Method for extracting extracts with different proportions from lindley eupatorium herb

Abstract

The invention discloses a method for extracting extracts with different proportions in lindley eupatorium lindley, which comprises the following steps: adding lindley eupatorium lindley powder into a eutectic solvent, and preparing the extracts with stable components and controllable component proportion by adjusting the proportion of the eutectic solvent and combining a constant-temperature oscillation method. The lindley eupatorium herb extract prepared by the method contains flavonoid and alkaloid compounds, the extraction rate is about two times that of a traditional water extraction method, and the ratio of the flavonoid compounds to the alkaloid compounds is (1-2): (1-2). The method is green and efficient, required equipment is simple, operation is feasible, production cost is low, and the method can be used for large-scale industrial production.

Description

Technical Field

[0001] The invention relates to a method for extracting extracts of different proportions from yema zhui, belonging to the technical field of traditional Chinese medicine extraction. Background Art

[0002] Eupatorium lindleyanum is a perennial herbaceous plant of the Asteraceae family, mainly distributed in Jiangsu, Gansu, Shandong, Hunan and other places in China. In traditional medicine, Eupatorium lindleyanum has the effects of expectoration and cough relief, clearing away heat and detoxification, diuresis and swelling, and lowering blood pressure. It has a long history of application in the treatment of respiratory diseases and blood pressure regulation. Modern research shows that the chemical components of Eupatorium lindleyanum include flavonoids, alkaloids, sesquiterpenes, triterpenes, volatile oils, organic acids and other components. In particular, flavonoids have the effects of anti-myocardial hypoxia, ischemia, anti-arrhythmia, lowering serum cholesterol, inhibiting platelet aggregation, anti-aging and enhancing immunity, and alkaloids have anti-hypertensive effects.

[0003] At present, most of the solvents used to prepare Chinese medicine extracts are organic solvents, which pose a risk of environmental pollution. Finding green and efficient extraction solvents has good application prospects. Low eutectic solvents are mixtures formed by hydrogen bond interactions between two or more components. They are usually made from renewable raw materials and have low toxicity and use costs. The extraction performance of low eutectic solvents can be optimized by adjusting the ratio of ingredients. In addition, low eutectic solvents can dissolve plant cell wall components more effectively, promote the release of compounds, and have higher extraction efficiency than traditional solvents.

[0004] The preparation method of Chinese herbal medicine extracts is related to the quality and safety of the final product, and is also closely related to the production cost. Common preparation methods for Chinese herbal medicine extracts include: decoction, maceration, reflux extraction, ultrasound-assisted method, etc. Among them, the extraction efficiency of decoction and maceration is low; extraction methods such as reflux extraction and ultrasound-assisted method require specific extraction equipment, parameter settings are affected by the outside world, and the operation steps are complicated, which can easily cause safety hazards due to improper operation. Therefore, efficient and stable preparation methods are the trend in preparing Chinese herbal medicine extracts. The constant temperature oscillation method can provide a stable experimental environment. It only needs to control two parameters, temperature and oscillation speed. The parameters can be set after placing the sample. It is simple to operate and helps to maintain quality stability between different batches, which is convenient for large-scale production. Summary of the invention

[0005] Purpose of the invention: The present invention aims to provide a green and efficient preparation method to address the shortcomings of traditional extraction technology, such as low efficiency, high cost, heavy environmental pollution, complex operation and possible impact on the stability of the extract, so as to improve the yield of flavonoids and alkaloids in the wild horsetail extract and obtain wild horsetail extracts with different component ratios.

[0006] Technical solution: The method for extracting extracts of different proportions from Mustard Seedlings of the present invention comprises the following steps: adding a low eutectic solvent to Mustard Seedlings powder, shaking at a constant temperature, centrifuging, and concentrating and freeze-drying to obtain the extracts.

[0007] Furthermore, the extracts include total flavonoids and total alkaloids in different proportions.

[0008] Further, the deep eutectic solvent is an aqueous solution of choline chloride and citric acid.

[0009] Furthermore, by adjusting the dosage ratio of choline chloride and citric acid in the deep eutectic solvent, extracts with different ratios can be obtained.

[0010] Furthermore, the molar ratio of choline chloride to citric acid is 1-2:1-4, and the mass fraction of water in the low eutectic solvent is 20-30%.

[0011] Furthermore, the molar ratio of choline chloride to citric acid is 1-2:1-4, and the mass ratio of total flavonoids to total alkaloids in the obtained extract is 1-2:1-2.

