A kind of acidic solid disinfectant and preparation method thereof

Abstract

The invention provides an acidic solid disinfectant and a preparation method thereof, and relates to the technical field of disinfectant preparation. The solid disinfectant comprises 50-55 parts of a hollyhock extract, 20-25 parts of a thorn bark extract, 5-9 parts of beta-glucosidase, and 11-25 parts of alginic acid. The acidic solid disinfectant has a rapid killing effect on Escherichia coli and Staphylococcus aureus, and has a significant effect.

Description

Technical Field

[0001] The invention relates to the technical field of disinfectant preparation, and in particular to an acidic solid disinfectant and a preparation method thereof. Background Art

[0002] Disinfectants are common disinfecting products and are widely used in animal husbandry. Current disinfectants are mainly chemical products, which can effectively produce antibacterial and bactericidal properties, but may have adverse effects on animals themselves. For example, sodium hypochlorite disinfectants can easily irritate the nasal cavity and eyes of animals, and sodium benzoate disinfectants can easily have harmful effects on animals, especially on cats. The adverse effects of chemical disinfectants have greatly limited their application.

[0003] However, in animal husbandry, in order to prevent harmful strains in the environment such as Escherichia coli and Staphylococcus aureus from spreading among animals, disinfection of the breeding environment is essential. Therefore, providing an animal-friendly and effective disinfectant is of great significance to the animal husbandry industry. Summary of the invention

[0004] In view of this, the present invention provides an acidic solid disinfectant and a preparation method thereof, wherein the solid disinfectant comprises an extract of Sijiqing, an extract of wormwood bark, beta-glucosidase and alginic acid. The acidic solid disinfectant has a rapid killing effect on Escherichia coli and Staphylococcus aureus, and has a significant effect.

[0005] The technical solution of the present invention is as follows:

[0006] An acidic solid disinfectant comprising the following components in parts by weight:

[0007] 50-55 parts of Ipomoea aquatica extract, 20-25 parts of Echinops chinensis bark extract, 5-9 parts of beta-glucosidase and 11-25 parts of alginic acid.

[0008] Preferably, the acidic solid disinfectant comprises the following components in parts by weight:

[0009] 52 parts of Ipomoea aquatica extract, 22 parts of Echinops chinensis bark extract, 7 parts of β-glucosidase and 19 parts of alginic acid.

[0010] Preferably, the preparation process of the Sijiqing extract is as follows:

[0011] S1, crushing the waste Evergreen trees obtained by pruning into 0.5-1 mm, and soaking them in ethanol to obtain slurry A;

[0012] S2, reflux and extract slurry A for 2-3 hours, filter while hot to obtain an extract;

[0013] S3, purifying the extract using macroporous resin D101, collecting ethanol eluate with a concentration of 80-85%, concentrating and drying to obtain the ivy extract.

[0014] Preferably, in step S1, the concentration of ethanol is 80-85%, and the weight ratio of evergreen waste to ethanol is 1:5-7.

[0015] Preferably, the preparation process of the thorn bark extract is as follows:

[0016] SS1, crush the bark of the Chinese thorn tree to 4-5 mm, add water, mix well and soak for 24-36 hours to obtain slurry B;

[0017] SS2, enzymatic hydrolysis, adjusting the pH of slurry B to 3.5-3.8 with formic acid, then adding hemicellulase and cellulase, the addition amount of hemicellulase is 3.5-4.0u / g, the addition amount of cellulase is 4.3-4.8u / g, and enzymatic hydrolysis is carried out at 45±0.5℃ for 2-2.5h to obtain enzymatic hydrolyzate A;

[0018] SS3, inactivate the enzyme in the enzymatic hydrolyzate A and filter, collect the filtrate; add ethyl acetate to the filtrate, extract three times, collect the ethyl acetate phase, concentrate and dry to obtain the extract of the thorn bark.

[0019] Preferably, in step SS1, the weight ratio of the wormwood bark to water is 1:10-15.

[0020] Preferably, in step SS2, the amount of hemicellulase added is 3.8u / g, and the amount of cellulase added is 4.5u / g.

[0021] The preparation method of the above-mentioned acidic solid disinfectant is as follows:

[0022] (1) According to the proportion of each group, prepare the extract of Sijiqing, the extract of wormwood bark and alginic acid respectively, mix them evenly, and obtain a mixture;

[0023] (2) dispersing β-glucosidase in water, adjusting the pH to 4.0-4.2 with formic acid, and obtaining an aqueous solution;

[0024] (3) Spray the aqueous solution on the mixture and, after drying, obtain a solid disinfectant.

