CN108514628B - Production process of oral liquid for enriching, tonifying middle-jiao and replenishing qi - Google Patents
Production process of oral liquid for enriching, tonifying middle-jiao and replenishing qi Download PDFInfo
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- CN108514628B CN108514628B CN201810530671.7A CN201810530671A CN108514628B CN 108514628 B CN108514628 B CN 108514628B CN 201810530671 A CN201810530671 A CN 201810530671A CN 108514628 B CN108514628 B CN 108514628B
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Abstract
The oral liquid has effects of invigorating middle warmer and replenishing qi, and can be used for treating tiredness, debilitation, and viscera prolapse. The invention discloses a novel anti-precipitation treatment process of a traditional Chinese medicine oral liquid, and preferably certain medicinal materials are modified by microorganisms, so that the prepared oral liquid has excellent quality and high active ingredients, and not only has the effects of tonifying middle-jiao and Qi, but also has the effects of strengthening brain, benefiting intelligence and resisting aging. Further, the tubular easy-open bottle filled with the liquid medicine is placed into a water bath heating tank, a steam valve is opened, the temperature is gradually increased to 100 ℃, and the temperature is maintained at 100 ℃ for 10-15 minutes. And taking out after the heat preservation time is up. The advantages are that: the oral liquid treated by the method meets the drug standard by microbial limit inspection, does not precipitate within 2 years, and ensures no loss of effective components and stable drug property.
Description
Technical Field
The invention relates to the technical field of medicine production, in particular to a production process of an oral liquid for enriching, tonifying middle-jiao and replenishing qi.
Background
Tonify middle-jiao and qi oral liquid, tonify middle-jiao and qi. Can be used for treating asthenia and viscera prolapse. The medicine comprises radix astragali (processed with Mel), radix Codonopsis, Glycyrrhrizae radix (processed with Mel), Atractylodis rhizoma (parched), radix Angelicae sinensis, cimicifugae rhizoma, bupleuri radix, and pericarpium Citri Tangerinae. A liquid with a brown character; sweet, slightly bitter and pungent. It is an over-the-counter drug for deficiency syndrome.
In the traditional process, a tubular pop-top bottle filled with liquid medicine is put into a motor-driven door oral liquid sterilizer, steam is slowly introduced, cold air in a cavity is exhausted through preheating, the temperature is gradually raised to the required sterilization temperature, and the temperature is kept at 113 ℃ for 35 minutes. And after the sterilization is finished, rinsing for 300 seconds according to set parameters, and taking out. Disadvantages are that: the oral liquid treated by the method meets the drug standard by the microbial limit inspection, but slight precipitation is generated after the oral liquid is placed for more than 2 days, the precipitation is more serious after the oral liquid is placed for a longer time, and even the oral liquid is adhered to the wall of a bottle and is not easy to shake, so that the drug effect is reduced.
The invention aims to provide a novel anti-precipitation treatment process for traditional Chinese medicine oral liquid, so that the curative effect is better, a novel curative effect is endowed, and the novel process can prevent medicine precipitation.
Disclosure of Invention
The basic invention aims to provide a novel anti-precipitation treatment process of the traditional Chinese medicine oral liquid, so that the curative effect is better and a new curative effect is endowed.
The technical scheme of the invention is as follows:
a novel anti-precipitation treatment process of traditional Chinese medicine oral liquid is characterized by comprising the following steps:
(1) processing the medicinal materials:
honey-fried astragalus root: removing impurities from the raw materials, cleaning, moistening, slicing or slicing, drying, sieving to remove ash, mixing with refined honey, parching, taking out, and spreading for cooling;
codonopsis pilosula: taking raw medicine, removing impurities, cleaning, slightly moistening, cutting into thick pieces or sections, and drying;
honey-fried licorice root: removing impurities such as residual stems on the ground, grading, washing with strong water, moistening, slicing into thick pieces, cutting into 6mm segments for small ones, drying, slicing and segmenting in production place, sieving to remove ash and bits, mixing with refined honey, slightly stewing, parching, taking out, and spreading for cooling;
chinese angelica: taking raw medicines, removing impurities, cleaning with water, moistening, softening, slicing, and drying at low temperature;
frying the bighead atractylodes rhizome: removing impurities from the raw materials, grading the size, slightly soaking, cleaning, moistening, slicing, drying, placing bran stir-fried with honey in a hot pot, turning over, adding Atractylodis rhizoma when it is smoking, quickly stir-frying to dark yellow surface, taking out, sieving to remove bran, and spreading to cool; cimicifugae foetidae: collecting raw materials, removing impurities, slightly soaking, cleaning, moistening, slicing, and drying;
bupleurum root: collecting raw materials, removing impurities such as residual stem, cleaning, moistening, slicing into thick pieces or segments, and drying;
dried orange peel: collecting raw materials, removing impurities, cyan, and mould black, cleaning with water, shredding, and drying at low temperature; ginger: collecting raw materials, removing impurities, cleaning, and slicing into thick pieces;
chinese date: collecting raw materials, removing impurities and mould and moth-eaten people, cleaning with water, and drying.
Preferably, the astragalus, the angelica and the white atractylodes rhizome are further subjected to microbial fermentation treatment:
(a) the astragalus, the angelica and the white atractylodes rhizome are taken, and purified water which is 0.3 time of the total weight of the three medicinal materials is uniformly sprayed on the medicinal materials in parts.
