CN112336780A

CN112336780A - Citrus bacteriostat and preparation method and application thereof

Info

Publication number: CN112336780A
Application number: CN201910723830.XA
Authority: CN
Inventors: 王农跃; 胡文革
Original assignee: Ningbo Hanwei International Trade Co ltd
Current assignee: Ningbo Hanwei International Trade Co ltd
Priority date: 2019-08-07
Filing date: 2019-08-07
Publication date: 2021-02-09

Abstract

The invention provides a citrus bacteriostatic agent and a preparation method and application thereof. The citrus bacteriostatic agent comprises citrus extract, aloe extract and a pH regulator, wherein the content of the citrus extract accounts for 0.005-5% of the weight of the citrus bacteriostatic agent, and the content of the aloe extract accounts for 0.01-85% of the weight of the citrus bacteriostatic agent. The citrus bacteriostatic agent has strong antibacterial effect, rapid onset of drug action, and high stability over time.

Description

Citrus bacteriostat and preparation method and application thereof

Technical Field

The invention relates to the field of disinfection and sterilization, in particular to a citrus bacteriostatic agent and a preparation method and application thereof.

Background

The medicinal value of the citrus has been discovered by people for two thousand years, the citrus is also recorded in detail in 'compendium of materia medica' of Li Shizhen in the Ming dynasty of China, and the citrus essence is widely and comprehensively applied to the traditional Chinese medicine treatment of daily life, folk diet therapy, health care, health preservation and the like of people. At the end of the last 60 s of the century, experts in the institute of microbiology and food, university of florida, usa, have begun to study citrus essence, a natural substance with antimicrobial physiological activity, which is present in the fruits of citrus plants, and the components of citrus essence are safe and non-toxic, and have been widely used in food, health products, cosmetics and pharmaceutical industries at present.

CN101874844A discloses a citrus essence antibacterial agent, which contains 0.010-0.02% of citrus essence, 0.003% of peppermint oil and distilled water. The orange essence antibacterial agent has low antibacterial rate and short antibacterial time.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide the citrus composite bacteriostatic agent which has high-efficiency sterilization and disinfection effects and long antibacterial time.

The invention also provides a preparation method of the citrus bacteriostat.

It is still another object of the present invention to provide a method for using the citrus bacteriostatic agent as a female antibacterial agent, a skin antibacterial agent, a cleansing agent, etc.

The inventor finds that the main defects of the citrus bacteriostatic agent in the prior art are that the citrus bacteriostatic agent has low bacteriostatic rate, slow onset speed and short antibacterial time; in addition, citrus bacteriostats are not highly stable over time. The citrus bacteriostat of the invention enhances the antibacterial effect of the citrus bacteriostat by adding a proper amount of aloe extract into the citrus plant extract, and has fast effect taking speed and high stability along with the change of time.

The invention relates to a citrus bacteriostatic agent, which comprises citrus extract, aloe extract and pH regulator, wherein the citrus extract accounts for 0.005-5% of the weight of the citrus bacteriostatic agent, and the aloe extract accounts for 0.01-85% of the weight of the citrus bacteriostatic agent.

The citrus plant extract of the present invention may be present in an amount ranging from any of the stated ranges of 0.005% to 5%, or any combination of the stated ranges, e.g., 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2.0%, 2.2%, 2.4%, 2.6%, 2.8%, 3.0%, 3.2%, 3.4%, 3.6%, 3.8%, 4.0%, 4.2%, 4.4%, 4.6%, 4.8%, 5.0%. In various embodiments of the present invention, the citrus extract is preferably present in an amount of 0.03% to 0.5%, more preferably 0.01% to 0.3% by weight.

