CN115607603B - Oral liquid for clearing away plague and toxic materials and preparation method thereof - Google Patents
Oral liquid for clearing away plague and toxic materials and preparation method thereof Download PDFInfo
- Publication number
- CN115607603B CN115607603B CN202211053465.4A CN202211053465A CN115607603B CN 115607603 B CN115607603 B CN 115607603B CN 202211053465 A CN202211053465 A CN 202211053465A CN 115607603 B CN115607603 B CN 115607603B
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- root
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- oral liquid
- plague
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Classifications
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Abstract
The invention discloses an oral liquid for clearing plague and detoxifying and a preparation method thereof, and the oral liquid comprises the following raw materials in parts by weight: 200-400 parts of coptis chinensis, 250-350 parts of radix isatidis, 180-220 parts of divaricate saposhnikovia root, 200-250 parts of radix scutellariae, 100-150 parts of honeysuckle, 130-170 parts of gypsum, 80-120 parts of poria cocos, 50-80 parts of honey-fried licorice root, 120-180 parts of radix scrophulariae, 5-10 parts of lotus seed and 5-10 parts of peppermint. The prescription of the invention has reasonable compatibility of the prescription and cooperatively plays roles of clearing heat and detoxicating, strengthening body resistance and strengthening the body, replenishing qi and strengthening the spleen and improving the immunity of organisms. Meanwhile, the preparation method provided by the invention can improve the content of active ingredients in the composition, and improve the extraction efficiency and the product quality.
Description
Technical Field
The invention belongs to the technical field of veterinary medicines, and in particular relates to a pestilence-clearing and detoxifying oral liquid and a preparation method thereof.
Background
The oral liquid for clearing away the plague and detoxifying is a traditional Chinese veterinary medicine compound preparation which is most commonly used in prevention and control of poultry diseases in recent years, has the efficacy of clearing away the heat-evil and expelling superficial evils and mainly treats exogenous fever of the poultry. The prescription consists of radix isatidis, radix scutellariae, fructus forsythiae, fructus gardeniae, radix scrophulariae, radix rehmanniae and other medicinal materials. Traditional Chinese medicine suggests that fowl biological diseases are usually caused by invasion of heat toxin from shallow to deep, from exterior to interior and from heart lung to lower body, and the disease is caused by excessive heat evil in qi system, and the heat evil in blood system is easy to damage body fluid, so animals are ill: the general manifestations are: thirst, difficulty in acting, and stiffness of the extremities. The morbidity of animals is mainly acute, and the mortality rate is relatively high. Radix Isatidis in the oral liquid prescription for clearing away heat and toxic materials can detoxify and moisten throat; radix Scutellariae has effects of clearing lung-heat and purging heat; fructus forsythiae has effects of clearing heat and resisting bacteria, and eliminating upper middle-upper coke gas; fructus Gardeniae is used as adjuvant medicine for eliminating triple energizer heat syndrome, purging pathogenic fire and removing toxic substances; the brown participation of Sheng Di Huang can cool blood and reduce fire, thereby making up the kidney, and being an auxiliary drug. Clinical application in many countries proves that the preparation has obvious curative effect on prevention and control of poultry virus and germ, but the traditional production process has the defects of low extraction rate of medicinal material active ingredients, high loss in impurity removal and concentration links, high energy efficiency and the like. Meanwhile, due to the long-term use of various chemicals, antibiotics and the like, animals are extremely easy to generate drug resistance, although the dosage is continuously increased, the curative effect is not obviously improved or even continuously reduced.
For example, the traditional process generally has the defects of low dissolution rate of active ingredients extracted from decoction pieces, high loss rate of active ingredients except heterocyclic joints, unstable thermosensitive ingredients in a concentrating link, high loss rate of ingredients and the like, and also has the defects of high resource loss, high energy consumption, serious environmental pollution, high production cost and the like, so that the clinical and efficient exertion and the market competitiveness of the prescription are limited.
Therefore, how to innovate and improve the formulation of the oral liquid for clearing plague, and change the traditional preparation process, so as to provide a novel preparation process of the veterinary drug with simple and convenient process, high active ingredient yield, short extraction time, energy conservation and environmental protection, so as to enhance the exertion of clinical efficacy, which is a problem to be solved urgently.
Disclosure of Invention
The invention aims to provide an oral liquid for clearing away plague and toxic materials and a preparation method thereof, which can solve the problem of poor curative effect of the existing oral liquid and improve the dissolution rate of active ingredients of medicinal materials.
The invention provides an oral liquid for clearing plague and detoxicating, which comprises the following raw materials in parts by weight: 200-400 parts of coptis chinensis, 250-350 parts of radix isatidis, 180-220 parts of divaricate saposhnikovia root, 200-250 parts of radix scutellariae, 100-150 parts of honeysuckle, 130-170 parts of gypsum, 80-120 parts of poria cocos, 50-80 parts of honey-fried licorice root, 120-180 parts of radix scrophulariae, 5-10 parts of lotus seed and 5-10 parts of peppermint.
Further, the material comprises the following raw materials in parts by weight: 250-350 parts of coptis chinensis, 300-320 parts of radix isatidis, 200-210 parts of divaricate saposhnikovia root, 220-230 parts of radix scutellariae, 120-140 parts of honeysuckle, 130-150 parts of gypsum, 90-100 parts of poria cocos, 50-60 parts of honey-fried licorice root, 140-150 parts of radix scrophulariae, 5-8 parts of lotus seeds and 5-8 parts of mint.
