CN111374991A - Oral liquid for treating brain and heart diseases and its preparation method - Google Patents
Oral liquid for treating brain and heart diseases and its preparation method Download PDFInfo
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Abstract
The invention discloses a Naoxinshu oral liquid and a preparation method thereof, wherein the preparation method comprises the following steps: s1, preparing an Armillaria mellea concentrated solution: s11, inoculating the armillaria mellea strain into the fermentation culture solution, and performing fermentation culture to obtain fermentation liquor; s12, inactivating the fermentation liquor obtained in the step S11, filtering and concentrating to obtain Armillaria mellea concentrated solution; s2, heating and refining honey, filtering, cooling, mixing with the ground royal jelly uniformly, sieving, adding the Armillaria mellea concentrated solution, the sodium benzoate aqueous solution and the vanilla essence ethanol solution prepared in the step S1 in sequence, mixing uniformly, adding purified water to a constant volume, stirring uniformly, and filtering to obtain the honey royal jelly; meanwhile, the oral liquid for soothing the brain and heart prepared by the method is protected. The invention has short fermentation period, good thallus growth, high polysaccharide content and stable quality, improves the drug effect and the quality stability of the product, can meet the clinical requirements, and simultaneously the prepared product has better effect of reducing blood sugar.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to a traditional Chinese medicine preparation Naoxinshu oral liquid with the functions of nourishing and strengthening, tranquilizing and allaying excitement and improving cardiovascular and cerebrovascular diseases and a preparation method thereof.
Background
The Naoxinshu oral liquid is a Chinese patent medicine preparation which is on the market and has the functions of nourishing, strengthening, tranquilizing and allaying excitement, and is used for people with weak bodies, uneasiness, insomnia, dreaminess, neurasthenia, headache and dizziness which belong to the symptoms. At present, the production enterprises have 110 families, and 110 approval document numbers are involved. The 1000ml prescription formula is as follows:
the preparation method comprises the following steps:
(1) the process of the armillaria concentrated solution comprises the following steps:
placing 60% culture medium in fermentation tank, heating and sterilizing for 30 min, inoculating three-stage seeds in seeding tank by pressure difference method, culturing at 26-28 deg.C for 4-6 days, transferring into fermentation tank, culturing at 26-28 deg.C for 6-7 days until fermentation liquid turns into purple brown, and taking out after thickening and thinning, partial hypha autolysis, hypha granule breaking, pH value lowering to 5.20-5.60, and brightness weakening. Continuously concentrating the fermented Armillaria mellea fermentation culture solution to relative density of 1.030-1.035(20 deg.C) to obtain Armillaria mellea concentrated solution.
(2)60L of seeding tank culture medium: 200g of edible wheat bran, 200g of edible soybean meal, 100g of starch, 100g of sucrose, 100g of peptone, 100g of high fructose corn syrup, 100ml of soybean oil, 20g of magnesium sulfate and 20g of monopotassium phosphate. Soaking edible testa Tritici and soybean meal in 5 times of water for 30-60 min, boiling for 1 hr, filtering, and cooling. Adding other raw materials into the edible testa Tritici and soybean cake extractive solution. Others are scaled up.
Controlling fermentation culture conditions by introducing compressed air at a ventilation rate of 1:0.3-1:0.5v/v.min, culturing at 26-28 deg.C for 4-6 days, and checking the pressure in the tank (0.4-0.5 × 10)5pa), temperature (26-28 ℃), pH value (60L pH value is 6.20-6.60, 200L pH value is 5.80-6.20, 2000L pH value is 5.20-5.60).
(3) The patent refers to the field of 'pharmaceutical preparations':
heating Mel 450g, refining (90-100 deg.C), filtering, cooling, mixing with ground Lac Regis Apis, adding Armillariella Mellea concentrate, sodium benzoate 3g and ethanol solution of essence, mixing, adjusting total amount to 1000ml, stirring, and filtering.
CN107213172A discloses a preparation method of a Armillaria mellea oral liquid, wherein the Armillaria mellea concentrated liquid is prepared by a three-stage fermentation method, polypeptide impurities can be reduced, and the content of an active ingredient N6- (5-hydroxy-2-pyridylmethylamino) -9- β -purine nucleoside is increased, so that the problems are solved, but the problems of long fermentation period, low fermentation liquid yield, low content of active ingredients such as polysaccharide and the like still exist, so that the Armillaria mellea oral liquid has poor drug effect and unstable quality, and the clinical requirements are difficult to meet.