(b) And (3) microbial fermentation treatment: spreading the medicinal materials in a sample tray, uniformly spraying the composite bacterial liquid on the medicinal materials, wherein the spraying amount is 0.05 times of the dry weight, and standing and fermenting at 38-40 deg.C for 10-12 hr;
the compound bacterial liquid is lactobacillus rhamnosus: lactobacillus brevis: the lactobacillus adolescentis is prepared from the following components in a ratio of 1: 0.2: 1.5, the total amount of the live bacteria is 1.3X1011cfu/ml；
(c) Freeze-drying: placing the fermented medicinal materials in a freeze dryer, controlling the freezing temperature to be-35 ℃, starting a vacuum pump to vacuumize until the pressure of a drying chamber is 60Pa after the required temperature is reached, adjusting the temperature of the drying chamber to be 50 ℃ for drying, and finally controlling the water content to be below 8%.
(2) Preparing materials: taking each batch of qualified decoction pieces which are produced by 80 ten thousand ml according to the production instruction, respectively weighing honey-fried astragalus, codonopsis pilosula, honey-fried licorice, angelica, fried bighead atractylodes rhizome, rhizoma cimicifugae, radix bupleuri, dried orange peel, ginger and Chinese date according to the prescription amount, and filling into a dry clean container;
(3) extraction: when the prescription materials are extracted, the name, the quantity and the like of the ten medicines are checked to be correct according to production instructions, half of the ten medicines are weighed respectively and put into 2 3m3 multifunctional extraction tanks respectively, the operations are carried out according to the standard operation rules of the multifunctional extraction tanks, ten times of water is added into the first juice, the first juice is soaked for one hour, the first juice is heated and decocted for 2 hours, the decocted liquid is filtered by a 100-mesh sieve to a liquid medicine storage tank, 10 times of water is added into the second juice, the second juice is heated and decocted for 1 hour, the decocted liquid is filtered by 100 meshes and combined with the first juice decocted liquid, the mixture is kept stand for more than 12 hours, the supernatant is put into a single-effect evaporation concentrator, the concentration is carried out according to the standard operation rules of the single-effect evaporation concentrator, the concentration is carried out until the relative density is 1.08-1.10 (90 +/-5 ℃);
the ratio of the extraction water (drinking water to purified water) (30-50)% to 70-50)%;
(4) alcohol treatment: taking the clear paste, checking the weight and the name of the clear paste to be correct, putting the clear paste into an alcohol precipitation tank to operate according to the standard operation rule of the alcohol precipitation tank, adding (60 +/-5)% of ethanol which is 1.2 times of the weight of the clear paste while stirring, fully and uniformly mixing, standing, precipitating for 48-72 h at the alcohol precipitation temperature of 0-12 ℃, fully precipitating impurities, opening a liquid outlet of a supernatant liquid to discharge liquid, filtering the supernatant liquid through a plate frame, putting the supernatant liquid into a multifunctional ethanol recovery concentrator to operate according to the standard operation rule of the multifunctional ethanol recovery concentrator, recovering ethanol, concentrating until the liquid medicine has no alcohol smell, sieving by a 100-mesh sieve, putting the supernatant liquid into a clean container, weighing and recording for later use.
(5) Preparation: taking the extract and 120kg of cane sugar according to a production instruction list, cleaning and wrapping the extract and the cane sugar, then entering a clean area, taking 120kg of purified water and 120kg of cane sugar, placing the purified water and the cane sugar in a preparation tank, operating according to standard operating rules of the preparation tank, stirring and heating to completely dissolve the cane sugar, continuously heating and boiling for 15 minutes, then adding the extract and fully stirring; and dissolving 1.6kg of sodium benzoate with a small amount of purified water, filtering, mixing with the above liquid, cooling to room temperature, and adding appropriate amount of purified water to make up to full dose. Considering that the precipitate is filtered out by a filtering process, a proper amount of purified water is actually added to reach 85 ten thousand ml; stirring for 20 minutes to fully stir, adding 20% sodium hydroxide solution (the preparation method comprises the steps of dissolving 650g of sodium hydroxide in 2600g of purified water) to adjust the pH value of the liquid medicine to 4.5-5.5, informing QA to sample, sending to QC for inspection, filtering with a 200-mesh sieve after passing, putting into a clean container for sealed storage, hanging a material label, timely transferring into a refrigeration house, and refrigerating for more than one week in the environment of 0-4 ℃.
(6) Filtering, filling and sealing, and sterilizing:
and (3) placing the tubular easy-to-draw bottle filled with the liquid medicine into a water bath heating tank, opening a steam valve, gradually heating to 100 ℃, and preserving the heat at 100 ℃ for 10-15 minutes. And taking out after the heat preservation time is up.
The advantages are that: the oral liquid treated by the method meets the drug standard by microbial limit inspection, does not precipitate within 2 years, and ensures no loss of effective components and stable drug property.
The technical key points in the process observation of the principal component of the monarch drug, namely the astragalus root and astragaloside in the formula are as follows:
1. control of extraction temperature (90 + -5 deg.C)
2. Control of ethanol concentration in alcohol precipitation (60. + -. 5)%
3. The alcohol precipitation time is 48-72 h
4. The alcohol precipitation temperature is 0-12 DEG C
5. The ratio of the water for extraction (drinking water/purified water) (30-50)%: 70-50)%)
The invention has the advantages that:
1. tonify middle-jiao and qi oral liquid, tonify middle-jiao and qi. Can be used for treating asthenia and viscera prolapse. The invention provides a production process of an oral liquid for tonifying middle-jiao and qi, preferably certain medicinal materials are modified by microorganisms, and the prepared oral liquid has excellent quality and high active ingredients, not only can tonify middle-jiao and qi, but also has the effects of tonifying brain and intelligence and resisting aging.
2. The oral liquid treated by the method meets the drug standard by microbial limit inspection, does not precipitate within 2 years, and ensures no loss of effective components and stable drug property.
