CN116687999A

CN116687999A - Preparation method of selfheal oral liquid

Info

Publication number: CN116687999A
Application number: CN202310732445.8A
Authority: CN
Inventors: 刘均正; 谭闽怀; 童猛辉; 廖园红
Original assignee: Jiangxi Heying Pharmaceuticals Co ltd
Current assignee: Jiangxi Heying Pharmaceuticals Co ltd
Priority date: 2023-06-20
Filing date: 2023-06-20
Publication date: 2023-09-05

Abstract

The invention discloses a preparation method of selfheal oral liquid. Carrying out supercritical extraction on the effective components in the selfheal by using the compounded entrainer; the drug effect of the selfheal is further improved through combined fermentation. The first aim of the invention is to promote the extraction of the effective components of the selfheal, thereby reducing the waste. The second purpose of the invention is to provide a preparation method of the selfheal oral liquid with better curative effect.

Description

Preparation method of selfheal oral liquid

Technical Field

The invention relates to the field of preparations, in particular to a preparation method of selfheal oral liquid.

Background

Spica Prunellae is a common medicinal plant belonging to Labiatae, prunella, under the academic name Prunella vulgaris L, also called Spica Prunellae, lythrum, etc. It is grown on road side, field, hillside and other places and distributed in most areas of China. Prunella vulgaris is annual or biennial herbaceous plant, and its stem is upright or inclined, and its height is generally 15-60 cm. The leaves are contra-living, usually in the shape of oval or oval lanceolate needles, with fine serrations on the edges. The inflorescences are purple or blue-purple cone inflorescences, and the flowers are small and dense. The whole plant has rich grass fragrance. The main chemical components of the selfheal include flavonoids, tannins, volatile oil, organic acid and the like. Wherein the flavonoids are one of the important effective components of the selfheal, and have various pharmacological effects. Prunella vulgaris also contains various amino acids and minerals such as glutamic acid, serine, calcium, iron, zinc, etc.

Prunella vulgaris is regarded as a precious traditional Chinese medicine for treating various diseases in traditional Chinese medicine, and the root, stem, leaf, flower and other parts of the Prunella vulgaris can be used as medicines. Modern medical research shows that selfheal has broad-spectrum bioactivity and can be used for treating cold, hepatitis, hypertension, diabetes and other diseases.

The common selfheal fruit-spike oral liquid is taken as an oral liquid preparation convenient to drink, and the retention of the effective components is always a problem worthy of research. In addition, how to improve the curative effect of the selfheal oral liquid preparation is a hot spot of current research.

Disclosure of Invention

The invention aims to solve the technical problems that:

the first aim of the invention is to promote the extraction of the effective components of the selfheal, thereby reducing the waste.

The second purpose of the invention is to provide a preparation method of the selfheal oral liquid with better curative effect.

In order to solve the technical problems, the technical scheme adopted by the invention comprises the following steps:

the method comprises the following specific steps of:

1) Pretreatment:

cleaning Prunellae Spica, oven drying at 50-60deg.C to constant weight, pulverizing, and sieving with 60 mesh sieve to obtain Prunellae Spica powder;

2) Supercritical extraction:

extracting the selfheal powder in a supercritical extraction device, adding entrainer into an entrainer tank, setting the extraction pressure to be 16-18Mpa, the extraction temperature to be 28-30 ℃, the CO2 flow to be 20-25L/h, and the extraction time to be 1-1.5h, and collecting the rest materials and the mixed extract respectively after the extraction is completed;

3) Separating:

placing the above mixed extract in a reduced pressure distillation device, setting pressure at 15mmHg and temperature at 70deg.C, and separating effective substances from Prunellae Spica to obtain pure extractive solution;

wherein, entrainer is: adding 1.2-1.6 parts of N-long-chain acyl alkylene betaine and 1 part of trityl fluorosilane into 4-6 parts of 70% ethanol solution, stirring and dispersing at 400r/min for 10 min.