Further, the material comprises the following raw materials in parts by weight: 300 parts of coptis chinensis, 300 parts of radix isatidis, 200 parts of divaricate saposhnikovia root, 200 parts of radix scutellariae, 120 parts of honeysuckle, 150 parts of gypsum, 90 parts of poria cocos, 60 parts of honey-fried licorice root, 150 parts of radix scrophulariae, 8 parts of lotus seeds and 5 parts of mint.
The invention also provides a preparation method of the oral liquid for clearing plague and detoxifying, which comprises the following steps:
(1) Mixing the components in proportion, crushing to 100-200 meshes, and then adding water with the weight 10-20 times of the weight of the mixture for heating and soaking;
(2) Filtering the mixture soaked in the step (1), heating by microwaves, heating to 60 ℃ firstly, then keeping for 10-30min, heating to 70 ℃ again, then keeping for 10-30min, and finally heating to 90 ℃ and keeping for 20-40min; separating the liquid and the solid;
(3) Soaking the solid in water, adjusting pH to 5.0, and adding cellulase, hemicellulase, pectase and amylase for enzymolysis;
(4) Heating to extract liquid medicine after enzymolysis, mixing the liquid medicine with the liquid separated in the step (2), decocting for 3-5 times, mixing the decoctions, centrifuging, and micro-filtering the supernatant.
Further, the heating temperature in the step (1) is 35-45 ℃, and the soaking time is 60-120min.
Further, in the step (3), the weight of the added water is 5-8 times of that of the solid, the mass ratio of the cellulase to the hemicellulase to the pectinase to the amylase is 1-2:1:1:3-5, the enzymolysis time is 30-40min, and the enzymolysis temperature is 35-45 ℃.
Further, the final temperature of the temperature rise in the step (4) is 75-85 ℃, and the extraction time is 45-55min.
Further, the step of microfiltration in the step (4) specifically includes: and (3) circularly micro-filtering the supernatant by using an inorganic ceramic membrane micro-filter in a cross-flow mode, adding distilled water with the amount of 2 times of that of the medicinal materials, continuing micro-filtering, and concentrating the micro-filtered liquid.
Further, the parameters of the microfiltration include: the aperture of the inorganic ceramic membrane is 0.2 mu m, the flow rate of the feed is 30-50m/s, and the concentration multiple is 3-5 times.
The Chinese medicinal materials in the invention comprise:
the modern pharmacological experiment research proves that the radix scutellariae contains a large amount of Huang Tongyan substances, so that the radix scutellariae has various effects of resisting inflammation, clearing heat and detoxicating, and the like, and has certain inhibition effects on the aspects of resisting virus, resisting tumor, resisting inflammation, resisting bacteria, protecting neuron cells, and the like. The in vitro experiment of locust proves that after decocting the radix scutellariae with water, the extract is taken to inhibit the reproduction of leptospira, the concentrated extract of the radix scutellariae in water has direct killing effect on some pathogenic microorganisms in human body, the radix scutellariae also has strong inhibiting and killing effect on some viruses, and the radix scutellariae also has strong inhibiting effect on respiratory viruses. The radix Scutellariae contains a large amount of coumarins, which are the main substances for inhibiting respiratory syncytial virus, and the radix Scutellariae extract contains a large amount of coumarins. Meanwhile, the radix scutellariae extract has a certain elimination effect on viruses such as respiratory syncytial virus, influenza virus and hepatitis B virus.
The isatis root contains a large amount of polysaccharide, starch, amino acid and other components, and in terms of antivirus, the water extract of the isatis root, the active component extract in the isatis root and the like are found to have stronger antivirus effect, and the alkaloid extract in the isatis root has disinfection and sterilization effects on influenza viruses. And the polysaccharide captured by the radix isatidis can strengthen the body fluid immunity function and the nonspecific immunity function of poultry and improve the capability of animals for resisting influenza viruses.
The radix scrophulariae has complex chemical components and mainly contains alkaloid, iridoids, volatile oil, phytosterol, phenylpropanoid glycosides, flavonoids, saccharides, organic acids, trace amount of monoterpene and diterpene, etc. The radix scrophulariae polysaccharide extract can obviously reduce blood lactic acid and serum urea content and increase liver glycogen content. In addition, radix scrophulariae has effects of dilating coronary artery, improving microcirculation and capillary permeability, improving endotoxemia, relieving pain, tranquilizing, and resisting convulsion.
Rhizoma Coptidis is rhizome of Coptidis rhizoma of Coptis genus of Ranunculaceae family, coptis deltoidea leaf or Coptis yunnanensis; contains alkaloid such as berberine, epiberberine, and phellodendron ketone; bitter and cold in nature and enters heart, liver, stomach and large intestine meridians; has effects in clearing away heat and toxic materials, purging pathogenic fire, and eliminating dampness; is mainly used for treating heat toxin sores and ulcers, furuncle, swelling and pain of gum, mouth and tongue sores and the like; modern pharmacology proves that coptis chinensis has the effects of resisting microorganisms, resisting protozoa, viruses, arrhythmia, ulcer, tumor, inflammation and the like.
Radix Saposhnikoviae has effects of inducing sweat, eliminating phlegm, dispelling pathogenic wind, relieving exterior syndrome, and relieving pain, and can be used for treating common cold, headache, joint pain, neuralgia, etc. Fang Feng is used as root drug. Pungent and sweet in flavor and warm in nature. Has effects in relieving exterior syndrome, inducing perspiration, dispelling pathogenic wind, and removing dampness, and can be used for treating common cold due to wind-cold, headache, fever, joint pain, and tetanus; in addition, the wind-proof blade and the wind-proof flower can also be used for medicine.