Detailed Description
Example 1:
a novel anti-precipitation treatment process of traditional Chinese medicine oral liquid is characterized by comprising the following steps:
(1) processing the medicinal materials:
honey-fried astragalus root: removing impurities from the raw materials, cleaning, moistening, slicing or slicing, drying, sieving to remove ash, mixing with refined honey, parching, taking out, and spreading for cooling;
codonopsis pilosula: taking raw medicine, removing impurities, cleaning, slightly moistening, cutting into thick pieces or sections, and drying;
honey-fried licorice root: removing impurities such as residual stems on the ground, grading, washing with strong water, moistening, slicing into thick pieces, cutting into 6mm segments for small ones, drying, slicing and segmenting in production place, sieving to remove ash and bits, mixing with refined honey, slightly stewing, parching, taking out, and spreading for cooling;
chinese angelica: taking raw medicines, removing impurities, cleaning with water, moistening, softening, slicing, and drying at low temperature;
frying the bighead atractylodes rhizome: removing impurities from the raw materials, grading the size, slightly soaking, cleaning, moistening, slicing, drying, placing bran stir-fried with honey in a hot pot, turning over, adding Atractylodis rhizoma when it is smoking, quickly stir-frying to dark yellow surface, taking out, sieving to remove bran, and spreading to cool; cimicifugae foetidae: collecting raw materials, removing impurities, slightly soaking, cleaning, moistening, slicing, and drying;
bupleurum root: collecting raw materials, removing impurities such as residual stem, cleaning, moistening, slicing into thick pieces or segments, and drying;
dried orange peel: collecting raw materials, removing impurities, cyan, and mould black, cleaning with water, shredding, and drying at low temperature; ginger: collecting raw materials, removing impurities, cleaning, and slicing into thick pieces;
chinese date: collecting raw materials, removing impurities and mould and moth-eaten people, cleaning with water, and drying.
(2) Preparing materials: taking each batch of qualified decoction pieces which are produced by 80 ten thousand ml according to the production instruction, respectively weighing honey-fried astragalus, codonopsis pilosula, honey-fried licorice, angelica, fried bighead atractylodes rhizome, rhizoma cimicifugae, radix bupleuri, dried orange peel, ginger and Chinese date according to the prescription amount, and filling into a dry clean container;
(3) extraction: when the prescription materials are extracted, the name, the quantity and the like of the ten medicines are checked to be correct according to production instructions, half of the ten medicines are weighed respectively and put into 2 3m3 multifunctional extraction tanks respectively, the operations are carried out according to the standard operation rules of the multifunctional extraction tanks, ten times of water is added into the first juice, the first juice is soaked for one hour, the first juice is heated and decocted for 2 hours, the decocted liquid is filtered by a 100-mesh sieve to a liquid medicine storage tank, 10 times of water is added into the second juice, the second juice is heated and decocted for 1 hour, the decocted liquid is filtered by 100 meshes and combined with the first juice decocted liquid, the mixture is kept stand for more than 12 hours, the supernatant is put into a single-effect evaporation concentrator, the concentration is carried out according to the standard operation rules of the single-effect evaporation concentrator, the concentration is carried out until the relative density is 1.08-1.10 (90 +/-5 ℃);
the ratio of the extraction water (drinking water to purified water) (30-50)% to 70-50)%;
(4) alcohol treatment: taking the clear paste, checking the weight and the name of the clear paste to be correct, putting the clear paste into an alcohol precipitation tank to operate according to the standard operation rule of the alcohol precipitation tank, adding (60 +/-5)% of ethanol which is 1.2 times of the weight of the clear paste while stirring, fully and uniformly mixing, standing, precipitating for 48-72 h at the alcohol precipitation temperature of 0-12 ℃, fully precipitating impurities, opening a liquid outlet of a supernatant liquid to discharge liquid, filtering the supernatant liquid through a plate frame, putting the supernatant liquid into a multifunctional ethanol recovery concentrator to operate according to the standard operation rule of the multifunctional ethanol recovery concentrator, recovering ethanol, concentrating until the liquid medicine has no alcohol smell, sieving by a 100-mesh sieve, putting the supernatant liquid into a clean container, weighing and recording for later use.
(5) Preparation: taking the extract and 120kg of cane sugar according to a production instruction list, cleaning and wrapping the extract and the cane sugar, then entering a clean area, taking 120kg of purified water and 120kg of cane sugar, placing the purified water and the cane sugar in a preparation tank, operating according to standard operating rules of the preparation tank, stirring and heating to completely dissolve the cane sugar, continuously heating and boiling for 15 minutes, then adding the extract and fully stirring; and dissolving 1.6kg of sodium benzoate with a small amount of purified water, filtering, mixing with the above liquid, cooling to room temperature, and adding appropriate amount of purified water to make up to full dose. Considering that the precipitate is filtered out by a filtering process, a proper amount of purified water is actually added to reach 85 ten thousand ml; stirring for 20 minutes to fully stir, adding 20% sodium hydroxide solution (the preparation method comprises the steps of dissolving 650g of sodium hydroxide in 2600g of purified water) to adjust the pH value of the liquid medicine to 4.5-5.5, informing QA to sample, sending to QC for inspection, filtering with a 200-mesh sieve after passing, putting into a clean container for sealed storage, hanging a material label, timely transferring into a refrigeration house, and refrigerating for more than one week in the environment of 0-4 ℃.
(6) Filtering, bottling, and sterilizing. And (3) placing the tubular easy-to-draw bottle filled with the liquid medicine into a water bath heating tank, opening a steam valve, gradually heating to 100 ℃, and preserving the heat at 100 ℃ for 10-15 minutes. And taking out after the heat preservation time is up.
