Preparation method of radix ophiopogonis oropharyngeal prescription preparation extract

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a preparation method of a fructus phyllanthi oropharyngeal prescription preparation extract. The preparation method of the phyllanthus emblica and radix rehmanniae oropharyngeal prescription preparation extract comprises the following steps: a, cutting phyllanthus emblica, radix rehmanniae and liquorice, selecting radix ophiopogonis, and crushing radix paeoniae rubra to obtain radix paeoniae rubra powder; b, mixing the medicinal materials in the step a, and performing enzymolysis; and c, after enzymolysis, adding an ethanol solution for extraction, and concentrating. According to the method disclosed by the invention, cellulose and hemicellulose of plant cell walls are specifically degraded by adopting cellulase, so that cell contents are exposed, and beta-glucosidase and pectinase are matched for use, so that disintegration of the cell walls is further assisted, and the porosity of the medicinal material powder is increased by degradation of the cell walls; active substances in the medicinal materials can be better dissolved in an extraction solvent, and the content of effective components is increased.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a method for preparing a Yumai oropharyngeal prescription preparation extract. Background Art

[0002] Yumai Kouyan is a compound preparation used in traditional Chinese medicine to improve discomfort in the mouth and throat. It is made from Phyllanthus emblica, Rehmannia glutinosa, Paeonia lactiflora, Ophiopogon japonicus, and Licorice. It has the effects of nourishing yin and reducing fire. It is often used to treat burning sensation, pain, local redness and swelling, irritability, dry mouth, yellow and red urine, and recurrent oral ulcers caused by yin deficiency and excessive fire, and virtual fire ascending.

[0003] Red peony root and emblica fruit are the main active ingredients in Yumai oral and pharyngeal mixture, which have the effects of clearing heat and cooling blood, dispersing blood stasis and relieving pain, promoting digestion and strengthening the stomach, and promoting fluid production and relieving cough. Among them, paeoniflorin can relieve fever, anti-inflammatory and anti-ulcer; gallic acid can promote fluid production and quench thirst, stop bleeding and cool blood; corilagin can resist bacteria, kill bacteria and reduce inflammation, and inhibit viruses; ellagic acid can resist oxidation, prevent infection and inhibit ulcers.

[0004] The existing method for preparing the oropharyngeal extract of yumai is generally a water decoction method. The contents of paeoniflorin, gallic acid, corilagin and ellagic acid in the oropharyngeal extract of yumai prepared by this method are low, which affects the efficacy of the preparation. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a method for preparing an extract of a Yumai oropharyngeal prescription preparation which can improve the extraction rate and content of paeoniflorin, gallic acid, corilagin and ellagic acid.

[0006] The present invention provides a method for preparing a Yumai oropharyngeal prescription preparation extract, comprising the following steps:

[0007] a. Cut Phyllanthus emblica, Rehmannia root and Licorice root, select Ophiopogon japonicus, and crush Red Peony Root to obtain Red Peony Root Powder;

[0008] b. Mixing the medicinal materials in step a and performing enzymatic hydrolysis;

[0009] c. After enzymatic hydrolysis, add ethanol solution for extraction and concentration.

[0010] Further, in step a, 720 parts of Phyllanthus emblica, 360 parts of Rehmannia glutinosa, 360 parts of Ophiopogon japonicus, 270 parts of Red Peony Root, and 90 parts of Licorice.

[0011] Furthermore, in step a, the red peony root is crushed and passed through a 10 mm sieve to obtain red peony root powder.

[0012] Furthermore, in step b, the enzymes used for enzymolysis are cellulase, pectinase and β-glucosidase.

[0013] Furthermore, in step b, the amount of cellulase added is 0.5% to 2.0% of the total medicinal material, the amount of pectinase added is 0.2% to 1% of the total medicinal material, and the amount of β-glucosidase added is 0.1% to 0.5% of the total medicinal material.

[0014] Furthermore, in step b, the enzymatic hydrolysis is carried out for 3 to 4 hours using citric acid buffer as solvent at a temperature of 45 to 55° C. and a pH value of 5.0 to 5.5.

[0015] Furthermore, the concentration of the citric acid buffer is 0.1 M; the amount of the citric acid buffer added is 8 to 10 times the total mass of the medicinal material.

[0016] The inventors have found through research that the enzymatic hydrolysis method can effectively promote the release of paeoniflorin, gallic acid, corilagin and ellagic acid.

[0017] Preferably, the specific enzyme activity of the cellulase is 10U / g to 20U / g; the specific enzyme activity of the β-glucosidase is 5U / g to 10U / g; and the specific enzyme activity of the pectinase is 8U / g to 10U / g.