Disclosure of Invention
In order to solve the technical problems, the invention provides the Naoxinshu oral liquid and the preparation method thereof, which shorten the fermentation period, have good thallus growth, high polysaccharide content and stable quality by adjusting the fermentation process of the Armillaria mellea, improve the drug effect and the quality stability of the product, can meet the clinical requirements, and are suitable for large-scale popularization and application.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of Naoxinshu oral liquid comprises the following steps:
s1, preparing an Armillaria mellea concentrated solution:
s11, inoculating the armillaria mellea strain into the fermentation culture solution, and performing fermentation culture to obtain fermentation liquor;
s12, inactivating the fermentation liquor obtained in the step S11, filtering and concentrating to obtain Armillaria mellea concentrated solution;
s2, heating and refining honey, filtering, cooling, mixing with the ground royal jelly uniformly, sieving, adding the Armillaria mellea concentrated solution, the sodium benzoate aqueous solution and the vanilla essence ethanol solution prepared in the step S1 in sequence, mixing uniformly, adding purified water to a constant volume, stirring uniformly, and filtering to obtain the honey beverage.
Preferably, the Armillaria mellea strain in the step S11 is purchased from China forestry microorganism culture Collection center, resource number BNCC 121102.
Preferably, the fermentation broth in step S11 includes: high fructose corn syrup 18-22g/L, sucrose 18-22g/L, vitamin B10.008-0.012g/L, magnesium sulfate 0.6-0.8g/L, potassium dihydrogen phosphate 1.2-1.6g/L, peptone 0.8-1.2g/L, silkworm chrysalis meal 6-10g/L and soybean oil 0.8-1.2 ml/L.
Further preferably, the mass volume ratio of the vitamin B1, the silkworm chrysalis powder and the soybean oil is 1:800: 100.
Further preferably, the fermentation culture solution also comprises 4-9g/L of mannitol and 0.01-0.05g/L of arginine.
Preferably, the fermentation culture in step S11 is carried out at 26-28 deg.C under 0.05-0.08 × 105Pa for 3-5 days.
Preferably, the inactivation in step S12 is: after fermentation, heating the tank to 95-100 deg.C and 0Pa, maintaining for 120min, and cooling to 60 deg.C.
Preferably, the filtering in step S12 is: and (3) coarsely filtering the inactivated fermentation liquor by using 120-mesh filter cloth, and finely filtering by using a plate frame machine to obtain filtrate.
Preferably, the concentration in step S12 is: concentrating the filtrate to relative density of 1.030-1.035(20 deg.C) to obtain Armillaria mellea concentrated solution.
Further preferably, the concentration is carried out under the vacuum degree of-0.09 to-0.02 MPa, the temperature of 50 to 90 ℃ and the steam pressure of less than or equal to 0.15 MPa.
Preferably, the heating refining in step S2 is: heating at 90-100 deg.C for 2-5 min; the filtration is to pass through a 100-mesh sieve; the cooling is to be cooled to below 60 ℃.
Preferably, the sieving in step S2 is 100 mesh sieving.
The invention also provides the oral liquid for treating the brain and heart diseases prepared by the preparation method.
The invention has the beneficial effects that:
(1) the invention relates to a method for preparing a medicinal composition, which is prepared from armillaria mellea concentrate, sucrose, vitamin B1, magnesium sulfate, potassium dihydrogen phosphate, peptone, silkworm chrysalis meal and soybean oil through fermentation, wherein the armillaria mellea concentrate is the main raw material of Naoxinshu oral liquid.
(2) Meanwhile, the mannitol and arginine added into the fermentation culture solution can promote the growth of hyphae and improve the yield of polysaccharide, and the prepared product has a good effect of reducing blood sugar.