The aloe extract of the present invention may be present in an amount ranging from any value or combination of values within the stated range from 0.01% to 85%, e.g., 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2.0%, 2.2%, 2.4%, 2.6%, 2.8%, 3.0%, 3.2%, 3.4%, 3.6%, 3.8%, 4.0%, 4.2%, 4.4%, 4.6%, 4.8%, 5.0%, 5.2%, 5.4%, 5.6%, 5.8%, 6.0%, 6.2%, 6.8%, 6%, 6.8%, 7.8%, 8%, 9.8%, 8%, 9.8%, 9%, 8%, 9.8%, 9%, 8%, 9.8%, 9%, 9.8%, 8%, 9.8%, 9%, 8%, 9%, 8%, 9.8%, 9%, 10.8%, 9%, 9.8%, 10, 14.0%, 14.5%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%, 18.5%, 19.0%, 19.5%, 20.0%, 20.5%, 21.0%, 22.0%, 22.5%, 23.0%, 23.5%, 24.0%, 24.5%, 25.0%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%, 28.5%, 29.0%, 29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%, 33.0%, 33.5%, 34.0%, 34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%, 38.0%, 38.5%, 39.0%, 40.0%, 40.5%, 41.0%, 41.5%, 42.0%, 43.5%, 43.0%, 54.0%, 54.5%, 54.0%, 58.0%, 54.0%, 54.5%, 41.0%, 41.5%, 42.0%, 43.0%, 52.0%, 54.0%, 58%, 52.0%, 54.0%, 58%, 55.0%, 54.0%, 58%, 55.0%, 60%, 54.0%, 58%, 54.0%, 67.0%, 68.0%, 69.0%, 70.0%, 71.0%, 72.0%, 73.0%, 74.0%, 75.0%, 76.0%, 77.0%, 78.0%, 79.0%, 80.0%, 81.0%, 82.0%, 83.0%, 84.0%, 85.0%. In various embodiments of the present invention, the aloe vera extract is preferably present in an amount of 0.5% to 5%, more preferably 0.2% to 3% by weight.

The pH regulator is an acidic substance for regulating the pH value of the citrus bacteriostatic agent, and can be one or the combination of citric acid, lactic acid and acetic acid, preferably lactic acid.

The citrus plant is one or the combination of citrus, navel orange, citrus reticulata, lemon or grapefruit.

In a specific embodiment of the present invention, the citrus extract comprises glycoside tissue in an amount of 20% to 60% by weight of the citrus extract; the fruit acid accounts for 10-20% of the weight of the citrus extract; pectin accounts for 3% -10% of the weight of the citrus extract; the vitamin C accounts for 1-5% of the weight of the citrus extract; the amino acids account for 3% -8% of the weight of the citrus extract.

Further, the glycoside tissue is one or a combination of tangerine, hesperidin and naringin.

Further, the fruit acid is citric acid.

Further, the pectin is one or a combination of citrus pectin, grapefruit pectin and lemon pectin.

In a specific embodiment of the present invention, the citrus extract is prepared by pressing citrus to residue, adding distilled water, filtering, and removing the residue; then adding the filtered filtrate into an alcohol solvent for extraction and layering; concentrating the organic layer, and drying to obtain Citrus plant extract.

Further, the alcohol solvent is ethanol or glycerol.

The citrus plant extract prepared by the method is detected by adopting liquid chromatography-mass spectrometry, and the composition and content of the citrus plant extract are determined.

In a specific embodiment of the present invention, the aloe extract is prepared by a method comprising: adding Aloe into distilled water, stirring, soaking, extracting, centrifuging, and filtering to obtain Aloe extract.

The invention also relates to a preparation method of the citrus bacteriostat, which comprises the steps of mixing the citrus extract, the aloe extract and distilled water, stirring uniformly, adding the pH regulator, and regulating the pH value, wherein the weight content of the citrus extract is 0.005-5%, and the weight content of the aloe extract is 0.01-85%.

Further, the pH adjuster is at least one of citric acid, lactic acid, and acetic acid, and lactic acid is preferable. Mixing the above citrus extract, aloe extract and distilled water, adding pH regulator, and regulating pH to 1-7, preferably 2-5.