Gypsum, mainly: muscle-relaxing and heat-clearing; relieving restlessness and quenching thirst. The main fever is strong and the heat does not return; restlessness and unconsciousness; delirium and mania; thirst and dry throat; dyspnea due to lung heat; heatstroke spontaneous perspiration; headache due to stomach fire; toothache due to stomach fire; excessive heat toxin; spot eruption; sores of the mouth and tongue; carbuncle, cellulitis and sore and ulcer; the mouth is not closed; soup fire scalds. Channel tropism: enter lung and stomach meridians.
In summary, the invention has the following advantages:
1. the invention adopts isatis root, baikal skullcap root, coptis root and divaricate saposhnikovia root as principal drugs, honeysuckle flower and figwort root as ministerial drugs, lotus seed and peppermint as adjuvant drugs, gypsum for clearing heat and purging fire, poria for enhancing immunity, and honey-fried licorice root for harmonizing the drugs, thereby not only maintaining the efficacy of strong fire in original place, but also overcoming the defect of insufficient antiviral capability, and achieving the purposes of clearing heat and expelling toxin, activating blood and dissolving stasis, nourishing yin and cooling blood, replenishing qi and strengthening spleen and treating both principal and secondary aspect of disease.
2. The oral liquid for clearing away the plague and toxic materials has reasonable formula, has the functions of resisting bacteria and viruses, strengthening body resistance, clearing away heat and toxic materials, enriching blood and activating blood, replenishing qi and strengthening spleen, stimulating appetite, promoting growth and improving body immunity, has unique curative effect on the plague and toxic materials of livestock, and has obvious curative effect on various diseases of livestock caused by viruses or bacteria.
3. In the preparation method provided by the invention, microwave three-section heating is adopted, the traditional Chinese medicine can be fully soaked to fully swell cells, the cell internal pressure is rapidly increased under the action of microwave high frequency, the cell wall is broken, and the internal active ingredients are released; then carrying out enzymolysis treatment on plant cells containing cell walls, and further destroying the cell walls and improving the dissolution rate of active ingredients in the medicinal materials by optimizing the compound enzyme with the proportion relation; the combined liquid medicine is decocted again, and the yield of the baicalin can be obviously improved by a microfiltration method, so that the loss of main active ingredients in the impurity removal process is avoided.
Detailed Description
The principles and features of the present invention are described below in connection with the following examples, which are set forth to illustrate, but are not to be construed as limiting the scope of the invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1
The embodiment provides an oral liquid for clearing plague and detoxifying, which comprises the following raw materials in parts by weight: 300 parts of coptis chinensis, 300 parts of radix isatidis, 200 parts of divaricate saposhnikovia root, 200 parts of radix scutellariae, 120 parts of honeysuckle, 150 parts of gypsum, 90 parts of poria cocos, 60 parts of honey-fried licorice root, 150 parts of radix scrophulariae, 8 parts of lotus seeds and 5 parts of mint.
The embodiment also provides a preparation method of the oral liquid for clearing plague and detoxifying, which comprises the following steps:
(1) Mixing the components in proportion, pulverizing to 100 meshes, adding water 10 times of the weight of the mixture, heating to 35 ℃, and soaking for 60min;
(2) Filtering the mixture soaked in the step (1), heating by microwaves, heating to 60 ℃ firstly, then keeping for 20min, heating to 70 ℃ again, then keeping for 20min, and finally heating to 90 ℃ and keeping for 20min; separating the liquid and the solid;
(3) Soaking the solid in water with the mass of 5 times of that of the solid, adjusting the pH to 5.0, and adding cellulase, hemicellulase, pectase and amylase accounting for 0.1% of the mass of the solid for enzymolysis at 45 ℃; the mass ratio of the cellulase to the hemicellulase to the pectase to the amylase is 1:1:1:3, the enzymolysis time is 30min, and the enzymolysis temperature is 45 ℃;
(4) Heating to 80 ℃ after enzymolysis to extract liquid medicine for 50min, mixing the liquid medicine with the liquid collected in the step (2), decocting for 3 times, combining the decoction, centrifuging, and micro-filtering the supernatant to obtain the product;
the step of microfiltration specifically comprises: and (3) circularly micro-filtering the supernatant by using an inorganic ceramic membrane micro-filter with a membrane aperture of 0.2 mu m in a cross-flow mode, wherein the flow rate of the feed is 30m/s, adding 2 times of distilled water into the medicinal materials for continuous micro-filtration, and concentrating the micro-filtered liquid for 3 times.
Example 2
The embodiment provides an oral liquid for clearing plague and detoxifying, which comprises the following components in parts by weight:
200 parts of coptis chinensis, 250 parts of radix isatidis, 180 parts of divaricate saposhnikovia root, 200 parts of radix scutellariae, 100 parts of honeysuckle, 130 parts of gypsum, 80 parts of poria cocos, 50 parts of honey-fried licorice root, 120 parts of radix scrophulariae, 5 parts of lotus seed and 5 parts of mint.