[0025] Compared with the prior art, the beneficial effects of the present invention are:

[0026] 1. The solid disinfectant provided by the present invention uses the waste of evergreen shrubs obtained by pruning and the bark of the thorny vine as raw materials. According to the characteristics of each raw material, different preparation methods are adopted to obtain the evergreen shrub extract and the bark of the thorny vine extract with high content of biologically active substances. The solid disinfectant is a plant-based product, has the advantages of being safe and non-toxic, and is suitable for use as a disinfectant product in animal breeding.

[0027] 2. The solid disinfectant provided by the present invention has a rapid killing effect on Escherichia coli and Staphylococcus aureus, and the effect is significant.

[0028] 3. In the solid disinfectant provided by the present invention, the biologically active substances in the Sijiqing extract and the biologically active substances in the Tussop bark extract work synergistically, and on the basis of the presence of β-glucosidase, can better remove odors in the breeding house and provide a good environment for animal growth. DETAILED DESCRIPTION

[0029] In order to enable those skilled in the art to better understand the technical solutions of the present invention, the technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0030] In the present invention, the raw materials, reagents, etc. used are all commercially available products.

[0031] Example 1

[0032] An acidic solid disinfectant comprising the following components by weight:

[0033] 52g of Ipomoea aquatica extract, 22g of thorn bark extract, 7g of β-glucosidase and 19g of alginic acid;

[0034] The preparation process of Sijiqing extract is as follows:

[0035] S1, crushing the waste Evergreen trees obtained by pruning into pieces of 0.5-1 mm, and soaking them in 83% ethanol for 3 hours to obtain slurry A;

[0036] The weight ratio of Evergreen waste to ethanol is 1:6;

[0037] S2, reflux and extract slurry A for 2.5 hours, filter while hot to obtain an extract;

[0038] S3, loading the macroporous resin D101 pretreated by the prior art into a column, and then adding the extract into the resin column, so that the resin adsorbs the glycoside compounds;

[0039] Use pure water to elute for 2 column volumes, then use ethanol with a concentration of 20%, 40% and 50% to elute for 1 column volume respectively, and finally elute with an ethanol solution with a concentration of 83% and collect, and after concentrating and drying, obtain the evergreen extract;

[0040] The preparation process of the extract of thorn bark is as follows:

[0041] SS1, crush the bark of the Chinese thorn tree to 4-5 mm, add water, mix well and soak for 30 hours to obtain slurry B;

[0042] The weight ratio of thorn bark to water is 1:12;

[0043] SS2, enzymatic hydrolysis, the pH of slurry B was adjusted to 3.6 with formic acid, and then hemicellulase and cellulase were added, the addition amount of hemicellulase was 3.8u / g, and the addition amount of cellulase was 4.5u / g, and enzymatic hydrolysis was carried out at 45°C for 2h to obtain enzymatic hydrolyzate A;

[0044] SS3, inactivate the enzyme in the enzymatic hydrolyzate A and filter it to collect the filtrate; add ethyl acetate to the filtrate, the volume ratio of the filtrate to ethyl acetate is 1:1, extract for 3 times, collect the ethyl acetate phase, concentrate and dry it to obtain the extract of the thorn bark;

[0045] The preparation method of the above-mentioned acidic solid disinfectant is as follows:

[0046] (1) According to the proportion of each group, prepare the extract of Sijiqing, the extract of wormwood bark and alginic acid respectively, mix them evenly, and obtain a mixture;

[0047] (2) dispersing β-glucosidase in water, wherein the weight ratio of β-glucosidase to water is 1:1, and adjusting the pH to 4.1 with formic acid to obtain an aqueous solution;

[0048] (3) Spray the aqueous solution on the mixture and, after drying, obtain a solid disinfectant.