[0012] Further, the preparation of the deep eutectic solvent comprises the following steps:

[0013] Heat and stir choline chloride and citric acid until a uniform, stable, clear and transparent solution is formed, cool, add water and mix evenly to obtain a low eutectic solvent.

[0014] Furthermore, the heating and stirring temperature is 80-120° C., and the heating and stirring time is 120-180 min.

[0015] Furthermore, the temperature of the constant temperature oscillation is 50-60° C., the frequency of the constant temperature oscillation is 160-200 rpm, and the time of the constant temperature oscillation is 60-90 min.

[0016] Furthermore, the material-liquid ratio of the Mustang powder to the low eutectic solvent is 1:20 to 1:30 g / mL.

[0017] Furthermore, the particle size of the Mustang powder is 30 to 50 meshes.

[0018] Furthermore, the constant temperature shaking device is a constant temperature water bath shaker.

[0019] Furthermore, the extraction was repeated 2 to 3 times at a constant temperature, followed by centrifugation, and the supernatants were combined, concentrated and freeze-dried to obtain the wild horsetail extract.

[0020] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0021] The invention provides a method for extracting extracts of different proportions from wild horsetail, wherein the extracts include flavonoids and alkaloids, and the ratio of the flavonoids to the alkaloids in the extracts is 1:1 or 1:2; the extraction rate of the low eutectic solvent-assisted constant temperature shaking preparation method is better than that of traditional solvents such as methanol, ethanol, and water; the preparation method is efficient, green and pollution-free, the extract has good stability, the component ratio is controllable, the equipment is simple to operate, and is suitable for large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 is the standard curve of total flavonoids;

[0023] Figure 2 This is the standard curve of total alkaloids. DETAILED DESCRIPTION

[0024] The technical solution of the present invention is further described below in conjunction with the accompanying drawings.

[0025] Example 1

[0026] Choline chloride and citric acid in a molar ratio of 1:2 were placed in a reaction bottle, heated to 90°C in a magnetic stirrer, and stirred continuously for 120 minutes until a uniform, stable, clear and transparent solution was formed. After cooling to room temperature, 30% of water was added to obtain a low eutectic solvent 1.

[0027] The dried wild horse herb was crushed and passed through a 40-mesh sieve. 1 g of wild horse herb powder was weighed and 20 mL of low eutectic solvent 1 was added. Then, the extract was extracted at a constant temperature of 50 ° C and 160 rpm for 60 min, and centrifuged at 5000 rpm for 10 min. The extraction was repeated 3 times and then centrifuged. The supernatant was combined and concentrated and freeze-dried to obtain wild horse herb extract A. The extraction rate was 5.43 ± 1.33 mg / g (RSD < 0.5). The total flavonoids kit and the total alkaloid content determination kit (Suzhou Ge Ruisi Biotechnology Co., Ltd.) were used to detect the yields of flavonoids and alkaloids in extract A. The results showed that the flavonoids in extract A were 3.41 ± 0.67 mg / g, the alkaloids were 3.11 ± 0.51 mg / g, and the mass ratio of flavonoids to alkaloids was about 1:1.

[0028] Example 2

[0029] Choline chloride and citric acid in a molar ratio of 2:1 were placed in a reaction bottle, heated to 90°C in a magnetic stirrer, and stirred continuously for 120 minutes until a uniform, stable, clear and transparent solution was formed. After cooling to room temperature, 30% of water was added to obtain a low eutectic solvent 2.

[0030] The dried wild horse herb was crushed, passed through a 40-mesh sieve, 1 g of wild horse herb powder was weighed, 20 mL of low eutectic solvent 2 was added, and then constant temperature shaking extraction was performed at 50 ° C and 160 rpm for 60 min, centrifuged at 5000 rpm for 10 min, and the extraction was repeated 3 times and then centrifuged. The supernatant was combined, concentrated and freeze-dried to obtain wild horse herb extract B, and the extraction rate was 5.21 ± 1.47 mg / g (RSD < 0.5). The total flavonoids kit and the total alkaloid content determination kit (Suzhou Ge Ruisi Biotechnology Co., Ltd.) were used to detect the yields of flavonoids and alkaloids in extract B. The results showed that the flavonoids in extract B were 3.94 ± 0.54 mg / g, the alkaloids were 7.34 ± 0.68 mg / g, and the mass ratio of flavonoids to alkaloids was about 1:2.