The invention also relates to the application of the citrus bacteriostat in female antibacterial agent, skin antibacterial agent, air and environment cleaning agent, etc.

Compared with the prior art, the citrus bacteriostatic agent disclosed by the invention utilizes the synergistic effect generated by the citrus plant extract and the aloe extract, so that the bacteriostatic effect is greatly improved, and the bacteriostatic rate reaches 99.9%; and has fast effect and high stability along with time change. The citrus bacteriostat is safe to skin, free of irritation and wide in application.

Detailed Description

The invention is further illustrated by the following specific examples, which are intended to be illustrative in more detail only and are not intended to be limiting in any way.

The preparation method of the aloe extract comprises the following steps: adding 10kg Aloe into distilled water, stirring, soaking, extracting, centrifuging, and filtering to obtain Aloe extract.

Example 1

Squeezing 10kg of orange into residue, adding distilled water, filtering, and removing residue; then adding 50% ethanol into the filtered filtrate for extraction, and layering; concentrating the organic layer, and drying to obtain citrus extract.

The liquid chromatography-mass spectrometry combined technology is adopted to detect the citrus extract, and the citrus extract comprises 25% of hesperidin, 15% of citric acid, 7% of citrus pectin, 1% of vitamin C and 3% of amino acid.

Example 2

Squeezing 10kg of navel orange into residue, adding distilled water, filtering, and removing residue; then adding 50% ethanol into the filtered filtrate for extraction, and layering; concentrating the organic layer, and drying to obtain fructus Citri Junoris extract.

The navel orange extract is detected by adopting a liquid chromatography-mass spectrometry combined technology, and comprises 50% of aurantiamarin, 10% of citric acid, 3% of citrus pectin, 5% of vitamin C and 8% of amino acid.

Example 3

Squeezing 10kg of pomelo into residue, adding distilled water, filtering, and removing residue; then adding 50% ethanol into the filtered filtrate for extraction, and layering; concentrating the organic layer, and drying to obtain fructus Citri Grandis extract.

The pomelo extract is detected by adopting a liquid chromatography-mass spectrometry technology, and the pomelo extract comprises 30% of naringin, 20% of citric acid, 10% of pomelo pectin, 1% of vitamin C and 6% of amino acid.

Example 4

Squeezing 10kg of lemon into residue, adding distilled water, filtering, and removing residue; then adding 80% glycerol into the filtered filtrate for extraction, and layering; concentrating the organic layer, and drying to obtain fructus Citri Limoniae extract.

The lemon extract is detected by adopting a liquid chromatography-mass spectrometry technology, and comprises the following components of hesperidin accounting for 40% of the weight of the citrus extract, citric acid accounting for 12% of the weight of the citrus extract, lemon pectin accounting for 8% of the weight of the citrus extract, vitamin C accounting for 2% of the weight of the citrus extract, and amino acid accounting for 7% of the weight of the citrus extract.

Example 5

Squeezing 10kg of Ponkan to obtain residue, adding distilled water, filtering, and removing residue; then adding 80% glycerol into the filtered filtrate for extraction, and layering; concentrating the organic layer, and drying to obtain Ponkan extract.

Detecting the citrus reticulata extract by adopting a liquid chromatography-mass spectrometry combined technology, wherein the citrus reticulata extract comprises 55% of hesperidin, 18% of citric acid, 5% of citrus pectin, 4% of vitamin C and 5% of amino acid.

Example 6

Squeezing 5kg of Ponkan and 5kg of navel orange to obtain residue, adding water, filtering, and removing residue; adding 50% ethanol into the filtrate, extracting, and layering; concentrating the organic layer, and drying to obtain Citrus plant extract.