[0018] Furthermore, in step c, extraction is performed twice using a 70% ethanol solution with a volume concentration of 2 times. The first time, 6 to 6.5 times the total mass of the medicinal material is added with a 70% ethanol solution and reflux extraction is performed for 2 hours. The second time, 4 to 4.5 times the total mass of the medicinal material is added with a 70% ethanol solution and reflux extraction is performed for 2 hours.

[0019] Furthermore, in step c, the collected extract is filtered through a plate and frame filter and then concentrated using a single-effect concentrator at 50-60° C. until the relative density of the extract is 1.17-1.20.

[0020] The beneficial effects of the present invention are:

[0021] The present invention provides a method for preparing an extract of a prescription preparation of yumai oropharyngeal preparation. The inventor uses cellulase, β-glucosidase and pectinase to enzymolyze the medicinal materials in yumai oropharyngeal preparation, and extracts with ethanol. Cellulase is used to specifically degrade the cellulose and hemicellulose of the plant cell wall, so that the cell contents are exposed, and then β-glucosidase and pectinase are used in combination to further assist the disintegration of the cell wall. The degradation of the cell wall increases the porosity of the medicinal material powder, so that the active substances in the medicinal material can be better dissolved in the extraction solvent, and the leaching rate of the effective ingredients in the medicinal material is increased. In addition, the content of paeoniflorin, gallic acid, corilagin and ellagic acid is increased. Compared with traditional methods such as hot reflux or Soxhlet extraction, the enzymolysis of the method of the present invention can significantly shorten the extraction cycle, pre-activate the active ingredients in the medicinal material to a certain extent, and reduce the energy consumption and time investment required for subsequent extraction. DETAILED DESCRIPTION

[0022] Example 1

[0023] Cut the emblica, rehmannia and liquorice, select ophiopogon, crush red peony root to obtain red peony powder, and pass through a 10mm sieve; evenly mix 720g of the processed emblica, 360g of ophiopogon, 270g of red peony root, 360g of rehmannia and 90g of liquorice, add 36g of cellulase (enzyme specific activity 20U / g), 18g of pectinase (enzyme specific activity 10U / g), and 9g of β-glucosidase (enzyme specific activity 10U / g), then add 0.1M citric acid buffer solution 10 times the total weight of the medicinal materials to completely immerse the medicinal materials, enzymolyze at 55°C and pH 5.5 for 3h, then add 70% ethanol solution 6.5 times the total weight of the medicinal materials, reflux and extract at 85°C for 2h, and after draining the extract, add 70% ethanol solution 4.5 times the total weight of the medicinal materials again, reflux and extract at 85°C for 2h;

[0024] The collected extract was filtered through a plate and frame filter and then concentrated at low temperature under vacuum using a single-effect concentrator at 60°C until the relative density of the extract was 1.20 and the amount of the extract was 2000 kg.

[0025] Example 2

[0026] Cut the emblica, rehmannia and liquorice, select ophiopogon, crush red peony root to obtain red peony powder, and pass through a 10mm sieve; evenly mix 720g of the processed emblica, 360g of ophiopogon, 270g of red peony root, 360g of rehmannia and 90g of liquorice, add 18g of cellulase (enzyme specific activity 15U / g), 14.4g of pectinase (enzyme specific activity 9U / g), 5.3g of β-glucosidase (enzyme specific activity 7U / g), and then add 0.1M citric acid buffer solution of 8 times the total weight of the medicinal materials to completely immerse the medicinal materials, enzymolyze at 50°C and pH 5.0 for 3h, then add 70% ethanol solution of 6 times the total weight of the medicinal materials, reflux and extract at 85°C for 2h, and after draining the extract, add 70% ethanol solution of 4.5 times the total weight of the medicinal materials again, reflux and extract at 85°C for 2h;

[0027] The collected extract was filtered through a plate and frame filter and then concentrated at low temperature under vacuum using a single-effect concentrator at 60°C until the relative density of the extract was 1.19 and the amount of the extract was 1940 kg.

[0028] Example 3

[0029] Cut the emblica, rehmannia and liquorice, select ophiopogon, crush red peony root to obtain red peony powder, and pass through a 10mm sieve; evenly mix 720g of the processed emblica, 360g of ophiopogon, 270g of red peony root, 360g of rehmannia and 90g of liquorice, add 18g of cellulase (enzyme specific activity 10U / g), 14.4g of pectinase (enzyme specific activity 8U / g), 5.3g of β-glucosidase (enzyme specific activity 5U / g), and then add 0.1M citric acid buffer solution of 8 times the total weight of the medicinal materials to completely immerse the medicinal materials, enzymolyze at 50°C and pH 5.0 for 3h, then add 70% ethanol solution of 6 times the total weight of the medicinal materials, reflux and extract at 85°C for 2h, and after draining the extract, add 70% ethanol solution of 4.5 times the total weight of the medicinal materials again, reflux and extract at 85°C for 2h;

[0030] The collected extract was filtered through a plate and frame filter and then concentrated at low temperature under vacuum using a single-effect concentrator at 60°C until the relative density of the extract was 1.17 and the amount of the extract was 1920 kg.

