CN107467045B - Preparation method of berberine aqueous solution taking biogas slurry as solvent - Google Patents
Preparation method of berberine aqueous solution taking biogas slurry as solvent Download PDFInfo
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- CN107467045B CN107467045B CN201710767432.9A CN201710767432A CN107467045B CN 107467045 B CN107467045 B CN 107467045B CN 201710767432 A CN201710767432 A CN 201710767432A CN 107467045 B CN107467045 B CN 107467045B
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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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
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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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
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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
- A01N61/00—Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action
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- Engineering & Computer Science (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
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Abstract
The invention discloses a preparation method of a berberine aqueous solution taking biogas slurry as a solvent. The method comprises the following process steps: (1) stirring, precipitating and filtering biogas slurry with magnet and gypsum, adding diatomite into the filtrate, stirring, precipitating and filtering to obtain a biogas slurry solvent; (2) mixing humic acid and berberine, pulverizing, and sieving with 200 mesh sieve to obtain berberine coated with humic acid; (3) adding berberine wrapped by humic acid into biogas slurry solvent, adding antifreeze solution and antiseptic, and performing ultrasonic treatment to obtain berberine aqua. The natural bactericide for preventing and treating the ring spot of the apples and the tea leaves, which is green and environment-friendly and has good bactericidal effect, is provided by the invention. The invention has rich raw material sources and low cost, adopts pure natural components to carry out harmless treatment on the biogas slurry, and comprehensively utilizes wastes.
Description
Technical Field
The invention relates to a preparation method of a berberine aqueous solution taking biogas slurry as a solvent, belonging to the field of recycling agriculture for recycling wastes and preventing and treating plant diseases and insect pests.
Background
A large amount of biogas slurry is generated by anaerobic fermentation of livestock and poultry feces, belongs to high pollutants and cannot be directly discharged into the environment, and the construction and operation cost is greatly increased due to the fact that standard treatment is carried out through post-construction treatment facilities, so that the serious problem of digestion exists. The biogas slurry contains rich nitrogen, phosphorus, potassium, amino acid, rich trace elements, B vitamins, various hydrolases, organic acid, humic acid and other bioactive substances, and can stimulate the growth of crops, enhance the stress resistance of the crops and improve the product quality. The biogas slurry can be used as an organic fertilizer, but has the problems of uneven utilization space-time distribution and economy such as insufficient digestion capacity of nearby farmlands, small utilization amount in winter, high long-distance transportation cost and the like, and simultaneously compared with chemical fertilizers or commercial organic fertilizers, the biogas slurry has low concentration of various nutrient elements, serious heavy metal pollution and limits the feasibility of agricultural digestion to a certain extent.
Currently, chemical pesticide bactericides such as mancozeb, dimoxystrobin, azoxystrobin, polyoxin, thiophanate-methyl and the like are commonly used for preventing and treating apple ring rot and tea leaf ring spot. The apple ring spot not only infects the branches, but also infects the fruits, and the important point is fruit damage in terms of loss, but the incidence of the branches and the incidence of the fruits are closely related, and the prevention and the treatment of the ring spot of the branches and the fruits are considered in the prevention and the treatment. The tea leaf ring spot disease is mainly caused by spraying benomyl, carbendazim, captafol, chlorothalonil, thiophanate-methyl and the like after tea picking or at the early stage of disease occurrence, and the problem of environmental pollution caused by using a large amount of chemical pesticides is serious day by day. Meanwhile, the chemical pesticide bactericide is adopted, so that pathogenic bacteria strains generate drug resistance and drug resistance, the difficulty of prevention and treatment is increased, and even the prevention and treatment failure is caused. In addition, chemical pesticides pose a threat to beneficial organisms because they are not selective. Moreover, many chemical pesticides are undegradable or difficult to degrade and tend to accumulate in animals and plants, resulting in an enrichment of pollutants. Along with the increasing public requirements on green food and pollution-free food, the market demand for pollution-free and naturally degradable ecological pesticides is increased gradually. The natural return and the pollution-free and agricultural zero-growth of pesticides in the aspect of pesticide development are inevitable trends of social and natural scientific development, so that the novel biological broad-spectrum bactericide has wide development and application prospects.
Based on the prior art, the invention is subsidized by the innovation capability promotion plan project of the university of the Beijing education Committee city.
