CN111567570B - Plant extract, disinfectant and application of disinfectant in disinfection of medical instruments - Google Patents
Plant extract, disinfectant and application of disinfectant in disinfection of medical instruments Download PDFInfo
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- CN111567570B CN111567570B CN202010408864.2A CN202010408864A CN111567570B CN 111567570 B CN111567570 B CN 111567570B CN 202010408864 A CN202010408864 A CN 202010408864A CN 111567570 B CN111567570 B CN 111567570B
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- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/24—Lauraceae [Laurel family], e.g. laurel, avocado, sassafras, cinnamon or camphor
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- 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
- A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
- A01N31/02—Acyclic compounds
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- 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
- A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
- A01N35/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
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- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- Health & Medical Sciences (AREA)
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- Wood Science & Technology (AREA)
- Agronomy & Crop Science (AREA)
- Environmental Sciences (AREA)
- Biotechnology (AREA)
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- Mycology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
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Abstract
The invention relates to the technical field of natural medicinal chemistry and disinfectants, and particularly discloses a plant extract, a disinfectant and application of the disinfectant in disinfection of medical instruments. The preparation method of the plant extract comprises the following steps: extracting seeds of lignum Cinnamomi Camphorae with mixed solvent of chloroform and petroleum ether. The plant extract has excellent MRSA inhibitory effect; can be used as bacteriostatic agent; meanwhile, the plant extract can be matched with a conventional disinfectant to further improve the sterilization and bacteriostasis effects of the MRSA.
Description
Technical Field
The invention relates to the technical field of natural medicinal chemistry and disinfectants, in particular to a plant extract, a disinfectant and application of the disinfectant in disinfection of medical instruments.
Background
Methicillin-resistant staphylococcus aureus (MRSA) is a clinically common bacterium with strong toxicity, and belongs to one of super bacteria; MRSA from the discovery to date infections are almost worldwide and have become one of the important pathogens of nosocomial and community infections. The conventional disinfectant such as disinfectant containing alcohol has the effects of killing and inhibiting MRSA, and can effectively control the infection of MRSA. But with the long-term use of the same disinfectant, superbacteria become resistant to the disinfectant. Therefore, the development of a novel bacteriostatic agent is of great significance.
Camphor tree seeds, also known as camphor tree seeds and camphor tree seeds, are the fruits of the Lauraceae plant, Camphor tree. The closest comparison document (Yuhai faithful, etc. Cinnamomum camphora seed ethanol extract component produced in northwest of Ebei and antibacterial activity research [ J ] agricultural science of southwest, 2010,23(4): 1094-; however, it does not disclose that the ethanol extract of cinnamomum camphora seeds has an MRSA inhibitory effect. Further, it is not disclosed how to prepare an effective fraction having an inhibitory effect on MRSA from cinnamomum camphora seeds.
Disclosure of Invention
The invention aims to solve the technical problem that in order to overcome the defect of research on the bacteriostatic action of camphor tree seeds in the prior art, firstly, a preparation method of a plant extract is provided, and the plant extract prepared by the method has the function of inhibiting MRSA.
In addition, the invention also provides application of the plant extract as a bacteriostatic agent. Also, a disinfectant containing the plant extract is provided.
The invention aims to solve the technical problems and is realized by the following technical scheme:
a method for preparing plant extract comprises extracting seeds of lignum Cinnamomi Camphorae with mixed solvent of chloroform and petroleum ether.
The inventor surprisingly discovers in the research that when the camphor tree seeds are extracted by using a mixed solvent consisting of chloroform and petroleum ether, the obtained extract shows stronger MRSA inhibition activity; however, the extract having a strong MRSA inhibitory activity cannot be obtained by extraction with a single chloroform, petroleum ether or other solvents, or even cannot be obtained by extraction. This indicates that chloroform and petroleum ether as a mixed solvent can extract MRSA inhibitory active ingredients from camphor tree seeds.