The embodiment also provides a preparation method of the oral liquid for clearing plague and detoxifying, which comprises the following steps:
(1) Mixing the components in proportion, pulverizing to 200 meshes, adding water with the weight 20 times of that of the mixture, heating to 35 ℃, and soaking for 120min;
(2) Filtering the mixture soaked in the step (1), heating by microwaves, heating to 60 ℃ firstly, then keeping for 30min, heating to 70 ℃ again, then keeping for 10min, and finally heating to 90 ℃ and keeping for 20min; collecting liquids and solids;
(3) Soaking the solid in water with the mass of 8 times of that of the solid, adjusting the pH to 5.0, and adding cellulase, hemicellulase, pectase and amylase accounting for 0.1% of the mass of the solid for enzymolysis at 45 ℃; the mass ratio of the cellulase to the hemicellulase to the pectase to the amylase is 2:1:1:5, and the enzymolysis time is 30min;
(4) Heating to 85 ℃ after enzymolysis to extract liquid medicine for 45min, mixing the liquid medicine with the liquid collected in the step (2), decocting for 3 times, combining the decoction, centrifuging, and micro-filtering the supernatant to obtain the product;
the step of microfiltration specifically comprises: and (3) circularly micro-filtering the supernatant by using an inorganic ceramic membrane micro-filter with a membrane aperture of 0.2 mu m in a cross-flow mode, wherein the flow rate of the feed is 30m/s, adding 2 times of distilled water into the medicinal materials for continuous micro-filtration, and concentrating the micro-filtered liquid for 3 times.
Example 3
The embodiment provides an oral liquid for clearing plague and detoxifying, which comprises the following components in parts by weight:
400 parts of coptis chinensis, 350 parts of isatis root, 220 parts of divaricate saposhnikovia root, 20 parts of radix scutellariae, 150 parts of honeysuckle, 170 parts of gypsum, 120 parts of poria cocos, 80 parts of honey-fried licorice root, 180 parts of radix scrophulariae, 10 parts of lotus seed and 10 parts of mint.
The embodiment also provides a preparation method of the oral liquid for clearing plague and detoxifying, which comprises the following steps:
(1) Mixing the components in proportion, pulverizing to 200 meshes, adding water 10 times of the weight of the mixture, heating to 45 ℃, and soaking for 60min;
(2) Filtering the mixture soaked in the step (1), heating by microwaves, heating to 60 ℃ firstly, then keeping for 30min, heating to 70 ℃ again, then keeping for 30min, and finally heating to 90 ℃ and keeping for 20min; collecting liquids and solids;
(3) Soaking the solid in water with the mass of 8 times of that of the solid, adjusting the pH to 5.0, and adding cellulase, hemicellulase, pectase and amylase accounting for 0.1% of the mass of the solid for enzymolysis at 45 ℃; the mass ratio of the cellulase to the hemicellulase to the pectase to the amylase is 2:1:1:3, and the enzymolysis time is 30min;
(4) Heating to 85 ℃ after enzymolysis to extract liquid medicine for 45min, mixing the liquid medicine with the liquid collected in the step (2), decocting for 3 times, combining the decoction, centrifuging, and micro-filtering the supernatant to obtain the product;
the step of microfiltration specifically comprises: and (3) circularly micro-filtering the supernatant by using an inorganic ceramic membrane micro-filter with a membrane aperture of 0.2 mu m in a cross-flow mode, wherein the feeding flow rate is 50m/s, adding distilled water with the amount of 2 times that of the medicinal materials for continuous micro-filtration, and concentrating the micro-filtered liquid for 3 times to obtain the medicine.
Comparative example 1
300 parts of coptis chinensis, 200 parts of divaricate saposhnikovia root, 200 parts of radix scutellariae, 120 parts of honeysuckle, 90 parts of poria cocos, 60 parts of honey-fried licorice root, 150 parts of radix scrophulariae, 8 parts of lotus seeds and 5 parts of mint.
This comparative example was prepared in the same manner as in example 1.
Comparative example 2
300 parts of coptis chinensis, 300 parts of isatis root, 200 parts of divaricate saposhnikovia root, 120 parts of honeysuckle, 150 parts of gypsum, 90 parts of poria cocos, 150 parts of radix scrophulariae, 8 parts of lotus seed and 5 parts of mint.
This comparative example was prepared in the same manner as in example 1.
Comparative example 3
The preparation method of the oral liquid for clearing plague and detoxicating is as follows in the second book of national standard of local standard liter of veterinary medicine:
taking 1000mL of oral liquid as an example, decocting 150g of rehmannia root, 250g of cape jasmine fruit, 225g of radix scutellariae, 200g of weeping forsythia, 150g of radix scrophulariae and 200g of radix isatidis with water twice, mixing the decoctions, standing, filtering, concentrating, adding a proper amount of sodium benzoate, adding water to 1000mL, filling and sterilizing.
Test example 1
The oral liquid for clearing away the plague and expelling the toxin prepared in examples 1-3 and comparative examples 1-3 was studied for its drug properties.
1.1 clinical trial:
1.1.1 laboratory animals: pigs are 300 pigs at 6-10 months of age and all suffer from pestilence and are randomly divided into 6 groups of 50 pigs each, namely three experimental groups and three control groups.
1.1.2 administration: the experimental group and the control group correspond to the oral liquid prepared in the examples 1-3 and the comparative examples 1-3 respectively. Every 1000mL of the oral liquid is added with 25kg of water, and the oral liquid is fed in drinking water for 5 days as one period and for 2 periods.
1.1.3 effect judgment:
(1) Decision criterion
And (3) curing: the pig has good status and strong ingestion ability.
Invalidation: the symptoms were not alleviated.
1.2 experimental results
After 2 cycles, the results of 300 pigs with disease are shown in table 1:
table 1 experimental results
| Cure only a few | Invalid count | |
| Example 1 | 49 | 1 |
| Example 2 | 48 | 2 |
| Example 3 | 48 | 2 |
| Comparative example 1 | 35 | 15 |
| Comparative example 2 | 40 | 10 |
| Comparative example 3 | 37 | 13 |
As shown in Table 1, the oral liquid for clearing away the plague and toxic materials prepared by the invention has unique curative effect and obvious curative effect on various diseases of livestock and poultry caused by viruses or bacteria.
