CN115177655A - Preparation method of licorice extract and application of licorice extract in antibacterial and anti-inflammatory traditional Chinese medicine composition - Google Patents
Preparation method of licorice extract and application of licorice extract in antibacterial and anti-inflammatory traditional Chinese medicine composition Download PDFInfo
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Abstract
The invention relates to the technical field of traditional Chinese medicine application, and provides a preparation method of a liquorice extract and application of the liquorice extract in an antibacterial and anti-inflammatory traditional Chinese medicine composition, wherein the preparation method of the liquorice extract comprises the steps of S1, weighing liquorice powder, adding ethanol and cedrol into the liquorice powder for extraction, centrifuging and collecting supernate; s2, adding a mixed solution of ammonia water and ethanol into the centrifuged precipitate for extraction, centrifuging and collecting the supernatant again; and S3, combining the supernatants, concentrating, purifying and drying in vacuum to obtain the licorice extract. Through the technical scheme, the problem of low content of liquiritin and glycyrrhizic acid in the licorice extract in the prior art is solved.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine application, in particular to a preparation method of a liquorice extract and application of the liquorice extract in an antibacterial and anti-inflammatory traditional Chinese medicine composition.
Background
The licorice is the root and stem of the perennial plant licorice of the leguminosae, has sweet taste and moderate nature, has long medicinal history and is called as 'aged people'. The traditional Chinese medicine considers that the liquorice is sweet and mild in taste and is a good medicine for tonifying spleen and qi, relieving cough and disease, relieving spasm and pain and harmonizing the medicines. The liquorice contains rich liquiritin, glycyrrhizic acid, alkaloid, polysaccharide and the like, and the content of the liquiritin and the glycyrrhizic acid in the liquorice can reach 11 to 19 percent. Licorice is a food and a medicine resource, and the American FDA has already listed the licorice extract as a safe and nontoxic substance, and scientific research proves that the licorice extract is an additive with multiple functions of bacteriostasis, inflammation diminishing, detoxification, deodorization and the like, and is mainly used for treating gastric ulcer, duodenal ulcer, diabetes insipidus, tuberculosis, dermatitis, bronchial asthma and the like clinically. The liquorice and the extract thereof have wide application in the industries of medicine, food, cosmetics and the like.
Patent application No. 202011291262.X adopts three kinds of alcoholic solutions to extract, has extracted liquiritin, has not extracted the glycyrrhizic acid in the licorice, and is one-step extraction, may cause and extract inadequately, leads to the reduction of active substance such as liquiritin, glycyrrhizic acid in the licorice extract.
Disclosure of Invention
The invention provides a preparation method of a licorice extract and application of the licorice extract in an antibacterial and anti-inflammatory traditional Chinese medicine composition, and solves the problem that the content of liquiritin and glycyrrhizic acid in the licorice extract is low in related technologies.
The technical scheme of the invention is as follows:
the preparation method of the licorice extract comprises the following steps:
s1, weighing licorice powder, adding ethanol and cedrol into the licorice powder for extraction, centrifuging and collecting supernatant;
s2, adding a mixed solution containing ammonia water and ethanol into the centrifuged precipitate for extraction, centrifuging and collecting the supernatant again;
and S3, combining the supernatants, concentrating, purifying and drying in vacuum to obtain the licorice extract.
As a further technical scheme, the licorice powder in the step S1 is obtained by crushing licorice and then sieving the licorice with a 80-mesh sieve.
As a further technical scheme, the mass ratio of the licorice powder to the ethanol in the step S1 is 1; the mass ratio of the licorice powder to the cedrol is 1:1-3:1.
As a further technical scheme, the ethanol is 60wt% ethanol.
As a further technical scheme, the extraction in the step S1 is ultrasonic extraction, the extraction temperature is 60 ℃, the extraction time is 60min, and the extraction power is 250W; the extraction in the step S2 is reflux extraction, and the extraction time is 60min.
As a further technical proposal, the mixed solution of ammonia water and ethanol in the step S2 contains 0.4 to 0.6 weight percent of ammonia water and 60 weight percent of ethanol.
As a further technical scheme, the addition amount of the mixed solution of ammonia and ethanol in the step S2 is 5-8 times of the mass of the licorice powder.