[0049] Comparative Example 1

[0050] The difference from Example 1 is that, referring to the method for preparing the test solution of Sijiqing and Tujingpi in the 2020 edition of the Chinese Pharmacopoeia, Sijiqing extract and Tujingpi extract were prepared respectively as follows:

[0051] Evergreen extract: Take the powder of this product (passed through a No. 4 sieve), add 80% methanol, the weight volume ratio of powder to methanol (g / mL) is 1:50, ultrasonically treat for 30 minutes at a power of 300 W and a frequency of 40 kHz, filter, collect the filtrate, and dry to obtain the evergreen extract;

[0052] Extract of the bark of thorny thorn: Take the powder of this product (passed through a No. 3 sieve), add methanol, the weight volume ratio of powder to methanol (g / mL) is 1:125, heat and reflux for 1 hour, filter, collect the filtrate, and dry to obtain the extract of the bark of thorny thorn;

[0053] The solid disinfectant was prepared by using the above extract, and the weight was as follows:

[0054] 52g of Ipomoea aquatica extract, 22g of thorn bark extract, 7g of β-glucosidase and 19g of alginic acid;

[0055] The preparation method of the solid disinfectant is the same as that of Example 1.

[0056] Comparative Example 2

[0057] The difference from Example 1 is that a solid disinfectant comprises the following components by weight:

[0058] 52g of Sijiqing extract, 22g of wormwood bark extract, and 26g of alginic acid;

[0059] The above-mentioned Ipomoea aquatica extract, Ipomoea scabra extract and alginic acid are mixed to obtain a solid disinfectant.

[0060] Comparative Example 3

[0061] The difference from Example 1 is that a solid disinfectant comprises the following components by weight:

[0062] 52g of Ipomoea aquatica extract, 22g of thorn bark extract, and 26g of β-glucosidase;

[0063] The preparation method of the solid disinfectant is the same as that of Example 1.

[0064] Example 2

[0065] An acidic solid disinfectant comprising the following components by weight:

[0066] 50g of Ipomoea aquatica extract, 20g of thorn bark extract, 5g of β-glucosidase and 25g of alginic acid;

[0067] The preparation process of Sijiqing extract is as follows:

[0068] S1, crushing the waste Evergreen trees obtained by pruning into pieces with a size of 0.5-1 mm, and soaking them in 80% ethanol for 4 hours to obtain slurry A;

[0069] The weight ratio of Evergreen waste to ethanol is 1:5;

[0070] S2, reflux and extract slurry A for 2 hours, filter while hot to obtain an extract;

[0071] S3, loading the macroporous resin D101 pretreated by the prior art into a column, and then adding the extract into the resin column, so that the resin adsorbs the glycoside compounds;

[0072] Use pure water to elute for 2 column volumes, then use ethanol with a concentration of 20%, 40% and 50% to elute for 1 column volume respectively, and finally elute with an ethanol solution with a concentration of 80% and collect, and after concentrating and drying, obtain the evergreen extract;

[0073] The preparation process of the extract of thorn bark is as follows:

[0074] SS1, crush the bark of the Chinese thorn tree to 4-5 mm, add water, mix well and soak for 24 hours to obtain slurry B;

[0075] The weight ratio of thorn bark to water is 1:10;

[0076] SS2, enzymatic hydrolysis, the pH of slurry B was adjusted to 3.5 with formic acid, and then hemicellulase and cellulase were added, the addition amount of hemicellulase was 3.5u / g, and the addition amount of cellulase was 4.3u / g, and the enzymatic hydrolysis was carried out at 45°C for 2.5h to obtain enzymatic hydrolyzate A;

[0077] SS3, inactivate the enzyme in the enzymatic hydrolyzate A and filter it to collect the filtrate; add ethyl acetate to the filtrate, the volume ratio of the filtrate to ethyl acetate is 1:1, extract for 3 times, collect the ethyl acetate phase, concentrate and dry it to obtain the extract of the thorn bark;

[0078] The preparation method of the above-mentioned acidic solid disinfectant is as follows:

[0079] (1) According to the proportion of each group, prepare the extract of Sijiqing, the extract of wormwood bark and alginic acid respectively, mix them evenly, and obtain a mixture;

[0080] (2) dispersing β-glucosidase in water, wherein the weight ratio of β-glucosidase to water is 1:1, and adjusting the pH to 4.0 with formic acid to obtain an aqueous solution;

[0081] (3) Spray the aqueous solution on the mixture and, after drying, obtain a solid disinfectant.