[0031] Comparative Example 1: Traditional hot water extraction

[0032] Cut the emblica, rehmannia and liquorice, select ophiopogon, crush red peony root to obtain red peony powder, and pass through a 10mm sieve; mix 720g of the processed emblica, 360g of ophiopogon, 270g of red peony root, 360g of rehmannia and 90g of liquorice evenly, boil in water, add 6.5 times the amount of process drinking water for the first time, soak for 2h, heat to boiling, and boil for 2h; add 4.5 times the amount of process drinking water for the second time, heat to boiling, and boil for 2h; filter the collected extract through a plate and frame filter, and then vacuum-low-temperature concentrate it, using a double-effect concentrator for concentration, with the first effect temperature at 70-80℃ and the second effect temperature at 50-60℃, and concentrate it until the relative density of the extract is 1.17 and the amount of the extract is 1700kg.

[0033] Comparative Example 2 Single Enzyme Hydrolysis

[0034] Cut the emblica, rehmannia and liquorice, select ophiopogon, crush red peony root to obtain red peony powder, and pass through a 10mm sieve; take 720g of the processed emblica, 360g of ophiopogon, 270g of red peony root, 360g of rehmannia and 90g of liquorice and mix them evenly. Add 60g of cellulase (enzyme activity ratio 15U / g), then add 0.1M citric acid buffer solution of 8 times the total weight of the medicinal materials, immerse the medicinal materials completely, and perform enzymolysis at 50℃ for 3h; after enzymolysis, add 6.5 times the total weight of the medicinal materials and 70% ethanol solution at 85℃ for 2h, drain the extract and add 4.5 times the total weight of the medicinal materials and 70% ethanol solution at 85℃ for 2h; filter the collected extract through a plate and frame filter and perform vacuum low-temperature concentration, and use a single-effect concentrator to concentrate at 60℃ until the relative density of the extract is 1.17 and the amount of the extract is 1820kg.

[0035] Comparative Example 3: Combination of Cellulase and Pectinase

[0036] Cut the emblica fructus, rehmannia root and liquorice, select the ophiopogon japonicus, crush the red peony root to obtain the red peony root powder, and pass it through a 10mm sieve; take 720g of the processed emblica fructus, 360g of ophiopogon japonicus, 270g of red peony root, 360g of rehmannia root and 90g of liquorice and mix them evenly. Add 36g of cellulase (enzyme activity ratio 15U / g) and 18g of pectinase (enzyme activity ratio 8U / g), and then add 0.1M citric acid buffer solution in an amount of 8 times the total weight of the medicinal material to completely immerse the medicinal material, and perform enzymolysis at 50°C for 3h; after enzymolysis, add 70% ethanol solution in an amount of 6.5 times the total weight of the medicinal material and perform reflux extraction at 85°C for 2h, and after draining the extract, add 70% ethanol solution in an amount of 4.5 times the total weight of the medicinal material again and perform reflux extraction at 85°C for 2h; filter the collected extract through a plate and frame filter and perform vacuum low-temperature concentration, and use a single-effect concentrator to concentrate at 60°C until the relative density of the extract is 1.17 and the amount of the extract is 1880kg.

[0037] Comparative Example 4: Enzyme specific activity is too low

[0038] Cut the emblica, rehmannia and liquorice, select ophiopogon, crush red peony root to obtain red peony powder, and pass through a 10mm sieve; evenly mix 720g of the processed emblica, 360g of ophiopogon, 270g of red peony root, 360g of rehmannia and 90g of liquorice, add 36g of cellulase (enzyme specific activity 8U / g), 18g of pectinase (enzyme specific activity 6U / g), and 9g of β-glucosidase (enzyme specific activity 4U / g), then add 0.1M citric acid buffer solution of 10 times the total weight of the medicinal materials to completely immerse the medicinal materials, and perform enzymolysis at 55°C and pH 5.5 for 3h, then add 70% ethanol solution of 6.5 times the total weight of the medicinal materials, reflux and extract at 85°C for 2h, and after draining the extract, add 70% ethanol solution of 4.5 times the total weight of the medicinal materials again, reflux and extract at 85°C for 2h;

[0039] The collected extract was filtered through a plate and frame filter and then concentrated at low temperature under vacuum using a single-effect concentrator at 60°C until the relative density of the extract was 1.20 and the amount of the extract was 1760 kg.