Further preferably, the camphor tree seeds are fresh camphor tree seeds.
Further studies by the inventors have shown that the MRSA inhibitory activity in fresh camphor tree seeds is higher than that in dried camphor tree seeds.
Most preferably, the camphor tree seeds are fresh camphor tree green seeds.
Further studies by the inventors have shown that the MRSA inhibitory activity in fresh camphor tree green seeds (i.e. cyan camphor tree seeds) is higher than the MRSA inhibitory activity in fresh camphor tree purple seeds (i.e. purple camphor tree seeds).
Preferably, the camphor tree seeds are camphor tree seeds.
Preferably, the volume ratio of the chloroform to the petroleum ether is 1-3: 1.
Further preferably, the volume ratio of chloroform to petroleum ether is 2: 1.
Preferably, the extraction refers to heating reflux extraction, and the extraction time is 40-90 min.
Most preferably, the extraction refers to heating reflux extraction, and the extraction time is 60 min.
Preferably, the camphor tree seeds are extracted by a mixed solvent consisting of chloroform and petroleum ether, and then the crude extract is further separated by silica gel column chromatography;
the separation condition of silica gel column chromatography comprises the steps of loading the crude extract into a silica gel column, eluting and removing impurities by using petroleum ether, and then performing separation by using a volume ratio of 97: 3-95: 5, a mixed solvent system consisting of petroleum ether and ethyl acetate; collecting petroleum ether and ethyl acetate elution parts, concentrating and drying to obtain the plant extract.
Further preferably, the silica gel column chromatography is performed under the separation condition that the crude extract is applied to a silica gel column, petroleum ether is used for elution and impurity removal, and then the volume ratio of the crude extract to the silica gel column is 96: 4, mixed solvent composed of petroleum ether and ethyl acetate is used for carrying out the system; collecting petroleum ether and ethyl acetate elution parts, concentrating and drying to obtain the plant extract.
Further research by the inventors shows that the extract with stronger MRSA inhibitory activity can be obtained under the above silica gel column chromatography conditions.
The invention also provides a plant extract prepared by the preparation method of the plant extract.
The invention also provides application of the plant extract in preparation of a bacteriostatic agent.
Preferably, the bacteriostatic agent is a bacteriostatic agent for methicillin-resistant staphylococcus aureus.
The invention also provides a disinfectant comprising a first active ingredient and a second active ingredient; the first active ingredient is the plant extract; the second active ingredient is a conventional disinfectant; the mass ratio of the first active ingredient to the second active ingredient is 1-10: 100-200.
Preferably, the conventional disinfection is one or more of ethanol, glutaraldehyde disinfectant, hydrogen peroxide solution, chlorine-containing disinfectant, iodophor and hand disinfectant.
The invention also provides an application of the disinfectant in disinfection of medical instruments.
Has the advantages that: the invention provides an extract with MRSA inhibition effect extracted from camphor tree seeds for the first time; research shows that the plant extract prepared by the method has excellent MRSA inhibition effect. Furthermore, the plant extract can be used as bacteriostatic agent; meanwhile, the plant extract can be matched with a conventional disinfectant to further improve the sterilization and bacteriostasis effects of the MRSA.
Detailed Description
The present invention is further explained below with reference to specific examples, which do not limit the scope of the present invention.
Example 1 preparation of plant extracts
Collecting fresh (within 3 days of picking) Cinnamomum camphora green seeds 500g, extracting with 4L mixed solvent of chloroform and petroleum ether at volume ratio of 2:1 under heating and refluxing, concentrating the extractive solution, and drying to obtain plant extract.
Example 2 preparation of plant extracts
Taking 500g of fresh (picked up within 3 days) cinnamomum camphora purple seeds, heating and refluxing the fresh (picked up) cinnamomum camphora purple seeds by using 4L of mixed solvent consisting of chloroform and petroleum ether with the volume ratio of 2:1, concentrating the extracting solution and drying the extracting solution to obtain the plant extract.