[0082] Example 3

[0083] An acidic solid disinfectant comprising the following components in parts by weight:

[0084] 55g of Sijiqing extract, 25g of thorn bark extract, 9g of β-glucosidase and 11g of alginic acid;

[0085] The preparation process of Sijiqing extract is as follows:

[0086] S1, crushing the waste Evergreen trees obtained by pruning into pieces with a size of 0.5-1 mm, and soaking them in 85% ethanol for 2 h to obtain slurry A;

[0087] The weight ratio of Evergreen waste to ethanol is 1-7;

[0088] S2, reflux and extract slurry A for 3 hours, filter while hot to obtain an extract;

[0089] S3, loading the macroporous resin D101 pretreated by the prior art into a column, and then adding the extract into the resin column, so that the resin adsorbs the glycoside compounds;

[0090] Using pure water to elute for 2 column volumes, then using ethanol with a concentration of 20%, 40% and 50% to elute for 1 column volume respectively, and finally eluting with an ethanol solution with a concentration of 85% and collecting, concentrating and drying, to obtain the evergreen extract;

[0091] The preparation process of the extract of thorn bark is as follows:

[0092] SS1, crush the bark of the Chinese thorn tree to 4-5 mm, add water, mix well and soak for 36 hours to obtain slurry B;

[0093] The weight ratio of thorn bark to water is 1:15;

[0094] SS2, enzymatic hydrolysis, the pH of slurry B was adjusted to 3.8 with formic acid, and then hemicellulase and cellulase were added, the addition amount of hemicellulase was 4.0u / g, and the addition amount of cellulase was 4.8u / g, and enzymatic hydrolysis was carried out at 45°C for 2h to obtain enzymatic hydrolyzate A;

[0095] SS3, inactivate the enzyme in the enzymatic hydrolyzate A and filter it to collect the filtrate; add ethyl acetate to the filtrate, the volume ratio of the filtrate to ethyl acetate is 1:1, extract for 3 times, collect the ethyl acetate phase, concentrate and dry it to obtain the extract of the thorn bark;

[0096] The preparation method of the above-mentioned acidic solid disinfectant is as follows:

[0097] (1) According to the proportion of each group, prepare the extract of Sijiqing, the extract of wormwood bark and alginic acid respectively, mix them evenly, and obtain a mixture;

[0098] (2) dispersing β-glucosidase in water, wherein the weight ratio of β-glucosidase to water is 1:1, and adjusting the pH to 4.2 with formic acid to obtain an aqueous solution;

[0099] (3) Spray the aqueous solution on the mixture and, after drying, obtain a solid disinfectant.

[0100] test

[0101] 1. Sterilization test

[0102] Take the cultured Escherichia coli and Staphylococcus aureus and dilute them with sterile saline to a concentration of 1.5×10 8 cfu / mL bacterial suspension;

[0103] According to the addition amount of 100mL / bottle, the bacterial suspension was poured into the shaking flask, and 1 bottle was kept as a blank control; then the disinfectants of Examples 1-3 and Comparative Examples 1-3 were added to the remaining shaking flasks, and the addition amount of the disinfectant was 5g. After stirring evenly, the mixture was allowed to stand at 37°C for 0.5h, 1h, 3h, 5h and 8h, and then 100μL was taken and spread on TSA culture medium, cultured for 48h, and bacteria were counted. A plate control with only culture medium was set. The results are shown in Tables 1 and 2, as follows:

[0104] Table 1 The sterilization results of disinfectants on Escherichia coli

[0105]

[0106] Table 2 Sterilization results of disinfectants on Staphylococcus aureus

[0107]

[0108] (II) Ammonia odor test

[0109] Odor source: Take 40mL of ammonia water, put it into 200mL of pure water, and mix well;

[0110] Prepare six sealed containers, labeled A, B, C, D, E, and F;

[0111] Place corn cob litter in A, B, C, D, E, and F respectively until the litter thickness is 3 cm, and spray the odor source in the sealed containers, with a spraying volume of 30 mL in each container;

[0112] The disinfectant of Example 1 was evenly sprinkled on the corn cob bedding of container A, with a usage amount of 5 g, and the container was sealed; after 5 minutes, the container was opened, and no obvious ammonia odor was found in the container A, indicating that the disinfectant of the present invention can effectively treat the ammonia odor in the environment; the container was sealed for another 2 hours, and no odor appeared in the container A;

[0113] The disinfectant of Example 2 was evenly sprinkled on the corn cob bedding of container B, with a usage of 5 g, and the container was sealed; after 6 minutes, the container was opened, and no obvious ammonia odor was found in container B, indicating that the disinfectant of the present invention can effectively treat the ammonia odor in the environment; the container was sealed for another 2 hours, and no odor appeared in container B;