As a further technical scheme, the concentration in the step S3 is that the combined supernatant is concentrated under reduced pressure at 65 ℃ until the density is 1.15g/cm 3 (ii) a Purifying by passing the concentrated solution through macroporous adsorbent resin column, eluting with 2-5 times of column volume of distilled water, eluting with 60-80% ethanol, and collecting ethanol eluate.
As a further technical scheme, the application of the licorice extract obtained by the preparation method of the licorice extract in an antibacterial and anti-inflammatory traditional Chinese medicine composition is characterized in that the traditional Chinese medicine composition comprises the following components in parts by weight: 20-25 parts of licorice extract, 10-15 parts of Chinese pulsatilla root, 10-15 parts of whiteflower hogfennel root, 10-15 parts of centella, 8-12 parts of pinellia ternate, 8-12 parts of Chinese angelica, 8-12 parts of astragalus, 5-8 parts of Chinese magnoliavine fruit and 5-8 parts of Chinese herbaceous peony.
As a further technical scheme, the preparation method of the traditional Chinese medicine composition comprises the following steps:
SS1, weighing the traditional Chinese medicine composition for later use;
SS2, grinding the Chinese pulsatilla root, the whiteflower hogfennel root, the centella, the pinellia tuber, the Chinese angelica, the astragalus root, the Chinese magnoliavine fruit and the Chinese herbaceous peony into powder, and then sieving the powder with a 80-mesh sieve to obtain mixed Chinese medicinal powder;
SS3, adding water into the mixed traditional Chinese medicine powder for decocting for 3 times, adding water with the weight 10 times of the traditional Chinese medicine powder into the traditional Chinese medicine powder for the first time, soaking for 30min, decocting, and pouring out supernatant; adding water 8 times the weight of the Chinese medicinal powder into the Chinese medicinal powder for the second time, decocting, and collecting supernatant; adding water 6 times the weight of the Chinese medicinal powder for the third time, decocting, and collecting supernatant; collecting supernatant, filtering, concentrating at 65 deg.C under reduced pressure to density of 1.15g/cm 3 To obtain mixed Chinese medicine extract;
SS4, dissolving the licorice extract with absolute ethyl alcohol, and mixing with the mixed traditional Chinese medicine extract to obtain the traditional Chinese medicine composition extract.
The working principle and the beneficial effects of the invention are as follows:
1. the invention can simultaneously improve the content of liquiritin and glycyrrhizic acid in the extract by a two-step extraction method, and the ethanol and the cedrol are compounded and extracted, thereby avoiding the dissolution of a large amount of water-soluble impurities such as sugar, starch and the like and improving the quality of the licorice extract.
2. In the preparation process of the licorice extract, the licorice is extracted in two steps, and in the first step, the licorice is subjected to ultrasonic extraction by adopting ethanol and cedrol, so that liquiritin and glycyrrhizic acid in the licorice can be promoted to be dissolved out; and the second step adopts ethanol and ammonia water for reflux extraction, on one hand, glycyrrhizic acid can generate ammonium salt with a small amount of ammonia water, the solubility of glycyrrhizic acid in water is increased, and further, the content of glycyrrhizic acid in the extract is improved, and on the other hand, the content of liquiritin in the licorice extract can be improved by two times of extraction.