Example 3 preparation of plant extracts
Taking 500g of fresh (picked up within 3 days) camphor tree green seeds, heating and refluxing the fresh camphor tree green seeds by using 4L of mixed solvent consisting of chloroform and petroleum ether with the volume ratio of 2:1, and concentrating and drying the extracting solution to obtain a crude extract;
loading the crude extract to a silica gel column (the silica gel column adopts 300-400 mesh silica gel, the weight of the silica gel is 30 times of that of the crude extract), eluting and removing impurities by using petroleum ether with 3 times of column volume, and then eluting and removing impurities by using petroleum ether with 5 times of column volume according to a volume ratio of 96: 4, mixed solvent composed of petroleum ether and ethyl acetate is used for carrying out the system; and (6) collecting 96: 4, concentrating and drying the petroleum ether and ethyl acetate elution part to obtain the plant extract.
Example 4 preparation of plant extracts
Taking 500g of fresh (picked up within 3 days) camphor tree green seeds, heating and refluxing the fresh camphor tree green seeds by using 4L of mixed solvent consisting of chloroform and petroleum ether with the volume ratio of 2:1, and concentrating and drying the extracting solution to obtain a crude extract;
loading the crude extract to a silica gel column (the silica gel column adopts 300-400 mesh silica gel, the weight of the silica gel is 30 times of that of the crude extract), eluting and removing impurities by using petroleum ether with 3 times of column volume, and then eluting and removing impurities by using petroleum ether with 5 times of column volume according to a volume ratio of 97: 3, a mixed solvent system consisting of petroleum ether and ethyl acetate; and (7) collecting: 3, concentrating and drying the petroleum ether and ethyl acetate elution part to obtain the plant extract.
Example 5 preparation of plant extracts
Taking 500g of fresh (picked up within 3 days) camphor tree green seeds, heating and refluxing the fresh camphor tree green seeds by using 4L of mixed solvent consisting of chloroform and petroleum ether with the volume ratio of 2:1, and concentrating and drying the extracting solution to obtain a crude extract;
loading the crude extract to a silica gel column (the silica gel column adopts 300-400 mesh silica gel, the weight of the silica gel is 30 times of that of the crude extract), eluting and removing impurities by using petroleum ether with 3 times of column volume, and then eluting and removing impurities by using petroleum ether with 5 times of column volume according to a volume ratio of 95: 5, a mixed solvent system consisting of petroleum ether and ethyl acetate; and (4) collecting 95: and 5, concentrating and drying the petroleum ether and ethyl acetate elution part to obtain the plant extract.
Comparative example 1 preparation of plant extract
Collecting fresh (within 3 days of picking) Cinnamomum camphora green seeds 500g, extracting with 4L chloroform under heating and refluxing, concentrating the extractive solution, and drying to obtain plant extract.
Comparative example 2 preparation of plant extract
Collecting fresh (within 3 days of picking) Cinnamomum camphora green seeds 500g, extracting with 4L petroleum ether under heating and refluxing, concentrating the extractive solution, and drying to obtain plant extract.
Comparative example 3 preparation of plant extract
Collecting fresh (within 3 days of picking) Cinnamomum camphora green seeds 500g, extracting with 4L anhydrous ethanol under heating and refluxing, concentrating the extractive solution, and drying to obtain plant extract.
Comparative example 4 preparation of plant extract
Taking 500g of fresh (picked up within 3 days) camphor tree green seeds, heating and refluxing the fresh camphor tree green seeds by using 4L of mixed solvent consisting of chloroform and petroleum ether with the volume ratio of 2:1, and concentrating and drying the extracting solution to obtain a crude extract;
loading the crude extract to a silica gel column (the silica gel column adopts 300-400 mesh silica gel, the weight of the silica gel is 30 times of that of the crude extract), eluting and removing impurities by using petroleum ether with 3 times of column volume, and then eluting and removing impurities by using petroleum ether with 5 times of column volume according to a volume ratio of 90: 10, a mixed solvent system consisting of petroleum ether and ethyl acetate; and (5) collecting 90: 10, concentrating and drying the elution part of petroleum ether and ethyl acetate to obtain the plant extract.
