; 2) Succinic acid and/or L-valine.
Preferably, the ingredients in the composition comprise: (1) malic acid; (2) hippuric acid; (3) optionally at least one group of weakly acidic components of: 1) Fumaric acid and/or glycine; 2) Succinic acid and/or L-valine.
According to a preferred embodiment, the first component, consisting of malic acid, hippuric acid, succinic acid, L-valine, is formulated together with the second component, consisting of fumaric acid or succinic acid, to form a composition having an inhibitory effect on the growth of microorganisms.
The technical scheme has the advantages that: compared with the application of single organic acid, the organic acid combination formed by multiple combinations can form better antibacterial effect (proved by antibacterial tests) compared with the single organic acid, and different flavors of the disinfection components which are suitable for food, makeup, oral cavity cleaning and the like and act on human bodies are formed by the combination of multiple organic acids, so that the aims of deodorization and refreshment are fulfilled. The formation and decomposition of organic acid are harmless to human body and environment, and the principle of bacteriostasis and sterilization is that small molecules enter the bacterial body and destroy the alkaline active environment in the bacterial body, thereby inhibiting the bacterial activity. The combination of the organic acids in various combinations forms a covalent structure with each other and forms bacteriostatic effects which are mutually increased, and the organic acids which are combined randomly in a single way or a plurality of ways and need to be added in a large dose are replaced by the composition with a smaller dose, so that the harm to human bodies caused by excessive intake of the food additive is reduced.
According to a preferred embodiment, said composition for acting on the human body or on the environment comprises malic acid, hippuric acid, glycine and citric acid, wherein the composition is capable of enhancing the fruity ingredient for acting on the environment in a buffered acidic organoleptic manner, based on the interaction of malic acid and citric acid.
According to a preferred embodiment, the composition comprises at least malic acid, hippuric acid, glycine and citric acid, wherein the components constituting said composition are applied to the skin surface in the form of a non-newtonian fluid.
According to a preferred embodiment, the composition comprises at least malic acid, hippuric acid, glycine and fumaric acid, wherein the components constituting the composition are applied to the skin surface in the form of a non-newtonian fluid.
The technical scheme has the advantages that: drugs and cosmetics used for repairing, treating or protecting skin surfaces cannot be stably hung on the face due to the fluidity of fluid in the process of applying, but paste drugs are not easy to be uniformly applied due to the viscosity. The invention adopts the component form of non-Newtonian fluid, reduces the fluidity of the non-Newtonian fluid and does not change the viscosity, thereby ensuring the stability of the components attached on the face and the skin.
According to a preferred embodiment, the composition is capable of inhibiting the division of escherichia coli and staphylococcus aureus in a synergistic manner.
A composition capable of forming, in the form of a dispersed phase, the cleaning component of an oral cleaning agent together with an abrasive, the composition being for use in inhibiting the growth of escherichia coli and staphylococcus aureus in the interior of the oral cavity, wherein the composition comprises one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine, and citric acid.
A composition which can inhibit the multiplication of Escherichia coli and Staphylococcus aureus on the surface of human skin or in food, wherein the composition comprises one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine and citric acid.
Antimicrobial use of a composition for application as an additive in a detergent or bactericide, said composition as an additive being provided with means for releasing H + And a second composition for increasing the kill effect and time. The first composition comprises: malic acid, hippuric acid, fumaric acidOne or more of glycine, succinic acid, L-valine and citric acid, and the second composition comprises hydrocolloid. Wherein the first composition intervenes in the second composition when the composition as an additive is applied, and acts on the interior of the bacterial body of Escherichia coli or Staphylococcus aureus accompanying with the coating of the second composition on Escherichia coli or Staphylococcus aureus carried by a target of detergent or bactericide application.
Antibacterial use of a composition for inhibiting the growth of bacteria in tissue inflammation caused by bacterial infection, said medicament comprising as essential components: one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine and citric acid.
Detailed Description
The following detailed description is made with reference to the accompanying drawings.
The invention relates to a bacteriostatic composition which can achieve the most effective inhibitory action on microorganisms represented by escherichia coli and staphylococcus aureus.