3. In the traditional Chinese medicine composition, the liquorice extract is a main medicine, the Chinese pulsatilla root, the radix peucedani and the centella are added as ministerial medicines, the Chinese pulsatilla root, the radix peucedani and the centella have good antibacterial and anti-inflammatory effects, the centella has the anti-inflammatory and antibacterial effects, the radix peucedani can obviously increase the secretion of respiratory mucus, has the effects of relieving cough and eliminating phlegm, and can assist the liquorice extract to better exert the effects of bacteriostasis, anti-inflammation, relieving cough and relieving asthma, but the three medicines are cold in nature, so that the warm-natured pinellia tuber, the angelica and the astragalus are used as adjuvant medicines to jointly restrict the cold and cold qi of the Chinese pulsatilla root, the radix peucedani and the centella, prevent the cold property from being too high, and further damage the stomach qi of a human body, wherein the pinellia tuber has the effects of drying dampness and eliminating phlegm, calming adverse-rising energy and relieving vomiting, and dispersing lumps; the Chinese angelica has the functions of resisting bacteria and inflammation, protecting the kidney, protecting the liver and relieving asthma, the schisandra and the peony are sour in taste, the schisandra has the efficacy of astringing the lung and relieving cough and asthma, and the schisandra and the peony are used as guiding medicines, so that the effects of dispersing and collecting medicines can be realized, and the whole formula can be prevented from being too warm and dispersing. The liquorice is warm in nature, has the effects of inhibiting bacteria, resisting inflammation, relieving cough, reducing phlegm, clearing heat, removing toxicity, blending various medicines as a guiding medicine, moderating the properties of other medicines, coordinating the properties of other medicines, and making various medicines harmonious with each other, thereby achieving the effect of removing sexual retention and better playing the effects of inhibiting bacteria, diminishing inflammation, relieving cough and relieving asthma, which cannot be achieved when a single medicine is used.
4. The traditional Chinese medicine composition prepared by the invention has better antibacterial and anti-inflammatory effects, and the anti-inflammatory effect of the traditional Chinese medicine composition is better than that of aspirin, so that the traditional Chinese medicine composition not only improves the anti-inflammatory effect, but also reduces the use of anti-inflammatory medicines.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any inventive step, are intended to be within the scope of the present invention.
Example 1
The preparation method of the licorice extract comprises the following steps:
s1, crushing liquorice, sieving the crushed liquorice with a 80-mesh sieve, weighing liquorice powder, adding 60wt% of ethanol and cedrol into the liquorice powder, carrying out ultrasonic extraction, centrifuging and collecting supernate; the mass ratio of the licorice powder to the ethanol is 1:8; the mass ratio of the licorice powder to the cedrol is 1:1.
S2, adding a mixed solution containing 0.6wt% of ammonia water and 60wt% of ethanol into the centrifuged precipitate, performing reflux extraction for 60min, centrifuging, and collecting the supernatant again; the addition amount of the mixed solution is 8 times of the weight of the licorice powder.
S3, combining the supernatants, concentrating at 65 ℃ under reduced pressure until the density is 1.15g/cm 3 Then passing the concentrated solution through macroporous adsorbent resin column, eluting with 5 times column volume of distilled water, eluting with 60wt% ethanol, collecting ethanol eluate, and vacuum drying to obtain Glycyrrhrizae radix extract.
The preparation method of the traditional Chinese medicine composition comprises the following steps:
SS1, weighing the traditional Chinese medicine components according to the weight parts in the table 1 for later use;
SS2, grinding the Chinese pulsatilla root, the whiteflower hogfennel root, the centella, the pinellia tuber, the Chinese angelica, the astragalus root, the Chinese magnoliavine fruit and the Chinese herbaceous peony into powder, and then sieving the powder with a 80-mesh sieve to obtain mixed Chinese medicinal powder;
SS3, adding water into the mixed traditional Chinese medicine powder for decocting for 3 times, adding water with the weight 10 times of the traditional Chinese medicine powder into the traditional Chinese medicine powder for the first time, soaking for 30min, decocting, and pouring out supernatant; adding water 8 times the weight of the Chinese medicinal powder into the Chinese medicinal powder for the second time, decocting, and pouring out supernatant; adding water 6 times the weight of the Chinese medicinal powder for the third time, decocting, and collecting supernatant; collecting supernatant, filtering, concentrating at 65 deg.C under reduced pressure to density of 1.15g/cm 3 To obtain mixed Chinese medicine extract;
SS4, mixing the licorice extract and the mixed traditional Chinese medicine extract to prepare the traditional Chinese medicine composition extract.
Example 2
The preparation method of the licorice extract comprises the following steps:
the process conditions of ultrasonic extraction and reflux extraction were the same as in example 1;
s1, crushing liquorice, sieving with a 80-mesh sieve, weighing liquorice powder, adding 60wt% of ethanol and cedrol into the liquorice powder, performing ultrasonic extraction, centrifuging, and collecting supernate; the mass ratio of the licorice powder to the ethanol is 1:9; the mass ratio of the licorice powder to the cedrol is 2:1;
s2, adding a mixed solution containing 0.5wt% of ammonia water and 60wt% of ethanol into the centrifuged precipitate for reflux extraction, wherein the addition amount of the mixed solution is 6 times of the mass of the licorice powder;
s3 is the same as step S3 in example 1.