[0031] Example 3

[0032] Choline chloride and citric acid in a molar ratio of 1:1 were placed in a reaction bottle, heated to 90°C in a magnetic stirrer, and stirred continuously for 120 minutes until a uniform, stable, clear and transparent solution was formed. After cooling to room temperature, 30% of water was added to obtain a low eutectic solvent 3.

[0033] The dried wild horse herb was crushed, passed through a 40-mesh sieve, 1 g of wild horse herb powder was weighed, 20 mL of low eutectic solvent 3 was added, and then constant temperature shaking extraction was performed at 50 ° C and 160 rpm for 60 min, centrifuged at 5000 rpm for 10 min, and the extraction was repeated 3 times and then centrifuged. The supernatant was combined, concentrated and freeze-dried to obtain wild horse herb extract C, and the extraction rate was 5.18 ± 1.61 mg / g (RSD < 0.5). The total flavonoids kit and the total alkaloid content determination kit (Suzhou Ge Ruisi Biotechnology Co., Ltd.) were used to detect the yields of flavonoids and alkaloids in extract C. The results showed that the flavonoids in extract C were 2.89 ± 0.82 mg / g, the alkaloids were 3.57 ± 0.77 mg / g, and the mass ratio of flavonoids to alkaloids was about 1:1.2.

[0034] Example 4

[0035] Choline chloride and citric acid in a molar ratio of 1:4 were placed in a reaction bottle, heated to 90°C in a magnetic stirrer, and stirred continuously for 120 minutes until a uniform, stable, clear and transparent solution was formed. After cooling to room temperature, 30% of water was added to obtain a low eutectic solvent 4.

[0036] The dried wild horse herb was crushed, passed through a 40-mesh sieve, 1 g of wild horse herb powder was weighed, 20 mL of low eutectic solvent 3 was added, and then constant temperature shaking extraction was performed at 50 ° C and 160 rpm for 60 min, centrifuged at 5000 rpm for 10 min, and the extraction was repeated 3 times and then centrifuged. The supernatant was combined, concentrated and freeze-dried to obtain wild horse herb extract D, and the extraction rate was 4.63 ± 1.13 mg / g (RSD < 0.5). The total flavonoids kit and the total alkaloid content determination kit (Suzhou Ge Ruisi Biotechnology Co., Ltd.) were used to detect the yields of flavonoids and alkaloids in extract D. The results showed that the flavonoids in extract D were 2.90 ± 0.42 mg / g, the alkaloids were 1.89 ± 0.48 mg / g, and the mass ratio of flavonoids to alkaloids was about 1.5:1.

[0037] Comparative Example 1

[0038] The experimental process is the same as in Example 1, except that ultrapure water is used instead of low co-solvent, the dried wild horse chase is crushed, passed through a 40-mesh sieve, 1g of wild horse chase powder is weighed, 20mL of ultrapure water is added, and then the temperature is 50°C, 160rpm, constant temperature shaking extraction is performed for 60min, 5000rpm centrifugation is performed for 10min, the extraction is repeated 3 times and then centrifuged, the supernatant is combined, and the concentration and freeze-drying are performed to obtain wild horse chase extract E, and the extraction rate is 2.89±0.99mg / g (RSD<0.5). The total flavonoids kit and the total alkaloid content determination kit (Suzhou Ge Ruisi Biotechnology Co., Ltd.) were used to detect the yields of flavonoids and alkaloids in extract E. The results showed that the flavonoids were 1.89±0.63mg / g and the alkaloids were 1.64±0.77mg / g. Although the mass ratio of flavonoids to alkaloids was about 1:1, the extraction rate of flavonoids was only 55% of that in Example 1, and the extraction rate of alkaloids was 52% of that in Example 1.

[0039] Comparative Example 2

[0040] The experimental process is the same as that of Comparative Example 1, except that ultramethanol is used instead of ultrapure water, the dried wild horse chase is crushed, passed through a 40-mesh sieve, 1g of wild horse chase powder is weighed, 20mL of methanol is added, and then the temperature is 50°C, 160rpm, constant temperature shaking extraction is performed for 60min, 5000rpm is centrifuged for 10min, the extraction is repeated 3 times and then centrifuged, the supernatant is combined, and the concentration and freeze-drying are performed to obtain wild horse chase extract F, and the extraction rate is 3.47±1.48mg / g (RSD<0.5). The total flavonoids kit and the total alkaloid content determination kit (Suzhou Ge Ruisi Biotechnology Co., Ltd.) were used to detect the yields of flavonoids and alkaloids in extract F. The results showed that the flavonoids were 1.47±0.89mg / g and the alkaloids were 1.91±0.91mg / g. Although the mass ratio of flavonoids to alkaloids was about 1:1, the extraction rate of flavonoids was only 43% of that in Example 1, and the extraction rate of alkaloids was 61% of that in Example 1.