Organic acids used for bacteriostasis include Fumaric acid (Fumaric acid), succinic acid (Succinic acid), L-Valine (L-Valine), malic acid (Malic acid), citric acid (Citric acid), glycine (Glycine), and Hippuric acid (Hippuric acid). The proportion of the extract is consistent with the content of squama Manis, wherein in the total extract (CV), fumaric acid accounts for 0.1%, succinic acid accounts for 0.4%, L-valine accounts for 1.3%, malic acid accounts for 1.2%, citric acid accounts for 0.2%, glycine accounts for 0.1%, and hippuric acid accounts for 0.3%. Preferably, the above ratio is adjustable and effective.
The microorganisms referred to herein can comprise gram-negative bacteria, gram-positive bacteria, or comprise a pathogenic staphylococcal representative, a conditional pathogenic bacterium, such as escherichia coli, or other pathogenic bacteria, such as mycobacterium tuberculosis.
The organic acids used in the experiments and constituting the components of the composition of the present invention are all artificially synthesized organic acids.
By the special content proportion of the composition, the better antibacterial effect of a single or few compositions with the same volume is achieved.
Example 1
The antibacterial metabolites were used for Minimum Inhibitory Concentrations (MIC) detection of Escherichia coli (E.coli; escherichia coli) and Staphylococcus aureus (S.aureus; staphylococcus aureus), pseudomonas aeruginosa (P.aeruginosa; pseudomonas aeruginosa) and Serratia marcescens (S.marcocens) respectively, based on the Minimum Inhibitory Concentration (MIC) test.
(1) Experimental materials:
1) Culture medium
A lysis Broth (LB; lysogeny Broth) medium containing 10g/L of peptone, 5g/L of yeast extract and 10g/L of sodium chloride was used.
2) Antibacterial drugs:
comprises the whole composition consisting of one or more of fumaric acid, succinic acid, L-valine, malic acid, citric acid, glycine and hippuric acid.
(2) The bacteriostatic test comprises the following steps:
1) Preparation of bacterial suspension
And simultaneously recovering the preserved escherichia coli representing gram-negative bacteria and staphylococcus aureus representing gram-positive bacteria. A single colony having a diameter of about 1mm was picked up by using an inoculating loop on a plate of the test bacterium which had been purified, inoculated in 2mL of LB broth, and cultured in a biochemical incubator at 37 ℃. The enriched logarithmic phase strain solution is adjusted to a concentration of 0.5 McLeod's turbidimetric standard (i.e., an absorbance at 625nm is 0.08-0.10) with LB broth, and contains about 1X 10 8 ～2×10 8 CFU·mL -1 。
2) Preparation of antibacterial drugs
Preparing the combined antibacterial agent with concentration of 4C, 2C, 0.5C, 0.25C, 0.125C, 0.0625C, 0.03125C by two-fold dilution method, numbering 8 sterile test tubes in sequence, and adding 1ml of sterile water into 2-8 test tubes. Prepare 2ml 4C's antibacterial agent in first test tube, get the solution in 1ml first test tube and add a second test tube, after the mixing, get the solution in 1ml second test tube again and add a third test tube, dilute in proper order.
3) MIC assay
200. Mu.L of LB broth was added to column 1 of a 96-well plate as a negative control, and 180. Mu.L of the prepared bacterial suspension was added to columns 2-10 to a final concentration of 1X 10 5 CFU·mL -1 . The 2 nd column is added with 20 μ L of sterile water as a positive control group, and the 3 rd to 10 th columns are respectively added with 20 μ L of drugs with final concentrations of 0.4, 0.2, 0.1, 0.05, 0.025, 0.0125, 0.00625 and 0.003125C. The respectively inoculated drug sensitive plates were placed in an incubator at 37 ℃ for 24 hours, MIC values were detected by observing turbidity of LB broth medium and detecting OD values, and the experiment was repeated 3 times.
The results are shown in Table 1, the minimum inhibitory concentration of each metabolite on Escherichia coli and Staphylococcus aureus, the summary of the independently existing concentrated antibacterial metabolites, and malic acid is the best inhibitory effect on Escherichia coli and Staphylococcus aureus.
Figure GDA0003655223650000061
TABLE 1
In order to verify the optimal effect of malic acid and the composition thereof, the experiment further performed an antibacterial test using malic acid and other metabolites as compositions, and the results represented by MIC values are shown in table 2.