The preparation method of the traditional Chinese medicine composition comprises the following steps:
SS1, weighing the traditional Chinese medicine components according to the weight parts in the table 1 for later use;
SS2 and SS3 are the same as SS2 and SS3 in example 1.
Example 3
The preparation method of the licorice extract comprises the following steps:
the process conditions of ultrasonic extraction and reflux extraction were the same as in example 1;
s1, crushing liquorice, sieving the crushed liquorice with a 80-mesh sieve, weighing liquorice powder, adding 60wt% of ethanol and cedrol into the liquorice powder, carrying out ultrasonic extraction, centrifuging and collecting supernate; the mass ratio of the licorice powder to the ethanol is 1; the mass ratio of the licorice powder to the cedrol is 3:1;
s2, adding a mixed solution containing 0.4wt% of ammonia water and 60wt% of ethanol into the centrifuged precipitate for reflux extraction, wherein the addition amount of the mixed solution is 5 times of the mass of the licorice powder;
s3 is the same as step S3 in example 1.
The preparation method of the traditional Chinese medicine composition comprises the following steps:
SS1, weighing the traditional Chinese medicine components according to the weight parts in the table 1 for later use;
SS2 and SS3 are the same as SS2 and SS3 in example 1.
Comparative example 1
Comparative example 1 compared to example 3, replacing cedrol with an equal amount of ethanol, otherwise the same as example 3.
Comparative example 2
Comparative example 2 compared with example 3, the mixed solution containing 0.4wt% of aqueous ammonia and 60wt% of ethanol was replaced with an equal amount of 60wt% ethanol solution, and the other was the same as example 3.
Comparative example 3
Comparative example 3 compared with example 3, except that step S2 was removed, the other example 3.
TABLE 1 Chinese medicinal composition comprising Chinese medicinal materials in parts by weight
Experimental example 1
And (3) determining the content of liquiritin and glycyrrhizic acid in the licorice extract: the content of glycyrrhizin and glycyrrhizic acid in liquorice is measured by referring to a method for measuring the content of glycyrrhizin and glycyrrhizic acid in liquorice in 'Chinese pharmacopoeia' 2015 edition.
TABLE 2 Glycyrrhiza glycoside and Glycyrrhizic acid content of Glycyrrhiza extract
| Glycyrrhizin (%) | Glycyrrhizic acid (%) | |
| Example 1 | 39.59 | 11.25 |
| Example 2 | 38.87 | 10.55 |
| Example 3 | 38.12 | 10.25 |
| Comparative example 1 | 36.12 | 10.05 |
| Comparative example 2 | 37.55 | 9.58 |
| Comparative example 3 | 32.25 | 8.21 |
As can be seen from table 2, in the present invention, compared with example 3, comparative example 1 replaced cedrol with equal amount of ethanol, and both the content of glycyrrhizin and glycyrrhizic acid in the obtained extract were reduced, indicating that compounding cedrol and ethanol promotes the dissolution of glycyrrhizin and glycyrrhizic acid; compared with the example 3, the comparative example 2 replaces the mixed solution containing 0.4wt% of ammonia water and 60wt% of ethanol with the same amount of 60wt% of ethanol solution, the content of glycyrrhizin and glycyrrhizic acid in the obtained extract is reduced, and the glycyrrhizic acid content is reduced more, which indicates that the content of glycyrrhizic acid cannot be increased by carrying out two times of extraction without adding ammonia water; comparative example 3 compared with example 3, only one extraction was performed, and glycyrrhizin and glycyrrhizic acid in the obtained licorice extract were significantly reduced.