The advantages are that: the oral liquid treated by the method meets the drug standard by microbial limit inspection, does not precipitate within 2 years, and ensures no loss of effective components and stable drug property.
Example 2:
a novel anti-precipitation treatment process of traditional Chinese medicine oral liquid is characterized by comprising the following steps:
(1) processing the medicinal materials:
honey-fried astragalus root: removing impurities from the raw materials, cleaning, moistening, slicing or slicing, drying, sieving to remove ash, mixing with refined honey, parching, taking out, and spreading for cooling;
codonopsis pilosula: taking raw medicine, removing impurities, cleaning, slightly moistening, cutting into thick pieces or sections, and drying;
honey-fried licorice root: removing impurities such as residual stems on the ground, grading, washing with strong water, moistening, slicing into thick pieces, cutting into 6mm segments for small ones, drying, slicing and segmenting in production place, sieving to remove ash and bits, mixing with refined honey, slightly stewing, parching, taking out, and spreading for cooling;
chinese angelica: taking raw medicines, removing impurities, cleaning with water, moistening, softening, slicing, and drying at low temperature;
frying the bighead atractylodes rhizome: removing impurities from the raw materials, grading the size, slightly soaking, cleaning, moistening, slicing, drying, placing bran stir-fried with honey in a hot pot, turning over, adding Atractylodis rhizoma when it is smoking, quickly stir-frying to dark yellow surface, taking out, sieving to remove bran, and spreading to cool; cimicifugae foetidae: collecting raw materials, removing impurities, slightly soaking, cleaning, moistening, slicing, and drying;
bupleurum root: collecting raw materials, removing impurities such as residual stem, cleaning, moistening, slicing into thick pieces or segments, and drying;
dried orange peel: collecting raw materials, removing impurities, cyan, and mould black, cleaning with water, shredding, and drying at low temperature; ginger: collecting raw materials, removing impurities, cleaning, and slicing into thick pieces;
chinese date: collecting raw materials, removing impurities and mould and moth-eaten people, cleaning with water, and drying.
Wherein, the astragalus, the angelica and the white atractylodes rhizome are further subjected to microbial fermentation treatment:
(a) the astragalus, the angelica and the white atractylodes rhizome are taken, and purified water which is 0.3 time of the total weight of the three medicinal materials is uniformly sprayed on the medicinal materials in parts.
(b) And (3) microbial fermentation treatment: independently and uniformly spreading the medicinal materials in a sample tray in a single layer manner, uniformly spraying the compound bacterial liquid on the medicinal materials, wherein the spraying amount is 0.05 time of the dry weight, and standing and fermenting for 10-12 hours at 38-40 ℃;
the compound bacterial liquid is lactobacillus rhamnosus: lactobacillus brevis: the lactobacillus adolescentis is prepared from the following components in a ratio of 1: 0.2: 1.5, the total amount of the live bacteria is 1.3X1011cfu/ml；
(c) Freeze-drying: placing the fermented medicinal materials in a freeze dryer, controlling the freezing temperature to be-35 ℃, starting a vacuum pump to vacuumize until the pressure of a drying chamber is 60Pa after the required temperature is reached, adjusting the temperature of the drying chamber to be 50 ℃ for drying, and finally controlling the water content to be below 8%.
(2) Preparing materials: taking each batch of qualified decoction pieces which are produced by 80 ten thousand ml according to the production instruction, respectively weighing honey-fried astragalus, codonopsis pilosula, honey-fried licorice, angelica, fried bighead atractylodes rhizome, rhizoma cimicifugae, radix bupleuri, dried orange peel, ginger and Chinese date according to the prescription amount, and filling into a dry clean container;
(3) extraction: when the prescription materials are extracted, the name, the quantity and the like of the ten medicines are checked to be correct according to production instructions, half of the ten medicines are weighed respectively, the ten medicines are respectively put into 2 3m3 multifunctional extraction tanks, the operations are carried out according to the standard operation rules of the multifunctional extraction tanks, ten times of water is added into the first juice, the first juice is soaked for one hour, the first juice is heated and decocted for 2 hours, the decocted liquid is filtered by a 100-mesh sieve to a liquid medicine storage tank, 10 times of water is added into the second juice, the second juice is heated and decocted for 1 hour, the decocted liquid is filtered by 100 meshes and combined with the first juice decocted liquid, the mixture is kept stand for more than 12 hours, the supernatant is put into a single-effect evaporation concentrator, the concentration is carried out according to the standard operation rules of the single-effect evaporation concentrator, the concentration is carried out until the relative density is 1.08-1.10 (;
the ratio of the extraction water (drinking water to purified water) (30-50)% to 70-50)%;
(4) alcohol treatment: taking the clear paste, checking the weight and the name of the clear paste to be correct, putting the clear paste into an alcohol precipitation tank to operate according to the standard operation rule of the alcohol precipitation tank, adding (60 +/-5)% of ethanol which is 1.2 times of the weight of the clear paste while stirring, fully and uniformly mixing, standing, precipitating for 48-72 h at the alcohol precipitation temperature of 0-12 ℃, fully precipitating impurities, opening a liquid outlet of a supernatant liquid to discharge liquid, filtering the supernatant liquid through a plate frame, putting the supernatant liquid into a multifunctional ethanol recovery concentrator to operate according to the standard operation rule of the multifunctional ethanol recovery concentrator, recovering ethanol, concentrating until the liquid medicine has no alcohol smell, sieving by a 100-mesh sieve, putting the supernatant liquid into a clean container, weighing and recording for later use.