Test example 2
The content detection of baicalin in the following test examples is a high performance liquid chromatography detection method, and comprises the following steps:
sample preparation: taking 1mL of the liquid medicine, placing in a 50mL volumetric flask, adding a proper amount of 50% methanol solution, performing ultrasonic treatment for 20min (100W, 40 KHz), standing to room temperature, fixing the volume to a scale with the 50% methanol solution, shaking uniformly, and filtering with a microporous filter membrane (0.45 μm).
Chromatographic conditions: kromasil C18 (250 mm. Times.4.6 mm,5 μm) was used as column, octadecylsilane chemically bonded silica (C18) was used as filler, and acetonitrile was used as mobile phase: 0.2% phosphoric acid solution (22:78).
The detection wavelength is 274nm, the flow rate is 1.0mL/min, the column temperature is 40 ℃, and the sample injection amount is 10 mu L.
The optimization experiment of the crushing and extraction process is carried out, and the experimental example carries out the research of the crushing pretreatment process on the medicinal materials of the prescription of the oral liquid for clearing plague and detoxicating, and comprehensively compares the influence of different crushing and extraction parameters on the yield of active ingredients of the medicinal liquid.
The method specifically comprises the following steps:
2.1 test methods
2.1.1 particle size of the crushed material
300g of coptis chinensis, 300g of isatis root, 200g of divaricate saposhnikovia root, 200g of radix scutellariae, 120g of honeysuckle, 150g of gypsum, 90g of poria cocos, 60g of honey-fried licorice root, 150g of radix scrophulariae, 8g of lotus seeds and 5g of peppermint are respectively and evenly mixed after being crushed, and are respectively screened by a 50-mesh sieve, a 80-mesh sieve, a 100-mesh sieve, a 120-mesh sieve and a 200-mesh sieve, 10 times of water is added, and the rest steps are the same as in example 1, and the content of baicalin is measured by a high performance liquid chromatograph after the oral liquid for clearing plague and detoxicating is prepared.
2.1.2 heating time
300g of coptis chinensis, 300g of isatis root, 200g of divaricate saposhnikovia root, 200g of radix scutellariae, 120g of honeysuckle, 150g of gypsum, 90g of poria cocos, 60g of honey-fried licorice root, 150g of radix scrophulariae, 8g of lotus seeds and 5g of peppermint are respectively and evenly mixed after being crushed, screened and sieved by a 100-mesh sieve, 10 times of water is added, soaked for 60min, then directly heated to 90 ℃ for extraction for 30min, 40min, 50min and 60min respectively, and the method provided in the embodiment 1 is adopted, firstly heated to 60 ℃, kept for 20min, then heated to 70 ℃ for 20min, finally heated to 90 ℃ and kept for 20min, after extraction is finished, other steps are the same as those in the embodiment 1, and after the oral liquid for clearing plague and detoxifying is prepared, the content of baicalin is measured by a high-performance liquid chromatograph.
2.1.3 extraction temperature
300g of coptis chinensis, 300g of isatis root, 200g of divaricate saposhnikovia root, 200g of radix scutellariae, 120g of honeysuckle, 150g of gypsum, 90g of poria cocos, 60g of honey-fried licorice root, 150g of radix scrophulariae, 8g of lotus seeds and 5g of peppermint are respectively and evenly mixed, screened and sieved by a 100-mesh sieve, 10 times of water is added, soaked for 60min, then respectively extracted for 60min at 60 ℃ and 70 ℃ and 90 ℃, and the method provided in the embodiment 1 is that the temperature is firstly heated to 60 ℃ and then kept for 20min, then heated to 70 ℃ and then kept for 20min, after the extraction is finished, the rest steps are the same as those in the embodiment 1, and the content of the baicalin is measured by a high-performance liquid chromatograph after the oral liquid for clearing plague and detoxification is prepared.
2.1.4 Water addition
300g of coptis chinensis, 300g of isatis root, 200g of divaricate saposhnikovia root, 200g of radix scutellariae, 120g of honeysuckle, 150g of gypsum, 90g of poria cocos, 60g of honey-fried licorice root, 150g of radix scrophulariae, 8g of lotus seeds and 5g of peppermint are crushed, uniformly mixed, screened and screened by a 100-mesh sieve, 200g of each part is respectively added with 600mL, 1000mL, 1600mL, 2000mL and 3000mL of water, soaked for 60min, and after extraction, the rest steps are the same as in example 1, and the content of baicalin is measured by a high-performance liquid chromatograph after the oral liquid for clearing plague and detoxicating is prepared.