Disclosure of Invention
The invention aims to provide a preparation method of a berberine aqueous solution taking biogas slurry as a solvent, which is used for preparing the berberine aqueous solution with environmental protection and good sterilization effect while comprehensively utilizing wastes.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of berberine aqueous solution comprises the following process steps:
(1) stirring, precipitating and filtering biogas slurry with magnet and gypsum, adding diatomite into the filtrate, stirring, precipitating and filtering to obtain a biogas slurry solvent;
(2) mixing humic acid and berberine, pulverizing, and sieving with 200 mesh sieve to obtain berberine coated with humic acid;
(3) adding berberine wrapped by humic acid into biogas slurry solvent, adding antifreeze solution and antiseptic, and performing ultrasonic treatment to obtain berberine aqua.
In the method, humic acid is macromolecular organic substance widely existing in nature, mainly plant remains, and organic substance which is caused and accumulated by a series of processes of microbial decomposition and transformation and geochemistry. In agriculture, humic acid fertilizer prepared by combining with elements such as nitrogen, phosphorus, potassium and the like has the functions of fertilizer synergism, soil improvement, crop growth stimulation, agricultural product quality improvement and the like. Berberine has broad-spectrum bactericidal effect, but berberine has poor water solubility, and solubility in water is only 0.5%. According to the invention, through pretreatment of the biogas slurry, the heavy metal content in the biogas slurry is obviously lower than the national standard, and the mutual solubility of the added humic acid and berberine is good.
In the step (1), the magnet and the gypsum are respectively crushed in advance and sieved by a 80-mesh sieve.
In the step (1), the input amount of the magnet is 3-30mg/L, and the input amount of the gypsum is 5-50 mg/L.
In the step (1), after the magnetite and the gypsum are put into the reaction kettle, the mixture is rapidly stirred for 15 to 50 seconds at the speed of 200-400r/min, and then slowly stirred for 15 to 30 minutes at the speed of 40 to 60r/min, and then is precipitated for 30 to 60 minutes.
In the step (1), 60-100g/L of 60-mesh diatomite is added, slowly stirred for 15-30min at 40-60r/min, and precipitated for 30-60 min.
In the step (2), the ratio (weight percentage) of the humic acid to the berberine to the biogas slurry is 1-5: 1: 1000.
In the step (3), the ratio of the total volume of the biogas slurry to the anti-freezing solution and the preservative is 1000: 1-5.
The invention has the advantages that:
the natural bactericide for preventing and treating the ring spot of the apples and the tea leaves, which is green and environment-friendly and has good bactericidal effect, is provided by the invention. The invention has rich raw material sources and low cost, adopts pure natural components to carry out harmless treatment on the biogas slurry, and comprehensively utilizes wastes.
The method can be used for preparing the berberine aqueous solution with high concentration, and greatly reduces the transportation cost of the berberine aqueous solution compared with the berberine aqueous solution with low concentration (0.5 percent). The berberine aqueous solution prepared by the invention has the effect of preventing and treating apple ring spot and tea leaf ring spot more than 97 percent, and can obviously improve the crop yield.
Detailed Description
The present invention is further illustrated by the following specific examples. The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the specific material ratios, process conditions and results thereof described in the examples are illustrative only and should not be taken as limiting the invention as detailed in the claims.
Example 1
Pulverizing Magnetitum and Gypsum Fibrosum respectively, and sieving with 80 mesh sieve. Taking 2L of biogas slurry stock solution, adding 6mg of magnet and 80mg of gypsum into biogas slurry, rapidly stirring at 400r/min for 50s, slowly stirring at 40r/min for 30min, precipitating for 50min, filtering, and collecting supernatant as solvent I containing biogas slurry. Adding 120g of diatomite into the solvent I containing the biogas slurry, slowly stirring at 45r/min for 20min, precipitating for 40min, filtering, and taking 843mL of supernatant as the solvent II containing the biogas slurry. Mixing humic acid and berberine at a ratio of 1: 1, adding into a superfine pulverizer to pulverize, sieving with 200 mesh sieve, and collecting 140g of humic acid-coated berberine. Slowly adding berberine wrapped by humic acid into solvent II, adding antifreezing solution glycerol 5mL and 95% antiseptic ethanol 3mL, ultrasonic power 180W, ultrasonic time 30min, and diluting to 1000mL with solvent II to obtain berberine aqua. The berberine is completely dissolved in the solvent, and the content of the berberine is 7 percent.