Figure GDA0003655223650000062
TABLE 2
Experimental results show that the MIC value of the two-component composition added with malic acid on escherichia coli and staphylococcus aureus is reduced, and the two-component composition has a higher inhibition effect on the escherichia coli and the staphylococcus aureus. Still further, random dispensing of the three compositions was performed and the bacteriostatic tests were performed as shown in table 3.
Figure GDA0003655223650000071
TABLE 3
The combination of malic acid and fumaric acid has a weak rebound of bacteriostatic effect when combined with L-valine or citric acid, and similarly, malic acid and L-valine have the same result when combined with glycine or hippuric acid, and have a weak inhibitory effect on Escherichia coli and Staphylococcus aureus, and the MIC value rises to 0.4C. The composition formed by malic acid, hippuric acid and citric acid has optimal inhibition effect on escherichia coli, and the MIC value is 0.1C.
The experiment also carried out bacteriostatic tests on four compositions consisting of malic acid and other metabolites distributed in the four compositions, wherein malic acid is selected as the main component, and the results are shown in Table 4.
Figure GDA0003655223650000081
TABLE 4
The results show that the combination of malic acid, fumaric acid, glycine and hippuric acid and the combination of malic acid, citric acid, glycine and hippuric acid are able to develop bacteriostatic effects with MIC as low as 0.1C.
The experiment also carried out bacteriostatic tests on five compositions consisting of malic acid and other metabolites distributed in the main component, and the results are shown in table 5.
Figure GDA0003655223650000082
Figure GDA0003655223650000091
TABLE 5
The results show that malic acid, L-valine, citric acid, glycine, hippuric acid; malic acid, fumaric acid, succinic acid, L-valine, hippuric acid; the three compositions of malic acid, fumaric acid, L-valine, glycine and hippuric acid can have an inhibitory effect on escherichia coli with an MIC value of 0.1C. The composition comprises malic acid, succinic acid, L-valine, glycine and hippuric acid; malic acid, fumaric acid, L-valine, citric acid, hippuric acid; malic acid, fumaric acid, citric acid, glycine, hippuric acid; malic acid, succinic acid, L-valine, citric acid, hippuric acid; any one group of malic acid, succinic acid, citric acid, glycine and hippuric acid can have an inhibitory effect on Escherichia coli and Staphylococcus aureus with an MIC value of 0.1C. Compared with the prior art in which a single organic acid or two organic acids are used as the additive, the bacteriostatic effect of the bacteriostatic additive cannot reach that of the bacteriostatic additive formed by combining more than four organic acids.
In the experiment, the results of bacteriostatic tests on six compositions and seven metabolites formed by distributing other metabolites and malic acid by selecting malic acid as a main component are shown in Table 6.
Figure GDA0003655223650000092
Figure GDA0003655223650000101
TABLE 6
The results show that even if six or more metabolites are mixed, the bacteriostatic effect on staphylococcus aureus and escherichia coli can reach an MIC value of 0.1C.
Further, the metabolite combinations comprising malic acid, hippuric acid, fumaric acid and glycine were tested for Escherichia coli, staphylococcus metalens, pseudomonas aeruginosa and Serratia marcescens, and the results are shown in Table 7:
Figure GDA0003655223650000102
TABLE 7
The results show that the organic acid metabolite composition consisting of malic acid, hippuric acid, fumaric acid and glycine has an MIC value of 0.1C for Escherichia coli and Staphylococcus aureus and an MIC value of 0.2C for Pseudomonas aeruginosa and Serratia marcescens. The organic acid metabolite composition comprising malic acid, hippuric acid, fumaric acid and glycine has growth inhibiting effect on bacteria represented by Escherichia coli, staphylococcus metalens, pseudomonas aeruginosa and Serratia marcescens, especially pathogenic bacteria.
Serratia marcescens, which is the smallest bacterium, belongs to gram-negative bacilli and is a conditional pathogen.
Pseudomonas aeruginosa is a gram-negative bacterium, and is an opportunistic infectious bacterium.