[0040] Comparative Example 5: Enzyme specific activity is too high

[0041] Cut the emblica, rehmannia and liquorice, select ophiopogon, crush red peony root to obtain red peony powder, and pass through a 10mm sieve; evenly mix 720g of the processed emblica, 360g of ophiopogon, 270g of red peony root, 360g of rehmannia and 90g of liquorice, add 36g of cellulase (enzyme specific activity 25U / g), 18g of pectinase (enzyme specific activity 12U / g), and 9g of β-glucosidase (enzyme specific activity 15U / g), then add 0.1M citric acid buffer solution of 10 times the total weight of the medicinal materials to completely immerse the medicinal materials, enzymolyze at 55°C and pH 5.5 for 3h, then add 70% ethanol solution of 6.5 times the total weight of the medicinal materials, reflux and extract at 85°C for 2h, and after draining the extract, add 70% ethanol solution of 4.5 times the total weight of the medicinal materials again, reflux and extract at 85°C for 2h;

[0042] The collected extract was filtered through a plate and frame filter and then concentrated at low temperature under vacuum using a single-effect concentrator at 60°C until the relative density of the extract was 1.20 and the amount of the extract was 1740 kg.

[0043] Table 1 Extraction rate of active ingredients in Examples and Comparative Examples

[0044] Element Paeoniflorin / % Gallic acid / % Korilagin / % Ellagic acid / % Example 1 89 80 30 27 Example 2 87 74 27 25 Example 3 85 72 25 22 Comparative Example 1 55 63 16 14 Comparative Example 2 63 65 18 16 Comparative Example 3 70 68 22 19 Comparative Example 4 60 65 20 18 Comparative Example 5 58 63 19 15

[0045] The detection method with application number CN202310122400.9 was used to detect the contents of the effective ingredients paeoniflorin, gallic acid, corilagin and ellagic acid in the present invention. The test results are shown in Table 2.

[0046] Table 2 Content of active ingredients in Examples and Comparative Examples

[0047] Element Paeoniflorin / mg / g Gallic acid / mg / g Corilagin / mg / g Ellagic acid / mg / g Example 1 4.59 6.31 2.16 0.49 Example 2 4.29 6.01 2.0 0.46 Example 3 4.11 5.91 1.88 0.41 Comparative Example 1 3.49 5.84 1.36 0.30 Comparative Example 2 3.74 5.63 1.42 0.32 Comparative Example 3 4.02 5.70 1.69 0.36 Comparative Example 4 3.68 5.82 1.64 0.37 Comparative Example 5 3.6 5.71 1.57 0.31

Claims

1. A method for preparing an extract of a prescription preparation of yumai oropharyngeal preparation, characterized in that: The steps include: a. Cut Phyllanthus emblica, Rehmannia root and Licorice root, select Ophiopogon japonicus, and crush Red Peony Root to obtain Red Peony Root Powder; b. Mixing the medicinal materials in step a and performing enzymatic hydrolysis; c. After enzymatic hydrolysis, add ethanol solution for extraction and concentration.

2. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 1, characterized in that: In step a, 720 parts of emblica fructus, 360 parts of rehmannia root, 360 parts of ophiopogon japonicus, 270 parts of red peony root, and 90 parts of liquorice.

3. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 1, characterized in that: In step a, the red peony root is crushed and passed through a 10 mm sieve to obtain red peony root powder.

4. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 1, characterized in that: In step b, the enzymes used for enzymolysis are cellulase, pectinase and β-glucosidase.

5. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 4, characterized in that: In step b, the added amount of the cellulase is 0.5% to 2.0% of the total medicinal material, the added amount of the pectinase is 0.2% to 1% of the total medicinal material, and the added amount of the β-glucosidase is 0.1% to 0.5% of the total medicinal material.

6. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 1, characterized in that: In step b, the enzymatic hydrolysis is carried out using citric acid buffer as solvent at a temperature of 45 to 55° C. and a pH value of 5.0 to 5.5 for 3 to 4 hours.

7. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 6, characterized in that: In step b, the amount of citric acid buffer added is 8 to 10 times the total mass of the medicinal materials.

8. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 4, characterized in that: In step b, the specific activity of the cellulase is 10U / g to 20U / g; the specific activity of the β-glucosidase is 5U / g to 10U / g; and the specific activity of the pectinase is 8U / g to 10U / g.

9. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 1, characterized in that: In step c, 70% ethanol solution with a volume concentration is used for extraction twice. The first time, 6 to 6.5 times the total mass of the medicinal material is added with 70% ethanol solution and reflux extraction is performed for 2 hours; the second time, 4 to 4.5 times the total mass of the medicinal material is added with 70% ethanol solution and reflux extraction is performed for 2 hours.

10. The method for preparing the extract of the oropharyngeal prescription preparation of Yumai according to claim 1, characterized in that: In step c, the relative density of the concentrate is 1.17 to 1.20.