Detailed Description
The embodiments of the present invention are described below in conjunction with specific embodiments, and before the embodiments of the present invention are further described, it is to be understood that the scope of the present invention is not limited to the specific embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
The source of the raw materials adopted by the invention is not limited, and the raw materials adopted by the invention are all common commercial products in the technical field if no special description is provided, wherein the Armillaria mellea strain is purchased from China forestry microorganism culture collection management center with resource number BNCC 121102.
Basic embodiment
A preparation method of Naoxinshu oral liquid comprises the following steps:
s1, preparing an Armillaria mellea concentrated solution:
s11, inoculating the Armillaria mellea strain into the fermentation culture solution, and maintaining the temperature at 26-28 deg.C and the tank pressure at 0.05-0.08 × 105Pa, fermenting and culturing for 3-5 days to obtain fermentation liquor;
the fermentation culture solution comprises: high fructose corn syrup 18-22g/L, sucrose 18-22g/L, vitamin B10.008-0.012g/L, magnesium sulfate 0.6-0.8g/L, potassium dihydrogen phosphate 1.2-1.6g/L, peptone 0.8-1.2g/L, silkworm pupa powder 6-10g/L and soybean oil 0.8-1.2 ml/L;
preferably, the fermentation culture solution also comprises 4-9g/L of mannitol and 0.01-0.05g/L of arginine;
s12, inactivating the fermentation liquor obtained in the step S11, filtering and concentrating to obtain Armillaria mellea concentrated solution;
the inactivation is as follows: after fermentation is finished, heating the interior of the tank to 95-100 ℃, keeping the pressure of the tank at 0Pa for 120min, and cooling to 60 ℃ after fermentation is finished;
the filtration is as follows: coarsely filtering the inactivated fermentation liquor by using 120-mesh filter cloth, and finely filtering by using a plate frame machine to obtain filtrate;
the concentration is as follows: concentrating the filtrate at a vacuum degree of-0.09 MPa to-0.02 MPa, a temperature of 50-90 ℃ and a steam pressure of less than or equal to 0.15MPa until the relative density is 1.030-1.035(20 ℃), thus obtaining an armillaria mellea concentrated solution;
s2, heating and refining honey at 90-100 ℃ for 2-5min, filtering through a 100-mesh sieve, cooling to below 60 ℃, uniformly mixing with ground royal jelly, sieving through the 100-mesh sieve, sequentially adding the Armillariella mellea concentrated solution, the sodium benzoate aqueous solution and the vanilla essence ethanol solution prepared in the step S1, uniformly mixing, adding purified water to a constant volume, uniformly stirring and filtering to obtain the honey beverage.
The components and amounts of the fermentation media of examples 1-7 are shown in Table 1, where "-" indicates absence.
TABLE 1 fermentation media Components and amounts used in examples 1-7
The preparation method of the oral liquid for relieving the brain and heart comprises the following steps:
s1, preparing an Armillaria mellea concentrated solution:
s11, inoculating the Armillaria mellea strain into the fermentation culture solution, and performing tank pressure of 0.07 × 10 at 27 deg.C5Fermenting and culturing for 3 days under the conditions of Pa and pH5.3-5.5 to obtain fermentation liquor;
s12, inactivating the fermentation liquor obtained in the step S11, filtering and concentrating to obtain Armillaria mellea concentrated solution;
the inactivation is as follows: after fermentation is finished, heating the interior of the tank to 100 ℃, keeping the pressure of the tank at 0Pa for 120min, and cooling to 60 ℃ after fermentation is finished;
the filtration is as follows: coarsely filtering the inactivated fermentation liquor by using 120-mesh filter cloth, and finely filtering by using a plate frame machine to obtain filtrate;
the concentration is as follows: concentrating the filtrate at vacuum degree of-0.05 MPa, temperature of 60 deg.C, and steam pressure of no more than 0.15MPa to relative density of 1.030-1.035(20 deg.C) to obtain Armillaria mellea concentrated solution;
s2, heating and refining honey at 95 ℃ for 3min, filtering through a 100-mesh sieve, cooling to below 60 ℃, uniformly mixing with the ground royal jelly, sieving through the 100-mesh sieve, sequentially adding the Armillariella mellea concentrated solution, the sodium benzoate aqueous solution and the vanilla essence ethanol solution prepared in the step S1, uniformly mixing, adding purified water to a constant volume, uniformly stirring, and filtering to obtain the honey beverage.