Escherichia coli, staphylococcus metal, pseudomonas aeruginosa and serratia marcescens are widely distributed in nature, can enter a human body through the positions of the mouth, the nose and the like of the human body, and cause the human body to generate pathogenic reactions such as inflammation and the like under certain conditions or environmental conditions. Therefore, the four strains are selected as test objects of the bacteriostasis test in the experiment.
Based on the experimental results, the composition consisting of malic acid, glycine and hippuric acid can form an effect of inhibiting the bacteria of staphylococcus aureus and escherichia coli to achieve an MIC value of 0.1C together with citric acid or fumaric acid.
The composition for the experiment can be prepared from 0.1% of fumaric acid, 0.4% of succinic acid, 1.3% of L-valine, 1.2% of malic acid, 0.2% of citric acid, 0.1% of glycine and 0.3% of hippuric acid according to the above proportions.
Preferably, the composition for reducing the tendency of microorganisms present on the surface of the human body or in the environment in which the human body is located to multiply comprises: 0.125 to 0.482 percent of malic acid, 0.135 to 0.422 percent of L-valine, 0.021 to 0.081 percent of citric acid, 0.01 to 0.04 percent of succinic acid, 0.031 to 0.121 percent of hippuric acid, 0.001 to 0.041 percent of glycine and 0.01 to 0.041 percent of fumaric acid.
Preferably, the ratio of the components in the composition can be: 1 to 30 percent of malic acid, 0.1 to 20 percent of L-valine, 0.1 to 20 percent of citric acid, 0.1 to 20 percent of succinic acid, 0.1 to 20 percent of hippuric acid, 0.1 to 20 percent of glycine and 0.1 to 20 percent of fumaric acid.
Preferably, the ratio of the components in the composition can be: malic acid accounts for 10-40% of the total composition, L-valine accounts for 0.01-10% of the total composition, citric acid accounts for 0.01-10% of the total composition, succinic acid accounts for 0.01-20% of the total composition, hippuric acid accounts for 0.01-20% of the total composition, glycine accounts for 0.01-20% of the total composition, and fumaric acid accounts for 0.01-10% of the total composition.
Preferably, the malic acid can be in the range of about 0.1% to about 20%, about 1% to about 10%, about 3% to about 8%, about 6% to about 12%, about 15% to about 18%, about 17% to about 20%, 20% to 60% by weight, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 30%, or about 40% by weight, including all ranges and subranges therebetween, in the total composition.
Preferably, the L-valine can range from about 0.01% to about 1%, about 1% to about 10%, about 3% to about 8%, about 6% to about 12%, about 15% to about 18%, about 17% to about 20% by weight, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 30%, or about 40% by weight, including all ranges and subranges therebetween, in the total composition.
Preferably, the citric acid can range from about 0.01% to about 2%, about 1% to about 10%, about 3% to about 8%, about 6% to about 12%, about 15% to about 18%, about 17% to about 20% by weight, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 30%, or about 40% by weight, including all ranges and subranges therebetween, in the total composition.
Preferably, the succinic acid can range from about 0.1% to about 20%, about 1% to about 10%, about 3% to about 8%, about 6% to about 12%, about 15% to about 18%, about 17% to about 20% by weight, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 30%, or about 40% by weight, including all ranges and subranges therebetween, in the total composition.
Preferably, the hippuric acid can range from about 0.1% to about 20%, about 1% to about 10%, about 3% to about 8%, about 6% to about 12%, about 15% to about 18%, about 17% to about 20%, 20% to 60% by weight, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 30%, or about 40% by weight of the total composition, including all ranges and subranges therebetween.
Preferably, the glycine can be in the range of about 0.1% to about 2%, about 1% to about 10%, about 3% to about 8%, about 6% to about 12%, about 15% to about 18%, about 17% to about 20% by weight, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 30%, or about 40% by weight, including all ranges and subranges therebetween, in the total composition.
Preferably, the fumaric acid can range from about 0.01% to about 20%, about 1% to about 10%, about 3% to about 8%, about 6% to about 12%, about 15% to about 18%, about 17% to about 20% by weight, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 30%, or about 40% by weight, including all ranges and subranges therebetween, in the total composition.
Preferably, the combined range of malic acid, L-valine, citric acid, succinic acid, hippuric acid, glycine and fumaric acid can include, but is not limited to, the above-mentioned ratio and combined range.