Experimental example 2
The method for measuring the bacteriostatic activity of the traditional Chinese medicine composition comprises the following steps: respectively and uniformly coating Escherichia coli and Staphylococcus aureus on a beef extract peptone solid culture medium by a cup butterfly method, placing 4 sterile Oxford cups at equal intervals, and respectively injecting 200L of Chinese medicinal composition into the cups, wherein the final concentrations of the Chinese medicinal compositions are 4, 6, 8, 16 and 24mg/mL respectively, and absolute ethyl alcohol is used as a reference. Each group was subjected to 3 parallel experiments, incubated at 37 ℃ in an incubator for 24 hours, and then the diameter of the zone of inhibition was measured with a ruler, and the average value was taken.
TABLE 3 diameter of zone of inhibition of Chinese medicinal composition on Staphylococcus aureus and Escherichia coli
Table 3 shows that the Chinese medicinal composition has an antibacterial effect on Staphylococcus aureus and Escherichia coli, and the Chinese medicinal composition can inhibit the growth of Staphylococcus aureus and Escherichia coli as shown in Table 3.
Experimental example 3
The method for measuring the anti-inflammatory effect of the traditional Chinese medicine composition comprises the following steps: the traditional Chinese medicine composition is prepared in the example 1, and the anti-inflammatory effect is observed by adopting a xylene-induced mouse auricle swelling test model;
100 male mice are randomly divided into five groups, namely a blank control group (given with equal amount of normal saline), a positive control group (0.2 g/kg aspirin), a low, medium and high dosage group of the traditional Chinese medicine composition are continuously given for 7 days according to 0.5g/10g, and after 30min of the last administration, xylene is coated on the two sides of the right ear according to 0.03 mL/one, and the left ear is used as a control. After 30min, cervical dislocation was sacrificed and ears were cut along the base line of the auricle. The ear pieces were removed from the symmetrical parts of the left and right ears by a punch having a diameter of 7mm and weighed. Observing the auricle swelling condition of the mouse, and calculating the swelling inhibition rate;
ear swelling degree (mg) = right auricle mass-left auricle mass;
swelling inhibition (%) = (average ear swelling degree of blank group-average ear swelling degree of administration group)/average ear swelling degree of blank group × 100%.
TABLE 4 Effect of xylene on ear swelling in mice
| Group of | Dosage (g/kg) | Swelling of the skinInhibition ratio (%) |
| Blank control group | - | - |
| Aspirin group | 0.2 | 49.8 |
| Low-dose group of traditional Chinese medicine composition | 1 | 40.2 |
| Medium-dosage group of traditional Chinese medicine composition | 1.5 | 43.5 |
| High dosage group of Chinese medicinal composition | 3 | 52.8 |
The anti-inflammatory test results in table 4 show that the traditional Chinese medicine composition can inhibit mouse auricle swelling induced by xylene, has a good anti-inflammatory effect, and the anti-inflammatory effect of a high-dose group of the traditional Chinese medicine composition is superior to that of aspirin.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. The preparation method of the licorice extract is characterized by comprising the following steps:
s1, weighing licorice powder, adding ethanol and cedrol into the licorice powder for extraction, centrifuging and collecting supernatant;
s2, adding a mixed solution of ammonia water and ethanol into the centrifuged precipitate for extraction, centrifuging and collecting the supernatant again;
and S3, combining the supernatants, concentrating, purifying and drying in vacuum to obtain the licorice extract.
2. The method for preparing a licorice extract according to claim 1, wherein the licorice powder in step S1 is obtained by pulverizing licorice and sieving the pulverized licorice through a 80-mesh sieve.
3. The method for preparing a licorice extract according to claim 1, wherein the mass ratio of licorice powder to ethanol in step S1 is 1; the mass ratio of the licorice powder to the cedrol is 1:1-3:1.
4. The method of claim 3, wherein the ethanol is 60wt% ethanol.
5. The method for preparing a licorice extract according to claim 1, wherein the extraction in step S1 is ultrasonic extraction, the extraction temperature is 60 ℃, the extraction time is 60min, and the extraction power is 250W; the extraction in the step S2 is reflux extraction, and the extraction time is 60min.
6. The method of preparing licorice extract according to claim 1, wherein the mixed solution of ammonia water and ethanol in step S2 contains 0.4-0.6wt% of ammonia water and 60wt% of ethanol.