(5) Preparation: taking the extract and 120kg of cane sugar according to a production instruction list, cleaning and wrapping the extract and the cane sugar, then entering a clean area, taking 120kg of purified water and 120kg of cane sugar, placing the purified water and the cane sugar in a preparation tank, operating according to standard operating rules of the preparation tank, stirring and heating to completely dissolve the cane sugar, continuously heating and boiling for 15 minutes, then adding the extract and fully stirring; and dissolving 1.6kg of sodium benzoate with a small amount of purified water, filtering, mixing with the above liquid, cooling to room temperature, and adding appropriate amount of purified water to make up to full dose. Considering that the precipitate is filtered out by a filtering process, a proper amount of purified water is actually added to reach 85 ten thousand ml; stirring for 20 minutes to fully stir, adding 20% sodium hydroxide solution (the preparation method comprises the steps of dissolving 650g of sodium hydroxide in 2600g of purified water) to adjust the pH value of the liquid medicine to 4.5-5.5, informing QA to sample, sending to QC for inspection, filtering with a 200-mesh sieve after passing, putting into a clean container for sealed storage, hanging a material label, timely transferring into a refrigeration house, and refrigerating for more than one week in the environment of 0-4 ℃.
(6) Filtering, bottling, and sterilizing.
And (3) placing the tubular easy-to-draw bottle filled with the liquid medicine into a water bath heating tank, opening a steam valve, gradually heating to 100 ℃, and preserving the heat at 100 ℃ for 10-15 minutes. And taking out after the heat preservation time is up.
The advantages are that: the oral liquid treated by the method meets the drug standard by microbial limit inspection, does not precipitate within 2 years, and ensures no loss of effective components and stable drug property.
Example 3: and (5) comparing the alcohol precipitation time for 18-36 h, and precipitating the alcohol at normal temperature. The alcohol precipitation temperature is 0-12 ℃, the alcohol precipitation time is 48-72 hours, and the precipitation phenomenon does not occur in the trial sample within 2 years.
Example 4:
comparative example 1, otherwise same as example 2, wherein microbial fermentation treatment of astragalus, angelica, atractylodes:
(a) the astragalus, the angelica and the white atractylodes rhizome are taken, and purified water which is 0.3 time of the total weight of the three medicinal materials is uniformly sprayed on the medicinal materials in parts.
(b) And (3) microbial fermentation treatment: independently and uniformly spreading the medicinal materials in a sample tray in a single layer manner, uniformly spraying a bacterial solution on the medicinal materials, wherein the spraying amount is 0.05 time of the dry weight, and standing and fermenting for 10-12 hours at 38-40 ℃;
the bacterial liquid is Lactobacillus brevis, and the total viable bacteria amount is 1.3X1011cfu/ml。
Example 5:
comparative example 1, otherwise same as example 2, wherein microbial fermentation treatment of astragalus, angelica, atractylodes:
(a) the astragalus, the angelica and the white atractylodes rhizome are taken, and purified water which is 0.3 time of the total weight of the three medicinal materials is uniformly sprayed on the medicinal materials in parts.
(b) And (3) microbial fermentation treatment: independently and uniformly spreading the medicinal materials in a sample tray in a single layer manner, uniformly spraying a bacterial solution on the medicinal materials, wherein the spraying amount is 0.05 time of the dry weight, and standing and fermenting for 10-12 hours at 38-40 ℃;
the bacterial liquid is lactobacillus adolescentis, and the total viable bacteria amount is 1.3X1011cfu/ml；
Example 6:
comparative example 1, otherwise same as example 2, wherein microbial fermentation treatment of astragalus, angelica, atractylodes:
(a) the astragalus, the angelica and the white atractylodes rhizome are taken, and purified water which is 0.3 time of the total weight of the three medicinal materials is uniformly sprayed on the medicinal materials in parts.
(b) And (3) microbial fermentation treatment: independently and uniformly spreading the medicinal materials in a sample tray in a single layer manner, uniformly spraying a bacterial solution on the medicinal materials, wherein the spraying amount is 0.05 time of the dry weight, and standing and fermenting for 10-12 hours at 38-40 ℃;
the bacterial liquid is lactobacillus rhamnosus, and the total viable bacteria amount is 1.3X1011cfu/ml；
Example 7:
(1) and (3) evaluating the normal pressure and oxygen deficiency resistance of the mice:
taking 60 male mice, averagely dividing the mice into 6 groups, continuously performing intragastric administration for 7 days, wherein the dosage is 150g/kg (purified water is infused into a control group), grouping the mice into 250ml wide-mouth bottles 30 minutes after the last intragastric administration, sealing, recording the anoxic death time taking the apnea as an index, and calculating the average survival time, wherein no significant difference exists among the groups; the experiment was repeated 5 times to take an average.
TABLE 1
Therefore, the oral liquid has the obvious effect of improving the hypoxia tolerance of mice, and the three microorganisms have the obvious synergistic effect in the co-fermentation.
(2) And (3) evaluating the anti-aging effect:
the D-galactose aging mouse model is a model for evaluating anti-aging drugs commonly used in the field, and has been disclosed, and the evaluation effect is stable. The invention fully refers to the judgment standard of the model when evaluating the anti-aging and intelligence-improving effects.