Example 2
Pulverizing Magnetitum and Gypsum Fibrosum respectively, and sieving with 80 mesh sieve. Taking 2L of biogas slurry stock solution, adding magnetite and gypsum powder into biogas slurry, wherein the adding amount of the magnetite is 60mg and the adding amount of the gypsum is 70mg, quickly stirring for 26s at 260r/min, then slowly stirring for 25min at 50r/min, precipitating for 60min, filtering, and taking supernatant as solvent I containing biogas slurry. Adding 140g of diatomite into the solvent I containing the biogas slurry, slowly stirring at 50r/min for 30min, precipitating for 35min, filtering, and taking 843mL of supernatant as the solvent II containing the biogas slurry. Mixing humic acid and berberine at a ratio of 2: 1, adding into a superfine pulverizer, pulverizing, sieving with 200 mesh sieve, and collecting 210g of humic acid coated berberine. Slowly adding berberine wrapped by humic acid into solvent II, adding 4mL of antifreeze solution glycerol and 3mL of 95% preservative ethanol, performing ultrasonic treatment with ultrasonic power of 180W for 30min, and diluting to 1000mL with solvent II to obtain berberine aqua. The berberine is completely dissolved in the solvent, and the content of the berberine is 7 percent.
Example 3
Pulverizing Magnetitum and Gypsum Fibrosum respectively, and sieving with 80 mesh sieve. Taking 2L of biogas slurry stock solution, adding magnetite and gypsum powder into biogas slurry, wherein the adding amount of the magnetite is 30mg and the adding amount of the gypsum is 100mg, quickly stirring for 30s at 300r/min, then slowly stirring for 28min at 46r/min, precipitating for 55min, filtering, and taking supernatant as solvent I containing biogas slurry. Adding 200g of diatomite into a solvent I containing biogas slurry, slowly stirring at 45r/min for 28min, precipitating for 43min, filtering, taking 843mL of supernatant as a solvent II containing biogas slurry, mixing humic acid and berberine according to a ratio of 3:1, adding into a superfine pulverizer, pulverizing, sieving with 200 meshes, and taking 280g of humic acid-coated berberine. Slowly adding berberine wrapped by humic acid into solvent II, adding 3mL of antifreeze solution glycerol and 5mL of 95% preservative ethanol, performing ultrasonic treatment with ultrasonic power of 180W for 30min, and diluting to 1000mL with solvent II to obtain berberine aqua. The berberine is completely dissolved in the solvent, and the content of the berberine is 7 percent.
Example 4 (conventional centrifugation Process)
Taking 2L of biogas slurry stock solution, placing on a centrifuge, centrifuging at 4000r/min for 30min, filtering, and taking supernatant. Pulverizing berberine in a superfine pulverizer, sieving with 200 mesh sieve, and dissolving berberine in the supernatant. The filter paper is dried (about 1 hour at 60 degrees celsius) to constant weight and cooled to the operating ambient temperature for use. Measuring 100mL of biogas slurry in a beaker by using a measuring cylinder, gradually adding berberine, heating to assist dissolution, continuously stirring until all berberine is dissolved, performing suction filtration while hot, washing residues with the biogas slurry for three times, and drying the filter paper after suction filtration at 50 ℃ to constant weight. And calculating to obtain the solubility of the berberine in the biogas slurry. The experimental data are shown in table 1.
TABLE 1 solubility of berberine in supernatant of biogas slurry (traditional centrifugation process)
Amount of undissolved berberine | Adding berberine in an amount | Berberine dissolving amount | Solubility of Berberine |
0.0018g | 0.0158g | 0.0140g | 0.014% |
As can be seen, the solubility of berberine in the conventional centrifugation process of example 4 is only 0.014%.
Example 5 (solubility of Berberine in Water)
Pulverizing berberine in 2L distilled water in a superfine pulverizer, sieving with 200 mesh sieve, and dissolving berberine in distilled water. The filter paper is dried (about 1 hour at 60 degrees celsius) to constant weight and cooled to the operating ambient temperature for use. Measuring 100mL of biogas slurry in a beaker by using a measuring cylinder, gradually adding berberine, heating to assist dissolution, continuously stirring until all berberine is dissolved, performing suction filtration while hot, washing residues with water for three times, and drying the filter paper after suction filtration at 50 ℃ to constant weight. And calculating to obtain the solubility of the berberine in the biogas slurry. The experimental data are shown in table 2.