2.2 test results
2.2.1 particle size of the crushed material
The yield of the active ingredients of the traditional Chinese medicinal materials is closely related to whether the traditional Chinese medicinal materials are crushed and extracted or not, the crushing meshes of the medicinal materials are different, and the yields of the active ingredients are also different, which is probably caused by the difference of the contact areas of the solvent and the medicinal materials. As the number of pulverized medicinal materials increases, the dissolution rate of active ingredients increases, because the solvent can be more fully contacted with the medicinal materials. The test results show that: when the crushing fineness is 100 meshes, the baicalin yield in the liquid medicine reaches the maximum value, but the granularity of the medicinal materials is too small, so that the problem of removing impurities in the later period is solved, and after comprehensive comparison, the crushing fineness of the medicinal materials is selected to be better than that of the medicinal materials passing through a 100-mesh sieve. The test results are shown in table 2:
TABLE 2 influence of crushing particle size on baicalin content
| Crushing granularity (mesh) | 50 | 80 | 100 | 120 | 200 |
| Content (mg/mL) | 6.79 | 6.95 | 6.98 | 6.82 | 6.55 |
2.2.2 extraction time
The extraction time in the extraction process of the traditional Chinese medicine is closely related to the yield of the active ingredients, and the longer the extraction time is, the longer the solvent is contacted with the medicine, the larger the dissolution rate of the active ingredients is, because the solvent can be contacted with the medicine more fully. Meanwhile, the cell walls can be broken gradually by adopting stage heating, the dissolution efficiency is improved, and the test result is obtained: when the extraction time is 60min and the temperature is raised in stages, the baicalin content of the obtained liquid medicine reaches the maximum value, and the test results are shown in the following table 3:
TABLE 3 influence of extraction time on baicalin content
| Extraction time (min) | 30 | 40 | 50 | 60 | Example 1 |
| Content (mg/mL) | 5.23 | 5.65 | 6.04 | 6.65 | 6.98 |
2.2.3 extraction temperature
The yield of the active ingredients extracted from the traditional Chinese medicine is closely related to the extraction time and the extraction temperature, and the more the extraction time is sufficient, the more the active ingredients are dissolved out at a certain temperature. Meanwhile, as the extraction temperature is increased, the dissolution rate of the active ingredients is gradually increased, because the higher the temperature is, the more active ingredients in the medicinal materials move more severely and are more easily dissolved. Meanwhile, when the temperature is raised to 80-90 ℃, the content change is not obvious, but a certain dissolution rate can be improved by adopting a stage temperature raising method, so that the test results show that: when the temperature reaches above 90 ℃, the yield of the active ingredient tends to be balanced, so that the extraction temperature of 90 ℃ is selected to be better. As shown in table 4 below:
TABLE 4 influence of extraction temperature on baicalin content
| Extraction temperature (. Degree. C.) | 60 | 70 | 80 | 90 | Example 1 |
| Content (mg/mL) | 5.69 | 5.88 | 6.44 | 6.65 | 6.98 |
2.2.4 Water addition
In the extraction process of the traditional Chinese medicinal materials, the water adding amount is extremely related to the yield of the active ingredients, and researches prove that the yield of the active ingredients is gradually increased along with the increase of the water adding amount, and the more the solvent amount is, the more the solvent amount is in full contact with the medicinal materials, so that the active ingredients are more soluble. As can be seen from the data shown in table 5 below, when the water addition amount reaches 10 times or more, the yield of the active ingredient tends to be maximum, and the factors such as energy consumption are comprehensively considered, and the test results show that: the water adding amount is 10 times of that of the medicinal materials, and is better.
TABLE 5 influence of the amount of added water on the baicalin content
| Water addition multiple (mL) | 600 | 1000 | 1600 | 2000 | 3000 |
| Content (mg/mL) | 4.98 | 5.24 | 5.68 | 6.15 | 6.23 |
The research shows that when plant cells are broken, the fiber structure is destroyed, the effective components in the cells can be fully exposed, and then the effective components are directly contacted with the organic reagent for extraction and separation, which shortens the extraction and separation time greatly compared with the extraction of Chinese medicinal decoction pieces, thereby improving the dissolution rate of the Chinese medicaments and reducing the resource loss.
In summary, the optimal technical parameters of the crushing and extracting process of the invention are as follows: pulverizing the medicinal materials, sieving with 100 mesh sieve, extracting for 60min with three-stage heating, extracting at 90deg.C with water amount 10 times of that of the medicinal materials. Under the condition of the technological parameters, the yield of the active ingredients in the obtained liquid medicine is highest.
Test example 3
The optimization experiment of the biological enzyme extraction process is carried out, and the experiment example comprehensively compares the influence of the type and the dosage of the biological enzyme on the yield of the active ingredients of the liquid medicine. The experimental example utilizes the principle of enzymolysis wall breaking of plant cell walls by biological enzymes, selects four enzymes of cellulase, hemicellulase, pectase, amylase and the like to form compound enzyme according to the characteristics of the medicinal ingredients of the composition, and screens out the optimal enzyme dosage and optimal enzyme reaction conditions.
3.1 test methods
3.1.1 enzyme dosage
300g of coptis chinensis, 300g of radix isatidis, 200g of divaricate saposhnikovia root, 200g of radix scutellariae, 120g of honeysuckle, 150g of gypsum, 90g of poria cocos, 60g of honey-fried licorice root, 150g of radix scrophulariae, 8g of lotus seed and 5g of peppermint are crushed and screened by a 100-mesh sieve for standby. Heating purified water 10 times of the weight of the medicinal materials to 35 ℃, and soaking for 60min at constant temperature; filtering the soaked mixture, heating by microwaves, heating to 60 ℃ firstly, then keeping for 20min, heating to 70 ℃ again, then keeping for 20min, and finally heating to 90 ℃ and keeping for 20min; collecting liquids and solids;
introducing condensed water into the solid, regulating the pH of the liquid medicine to 5.0 by using 40% citric acid aqueous solution, respectively adding cellulase, hemicellulase, pectase and amylase which are 0.1% in total, and designing the following different proportions for experiments: 1:0.5:1:5, 1:1:2:2, 1:1:2.5:1 and 1:1:1:3, stirring uniformly, performing enzymolysis at 45 ℃ for 30min, performing the rest steps in the same way as in example 1, and checking the content of baicalin after preparing the oral liquid for clearing plague and detoxicating.