Detecting the citrus extract by adopting a liquid chromatography-mass spectrometry combined technology, wherein the content of the hesperidin in the citrus extract is 35%, the content of the citric acid in the citrus extract is 16%, the content of the citrus pectin in the citrus extract is 9%, the content of the vitamin C in the citrus extract is 4%, and the content of the amino acid in the citrus extract is 6%.

Example 7

Squeezing 5kg of orange and 5kg of navel orange to obtain residue, adding distilled water, filtering, and removing residue; then adding 50% ethanol into the filtered filtrate for extraction, and layering; concentrating the organic layer, and drying to obtain Citrus plant extract.

Detecting the citrus extract by adopting a liquid chromatography-mass spectrometry combined technology, wherein the content of the hesperidin in the citrus extract is 45 wt%, the content of the citric acid in the citrus extract is 10 wt%, the content of the citrus pectin in the citrus extract is 8 wt%, the content of the vitamin C in the citrus extract is 3 wt%, and the content of the amino acid in the citrus extract is 7 wt%.

Example 8

Squeezing 5kg of Ponkan and 5kg of Mandarin orange into residue, adding distilled water, filtering, and removing residue; adding ethanol into the filtrate for extraction, and layering; concentrating the organic layer, and drying to obtain citrus extract.

The liquid chromatography-mass spectrometry technology is adopted to detect the citrus extract, and the citrus extract comprises 58% of hesperidin, 14% of citric acid, 4% of citrus pectin, 4% of vitamin C and 8% of amino acid.

Example 9

Mixing the citrus extract 0.05g and aloe extract 0.1g of example 1 with 1000ml of distilled water, stirring well, adding lactic acid, and adjusting pH to 2 to obtain the citrus bacteriostatic agent.

Example 10

Mixing the navel orange extract 3g and aloe extract 5g in example 2 with 1000ml of distilled water, stirring well, adding lactic acid, and adjusting pH to 5 to obtain the citrus bacteriostat.

Example 11

Mixing the grapefruit extract 5g and the aloe extract 50g in the example 3 with 1000ml of distilled water, stirring uniformly, adding citric acid, and adjusting the pH to 3 to obtain the citrus bacteriostat.

Example 12

0.1g of lemon extract and 30g of aloe extract obtained in example 4 are mixed with 1000ml of distilled water, stirred uniformly, added with citric acid and adjusted to pH 4 to obtain the citrus bacteriostat.

Example 13

Mixing 10g of Ponkan extract and 100g of Aloe extract obtained in example 5 with 1000ml of distilled water, stirring, adding acetic acid, and adjusting pH to 2 to obtain citrus bacteriostatic agent.

Example 14

Mixing 30g of the extract of Poncirus trifoliata and navel orange and 500g of the extract of Aloe in example 6 with 1000ml of distilled water, stirring, adding acetic acid, and adjusting pH to 3 to obtain the citrus bacteriostatic agent.

Example 15

Mixing the citrus and navel orange extract 50g and aloe extract 850g of example 7 with 1000ml of distilled water, stirring well, adding lactic acid, and adjusting pH to 4 to obtain the citrus bacteriostatic agent.

Example 16

Mixing 20g of Ponkan and Citrus reticulata extract and 2g of Aloe extract from example 8 with 1000ml of distilled water, stirring, adding citric acid, and adjusting pH to 3 to obtain Citrus bacteriostatic agent.

Comparative example 1

A method for preparing citrus essence comprises cleaning citrus fruit with water, removing water, and crushing into emulsion with a press mill; drying the emulsion to one eighth of the original weight by using centrifugal drying equipment; adding 30% glycerol by weight into the concentrated emulsion, mixing, standing for 72 hr, and randomly shaking for 10 min; centrifuging with 120 mesh sieve, filtering, concentrating the emulsion, and removing residue; sufficient lactic acid is added to the filtered cleaning solution to adjust the pH to 2.2-3.2. Through detection, the citrus essence comprises 47.6% of citrus fruit essence by weight; 7.0% of citric acid; citrus pectin 5.0%; 5.0 percent of vitamin C; 0.2% of amino acid; 10% of glyceride; 25.2 percent of glycerin.