TABLE 2 solubility of Berberine in distilled Water
Amount of undissolved berberine | Adding berberine in an amount | Berberine dissolving amount | Solubility of Berberine |
0.044g | 0.547g | 0.503g | 0.503% |
Examples 1-3 berberine treated by the invention can be prepared into 7% aqueous solution in biogas slurry, and the concentration is obviously improved. Examples 1-5 inhibition of apple ring rot by aqueous berberine preparations
The berberine aqueous solutions of examples 1-5 are respectively prepared into drug-carrying culture media with berberine final concentrations of 32 mug/mL, 64 mug/mL, 128 mug/mL, 256 mug/mL and 512 mug/mL, sterilized, punched on the activated apple ring rot disease bacteria by an 8mm puncher, hypha is downward, the bacterial cake is inoculated to the center of a flat plate, the flat plate is placed in an incubator at 37 ℃, and the inverted culture is carried out. After culturing for 96h, measuring the colony diameter (cross method), and calculating formulas are shown as formula 1-1 and formula 1-2.
Formula 1-1: colony diameter (mm) — average value of colony diameters (mm) measured-8
Formulas 1 to 2: the inhibition rate (%) of berberine on apple ring spot bacteria is 100% (blank control group colony diameter-area medicine group colony diameter)/blank control group colony diameter%
Calculation of apple ring lines for each example by the virulence regression equationHalf inhibitory concentration EC of pathogenic bacteria50See table 3.
TABLE 3 median inhibitory concentration EC of berberine aqua prepared in different examples on apple ring rot pathogen50
Examples | EC50μg/mL |
1 | 12.6 |
2 | 14.9 |
3 | 13.2 |
4 | 163.2 |
Examples 1-4 Effect of berberine aqua on tea leaf alternaria leaf Spot
The test field is selected in a Wuyi mountain Wansheng tea field, each treatment is carried out for 3 times in each example, the berberine aqua prepared in 5 examples is sprayed for 1 time in the middle and the last ten days of 6 months by using the berberine aqueous solution with the effective component concentration of 0.28mg/mL and without medicine blank control, the pesticide is sprayed at the same time in each test point at the pesticide application interval of 14 days, and the round spot disease is investigated at the interval of 18 days after the last treatment. And randomly checking 30 leaves of the middle plant every time, and observing and recording the number of the diseased leaves.
TABLE 4 influence of aqueous berberine preparations of different examples on tea leaf verticillium (concentration 0.28mg/mL)
Treatment of | Index of disease condition | Control effect% |
CK | 29.32 | 0.0000 |
1 | 1.01 | 96.56 |
2 | 1.23 | 95.80 |
3 | 0.62 | 97.89 |
4 | 14.24 | 51.43 |
The control effect of example 4 is significantly lower than that of examples 1-3.
Claims (5)
1. A preparation method of berberine aqueous solution taking biogas slurry as solvent is characterized by comprising the following process steps:
(1) stirring, precipitating and filtering biogas slurry with magnet and gypsum, adding diatomite into the filtrate, stirring, precipitating and filtering to obtain a biogas slurry solvent; the input amount of the magnet is 3-30mg/L, and the input amount of the gypsum is 35-50 mg/L; the input amount of the diatomite is 60-100 g/L;
(2) mixing humic acid and berberine, pulverizing, and sieving with 200 mesh sieve to obtain berberine coated with humic acid, wherein the ratio of humic acid to berberine is 1-3: 1;
(3) adding 140 and 280g of humic acid-coated berberine into a biogas slurry solvent, adding an antifreezing solution and a preservative, performing constant volume to 1000mL by using the biogas slurry solvent, and performing ultrasonic treatment to obtain a berberine aqueous solution, wherein the berberine content is 7%.
2. The production method according to claim 1, wherein in the step (1), the magnetite and the gypsum are separately pulverized in advance and sieved with a 80-mesh sieve.
3. The method as claimed in claim 1, wherein in the step (1), after the magnetite and gypsum are added, the mixture is rapidly stirred for 15-50s at a speed of 200-400r/min, and then slowly stirred for 15-30min at a speed of 40-60r/min, and then precipitated for 30-60 min.
4. The method according to claim 1, wherein in the step (1), 60-mesh diatomaceous earth is added, and the mixture is slowly stirred at 40-60r/min for 15-30min and then precipitated for 30-60 min.
5. The preparation method according to claim 1, wherein in the step (3), the ratio of the total volume of the biogas slurry to the anti-freezing solution and the preservative is 1000:1-5: 1-5.
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CN110447643B (en) * | 2019-09-03 | 2021-09-28 | 北京联合大学 | Berberine smoke agent and application thereof |
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