3.1.2 enzyme reaction time
300g of coptis chinensis, 300g of radix isatidis, 200g of divaricate saposhnikovia root, 200g of radix scutellariae, 120g of honeysuckle, 150g of gypsum, 90g of poria cocos, 60g of honey-fried licorice root, 150g of radix scrophulariae, 8g of lotus seed and 5g of peppermint are crushed and screened by a 100-mesh sieve for standby. Heating purified water 10 times of the weight of the medicinal materials to 35 ℃, and soaking for 60 minutes at constant temperature; filtering the soaked mixture, heating by microwaves, heating to 60 ℃ firstly, then keeping for 20min, heating to 70 ℃ again, then keeping for 20min, and finally heating to 90 ℃ and keeping for 20min; collecting liquids and solids;
introducing condensed water into the solid, regulating the pH of the liquid medicine to 5.0 by using 40% citric acid aqueous solution, adding cellulase, hemicellulase, pectase and amylase with the mass ratio of 1:1:1:3, respectively carrying out enzymolysis for 10, 20, 30 and 60min at 45 ℃, and then heating to 80 ℃ for enzyme inactivation treatment; extracting for 50min, and checking the content of baicalin after preparing the oral liquid for clearing plague and detoxicating by the rest steps as in the example 1.
3.1.3 optimal temperature for enzymatic reaction
300g of coptis chinensis, 300g of radix isatidis, 200g of divaricate saposhnikovia root, 200g of radix scutellariae, 120g of honeysuckle, 150g of gypsum, 90g of poria cocos, 60g of honey-fried licorice root, 150g of radix scrophulariae, 8g of lotus seed and 5g of peppermint are crushed and screened by a 100-mesh sieve for standby. Heating purified water 10 times of the weight of the medicinal materials to 35 ℃, and soaking for 60 minutes at constant temperature; filtering the soaked mixture, heating by microwaves, heating to 60 ℃ firstly, then keeping for 20min, heating to 70 ℃ again, then keeping for 20min, and finally heating to 90 ℃ and keeping for 20min; collecting liquids and solids;
introducing condensed water into the solid, regulating the pH of the liquid medicine to 5.0 by using 40% citric acid aqueous solution, adding cellulase, hemicellulase, pectase and amylase with the mass ratio of 1:1:1:3, respectively carrying out enzymolysis at 20, 30, 40 and 45 ℃ for 30min, and then heating to 80 ℃ for enzyme inactivation treatment; extracting for 50min, and checking the content of baicalin after preparing the oral liquid for clearing plague and detoxicating by the rest steps as in the example 1.
3.1.4 optimal pH for enzymatic reaction
300g of coptis chinensis, 300g of radix isatidis, 200g of divaricate saposhnikovia root, 200g of radix scutellariae, 120g of honeysuckle, 150g of gypsum, 90g of poria cocos, 60g of honey-fried licorice root, 150g of radix scrophulariae, 8g of lotus seed and 5g of peppermint are crushed and screened by a 100-mesh sieve for standby. Heating purified water 10 times of the weight of the medicinal materials to 35 ℃, and soaking for 60 minutes at constant temperature; filtering the soaked mixture, heating by microwaves, heating to 60 ℃ firstly, then keeping for 20min, heating to 70 ℃ again, then keeping for 20min, and finally heating to 90 ℃ and keeping for 20min; collecting liquids and solids;
introducing condensed water into the solid, regulating the pH of the liquid medicine to 4.0, 4.5, 5.0 and 5.5 by using 40% citric acid aqueous solution, adding cellulase, hemicellulase, pectase and amylase in a mass ratio of 1:1:1:3, carrying out enzymolysis for 30min at 45 ℃, and then heating to 80 ℃ for enzyme inactivation treatment; extracting for 50min, and checking the content of baicalin after preparing the oral liquid for clearing plague and detoxicating by the rest steps as in the example 1.
3.2 experimental results
3.2.1 enzyme dosage
The content of baicalin is detected by HPLC, and the detection results are shown in the following table, and the data statistics result shows that when the total content of cellulase, hemicellulase, pectase and amylase is 0.1%, the extraction rate of main active ingredients reaches the maximum value, and the four enzymes are different in dosage and proportion, which is caused by the compatibility characteristics of the medicinal materials in the prescription in the oral liquid for clearing plague and detoxicating, and the cell wall structures of the medicinal materials in different rhizomes and fruits are different, so that the comprehensive research result shows that the following table 6 shows that the four different enzymes are selected in the research to have the proportion of 1:1:1:3.
TABLE 6 influence of enzyme dosage on baicalin content
| Enzyme ratio | 1:0.5:1:5 | 1:1:2:2 | 1:1:2.5:1 | 1:1:1:3 |
| Baicalin (mg/mL) | 6.03 | 5.98 | 6.31 | 6.98 |
3.2.2 enzyme reaction time
The enzyme reaction was examined by designing different time gradients and detecting baicalin content by HPLC method, and the results are shown in table 7 below, which shows that when the extraction time is equal to or higher than 30min, the enzyme reaction reached the maximum, and the main active ingredient had reached the maximum, so the experiment was selected to be the optimal time for the enzyme reaction for 30 min.
TABLE 7 influence of extraction time on baicalin content
| Extraction time (min) | 10 | 20 | 30 | 60 |
| Baicalin (mg/mL) | 5.56 | 6.55 | 6.98 | 7.01 |
3.2.3 optimal temperature for enzymatic reaction
The baicalin yield was examined by designing different enzyme reaction temperature gradients and detecting the baicalin content by HPLC method, as shown in table 8 below, when the enzymolysis temperature was 45 ℃, the main active ingredient reached the maximum value, and the temperature was lower or higher than this value, which affects the enzyme reaction efficiency, which was related to the optimum reaction temperatures of the four enzymes, thereby determining that 45 ℃ is the optimum temperature for the enzyme reaction.