Comparative example 1
This comparative example differs from example 3 in that: the vitamin B1 is 0.015g/L, the silkworm chrysalis powder is 5.5g/L, and the soybean oil is 0.7 ml/L.
Comparative example 2
This comparative example differs from example 3 in that: the vitamin B1 is 0g/L, the silkworm chrysalis powder is 8g/L and the soybean oil is 1.0 ml/L.
Comparative example 3
This comparative example differs from example 7 in that arginine is not present.
Comparative example 4
This comparative example differs from example 7 in that histidine is used instead of arginine.
Experimental example 1
1. Extraction and determination of polysaccharide in fermentation liquor
Centrifuging a certain volume of fermentation liquor at 3000r/min for 10min, taking 20mL of supernatant, concentrating to 10mL by blowing at 50 ℃, adding 3 times of volume of ethanol for precipitation for 12h, centrifuging at 3000r/min for 15min, washing the precipitate with 75% ethanol until no reducing sugar reaction occurs, centrifuging, fully dissolving the precipitate with 60 ℃ hot water, centrifuging to remove the precipitate, and putting the supernatant into a 100mL volumetric flask. And measuring the content of the polysaccharide by a phenol-sulfuric acid method.
2. Measurement of Dry weight of hyphae in fermentation broth
Filtering the fermented product with two layers of gauze, and air drying the mycelia at 45 deg.C to constant weight, and converting into dry weight of mycelia in each liter of fermentation liquid.
The results are shown in Table 2.
TABLE 2
| Dry weight of hyphae g/L | Polysaccharide content mg/mL | |
| Example 1 | 25.0 | 0.558 |
| Example 2 | 24.5 | 0.587 |
| Example 3 | 26.2 | 0.616 |
| Example 4 | 25.4 | 0.601 |
| Example 5 | 29.4 | 0.659 |
| Example 6 | 28.3 | 0.684 |
| Example 7 | 30.4 | 0.727 |
| Comparative example 1 | 22.3 | 0.493 |
| Comparative example 2 | 21.2 | 0.511 |
| Comparative example 3 | 27.5 | 0.628 |
| Comparative example 4 | 27.8 | 0.622 |
From the above table, the fermentation period of the Armillaria mellea fermentation method is short, the thallus grows well, the polysaccharide content in the fermentation liquid is high, and when the mass-volume ratio of the vitamin B1, the silkworm chrysalis powder and the soybean oil in the fermentation medium is 1:800:100, the effect is better; meanwhile, experiments show that when mannitol and arginine are added into the culture solution at the same time, the growth of hyphae can be promoted, and the yield of polysaccharide is improved.
Experimental example II evaluation of blood sugar lowering function
db/db diabetic mice: 5 weeks old, male, purchased at the university of Nanjing model animal institute. Measuring body weight and random blood sugar, and removing mice with abnormal body weight and abnormal blood sugar, wherein the mice are randomly distributed into 12 groups, and each group comprises 8 mice; the Naoxinshu oral liquids prepared in examples 1-7 and comparative examples 1-4 were administered to db/db diabetic mice by gavage administration at a dose of 1.08g/kg/d, once a day for 4 weeks, and the model control group was administered with the same dose of physiological saline. Fasting blood glucose was monitored weekly, i.e., one night before blood glucose was measured, mice were fasted and tail vein blood was taken the next day for testing. When measuring blood sugar, the mouse was treated gently, and the mouse was not stressed as much as possible, so as not to affect the accuracy, and when repeatedly measuring blood sugar of the same mouse, the blood stain before wiping off with an alcohol sheet was required, and the results are shown in table 3.