The model of aging caused by D-galactose is an animal model of senilism and old people, which is provided by many scholars at home and abroad, and all indexes of the model have the characteristics of old mice, and previous experiments also fully prove that the model of aging caused by D-galactose is a stable and reliable model of aging. At present, D-galactose is widely used for duplicating experimental animal models such as brain aging, cataract and the like, and under the condition of long-term mass injection of D-galactose, a large amount of superoxide anion free radicals are generated in vivo, so that peroxidation damage is generated on tissues, the immunologic function is low, the gene expression and regulation are abnormal, the cell growth and reproduction capability is reduced, cells generate degenerative changes, the learning and memory behaviors of animals are obviously damaged, the learning and memory disorder characteristics similar to aging are realized, and the changes of biochemistry and tissue structures are basically consistent with the natural aging performance.
Therefore, the invention selects the D-galactose to replicate the aging mouse model, the model has simple and convenient manufacturing method, obvious aging characteristics and lower experimental cost, is one of the aging models commonly used at home and abroad at present, and has very wide application in researching the aging mechanism and the anti-aging drug action mechanism.
There is increasing evidence that aging and neurodegenerative diseases caused by aging can lead to a decline in learning cognitive ability, particularly spatio-memory ability. At present, many experimental models for studying learning and memory abilities of animals in an overall level are available, and a diving platform experiment is one of the common methods. The mouse is electrified in the training period to obtain memory in the process of being continuously shocked, and the memory retention capacity of the mouse is judged according to the time of the mouse jumping off the platform for the first time and the frequency of jumping off the platform within 5min during testing. The mouse escapes to a safe position immediately after receiving the electric shock, the mouse forms memory after multiple electric shocks within 5min, and the frequency (error frequency) of the electric shock received by the mouse within 5min and the time (escape latency) of the first error making are recorded in a test after 24h to reflect the memory consolidation and reproduction capability of the mouse. Therefore, the invention selects a diving tower experiment and a dark avoidance experiment to observe the influence of the oral liquid for tonifying middle-jiao and Qi on the passive learning and memory ability of the mice aged by the D-galactose.
Research shows that the learning and memory ability is closely related to the cholinergic nervous system, and the phthalein choline (Ach) is an important nerve conduction medium of the system, is catalyzed and synthesized by Choline Acetyltransferase (CAT), acts on an M receptor after being released, is catalyzed and decomposed by phthalein cholinesterase, is transported to a pre-protrusion membrane by a vesicle, is released to a protrusion gap, and acts on a receptor on a post-synaptic membrane to play a biological effect. Research shows that in brain tissue of senile dementia patient, the activity of choline acetylphthalein transferase is obviously reduced, while the activity of acetylphthalein cholinesterase is obviously increased, which results in insufficient content of acetylphthalein choline in brain, which results in dysfunction of central nervous system, and the decline of learning-memory ability, learning memory and cognitive ability, and is the main clinical manifestation of brain aging. Research shows that half of lactose model mice brain tissue B phthalein cholinesterase (AChE) activity is obviously improved, monoamine oxidase activity is obviously reduced, a cholinergic system is closely related to learning and memory, B phthalein choline (Ach) is a neurotransmitter for promoting learning and memory, B phthalein cholinesterase is an enzyme for degrading B phthalein choline, and B phthalein cholinesterase activity is improved, so that cholinergic function is reduced indirectly, and the increase of B phthalein cholinesterase activity of D-galactose-induced aging mice brain tissue indicates that the central cholinergic function is disordered, so that the learning ability of the D-galactose-induced aging mice is reduced. According to the invention, the activity of the phthalein cholinesterase in brain tissues of the D-galactose aging mice is obviously increased, and the spleen-stomach invigorating and qi tonifying oral liquid under a certain dosage is observed to obviously reduce the activity of the phthalein cholinesterase, so that the oral liquid has a certain protection effect on a cholinergic system, and the action mechanism of the oral liquid for enhancing the learning and memory of the D-galactose aging model mice is probably related to the inhibition of cholinesterase activity and the reduction of decomposition of the phthalein cholinesterase.
The passive learning memory ability of the mouse is tested by adopting a diving platform method experiment, the experimental device is a mouse diving platform reaction box with 10cmx10cmx30cm, the test box is composed of 5 small rooms, 5 animals can be simultaneously tested at one time, a copper grid which can be continuously electrically stimulated by 36V is arranged at the bottom of the box, a rubber pad with the diameter and the height of 4.SCm is arranged at the right rear corner of each time, the rubber pad is used as a safety area for the mouse to avoid electric shock, the mouse is firstly placed in a diving platform instrument to adapt to the environment for 3min when the experiment is carried out, the power supply is turned on, the time is set for 5min, the voltage is 36V, the power supply of the test box is turned on to electrify the copper grid at the bottom, the experiment is started by pressing a 'start/stop' key of the diving platform tester, and the diving platform tester starts to record the number of times. After 24h, the mouse is placed in a platform jumping instrument to adapt for 3min, then the mouse is placed on a rubber pad, a copper grid at the bottom is electrified, and at the moment, the tester records the time of the mouse jumping off the platform for the first time, namely the latency period, and the times of jumping off the rubber pad within 5min, namely the error times of the test period, and the times are used as the memory retention performance.
The dark avoiding box is designed by utilizing the habit of darkness of rats, a box body is provided with a light and dark space and is inserted and separated by a manual operation door, when an animal enters the dark space, a floor grid generates current, the mouse escapes to a safe and bright part immediately after being shocked, the mouse forms memory after being shocked for a plurality of times within 5min, and the number of shocked times (error number) of the mouse within 5min and the time (escape latency) of first mistake making during testing for 24h are recorded to reflect the memory consolidation and reproduction capability of the mouse. Before dark avoidance experiment training, the mouse head is placed in a bright room with a hole on the back, the environment is adapted for 3min, then a 36V current is conducted to a copper grid in the dark room, the mouse is shocked once entering the dark room, the correct reaction is that the mouse returns to the bright room, and the copper grid is electrified for 5min, which is the training process. And (3) carrying out memory test on the mice after 24h, recording the time when the mice enter the darkroom for the first time, namely the dark avoidance latency, and recording the times of the mice entering the darkroom within s/min (namely the times of dark avoidance errors), wherein the latency of the mice which do not enter the darkroom within s/min is calculated according to 300 s.