[0114] The disinfectant of Example 3 was evenly sprinkled on the corn cob bedding of container C, with a usage amount of 5 g, and the container was sealed; after opening at 7 minutes, there was no obvious ammonia odor in container C, indicating that the disinfectant of the present invention can effectively treat the ammonia odor in the environment; the container was sealed for 2 hours, and no odor appeared in container C;

[0115] The disinfectant of comparative example 1 was evenly sprinkled on the corn cob bedding of container D, the usage amount was 5g, and the container was closed; after 5 minutes, the container D still had a strong odor, and after 70 minutes, the container D had no obvious odor;

[0116] The disinfectant of comparative example 2 was evenly sprinkled on the corn cob bedding of container E, the usage amount was 5g, and the container was closed; after 5 minutes, the container E had a pungent odor, and after 2.5 hours, the container E still had a odor;

[0117] The disinfectant of comparative example 3 was evenly sprinkled on the corn cob bedding of container F, with a usage amount of 5 g, and then sealed; when the container was opened after 5 minutes, there was a pungent odor in container F; when the container was opened after 110 minutes of sealing, there was no obvious odor in container F.

[0118] It can be seen that the disinfectant provided by the present invention not only has good bactericidal performance, but can also effectively remove ammonia odor in the breeding house, with significant effect.

[0119] Although the present invention has been described in detail by reference to the preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and these modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily think of changes or substitutions within the technical scope disclosed by the present invention, and these shall be within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be subject to the scope of protection of the claims.

Claims

1. An acidic solid disinfectant, characterized in that: The composition comprises the following components in parts by weight: 50-55 parts of Sijiqing extract, 20-25 parts of wormwood bark extract, 5-9 parts of β-glucosidase and 11-25 parts of alginic acid; The preparation process of the Sijiqing extract is as follows: S1, crushing the waste Evergreen trees obtained by pruning into pieces of 0.5-1 mm, and soaking them in ethanol to obtain slurry A; S2, reflux and extract slurry A for 2-3 hours, filter while hot to obtain an extract; S3, purifying the extract using macroporous resin D101, first eluting with pure water for 2 column volumes, then eluting with ethanol with a concentration of 20%, 40% and 50% for 1 column volume each, and finally eluting with an ethanol solution with a concentration of 80-85% and collecting, concentrating and drying, to obtain an Evergreen extract; The preparation process of the thorn bark extract is as follows: SS1, crush the bark of the Chinese thorn tree to 4-5 mm, add water, mix well and soak for 24-36 hours to obtain slurry B; SS2, enzymatic hydrolysis, adjusting the pH of slurry B to 3.5-3.8 with formic acid, then adding hemicellulase and cellulase, the addition amount of hemicellulase is 3.5-4.0u / g, the addition amount of cellulase is 4.3-4.8u / g, and enzymatic hydrolysis is carried out at 45±0.5℃ for 2-2.5h to obtain enzymatic hydrolyzate A; SS3, inactivate the enzyme in the enzymatic hydrolyzate A and filter, collect the filtrate; add ethyl acetate to the filtrate, extract three times, collect the ethyl acetate phase, concentrate and dry to obtain the extract of the thorn bark.

2. The acidic solid disinfectant according to claim 1, characterized in that The acidic solid disinfectant comprises the following components in parts by weight: 52 parts of Ipomoea aquatica extract, 22 parts of Echinops chinensis extract, 7 parts of β-glucosidase and 19 parts of alginic acid.

3. The acidic solid disinfectant according to claim 1, characterized in that In step S1, the concentration of ethanol is 80-85%, and the weight ratio of evergreen waste to ethanol is 1:5-7.

4. The acidic solid disinfectant according to claim 1, characterized in that In step SS1, the weight ratio of the thorn bark to water is 1:10-15.

5. The acidic solid disinfectant according to claim 1, characterized in that In step SS2, the amount of hemicellulase added was 3.8 u / g, and the amount of cellulase added was 4.5 u / g.

6. The method for preparing the acidic solid disinfectant according to claim 1, characterized in that: The process is as follows: (1) According to the proportion of each group, prepare the extract of Sijiqing, the extract of thorn bark and alginic acid respectively, mix them evenly, and obtain a mixture; (2) dispersing β-glucosidase in water, adjusting the pH to 4.0-4.2 with formic acid, and obtaining an aqueous solution; (3) Spray the aqueous solution on the mixture and, after drying, obtain a solid disinfectant.

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