The composition having an inhibitory effect against escherichia coli with an MIC value of 0.1C is represented by a composition of malic acid, hippuric acid and citric acid, while the composition having an inhibitory effect against escherichia coli and staphylococcus aureus with an MIC value of 0.1C is represented by malic acid, fumaric acid, glycine, hippuric acid; malic acid, citric acid, glycine, hippuric acid. Since all the components forming the composition are weakly acidic components and can be absorbed by the human body as food additives, the above-mentioned multi-component compositions can be applied to various fields as components harmless to the human body. In the prior art, the methods and cognition using organic acids are: any organic acid has the bacteriostatic action, and the organic acid can be added into food or other disinfecting liquid contacting with human body in large dose within the legal allowable range for bacteriostasis. However, when used in large quantities as an additive, organic acids generate conversion products in the body, a small amount of the organic acids depends on metabolism, but a large amount of the organic acids cannot ensure that all the products can be metabolized out of the body in time, so that the large amount of the organic acids can cause harm to the body of the human body within a certain period of time. According to the invention, the optimal bacteriostatic composition is obtained by carrying out bacteriostatic proportioning on the organic acid which can effectively inhibit bacteria, and the addition amount of the edible organic acid can be reduced by increasing the variety of the edible organic acid, so that the food or other food is helped to influence the health of a human body while the preservative effect is ensured.
Example 2
This embodiment is a further improvement of embodiment 1, and repeated contents are not described again.
Take the Liyan tablet for chronic pharyngitis treatment at present as an example. Aiming at chronic pharyngitis caused by bacterial infection, the sore throat relieving tablet can have a good effect on the disease. The ingredients of the sore throat relieving tablet comprise pangolin (prepared), ground beeltle, stiff silkworm, oyster (calcined) and figwort, and the pangolin (prepared) in the ingredients is a forbidden medicine because the pangolin (prepared) is taken from endangered organisms, so that the sore throat relieving tablet can not be used as a prescription medicine or a non-prescription medicine for treating patients. According to the invention, by researching the inhibition effect on flora in squama Manis, especially on Escherichia coli and Staphylococcus aureus, one or more groups of compositions with effective inhibition effect on Escherichia coli and Staphylococcus aureus are found. The main components of the medicine comprise: one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine and citric acid. When the pangolin scales of the liyan tablet are replaced, on one hand, the replacement component can adopt biological tissue extract with similar components, and on the other hand, the replacement component can also be a metabolite substitute component which is synthesized by people and effectively inhibits bacteria. Preferably, when an artificially synthesized compound is used as the substitute component, it can be a composition designed in this experiment to have one of the most excellent inhibitory effects on gram-negative bacteria represented by escherichia coli and gram-positive bacteria represented by staphylococcus aureus: malic acid, fumaric acid, succinic acid, L-valine, citric acid and glycine. The organic acid can bring fruity taste to the tablets, which is popular with children, thereby increasing the enthusiasm of infants to take medicine.
Example 3
This embodiment is a further improvement of embodiment 1 or embodiment 2, and repeated contents are not described again.
The composition can be used as additive composition in various foods, and is effective in inhibiting Escherichia coli and Staphylococcus aureus in foods. For example, the additive is added into some natural fruit juice, namely the natural flavor is increased through the proportion of malic acid and citric acid, and the preservative effect is realized by inhibiting the growth of escherichia coli and staphylococcus aureus through being matched with the fruit juice in vacuum or sealed package. The main components of the additive comprise: one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine and citric acid. The presence of malic acid and citric acid, separately or in combination, results in an additive having a fruity flavor.
Example 4
This embodiment is a further improvement on embodiment 1, embodiment 2, or embodiment 3, and repeated details are not repeated.
The composition can also be used for killing bacteria at home or under ordinary living conditions. For example, in the domestic life, bacteriostatic bacteria maintenance of the environment or bacteriostatic washing of clothes, especially staphylococcus aureus, which is harmful to the human body, is required. The disinfection of alcohol or 84 disinfectant is also harmful to human body and is not suitable for families with pregnant women or children. The invention can form the liquid agent for killing bacteria, which is suitable for families with children or pregnant women, based on the composition of the weakly acidic components and matched with other components which do not harm the bacteria.