After the diving platform and dark-avoiding experiment is finished, fasting is carried out for 24h, the animals are deeply anesthetized by using ether, the brains of the mice are quickly stripped on a belt, blood stains on the surfaces of the mice are washed by 0.9% ice-cold physiological saline, and the mice are weighed. A10% brain homogenate was prepared with ice-cold physiological saline, centrifuged at low temperature (400Orpm) for 15min, and the supernatant was collected for assay. Measurement of phthalein cholinesterase was performed according to a method conventional in the art.
The D-galactose model mice causing aging are divided into 6 groups, each group comprises 20 mice, 1 to 5 groups are respectively administered with the oral liquid (gavage, the dose is the same as the above embodiment) of the invention in the examples 1 to 5, one dose is taken every day, the control group is administered with the same dose of normal saline, one dose is taken every day, 0.5 g is taken every time, and 15 days is a course of treatment. The experiment was performed after 3 treatment periods. The experiment was repeated 3 times to take an average.
The study and memory ability of each group of mice is detected by a diving platform method, and the result shows that the response time of the mice in the control group is obviously prolonged compared with the treatment group and the error times are obviously increased compared with the blank control group in the study and training on the 1 st day.
TABLE 2 influence of the diving platform experiment on D-galactose-induced aging mice
| Group of | Reaction time(s) | Error frequency/5 min |
| Control group | 9.67 | 5.4 |
| Treatment group 1 | 6.31 | 4.1 |
| Treatment group 2 | 3.82 | 2.3 |
| Treatment group 3 | 6.56 | 4.1 |
| Treatment group 4 | 5.21 | 3.9 |
| Treatment group 5 | 6.32 | 3.8 |
The learning and memory abilities of all groups of mice are detected by a dark method, and the results show that the treatment groups with wrong times are obviously reduced in the learning scores of the mice. The results are shown in Table 2.
TABLE 3 influence of the darkening-avoidance experiment in D-galactose-induced aging mice
| Group of | Learning error times/5 min | Error frequency/5 min |
| Control group | 2.72 | 1.35 |
| Treatment group 1 | 2.52 | 1 |
| Treatment group 2 | 1.09 | 0 |
| Treatment group 3 | 2.51 | 1 |
| Treatment group 4 | 2.37 | 1 |
| Treatment group 5 | 2.55 | 1.3 |
Therefore, the oral liquid can obviously improve the memory and learning ability of mice and resist aging, and the three microorganisms have obvious synergistic effect in joint fermentation.
(3) And (3) biochemical level evaluation:
telomeres are natural ends of chromosomes, have the functions of protecting the integrity and stability of the chromosomes and preventing the chromosomes from being degraded, fused and recombined, so that the integrity of genetic information is ensured, and due to the fact that the end replication problem exists, telomere DNA is progressively shortened along with the increase of cell division times, and after the telomere DNA is shortened to a certain limit, cells lose the division and proliferation capacity and die due to senescence. The function of telomerase is just the extension of telomere DNA and the repair of broken chromosomes, so that the cell senescence can be delayed by regulating the activity of telomerase.
The telomerase activity of the brain tissue of the mouse is tested by a conventional method, and the result shows that: the mean telomerase activity in the control group was 78.53 (OD 450), which was significantly lower than that in normal mice 97.31 (OD 450). The mean telomerase activities of treatment groups 1-5 were 93.61,96.83,97.32, 97.52, 97.22 (OD 450), respectively.
The oral liquid can improve the telomerase activity of the brain tissue of the model mouse with the aging caused by the D-galactose, thereby delaying the aging of brain cells, and the three microorganisms have obvious synergistic effect when being fermented together.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (2)
1. The production method of the oral liquid for enriching, tonifying middle-jiao and replenishing qi is characterized by comprising the following steps:
(1) processing the medicinal materials:
honey-fried astragalus root: removing impurities from the raw materials, cleaning, moistening, slicing or slicing, drying, sieving to remove ash, mixing with refined honey, parching, taking out, and spreading for cooling;
codonopsis pilosula: taking raw medicine, removing impurities, cleaning, slightly moistening, cutting into thick pieces or sections, and drying;
honey-fried licorice root: removing impurities such as residual stems on the ground, grading, washing with strong water, moistening, slicing into thick pieces, cutting into 6mm segments for small ones, drying, slicing and segmenting in production place, sieving to remove ash and bits, mixing with refined honey, slightly stewing, parching, taking out, and spreading for cooling;
chinese angelica: taking raw medicines, removing impurities, cleaning with water, moistening, softening, slicing, and drying at low temperature;
frying the bighead atractylodes rhizome: removing impurities from the raw materials, grading the size, slightly soaking, cleaning, moistening, slicing, drying, placing bran stir-fried with honey in a hot pot, turning over, adding Atractylodis rhizoma when it is smoking, quickly stir-frying to dark yellow surface, taking out, sieving to remove bran, and spreading to cool; cimicifugae foetidae: collecting raw materials, removing impurities, slightly soaking, cleaning, moistening, slicing, and drying;
bupleurum root: collecting raw materials, removing impurities such as residual stem, cleaning, moistening, slicing into thick pieces or segments, and drying;
dried orange peel: collecting raw materials, removing impurities, cyan, and mould black, cleaning with water, shredding, and drying at low temperature; ginger: collecting raw materials, removing impurities, cleaning, and slicing into thick pieces;
chinese date: taking raw medicine, removing impurities and moldy and moth-eaten people, cleaning with water, and drying;