Comparative example 5 preparation of plant extract
Taking 500g of fresh (picked up within 3 days) camphor tree green seeds, heating and refluxing the fresh camphor tree green seeds by using 4L of mixed solvent consisting of chloroform and petroleum ether with the volume ratio of 2:1, and concentrating and drying the extracting solution to obtain a crude extract;
applying silica gel column (silica gel with 300-400 meshes is adopted in the silica gel column, the weight of the silica gel is 30 times of that of the crude extract), and firstly applying a silica gel column with 3 times of column volume and the volume ratio of 95: 5, eluting and removing impurities by using a mixed solvent consisting of petroleum ether and ethyl acetate, and then performing elution by using a mixed solvent with the volume ratio of 5 times of column volume of 90: 10, a mixed solvent system consisting of petroleum ether and ethyl acetate; and (5) collecting 90: 10, concentrating and drying the elution part of petroleum ether and ethyl acetate to obtain the plant extract.
Examples of the experiments
The method for testing the minimum inhibitory concentration comprises the following steps: the drug to be tested was diluted with chloroform by a double dilution method to give solutions of 640mg/mL, 320mg/mL, 160mg/mL, 80mg/mL, 40mg/mL, 20mg/mL, 10mg/mL, 5mg/mL, and 2.5mg/mL, respectively. 1mL of each gradient of the liquid medicine and 19mL of the molten solid medium (beef extract peptone medium) were added to the petri dish and mixed uniformly. After the culture medium is cooled and solidified, respectively photocopying the strains to be detected in culture dishes containing medicines with different concentrations by using a section of capillary tube by adopting a photocopying method, culturing for 24 hours in a thermostat at 37 ℃, observing the growth condition of the strains, and recording the concentration of the strains which do not grow as the minimum inhibitory concentration; the specific experimental results are shown in table 1.
The drugs to be tested are: the antibacterial compositions prepared in examples 1 to 5 and the antibacterial compositions prepared in comparative examples 1 to 5.
The strain to be detected is methicillin-resistant staphylococcus aureus (MRSA); before experiment, each strain was prepared to have a concentration of 1X 107The bacterial liquid of (4).
TABLE 1 minimum inhibitory concentration (mg/mL) of the plant extract of the present invention
As can be seen from Table 1, the bacteriostatic test data of the plant extracts of examples 1 and 2 can be reflected in that the extract with MRSA inhibitory effect can be extracted from the seeds of Cinnamomum camphora tree by using chloroform and petroleum ether; and the extract extracted from the green seeds of the camphor tree has greater MRSA inhibitory activity than the extract extracted from the purple seeds of the camphor tree.
As can be seen from the bacteriostatic test data of the plant extracts in examples 3-5, the crude extract can be further subjected to the silica gel column chromatography method disclosed by the invention to obtain an active site with higher activity and MRSA (methicillin resistant Staphylococcus aureus) inhibition effect.
The experiments show that the plant extract obtained by the extraction method has excellent MRSA inhibition effect; it can be used as a brand new bacteriostatic agent.
As can be seen from the bacteriostatic test data of the plant extracts of comparative examples 1 and 2, the bacteriostatic activity is much less than that of example 1, which indicates that chloroform or petroleum ether alone cannot extract a large amount of effective components having MRSA inhibitory effect from the seeds of Cinnamomum camphora.
The plant extract from comparative example 3 did not show MRSA bacteriostatic activity in the above experimental concentration range. This indicates that the effective component having an MRSA inhibitory effect cannot be extracted effectively by using absolute ethanol.