Mixing 0.1g of citrus essence with 1L of water, stirring, adding lactic acid, adjusting pH to 4, and adding 0.03g of peppermint oil to obtain the citrus antibacterial agent.

Comparative example 2

Mixing 0.05g of the citrus extract obtained in example 1 with 1000ml of distilled water, stirring uniformly, adding lactic acid, and adjusting the pH to 2 to obtain the citrus bacteriostatic agent.

Comparative example 3

Mixing 0.1g Aloe extract with 1000ml distilled water, stirring, adding lactic acid, and adjusting pH to 2 to obtain citrus bacteriostatic agent.

Comparative example 4

Mixing the citrus extract 60g and aloe extract 0.08g of example 1 with 1000ml of distilled water, stirring well, adding lactic acid, and adjusting pH to 2 to obtain the citrus bacteriostatic agent.

Comparative example 5

Mixing the citrus extract 55g and aloe extract 0.5g of example 1 with 1000ml of distilled water, stirring well, adding lactic acid, and adjusting pH to 2 to obtain the citrus bacteriostatic agent.

Comparative example 6

1g of the citrus extract and 0.08g of the aloe extract obtained in example 8 were mixed with 1000ml of distilled water, stirred uniformly, added with citric acid, and adjusted to pH 3 to obtain a citrus antimicrobial agent.

Bacteriostatic rate experiments and stability experiments were performed on the citrus bacteriostatic agents prepared in examples 9 to 16 and comparative examples 1 to 6, respectively; the citrus bacteriostats prepared in examples 9 to 16 were subjected to skin irritation test and skin allergy test.

First, bacteriostasis rate experiment

Clinical isolated strains of escherichia coli, staphylococcus aureus and candida albicans are respectively cultured in a culture medium, the citrus bacteriostats prepared in examples 9-16 and comparative examples 1-5 are used as test objects, 5ml of the citrus bacteriostats are respectively acted for 1min, 2min, 5min and 10min for carrying out antibacterial performance tests, the test objects are operated according to the national standard GB15979-2002, and the results are shown in table 1, wherein the table 1 is the test results of the antibacterial performance of the citrus bacteriostats of examples 1-6 and comparative examples 1-5 of the invention.

TABLE 1 antimicrobial Performance test results for the citrus antimicrobial agents of examples 9-16 and comparative examples 1-5

As can be seen from table 1, the citrus bacteriostats of examples 9 to 16 have bacteriostasis rates of 99.9% for 1 minute against escherichia coli, staphylococcus aureus and candida albicans, so that the citrus bacteriostat prepared by the examples of the present invention has faster onset speed.

Second, stability test

The citrus bacteriostats prepared in examples 9-16 and comparative examples 1-5 were stored at 37 c and 75% relative humidity for 90 days. According to the bacteriostasis rate experiment method of the embodiment of the invention, the bacteriostasis rates of the citrus bacteriostat on escherichia coli, staphylococcus aureus and candida albicans are tested. The results of the experiments are shown in table 2,

table 2 results of antibacterial property test of the citrus bacteriostatic agents of examples 9 to 16 and comparative examples 1 to 5

As can be seen from Table 2, the citrus bacteriostats prepared in examples 9-16 have 99% inhibition rate against Escherichia coli, Staphylococcus aureus and Candida albicans after being stored under the above conditions.

Three, one complete skin irritation experiment

1. The test substance: citrus bacteriostats prepared in examples 9-16

2. Experimental animals: selecting 16 healthy rabbits with female and male bodies respectively, and the weight of the rabbits is 2.0-2.5kg.