TABLE 8 influence of enzyme reaction temperature on baicalin content
| Enzyme reaction temperature (. Degree. C.) | 20 | 30 | 40 | 45 |
| Baicalin (mg/mL) | 4.43 | 5.98 | 6.63 | 6.98 |
3.2.4 optimal pH for enzymatic reactions
The first yield of baicalin is examined by designing different pH gradients, and the baicalin content is detected by adopting an HPLC method, the test results are shown in table 9, when the pH reaches 5.0, the extraction rate of baicalin components reaches the maximum value, and 5.0 is selected as the optimal pH for enzymolysis due to the specificity of enzyme reaction to the pH.
TABLE 9 influence of pH on baicalin content
| pH | 4 | 4.5 | 5 | 5.5 |
| Baicalin (mg/mL) | 6.49 | 6.75 | 6.98 | 6.72 |
The research results of the test example show that: the optimal technical parameters of the biological enzymolysis wall-breaking extraction process are as follows: the optimal acting dosage of the compound enzyme is 0.1%, the proportion of cellulase, hemicellulase, pectase and amylase is 1:1:1:3, the pH is 5.0, the reaction time is 30min, and the reaction temperature is 45 ℃.
In summary, the optimal proportion relation of the invention is obtained by sieving the composition components, the crushing granularity, the heating time, the extraction temperature, the water addition amount, the enzyme dosage, the enzyme reaction time, the enzyme reaction optimal temperature and the enzyme reaction optimal pH value, so that the dissolution rate of the active ingredients is greatly improved, and the quality of the product is improved.
While specific embodiments of the invention have been described in detail, it should not be construed as limiting the scope of the patent. Various modifications and variations which may be made by those skilled in the art without the creative effort are within the scope of the patent described in the claims.
Claims (8)
1. The oral liquid for clearing away plague and toxic materials is characterized by being prepared from the following raw materials in parts by weight: 200-400 parts of coptis chinensis, 250-350 parts of radix isatidis, 180-220 parts of divaricate saposhnikovia root, 200-250 parts of baical skullcap root, 100-150 parts of honeysuckle, 130-170 parts of gypsum, 80-120 parts of Indian buead, 50-80 parts of honey-fried licorice root, 120-180 parts of figwort root, 5-10 parts of lotus seed and 5-10 parts of peppermint.
2. The oral liquid for clearing away plague and toxic materials according to claim 1, which is characterized by being prepared from the following raw materials in parts by weight: 250-350 parts of coptis chinensis, 300-320 parts of radix isatidis, 200-210 parts of divaricate saposhnikovia root, 220-230 parts of baical skullcap root, 120-140 parts of honeysuckle, 130-150 parts of gypsum, 90-100 parts of poria cocos, 50-60 parts of honey-fried licorice root, 140-150 parts of figwort root, 5-8 parts of lotus seed and 5-8 parts of peppermint.
3. The oral liquid for clearing away plague and toxic materials according to claim 1, which is characterized by being prepared from the following raw materials in parts by weight: 300 parts of coptis chinensis, 300 parts of isatis root, 200 parts of divaricate saposhnikovia root, 200 parts of baical skullcap root, 120 parts of honeysuckle, 150 parts of gypsum, 90 parts of Indian buead, 60 parts of honey-fried licorice root, 150 parts of figwort root, 8 parts of lotus seed and 5 parts of mint.
4. A method for preparing an oral liquid for clearing plague and detoxicating according to any one of claims 1-3, comprising the steps of:
(1) Mixing the components in proportion, crushing to 100-200 meshes, and then adding water with the weight 10-20 times of the weight of the mixture for heating and soaking;
(2) Filtering the mixture soaked in the step (1), heating by microwaves, heating to 60 ℃ firstly, then keeping for 10-30min, heating to 70 ℃ again, then keeping for 10-30min, and finally heating to 90 ℃ and keeping for 20-40min; separating the liquid and the solid;
(3) Soaking the solid in water, adjusting pH to 5.0, and adding cellulase, hemicellulase, pectase and amylase for enzymolysis; the weight of the added water is 5-8 times of that of the solid, the mass ratio of the cellulase to the hemicellulase to the pectase to the amylase is 1-2:1:1:3-5, the enzymolysis time is 30-40min, and the enzymolysis temperature is 35-45 DEG C
(4) Heating to extract liquid medicine after enzymolysis, mixing the liquid medicine with the liquid separated in the step (2), decocting for 3-5 times, mixing the decoctions, centrifuging, and micro-filtering the supernatant.
5. The method for preparing oral liquid according to claim 4, wherein the heating temperature in the step (1) is 35-45 ℃ and the soaking time is 60-120min.
6. The method for preparing oral liquid according to claim 4, wherein the final temperature of the heating in the step (4) is 75-85 ℃, and the extraction time is 45-55min.
7. The method for preparing the oral liquid for clearing away heat and toxic materials according to claim 4, wherein the step of microfiltration in the step (4) specifically comprises: and (3) circularly micro-filtering the supernatant by using an inorganic ceramic membrane micro-filter in a cross-flow mode, adding distilled water with the amount of 2 times of that of the medicinal materials, continuing micro-filtering, and concentrating the micro-filtered liquid.
8. The method for preparing the oral liquid for clearing away heat and toxic materials according to claim 7, wherein the parameters of the microfiltration comprise: the aperture of the inorganic ceramic membrane is 0.2 mu m, the flow rate of the feed is 30-50m/s, and the concentration multiple is 3-5 times.
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