[0041] Comparative Example 3

[0042] The experimental process is the same as that of Comparative Example 1, except that ultrasonic extraction is used instead of constant temperature shaking extraction, choline chloride and citric acid in a molar ratio of 1:2 are placed in a reaction bottle, heated to 90°C in a magnetic stirrer, and stirred continuously for 120 minutes until a uniform, stable, clear and transparent solution is formed. After cooling to room temperature, 30% of water is added to obtain a low eutectic solvent 1.

[0043] The dried wild horse chase was crushed, passed through a 40-mesh sieve, 1 g of wild horse chase powder was weighed, 20 mL of low eutectic solvent 1 was added, and then ultrasonic extraction was performed at a temperature of 50 ° C for 60 min, centrifuged at 5000 rpm for 10 min, and the extraction was repeated 3 times and then centrifuged. The supernatant was combined and concentrated and freeze-dried to obtain wild horse chase extract G, and the extraction rate was 3.27 ± 6.56 mg / g (RSD> 1.5). The total flavonoids kit and the total alkaloid content determination kit (Suzhou Ge Ruisi Biotechnology Co., Ltd.) were used to detect the yields of flavonoids and alkaloids in extract E. The results showed that the flavonoids in extract G were 1.54 ± 2.79 mg / g, the alkaloids were 1.39 ± 2.61 mg / g, and the mass ratio of flavonoids to alkaloids was about 1: 1. Compared with Example 1, although the mass ratio of flavonoids to alkaloids was close, the extraction rates of flavonoids and alkaloids were only 45% of that in Example 1.

Claims

1. A method for extracting extracts of different proportions from yamachus chinensis, characterized in that: The following steps are involved: A low eutectic solvent is added to the Mustangui powder, the mixture is shaken at a constant temperature, centrifuged, concentrated and freeze-dried to obtain the product.

2. The method for extracting different proportions of extracts from Ophiopogon japonicus according to claim 1, characterized in that: The extracts included total flavonoids and total alkaloids in different proportions.

3. The method for extracting different proportions of extracts from Ophiopogon japonicus according to claim 1, characterized in that: The deep eutectic solvent is an aqueous solution of choline chloride and citric acid.

4. The method for extracting different proportions of extracts from Ophiopogon japonicus according to claim 3, characterized in that: Extracts with different ratios can be obtained by adjusting the dosage ratio of choline chloride and citric acid in the low eutectic solvent.

5. The method for extracting different proportions of extracts from Ophiopogon japonicus according to claim 4, characterized in that: The molar ratio of choline chloride to citric acid is 1-2:1-4, the mass fraction of water in the low eutectic solvent is 20-30%, and the mass ratio of total flavonoids to total alkaloids in the extract is 1-2:1-2.

6. The method for extracting extracts of different proportions from Ophiopogon japonicus according to claim 1, characterized in that: The preparation of the deep eutectic solvent comprises the following steps: Heat and stir choline chloride and citric acid to form a clear and transparent solution, cool it, add water and mix well to obtain a low eutectic solvent.

7. The method for extracting different proportions of extracts from Ophiopogon japonicus according to claim 6, characterized in that: The heating and stirring temperature is 80-120°C, and the heating and stirring time is 120-180 minutes.

8. The method for extracting extracts of different proportions from Ophiopogon japonicus according to claim 1, characterized in that: The temperature of the constant temperature oscillation is 50-60°C, the frequency of the constant temperature oscillation is 160-200 rpm, and the time of the constant temperature oscillation is 60-90 min.

9. The method for extracting extracts of different proportions from Ophiopogon japonicus according to claim 1, characterized in that: The material-liquid ratio of the wild horsetail powder to the low eutectic solvent is 1:20-1:30 g / mL, and the particle size of the wild horsetail powder is 30-50 meshes.

10. The method for extracting extracts of different proportions from Ophiopogon japonicus according to claim 1, characterized in that: The constant temperature shaking was repeated for 2 to 3 times, followed by centrifugal separation, the supernatants were combined, concentrated and freeze-dried to obtain the wild horsetail extract.

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