TABLE 3 Effect of Naoxinshu oral liquid on blood glucose in mice
| Mg/dl for 0 week | Mg/dl for 1 week | Mg/dl for 2 weeks | Mg/dl for 3 weeks | Mg/dl for 4 weeks | |
| Example 1 | 303 | 251 | 246 | 248 | 254 |
| Example 2 | 302 | 247 | 240 | 243 | 249 |
| Example 3 | 304 | 230 | 218 | 220 | 228 |
| Example 4 | 300 | 235 | 227 | 228 | 230 |
| Example 5 | 302 | 228 | 210 | 208 | 206 |
| Example 6 | 304 | 230 | 211 | 210 | 207 |
| Example 7 | 302 | 218 | 194 | 188 | 178 |
| Comparative example 1 | 299 | 262 | 246 | 253 | 276 |
| Comparative example 2 | 301 | 259 | 246 | 256 | 280 |
| Comparative example 3 | 300 | 223 | 209 | 213 | 218 |
| Comparative example 4 | 298 | 225 | 208 | 210 | 215 |
| Model control group | 301 | 310 | 405 | 448 | 504 |
As can be seen from the above table, the blood glucose level of the mouse in the model control group gradually increases with the increase of time, but the blood glucose content of the oral liquid for treating cardio-cerebral vascular disease prepared in the embodiments 1 to 7 of the present invention is significantly reduced after 4 weeks of administration, and compared with the model control group, the oral liquid for treating cardio-cerebral vascular disease shows a hypoglycemic activity (P <0.01), which indicates that the oral liquid for treating cardio-cerebral vascular disease prepared in the present invention has a significant hypoglycemic effect, and it is found that the addition of mannitol and arginine to the fermentation culture solution can significantly improve the hypoglycemic effect of the oral liquid for treating cardio-cerebral vascular disease.
The present invention has been further described with reference to specific embodiments, which are only exemplary and do not limit the scope of the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.
Claims (10)
1. The preparation method of the oral liquid for soothing the brain and heart is characterized by comprising the following steps of:
s1, preparing an Armillaria mellea concentrated solution:
s11, inoculating the armillaria mellea strain into the fermentation culture solution, and performing fermentation culture to obtain fermentation liquor;
s12, inactivating the fermentation liquor obtained in the step S11, filtering and concentrating to obtain Armillaria mellea concentrated solution;
s2, heating and refining honey, filtering, cooling, mixing with the ground royal jelly uniformly, sieving, adding the Armillaria mellea concentrated solution, the sodium benzoate aqueous solution and the vanilla essence ethanol solution prepared in the step S1 in sequence, mixing uniformly, adding purified water to a constant volume, stirring uniformly, and filtering to obtain the honey royal jelly;
the fermentation culture solution in the step S11 comprises: high fructose corn syrup 18-22g/L, sucrose 18-22g/L, vitamin B10.008-0.012g/L, magnesium sulfate 0.6-0.8g/L, potassium dihydrogen phosphate 1.2-1.6g/L, peptone 0.8-1.2g/L, silkworm chrysalis meal 6-10g/L and soybean oil 0.8-1.2 ml/L.
2. The preparation method of claim 1, wherein the mass-to-volume ratio of the vitamin B1, the silkworm chrysalis powder and the soybean oil in the step S11 is 1:800: 100.
3. The method according to claim 2, wherein the fermentation broth in step S11 further comprises 4-9g/L mannitol and 0.01-0.05g/L arginine.
4. The process according to claim 1, wherein the fermentation in step S11 is carried out at a temperature of 26-28 ℃ under a pot pressure of 0.05-0.08 × 105Pa, culturing for 3-5 days.
5. The method according to claim 1, wherein the inactivation in step S12 is: after fermentation, heating the tank to 95-100 deg.C and 0Pa, maintaining for 120min, and cooling to 60 deg.C.
6. The method according to claim 1, wherein the filtering in step S12 is: and (3) coarsely filtering the inactivated fermentation liquor by using 120-mesh filter cloth, and finely filtering by using a plate frame machine to obtain filtrate.
7. The method according to claim 1, wherein the concentration in step S12 is: concentrating the filtrate to relative density of 1.030-1.035 to obtain Armillaria mellea concentrated solution.
8. The method of claim 7, wherein the concentration conditions are: the vacuum degree is-0.09 to-0.02 MPa, the temperature is 50 to 90 ℃, and the steam pressure is less than or equal to 0.15 MPa.
9. The method according to claim 1, wherein the heating refining in step S2 is: heating at 90-100 deg.C for 2-5 min; the filtration is to pass through a 100-mesh sieve; the cooling is to be cooled to below 60 ℃.
10. An oral liquid for treating cerebral and heart diseases, prepared by the preparation method according to any one of claims 1 to 8.
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