The method comprises the following specific steps of fermenting the extracted residual materials:

a. washing the residual materials collected after extraction with deionized water for 4-6 times, draining off water, and drying at 50-60 ℃ until the weight is constant to obtain a dry material;

b. respectively taking bare stems Jin Yaozi and madder which have the same weight as the dried materials, uniformly mixing the bare stems Jin Yaozi and the madder with the dried materials, crushing the mixture, and sieving the crushed mixture with a 60-mesh sieve to obtain a fermented material;

c. adding the fermentation materials into a fermentation tank, adding a compound bacterial liquid accounting for 1-3% of the mass of the fermentation materials, adding corn flour accounting for 5-6% of the mass of the fermentation materials as a carbon source, finally adding deionized water with the mass-volume ratio of 2:1 to the fermentation materials, stirring uniformly, starting fermentation, fermenting for 3-5 days, and filtering and sterilizing to obtain a fermentation filtrate;

the preparation method of the compound bacterial liquid comprises the following steps: adding 2 parts of bacillus subtilis, 1 part of bacillus licheniformis, 1 part of lactobacillus and 1 part of saccharomycete into 5 parts of 10% brown sugar water, uniformly stirring, and standing for 12 hours at the temperature of 32-36 ℃ to obtain the compound bacterial liquid.

Wherein the pure extract obtained by extraction and the fermented filtrate obtained by fermentation are mixed to prepare the oral liquid preparation acceptable by medicine.

The invention is characterized in that:

1) Supercritical extraction is an effective low-temperature extraction means and relies on specific entrainers to enhance extraction efficiency. The principle of the entrainer is that the entrainer can influence the solubility and selectivity of solute in supercritical fluid, in order to improve the content of flavonoids and phenolic substances in the extract, N-long-chain acyl alkylene betaine and trityl fluorosilane are compounded as the entrainer, in the extraction process of the effective components of the traditional Chinese medicine, alkaloid can be used as an extraction solvent, mainly for the purpose that the alkaloid is purely natural and relatively safe to extract, but the conventional alkaloid has good extraction effect on flavonoids and is not obvious in extraction effect on phenols and triterpenes, the N-long-chain acyl alkylene betaine is selected, alkylene on N-long-chain acyl alkylene betaine molecules can enhance the wettability of molecules, so that the N-long-chain acyl betaine can better disperse, the polarity of the molecules can be enhanced, and the long-chain betaine has better wettability and emulsifying property compared with the short-chain betaine, so that the extraction effect on various effective components in the selfheal can be enhanced; the trityl fluorosilane is used as a surfactant, is more stable than the conventional surfactant, can regulate the polarity of carbon dioxide, is beneficial to the formation of supercritical fluid microemulsion, is beneficial to the increase of the permeability of cell membranes, is beneficial to the release of flavonoids, phenols and triterpenes, contains three benzyl groups in the trityl fluorosilane molecule, can play a role in protecting other functional groups, and can prevent other functional groups from participating in unwanted reactions, and the N-long-chain acyl alkylene betaine and the trityl fluorosilane are used together as entrainers to have a synergistic effect.

2) The invention uses the bare stem Jin Yaozi and the madder to carry out combined fermentation, and the fermented raw materials can promote the repair of abnormal activation of the pi3k/akt channel by the selfheal oral liquid, so that the proliferation and the apoptosis of cells are balanced, and the efficacy of resolving hard mass and detumescence is improved. In addition, the naked stem golden kidney has various pharmacological effects of resisting oxidation, reducing blood pressure, resisting tumor, regulating immunity and the like, and has certain auxiliary effects on preventing and treating cardiovascular and cerebrovascular diseases, hepatitis, tumor and other diseases. The bare stem Jin Yaozi and the selfheal can be fermented in a combined way, so that the bitter taste of the selfheal can be improved, the selfheal can be absorbed more easily, and the effects of clearing heat and detoxicating, promoting urination and detumescence, reducing blood pressure and the like are enhanced. Wu Qian as a medicinal material for clearing heat and detoxicating has synergistic effect on Prunellae Spica, and the fermentation of radix Rubiae increases vitamins and amino acids in the fermentation liquid, so that the taste of the medicinal liquid is soft.