7. The method of claim 1, wherein the amount of the mixed solution of ammonia and ethanol added in step S2 is 5-8 times the weight of the licorice powder.
8. The method of claim 1, wherein the step S3 is carried out by concentrating the extract to obtain a concentrated extractConcentrating the combined supernatant at 65 deg.C under reduced pressure to density of 1.15g/cm 3 (ii) a Purifying by passing the concentrated solution through macroporous adsorbent resin column, eluting with 2-5 times of column volume of distilled water, eluting with 60-80% ethanol, and collecting ethanol eluate.
9. The application of the licorice extract obtained by the preparation method of the licorice extract according to any one of claims 1-8 in an antibacterial and anti-inflammatory traditional Chinese medicine composition, wherein the traditional Chinese medicine composition comprises the following components in parts by weight: 20-25 parts of licorice extract, 10-15 parts of Chinese pulsatilla root, 10-15 parts of whiteflower hogfennel root, 10-15 parts of centella, 8-12 parts of pinellia tuber, 8-12 parts of Chinese angelica, 8-12 parts of astragalus root, 5-8 parts of Chinese magnoliavine fruit and 5-8 parts of Chinese herbaceous peony.
10. The application of the licorice extract obtained by the preparation method of the licorice extract according to claim 9 in an antibacterial and anti-inflammatory traditional Chinese medicine composition, wherein the preparation method of the traditional Chinese medicine composition comprises the following steps:
SS1, weighing the traditional Chinese medicine composition as claimed in claim 9 for later use;
SS2, grinding the Chinese pulsatilla root, the whiteflower hogfennel root, the centella, the pinellia tuber, the Chinese angelica, the astragalus root, the Chinese magnoliavine fruit and the Chinese herbaceous peony into powder, and then sieving the powder with a 80-mesh sieve to obtain mixed Chinese medicinal powder;
SS3, adding water into the mixed traditional Chinese medicine powder for decocting for 3 times, adding water with the weight 10 times of the traditional Chinese medicine powder into the traditional Chinese medicine powder for the first time, soaking for 30min, decocting, and pouring out supernatant; adding water 8 times the weight of the Chinese medicinal powder into the Chinese medicinal powder for the second time, decocting, and pouring out supernatant; adding water 6 times the weight of the Chinese medicinal powder for the third time, decocting, and collecting supernatant; collecting supernatant, filtering, concentrating at 65 deg.C under reduced pressure to density of 1.15g/cm 3 To obtain mixed Chinese medicine extract;
SS4, dissolving the licorice extract with absolute ethyl alcohol, and mixing with the mixed traditional Chinese medicine extract to obtain the traditional Chinese medicine composition extract.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005089402A (en) * | 2003-09-19 | 2005-04-07 | Ogawa & Co Ltd | External anti-pruritic agent |
| WO2014115726A1 (en) * | 2013-01-25 | 2014-07-31 | 株式会社ダイセル | Antibacterial composition |
| CN105311131A (en) * | 2015-06-16 | 2016-02-10 | 厦门医学高等专科学校 | Traditional Chinese medicine composition with antibacterial and anti-inflammation functions and preparation method and application of traditional Chinese medicine composition with antibacterial and anti-inflammation functions |
| CN106109644A (en) * | 2016-03-04 | 2016-11-16 | 广西梧州制药（集团）股份有限公司 | A kind of Chinese medicine composition of antipyretic and anti-inflammatory and preparation method thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005089402A (en) * | 2003-09-19 | 2005-04-07 | Ogawa & Co Ltd | External anti-pruritic agent |
| WO2014115726A1 (en) * | 2013-01-25 | 2014-07-31 | 株式会社ダイセル | Antibacterial composition |
| CN105311131A (en) * | 2015-06-16 | 2016-02-10 | 厦门医学高等专科学校 | Traditional Chinese medicine composition with antibacterial and anti-inflammation functions and preparation method and application of traditional Chinese medicine composition with antibacterial and anti-inflammation functions |
| CN106109644A (en) * | 2016-03-04 | 2016-11-16 | 广西梧州制药（集团）股份有限公司 | A kind of Chinese medicine composition of antipyretic and anti-inflammatory and preparation method thereof |
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