As can be seen from the bacteriostatic test data of the plant extract in the comparative example 4, the bacteriostatic activity is much less than that of the plant extract in the example 1; comparative example 5 the plant extract did not show MRSA bacteriostatic activity in the above experimental concentration range; this shows that the silica gel column chromatography method has an important influence on further obtaining the effective component having the MRSA inhibitory action, and the effective component having the excellent MRSA inhibitory activity can be obtained only under the silica gel column chromatography condition described in the present invention.
EXAMPLE 6A disinfectant
The disinfectant with the brand new composition is obtained by adding the plant extract of any one of the embodiments 1-5 with the mass of 2% of ethanol into 75% of ethanol.
The disinfectant can further effectively kill and inhibit MRSA by adding the plant extract; it can be used for sterilizing medical instruments in hospitals to greatly reduce MRSA infection.
EXAMPLE 7A disinfectant
The disinfectant with the brand new composition is obtained by adding the plant extract of any one of the embodiments 1-5 with the mass of 5% of ethanol into glutaraldehyde disinfectant.
The disinfectant can further effectively kill and inhibit MRSA by adding the plant extract; it can be used for sterilizing medical instruments in hospitals to greatly reduce MRSA infection.
Claims (4)
1. The application of the plant extract in preparing the bacteriostatic agent is characterized in that the bacteriostatic agent is a bacteriostatic agent for methicillin-resistant staphylococcus aureus;
the plant extract is prepared by the following method: extracting seeds of camphor trees with a mixed solvent consisting of chloroform and petroleum ether;
wherein the volume ratio of chloroform to petroleum ether is 1-3: 1; the extraction refers to heating reflux extraction, and the extraction time is 40-90 min.
2. Use according to claim 1, wherein the volume ratio of chloroform to petroleum ether is 2: 1.
3. The use of claim 1, wherein the crude extract is further separated by silica gel column chromatography after the camphor tree seeds are extracted by the mixed solvent of chloroform and petroleum ether;
the separation condition of silica gel column chromatography comprises the steps of loading the crude extract into a silica gel column, eluting and removing impurities by using petroleum ether, and then performing separation by using a volume ratio of 97: 3-95: 5, a mixed solvent system consisting of petroleum ether and ethyl acetate; collecting petroleum ether and ethyl acetate elution parts, concentrating and drying to obtain the plant extract.
4. The use of claim 3, wherein the silica gel column chromatography is performed under the conditions that the crude extract is applied to a silica gel column, and is eluted with petroleum ether to remove impurities, and then the volume ratio of the crude extract to the petroleum ether is 96: 4, mixed solvent composed of petroleum ether and ethyl acetate is used for carrying out the system; collecting petroleum ether and ethyl acetate elution parts, concentrating and drying to obtain the plant extract.
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| CN111567570B (en) * | 2020-05-14 | 2021-07-27 | 中山市沙溪镇食品药品监督所 | Plant extract, disinfectant and application of disinfectant in disinfection of medical instruments |
| CN113881393B (en) * | 2021-11-03 | 2023-01-17 | 东莞市山力高分子材料科研有限公司 | Silicone adhesive with mildew-proof effect |
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| CN103211734A (en) * | 2013-05-08 | 2013-07-24 | 汪洪涛 | Hand cleaner containing multiple natural antibacterial components and preparation technology |
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| CN111567570A (en) * | 2020-05-14 | 2020-08-25 | 中山市沙溪镇食品药品监督所 | Plant extract, disinfectant and application of disinfectant in disinfection of medical instruments |
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| CN101548687A (en) * | 2009-04-30 | 2009-10-07 | 马中华 | Method for extracting sterilization original liquid from broad leaf trees |
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| CN109966410A (en) * | 2019-05-08 | 2019-07-05 | 中山市沙溪镇食品药品监督所 | A kind of bactericidal composition and the wound dressing containing the composition |
| CN111567570A (en) * | 2020-05-14 | 2020-08-25 | 中山市沙溪镇食品药品监督所 | Plant extract, disinfectant and application of disinfectant in disinfection of medical instruments |
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