Description of the embodiments

The present invention will be described in further detail with reference to examples.

The N-long chain acyl alkylene betaine used in the embodiment of the invention is purchased by Shanghai Chuxing chemical industry Co., ltd, the selfheal is a Jiangxi local medicinal material, and the rest raw materials or chemical reagents are obtained through conventional commercial paths if no special description exists.

Example 1

The embodiment comprises the following steps:

1) Preparing an entrainer:

adding 1.4kg of N-long-chain acyl alkylene betaine and 1kg of trityl fluorosilane into 5L of 70% ethanol solution, stirring and dispersing at the speed of 400r/min for 10min to obtain the catalyst;

2) Extraction:

cleaning 800g of selfheal, drying at 55 ℃ to constant weight, crushing and sieving with a 60-mesh sieve to obtain selfheal powder; extracting the Prunella vulgaris powder in supercritical extraction device, adding 400ml entrainer into entrainer tank, setting extraction pressure to 17Mpa, and extracting at 29 deg.C, and CO ₂ The flow is 23L/h, the extraction time is 1.3h, and the residual materials and the mixed extract are respectively collected after the extraction is completed; placing the above mixed extract in a reduced pressure distillation device, setting pressure at 15mmHg and temperature at 70deg.C, and separating effective substances from Prunellae Spica to obtain pure extractive solution;

example 2

The embodiment comprises the following steps:

1) Preparing an entrainer:

adding 1.2kg of N-long-chain acyl alkylene betaine and 1kg of trityl fluorosilane into 4L of 70% ethanol solution, stirring and dispersing at the speed of 400r/min for 10min to obtain the catalyst;

3) Extraction:

cleaning 800g of selfheal, drying at 50 ℃ to constant weight, crushing and sieving with a 60-mesh sieve to obtain selfheal powder; extracting the Prunella vulgaris powder in supercritical extraction device, adding 300ml entrainer into entrainer tank, setting extraction pressure to 16Mpa, and extracting at 28deg.C, CO ₂ The flow is 20L/h, the extraction time is 1h, and the residual materials and the mixed extract are respectively collected after the extraction is completed; placing the above mixed extract in a reduced pressure distillation device, setting pressure at 15mmHg and temperature at 70deg.C, and separating effective substances from Prunellae Spica to obtain pure extractive solution;

example 3

The embodiment comprises the following steps:

1) Preparing an entrainer:

adding 1.6kg of N-long-chain acyl alkylene betaine and 1kg of trityl fluorosilane into 6L of 70% ethanol solution, stirring and dispersing at the speed of 400r/min for 10min to obtain the catalyst;

3) Extraction:

cleaning 800g of selfheal, drying at 60 ℃ to constant weight, crushing and sieving with a 60-mesh sieve to obtain selfheal powder; extracting the Prunella vulgaris powder in supercritical extraction device, adding 400ml entrainer into entrainer tank, setting extraction pressure at 18Mpa, and extracting at 30deg.C, and CO ₂ The flow is 25L/h, the extraction time is 1.5h, and the residual materials and the mixed extract are respectively collected after the extraction is completed; placing the above mixed extract in a reduced pressure distillation device, setting pressure at 15mmHg and temperature at 70deg.C, and separating effective substances from Prunellae Spica to obtain pure extractive solution;

comparative example 1

The difference between this comparative example and example 1 is that:

1) Preparing an entrainer:

adding 1.4kg of N-long-chain acyl alkylene betaine into 5L of 70% ethanol solution, stirring and dispersing at 400r/min for 10min to obtain the final product;

the procedure is as in example 1.

Comparative example 2

The difference between this comparative example and example 1 is that:

1) Preparing an entrainer:

adding 1kg of trityl fluorosilane into 5L of 70% ethanol solution, stirring and dispersing at the speed of 400r/min for 10min to obtain the preparation;

the procedure is as in example 1.

Comparative example 3

The difference between this comparative example and example 1 is that:

1) Preparing an entrainer:

adding 1kg of N-long-chain acyl alkylene betaine and 1.4kg of trityl fluorosilane into 5L of 70% ethanol solution, stirring and dispersing at the speed of 400r/min for 10min to obtain the preparation;

the procedure is as in example 1.