The composition of organic acids can be applied as an additive in detergents or bactericides. The composition as additive is used for clothes or environmental bacteriostasis and deodorization. The composition is provided with a first composition comprising a PH modifier and a second composition for increasing the disinfecting effect and time. The first composition comprises any one of two combinations: malic acid, glycine, hippuric acid and citric acid; malic acid, glycine, hippuric acid and fumaric acid. The second composition comprises a hydrocolloid.
Hydrocolloids, which contain polysaccharides and proteins, form a gel network. When the gel network wraps escherichia coli or staphylococcus aureus carried by a target applied by detergent or bactericide, the first composition existing in the hydrocolloid enters the thallus from the thallus based on the pressure of the gel network on the thallus, the first composition and the second composition act together to form organic acid with synergistic effect, and H is released inside the thallus + Thus, the alkaline environment inside the cells is destroyed, and the activity of the cells is suppressed. The first composition intervenes in the hydrocolloid and acts inside the thallus of the escherichia coli or the staphylococcus aureus along with the wrapping of the hydrocolloid on the escherichia coli or the staphylococcus aureus carried by a target applied by the detergent or the bactericide.
The molecular diameter of the organic acid used to act on the intracellular environment of the bacterium can be smaller than the diameter of the cell membrane channel of escherichia coli and staphylococcus aureus so that the organic acid molecule can enter the interior of the bacterium from the cell membrane channel and destroy the alkaline environment of the interior of the bacterium to inhibit the activity of the bacterium cell.
The joint inhibition effect of the escherichia coli and the staphylococcus aureus is realized through the synergistic effect of a first composition consisting of malic acid, glycine and hippuric acid and a second composition consisting of citric acid or fumaric acid.
Example 5
This embodiment is a further improvement on embodiment 1, embodiment 2, embodiment 3, or embodiment 4, and repeated details are not repeated.
The present invention contemplates a composition for spreading onto a target surface and killing and inhibiting escherichia coli and staphylococcus aureus bacteria by spraying, atomizing or otherwise forming a small particle gas or liquid.
The composition consists of combined liquid or gas particles. The liquid or gas particles comprise a particle core composed of organic acid with high bacteriostasis and an outer coating film which is covered on the particle core and is composed of zirconium oxide-tin oxide complex with nanometer inorganic micropore shape neutralized by silane coupling agent and used for protecting and increasing the adhesive capacity.
By dispersing the zirconium oxide-tin oxide complex in the form of nano-scale inorganic micropores neutralized by the silane coupling agent wrapping the organic acid in the manner of aerosol, aerosol or small-particle liquid in clothes and living environment, when the zirconium oxide-tin oxide complex in the form of nano-scale inorganic micropores neutralized by the silane coupling agent wrapping the organic acid touches escherichia coli and staphylococcus aureus on the target surface, the outer membrane releases the organic acid under pressure and allows the organic acid to enter the bacteria body through the cell membrane on the bacteria surface, thereby inhibiting the bacterial activity. The nano-scale inorganic micropores formed by the zirconia-tin oxide composite can allow organic acid molecules to enter and exit and limit the obstruction of the decomposition and attachment of the particles by large-volume particles. The zirconia-tin oxide composite can effectively maintain the hardness and the toughness of the outer coating film for preventing the collapse of nano-micropores when the outer coating film of particles existing in the forms of aerosol, liquid or gas is formed.
Preferably, the ratio of the silane coupling agent to the nano-scale inorganic microporous form of the zirconia-tin oxide composite is from 1. The ratio of the nano-inorganic microporous zirconium oxide-tin oxide composite to the organic acid is 1 to 1.
Microparticles composed of organism-inorganic bodies coupled with a silane coupling agent can move toward a target surface in a state of a diameter of at least less than 10 μm. The above microparticles have excellent dispersion stability in an aqueous phase for dilution or homogeneous liquid phase, a high adhesion rate of the organic acid carried on the target surface, and a high volatility of the organic acid core reduced by the coating of the outer membrane. The organic-inorganic hybrid membrane is characterized in that an intermediate phase which is low in cost and can stably couple an internal organic layer and an external inorganic layer is utilized, the inorganic layer which is coupled with the organic layer and does not generate other chemical reactions is added under the condition of not damaging the dissociation of hydrogen ions of the organic layer, and the problems of poor adhesion, no effect and volatility of the organic layer on a target surface are solved through the property of the inorganic layer. Preferably, the silane coupling agent coupling can be an aqueous silane coupling agent coupling.