3. The experimental method comprises the following steps: the hair on both sides of the spine of the rabbit was removed 24h before the start of the experiment with depilatory, and the size of the depilated area was 3cm × 3 cm. The next day, 2 female rabbits and 2 male rabbits were grouped into 8 groups, and 32 rabbits were divided into 8 groups. The citrus bacteriostatic agent prepared in example 9 was directly dropped on the depilatory area of one side of the spinal column of the first group of rabbits, the size of the sample application area was 2.5cm × 2.5cm, and then two layers of gauze were applied to cover the sample application area, which was then fixed with a non-irritating adhesive tape, and the other side of the citrus bacteriostatic agent was dropped on the depilatory area of the other side of the group of rabbits, the size of the sample application area was 2.5cm × 2.5cm, and then two layers of gauze were applied to cover the sample application area, which was then fixed with a non-irritating adhesive tape. After the application for 4 hours, the gauze is taken down, the residual test object is washed by warm water, and the local skin reaction of the tested part is observed after 1 hour, 24 hours, 48 hours and 72 hours after the test object is removed. Examples 10-16 were treated with 7 additional groups of rabbits according to the procedure described above.

The results of one complete skin irritation experiment performed in examples 9-16 are shown in table 3, and table 3 is the skin irritation response score results of one complete skin irritation experiment performed in examples 9-16.

TABLE 3 results of one complete skin irritation experiment performed in examples 9-16

As can be seen from table 3, the skin irritation response scores for one complete skin irritation experiment were all zero for the citrus bacteriostatic agents prepared in examples 9-16, indicating that the citrus bacteriostatic agents prepared in examples 9-16 were non-irritating to one complete skin.

Four, one-time damaged skin irritation experiment

1. The test substance: citrus bacteriostats prepared in examples 9-16

3. The experimental method comprises the following steps: the hair on both sides of the spine of the rabbit was removed 24h before the start of the experiment with depilatory, and the size of the depilated area was 3cm × 3 cm. The next day, 2 female rabbits and 2 male rabbits were grouped into 8 groups, and 32 rabbits were divided into 8 groups. Dehaired skins of all rabbits, 2.5cm by 2.5cm in size, were sanitized with 75% alcohol. And a 'well' -shaped damaged wound is scratched in the skin area by using a sterilizing blade or an injection needle in the sterilized area, and the damaged skin only reaches the epidermis without damaging the dermis. The citrus bacteriostatic agent prepared in example 9 was directly dropped on the depilatory area of one side of the spinal column of the first group of rabbits, the size of the sample application area was 2.5cm × 2.5cm, and then two layers of gauze were applied to cover the sample application area, which was then fixed with a non-irritating adhesive tape, and the other side of the citrus bacteriostatic agent was dropped on the depilatory area of the other side of the group of rabbits, the size of the sample application area was 2.5cm × 2.5cm, and then two layers of gauze were applied to cover the sample application area, which was then fixed with a non-irritating adhesive tape. After the application for 4 hours, the gauze is taken down, the residual test object is washed by warm water, and the local skin reaction of the tested part is observed after 1 hour, 24 hours, 48 hours and 72 hours after the test object is removed. Examples 10-16 were treated with 7 additional groups of rabbits according to the procedure described above.

The results of the primary damaged skin irritation test performed in examples 9-16 are shown in table 4, and table 4 shows the skin irritation response score results of the primary damaged skin irritation test performed in examples 9-16.

TABLE 4 results of one-time damaged skin irritation experiment conducted in examples 9-16

As can be seen from table 4, the skin irritation response scores of the citrus bacteriostatic agents prepared in examples 9-16 were all zero in the primary damaged skin irritation test, indicating that the citrus bacteriostatic agents prepared in examples 9-16 were non-irritating to primary damaged skin.

Fifth, skin allergy test

1. The test substance: citrus bacteriostats prepared in examples 9-16

2. Experimental animals: 48 healthy white guinea pigs with a body weight of 200-300 g.

3. The experimental method comprises the following steps: the citrus bacteriostats prepared in examples 9-16 were used as the citrus bacteriostat at the induced concentration and the citrus bacteriostat at the excited concentration, and the 2, 4-dinitrochlorobenzene solution at the induced concentration and the 2, 4-dinitrochlorobenzene solution at the excited concentration were prepared. For the regulations on solutions of induced and excited concentrations, see "general requirements for toxicology of sterilized products" in "Sterilization Specification (2002)".