Experiment 1:

1) Total flavone content: spectrophotometry is adopted to determine the ratio of total flavonoids in each group of samples to the raw materials, and the ratio is shown in table 1;

2) Total phenol content: the ratio of total phenols in each group of samples to the raw materials is determined by adopting a Fu Lin Fen method, and is shown in table 1;

3) Total triterpene content: determining the ratio of total triterpene in each group of samples to the crude drug by high performance liquid chromatography, see table 1;

TABLE 1

Project	Total flavone, percent	Total phenols, percent	Total triterpene, percent
				Example 1	6.31	3.89	4.12
Example 2	6.46	3.86	4.18
				Example 3	6.25	3.84	4.17
Comparative example 1	5.97	2.06	2.15
				Comparative example 2	3.83	1.95	1.89
Comparative example 3	5.92	3.76	4.03

Example 4

1) Preparing a compound bacterial liquid:

adding 200g of bacillus subtilis, 100g of bacillus licheniformis, 100g of lactic acid bacteria and 100g of saccharomycetes into 5L of 10% brown sugar water, uniformly stirring, and standing for 12 hours at the temperature of 32-36 ℃ to obtain compound bacteria liquid;

2) Fermentation:

extracting by adopting the same method as in example 1, collecting pure extracting solution and residual materials, washing the residual materials with deionized water for 5 times, draining off water, and drying at 55 ℃ to constant weight to obtain 721g of dried materials; then 721g of bare stems Jin Yaozi and 721g of madder are added, and the mixture is uniformly mixed with the dried material, crushed and sieved by a 60-mesh sieve to obtain a fermentation material; putting the fermentation materials into a fermentation tank, adding 43ml of compound bacteria liquid, adding 119g of corn meal as a carbon source, finally adding 1.1L of deionized water, uniformly stirring, starting fermentation, and filtering and sterilizing after 4d fermentation to obtain a fermentation filtrate;

3) Preparation:

mixing the pure extractive solution obtained by extraction with fermented filtrate obtained by fermentation, concentrating to about 800mL, standing for 24 hr, filtering, adding stevioside 2g and sodium benzoate 3g into the filtrate, heating to dissolve, adding water to adjust total volume to 1000mL, stirring, refrigerating for 24 hr, filtering, bottling, and sterilizing.

Example 5

1) Preparing a compound bacterial liquid:

2) Fermentation:

collecting pure extract and residual materials after extraction by the same method as in example 2, cleaning the residual materials with deionized water for 4 times, draining off water, and drying at 50deg.C to constant weight to obtain 732g of dried materials; then 732g of bare stems Jin Yaozi and 732g of madder are added, evenly mixed with the dried material, crushed and sieved by a 60-mesh sieve to obtain a fermentation material; adding the fermentation materials into a fermentation tank, adding 22ml of compound bacteria liquid, adding 110g of corn meal as a carbon source, finally adding 1.1L of deionized water, uniformly stirring, starting fermentation, and filtering and sterilizing after 3d fermentation to obtain a fermentation filtrate;

3) Preparation:

Example 6

1) Preparing a compound bacterial liquid:

2) Fermentation:

extracting by adopting the same method as in example 3, collecting pure extracting solution and residual materials, cleaning the residual materials with deionized water for 6 times, draining off water, and drying at 60 ℃ to constant weight to obtain 729g of dried materials; adding 729g of bare stems Jin Yaozi and 729g of radix Rubiae, uniformly mixing with the dried material, crushing, and sieving with a 60-mesh sieve to obtain a fermentation material; putting the fermentation materials into a fermentation tank, adding 65.6ml of compound bacteria liquid, adding 131.2g of corn meal as a carbon source, finally adding 1.1L of deionized water, uniformly stirring, starting fermentation, and filtering and sterilizing after fermentation for 5d to obtain a fermentation filtrate;

3) Preparation:

Comparative example 4

The comparative example comprises the following steps:

1) Water extraction:

extracting 800g of bare stems Jin Yaozi and 800g of radix Rubiae with water respectively, and mixing to obtain water extracts;

2) Preparation:

extracting by the same method as in example 1, collecting pure extract, mixing the pure extract with water extract, concentrating to about 800mL, standing for 24 hr, filtering, adding stevioside 2g and sodium benzoate 3g into filtrate, heating to dissolve, adding water to adjust total volume to 1000mL, stirring, refrigerating for 24 hr, filtering, bottling, and sterilizing.