in the step (1), the astragalus, the angelica and the white atractylodes rhizome are further subjected to microbial fermentation treatment:
(a) uniformly spraying purified water which is 0.3 time of the total weight of the three medicinal materials on the medicinal materials by times;
(b) and (3) microbial fermentation treatment: independently and uniformly spreading the medicinal materials in a sample tray in a single layer manner, uniformly spraying the compound bacterial liquid on the medicinal materials, wherein the spraying amount is 0.05 time of the dry weight, and standing and fermenting for 10-12 hours at 38-40 ℃;
the compound bacterial liquid is lactobacillus rhamnosus: lactobacillus brevis: the lactobacillus adolescentis is prepared from the following components in a ratio of 1: 0.2: 1.5, the total amount of the live bacteria is 1.3X1011cfu/ml；
(c) Freeze-drying: placing the fermented medicinal materials in a freeze dryer, controlling the freezing temperature to be-35 ℃, starting a vacuum pump to pump vacuum until the pressure of a drying chamber is 60Pa after the required temperature is reached, adjusting the temperature of the drying oven to be 50 ℃ for drying, and finally controlling the water content to be below 8%;
(2) preparing materials: taking each batch of qualified decoction pieces which are produced by 80 ten thousand ml according to a production instruction, respectively weighing honey-fried astragalus, codonopsis pilosula, honey-fried liquorice, angelica, fried bighead atractylodes rhizome, cimicifuga foetida, radix bupleuri, dried orange peel, ginger and Chinese date, and filling into a dry clean container;
(3) extraction: when the prescription materials are extracted, the name, the quantity and the like of the ten medicines are checked to be correct according to production instructions, half of the ten medicines are weighed respectively and put into 2 3m3 multifunctional extraction tanks respectively, the operations are carried out according to the standard operation rules of the multifunctional extraction tanks, ten times of water is added into the first juice, the first juice is soaked for one hour, the first juice is heated and decocted for 2 hours, the decocted liquid is filtered by a 100-mesh sieve to a liquid medicine storage tank, 10 times of water is added into the second juice, the second juice is heated and decocted for 1 hour, the decocted liquid is filtered by 100 meshes and combined with the first juice decocted liquid, the mixture is kept stand for more than 12 hours, the supernatant is put into a single-effect evaporation concentrator, the concentration is carried out according to the standard operation rules of the single-effect evaporation concentrator, the concentration is carried out until the relative density is 1.08-1.10 (90 +/-5 ℃); the ratio of drinking water to purified water in the extraction water is as follows: (30-50)%: (70-50)%;
(4) alcohol treatment: taking the concentrated clear paste obtained in the step (3), checking the weight and the name of the clear paste to be correct, putting the clear paste into an alcohol precipitation tank to operate according to the standard operation procedure of the alcohol precipitation tank, adding (60 +/-5)% of ethanol which is 1.2 times of the weight of the clear paste, stirring while adding, fully mixing, standing, precipitating for 48-72 h at the alcohol precipitation temperature of 0-12 ℃, fully precipitating impurities, opening a liquid outlet of a supernatant to discharge liquid, filtering the supernatant by a plate frame, putting the supernatant into a multifunctional ethanol recovery concentrator to operate according to the standard operation procedure of the multifunctional ethanol recovery concentrator, recovering ethanol and concentrating until the liquid medicine has no alcohol smell, sieving by a 100-mesh sieve, putting the liquid medicine into a clean container, weighing and recording for later use;
(5) preparation: taking the extract and 120kg of cane sugar according to a production instruction list, cleaning and wrapping the extract and the cane sugar, then entering a clean area, taking 120kg of purified water and 120kg of cane sugar, placing the purified water and the cane sugar in a preparation tank, operating according to standard operating rules of the preparation tank, stirring and heating to completely dissolve the cane sugar, continuously heating and boiling for 15 minutes, then adding the extract and fully stirring; dissolving sodium benzoate 1.6kg with a small amount of purified water, filtering, mixing with the liquid, cooling to room temperature, and adding purified water to full volume; considering that the precipitate is filtered out by a filtering process, a proper amount of purified water is actually added to reach 85 ten thousand ml; then stirring for 20 minutes to fully stir, and adding 20% sodium hydroxide solution; adjusting the pH value of the liquid medicine to 4.5-5.5, informing QA sampling to send QC inspection, filtering with a 200-mesh sieve after passing, placing in a clean container for sealed storage, hanging a material label, timely transferring into a refrigeration house, and refrigerating for more than one week in the environment of 0-4 ℃; the preparation method of the 20% sodium hydroxide solution comprises the following steps: 650g of sodium hydroxide was dissolved in 2600g of purified water;
(6) filtering, filling and sealing, and sterilizing: placing the tube-made easy-open bottle filled with the liquid medicine into a water bath heating tank, opening a steam valve, gradually heating to 100 ℃, and preserving heat for 10-15 minutes at 100 ℃; and taking out after the heat preservation time is up.
2. The method of claim 1, wherein:
the step (2) comprises the following steps: taking each batch of qualified decoction pieces which are produced by 80 ten thousand ml according to the production instruction, respectively weighing honey-fried astragalus, codonopsis pilosula, honey-fried licorice, angelica, fried bighead atractylodes rhizome, rhizoma cimicifugae, radix bupleuri, dried orange peel, ginger and Chinese date according to the prescription amount, and filling into a dry clean container.
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