The hair was removed from the left side of the guinea pig back 24h before the start of the experiment using depilatory, the dimensions of the depilatory area being 3cm x 3 cm. Of the 48 guinea pigs, 16 guinea pigs were used as an experimental group, 16 of the guinea pigs were used as a positive control group, and the remaining 16 guinea pigs were used as a negative control group. For an experimental group, 0.5ml of citrus bacteriostatic agent with induction concentration is directly coated on a depilation area on the left side of the back of a guinea pig in the experimental group, the size of a sample application area is 2cm multiplied by 2cm, then a layer of non-irritating plastic film is coated on the sample application area, and the non-irritating plastic film is fixed on the guinea pig by using a non-irritating adhesive tape. When applied for up to 6 hours, the non-irritating plastic film was removed and the remaining sample was removed with warm water. This was then repeated in the same manner on day 7 and 14 after the first application. On the 28 th day after the first application, 0.5ml of citrus bacteriostatic agent at an excitation concentration was directly applied to the application area, and then a non-irritating plastic film was applied to cover the application area, and the non-irritating plastic film was fixed to the guinea pig body with a non-irritating adhesive tape. When the application time reaches 6h, the non-irritating plastic film is taken down and the residual sample is removed with warm water. The skin reactions were observed again at 24h and 48h after removal of the residual sample.

For positive control guinea pigs, induction and challenge treatments were performed using 2, 4-dinitrochlorobenzene as described above.

For the negative control guinea pigs, only on day 28, 0.5ml of citrus bacteriostatic agent at an excitation concentration was applied directly to the application area, a non-irritating plastic film was then applied over the application area, and the non-irritating plastic film was secured to the guinea pigs using a non-irritating adhesive tape. When the application time reaches 6h, the non-irritating plastic film is taken down and the residual sample is removed with warm water. The skin reactions were observed again at 24h and 48h after removal of the residual sample.

The results of skin allergy experiments using the citrus agents prepared in examples 9 to 16 of the present invention are shown in table 5, and table 5 shows the results of sensitization intensity scoring in the experimental guinea pigs, positive control guinea pigs, and negative control guinea pigs.

TABLE 5 results of skin allergy experiments with citrus bacteriostats prepared in examples 9-16

Note: the sensitization intensity scoring method is described in "technical Specification for toxicological experiments on sterilized products" in "technical Specification for Disinfection (2002)".

As can be seen from Table 5, the guinea pigs in the experimental groups corresponding to examples 9-16 all had a zero sensitization intensity score, and therefore, the citrus bacteriostatic agent prepared by the examples of the present invention did not produce sensitization.

VI, vagina mucosa stimulation experiment

Taking 16 healthy female New Zealand fig. s, weighing 2-2.5 kg, randomly dividing into 8 groups, wherein the experimental group is the citrus bacteriostat prepared in the embodiments 9-16 of the invention, and the control group is sterilized normal saline. The animals were examined before the experiment for the presence of secretions, congestion, edema and other conditions at the vaginal orifice. Fixing the animal on the back, exposing the vaginal opening, wetting the catheter with normal saline, gently inserting the catheter into the vagina (4-5 cm), slowly injecting 2ml of a test object with an injector, and drawing out the catheter to finish the contamination. After 24 hours, the animals were sacrificed by air embolism, the vagina was completely cut off and longitudinally incised, and the presence of congestion, edema and the like was visually observed.

The results show that: the stimulation index of the tested substance to the vaginal mucosa of the rabbit is 0.22, and the tested substance is nonirritating.