Comparative example 5

The comparative example comprises the following steps:

1) Preparing a compound bacterial liquid:

2) Fermentation

Extracting by adopting the same method as in the embodiment 1, collecting pure extracting solution and residual materials, cleaning the residual materials by deionized water for 5 times, draining off water, and drying at 55 ℃ until the weight is constant to obtain 722g of dried materials; then adding 1444g of bare stems Jin Yaozi, uniformly mixing with the dried materials, crushing and sieving with a 60-mesh sieve to obtain fermentation materials; putting the fermentation materials into a fermentation tank, adding 43ml of compound bacteria liquid, adding 119g of corn meal as a carbon source, finally adding 1.1L of deionized water, uniformly stirring, starting fermentation, and filtering and sterilizing after 4d fermentation to obtain a fermentation filtrate;

3) Formulations

Comparative example 6

The comparative example comprises the following steps:

1) Preparing a compound bacterial liquid:

2) Fermentation

Extracting by adopting the same method as in the embodiment 1, collecting pure extracting solution and residual materials, cleaning the residual materials by deionized water for 5 times, draining off water, and drying at 55 ℃ until the weight is constant to obtain 722g of dried materials; adding 1444g of radix Rubiae, mixing with the dried material, pulverizing, and sieving with 60 mesh sieve to obtain fermented material; putting the fermentation materials into a fermentation tank, adding 43ml of compound bacteria liquid, adding 119g of corn meal as a carbon source, finally adding 1.1L of deionized water, uniformly stirring, starting fermentation, and filtering and sterilizing after 4d fermentation to obtain a fermentation filtrate;

3) Formulations

Experiment 2:

pharmacodynamic experiments:

1) Selecting Wistar rats purchased by Guangzhou southern medical laboratory animal science and technology development limited company, wherein the rats are 7-8 weeks old and have weights of 180-200 g;

2) The Prunella spike oral liquid prepared in examples 4-6 and comparative examples 4-6 was used as a test drug;

3) At the beginning of the experiment, bovine thyroglobulin is firstly adopted to immunize an experimental rat, after modeling is successful, experimental small groups are divided into 7 groups, wherein 1 group is taken as a blank control group to be given with distilled water, 6 groups are respectively given with the tested drugs, the stomach of all groups is irrigated according to the proportion of 1.05ml/kg, the administration is carried out 2 times a day at intervals of 8 hours, and the administration is sacrificed after 12 d. Detecting serum thyroglobulin antibody (TGAb), thyroperoxidase antibody (TPOAb); thyroid tissue was taken, pathological sections were prepared, and morphological examination was performed, and the results are shown in Table 2.

TABLE 2

Claims

1. A preparation method of selfheal oral liquid is characterized in that the selfheal oral liquid is prepared into a medical acceptable oral liquid preparation through extraction, fermentation and liquid preparation, and is characterized in that: the extraction comprises the following steps:

1) Pretreatment:

2) Supercritical extraction:

3) Separating:

the entrainer is as follows: adding 1.2-1.6 parts of N-long-chain acyl alkylene betaine and 1 part of trityl fluorosilane into 4-6 parts of 70% ethanol solution, stirring and dispersing at 400r/min for 10 min.

2. The method for preparing the selfheal oral liquid according to claim 1, which is characterized in that: the fermentation comprises the following steps:

3. The method for preparing the selfheal oral liquid according to claim 1 or 2, which is characterized in that: the liquid preparation method comprises the following steps:

mixing the pure extractive solution obtained by extraction with the fermented filtrate obtained by fermentation, and making into pharmaceutically acceptable oral liquid preparation.