The preparation method of the composition comprises the following steps:
(1) Weighing: weighing the silane coupling agent coupling and zirconia-tin oxide composite (which can be present in the form of a sol) in proportion by weight, and weighing the organic acid composition in terms of volume;
(2) Mixing: and (3) putting the zirconium oxide and tin oxide complex into organic acid in batches, uniformly mixing, slowly pouring a silane coupling agent along with stirring action for coupling, and stirring until three phases are uniform to obtain the concentrated composition.
The concentrated composition can be diluted with water before use, and sprayed to the target surface to be disinfected such as environment or clothes by using equipment such as an aerosol sprayer and a spray can.
The organic acid in the particle core can be combined with the organic acid which is related to the embodiment and can achieve the MIC of 0.1C on the antibacterial effect of escherichia coli and staphylococcus aureus. The organic acid composition wrapped by the outer involucra has good stability in water and has high-efficiency inhibition effect on escherichia coli and staphylococcus aureus.
Example 6
This embodiment is a further improvement on embodiment 1, embodiment 2, embodiment 3, embodiment 4, or embodiment 5, and repeated contents are not described again.
Aiming at the organic acid composition which can be permanently attached to the surface and acts on bacteria, the invention can design an antibacterial ointment or spray which contains one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine and citric acid and can act on the skin surface or mucous membrane of the face, arms, important organs and the like. The organic acid composition has obvious bacteriostatic action on the propagation of various pathogenic skin bacteria such as gram-negative bacteria, gram-positive bacteria and the like represented by escherichia coli and staphylococcus aureus.
Preferably, the antibacterial ointment comprises traditional Chinese medicine ointment, traditional Chinese medicine powder, mussel mucin and an organic acid composition containing one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine and citric acid.
Preferably, the bacteriostatic spray comprises pentanediol, 1,3 propanediol and an organic acid composition containing one or more of malic acid, hippuric acid, fumaric acid, glycine, succinic acid, L-valine and citric acid.
It should be noted that the above-mentioned embodiments are exemplary, and that those skilled in the art, having benefit of the present disclosure, may devise various arrangements that are within the scope of the present disclosure and that fall within the scope of the invention. It should be understood by those skilled in the art that the present specification and figures are illustrative only and are not limiting upon the claims. The scope of the invention is defined by the claims and their equivalents. The present description contains a plurality of inventive concepts such as "preferably", "according to a preferred embodiment" or "optionally" each indicating that the respective paragraph discloses a separate concept, the applicant reserves the right to apply for divisional applications according to each inventive concept. Throughout this document, the features referred to as "preferably" are only an optional feature and should not be understood as necessarily requiring that such applicant reserves the right to disclaim or delete the associated preferred feature at any time.

Claims (9)

1. A composition for spraying onto a human body surface or onto the environment in which the human body is located, comprising: 1.2 percent of malic acid, 0.3 percent of hippuric acid, 0.2 percent of citric acid, 0.1 percent of glycine and the balance of water, and the composition cleans a human body or the environment where the human body is positioned in a mode of inhibiting the propagation of escherichia coli and staphylococcus aureus.
2. The composition of claim 1, wherein the composition further comprises: the mass percent of the L-valine is 1.3 percent.
3. The composition according to claim 1 or 2, characterized in that it further comprises: the mass percent is 0.4 percent of succinic acid.
4. Use of the composition according to claim 1 for the preparation of a detergent, an antibacterial drug, a bactericide or a food additive.
5. A composition, comprising: 0.1 percent of fumaric acid, 1.2 percent of malic acid, 0.3 percent of hippuric acid, 0.1 percent of glycine and the balance of water.
6. The composition of claim 5, wherein the composition further comprises: 1.3% L-valine.
7. The composition according to claim 5 or 6, characterized in that it further comprises: 0.2% citric acid.
8. Use of a composition according to claim 5 in the preparation of an oral cleanser.
9. Use of the composition according to claim 5 for the preparation of a detergent, an antibacterial drug, a bactericide or a food additive.
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