Experiment in air environment

1. Sample name: citrus bacteriostat prepared in examples 9-16 of the present invention

2. The adopted places are as follows: school

3. The detection basis is as follows: sanitary standard for disposable sanitary articles GB15979-2002

The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the invention is not limited to the embodiments described above, which are described in the specification only to illustrate the principles of the invention. The invention also includes various insubstantial changes and modifications within the spirit of the invention, as claimed by those skilled in the art.

Claims

1. The citrus bacteriostatic agent comprises citrus extract, aloe extract and pH regulator, wherein the content of the citrus extract accounts for 0.005-5% of the weight of the citrus bacteriostatic agent, and the content of the aloe extract accounts for 0.01-85% of the weight of the citrus bacteriostatic agent.

2. A citrus bacteriostatic agent according to claim 1, wherein the citrus extract is contained in an amount of 0.03-0.5% by weight of the citrus bacteriostatic agent, preferably 0.01-0.3% by weight of the citrus bacteriostatic agent.

3. A citrus bacteriostatic agent according to claim 1, wherein the aloe extract is contained in an amount of 0.5-5% by weight of the citrus bacteriostatic agent, preferably 0.2-3% by weight of the citrus bacteriostatic agent.

4. A citrus bacteriostatic agent according to claim 1, wherein the pH adjusting agent is at least one of citric acid, lactic acid and acetic acid, preferably lactic acid.

5. The citrus bacteriostatic agent according to claim 1 or 2, wherein the citrus extract comprises glycoside tissue, fruit acid, pectin, vitamin C and amino acid, wherein the glycoside tissue accounts for 20-60% of the weight of the citrus extract; the fruit acid accounts for 10-20% of the weight of the citrus extract; pectin accounts for 3% -10% of the weight of the citrus extract; the vitamin C accounts for 1-5% of the weight of the citrus extract; the amino acids account for 3% -8% of the weight of the citrus extract.

6. A citrus bacteriostatic agent according to claim 1 or 5, characterized in that the citrus plant is one or the combination of citrus, navel orange, mandarin orange, lemon or pomelo.

7. The citrus bacteriostatic agent according to claim 5, wherein the glycoside tissue is one or a combination of hesperidin, naringin and hesperidin.

8. A citrus bacteriostatic agent according to claim 5, wherein the fruit acid is citric acid.

9. The citrus bacteriostatic agent according to claim 5, wherein the pectin is one or the combination of citrus pectin, lemon pectin and grapefruit pectin.

10. A method of preparing a citrus bacteriostatic agent according to claims 1-9, comprising mixing citrus extract and aloe extract with water, stirring uniformly, adding pH regulator, and adjusting pH, wherein the weight content of citrus extract is 0.005-5%, and the weight content of aloe extract is 0.01-85%.

11. The method of making a citrus bacteriostatic agent according to claim 10, wherein the citrus extract is contained in an amount of 0.03 to 0.5% by weight, preferably in an amount of 0.01 to 0.3% by weight.

12. A method of producing a citrus bacteriostatic agent according to claim 10, wherein the aloe extract is contained in an amount of 0.5-5% by weight, preferably 0.2-3% by weight.

13. A method of producing a citrus bacteriostatic agent according to claim 10, wherein the pH adjusting agent is at least one of citric acid, lactic acid and acetic acid, preferably lactic acid.

14. A method of producing a citrus bacteriostatic agent according to claim 10, wherein the pH is 1 to 7, preferably 2 to 5.

15. The method for preparing a citrus bacteriostatic agent according to claim 10, wherein the citrus plant is one or a combination of citrus, navel orange, mandarin orange, lemon or grapefruit.

16. A citrus bacteriostatic agent according to any one of claims 1 to 9 for use as a skin antiseptic.

17. A citrus bacteriostatic agent according to any one of claims 1 to 9 for use as a feminine antimicrobial agent.

18. A citrus bacteriostatic agent according to any one of claims 1 to 9 for use in a detergent.