CN115137766B

CN115137766B - Preparation method of anti-rheumatism traditional Chinese medicine preparation, traditional Chinese medicine preparation and application

Info

Publication number: CN115137766B
Application number: CN202210963560.1A
Authority: CN
Inventors: 兰卫; 李建梅
Original assignee: Xinjiang Medical University
Current assignee: Xinjiang Medical University
Priority date: 2022-08-11
Filing date: 2022-08-11
Publication date: 2023-07-07
Anticipated expiration: 2042-08-11
Also published as: CN115137766A

Abstract

The invention provides a preparation method of an antirheumatic traditional Chinese medicine preparation, a traditional Chinese medicine preparation and application, wherein the traditional Chinese medicine preparation comprises 14-16 parts of geranium, 5-7 parts of liquorice, 11-13 parts of radix angelicae pubescentis, 11-13 parts of gentiana macrophylla, 9-11 parts of elder, 9-11 parts of rhizoma corydalis, 9-11 parts of angelica sinensis and 7-9 parts of chamomile according to parts by weight, and has the effects of dispelling wind-cold dampness and clearing and activating the channels and collaterals to relieve pain.

Description

Preparation method of anti-rheumatism traditional Chinese medicine preparation, traditional Chinese medicine preparation and application

Technical Field

The invention relates to the technical field of traditional Chinese medicines, in particular to a preparation method of an antirheumatic traditional Chinese medicine preparation, a traditional Chinese medicine preparation and application.

Background

Rheumatoid arthritis is an autoimmune arthritis, often causes joint destruction and deformity, is easy to cause various complications, seriously affects the life quality of patients, has high morbidity and disability rate, has long disease course, and has great harm to physical and mental health of patients. The main purpose of rheumatoid arthritis treatment is to reduce joint inflammation, inhibit lesion development and irreversible bone destruction, protect functions of joints and muscles as much as possible, and finally achieve the aim of completely relieving illness state or reducing illness activity. At present, non-steroidal anti-inflammatory drugs, glucocorticoids, biological agents and the like are mostly adopted for Western medicine treatment, but the drugs can only be used as auxiliary treatment, and long-term use of the drugs can not improve disease development and can also cause serious side effects. One trend in current rheumatoid therapy is to shift to biological antirheumatic drug therapy and traditional drug therapy to prevent and delay progression of joint damage in patients. The Chinese medicine has the functions of multiple layers, multiple components and multiple targets, so that the Chinese patent medicine has certain advantages in treating the disease.

The compound geranium wilfordii recipe is a traditional meridian recipe for treating rheumatism by Kazakhstan, and has the effects of dispelling wind-cold-dampness, dredging meridian passage and relieving pain. The traditional Chinese medicine compound is inconvenient to carry and use, has unstable drug effect and other technical problems, and causes great barriers to the use and popularization of the compound geranium formula, and the research on the process of the compound geranium formula preparation is not reported in the prior art. Although the prior art has the technical proposal of the extraction process of the traditional Chinese medicine decoction pieces, a great amount of impurities and macromolecular substances are usually accompanied while the ingredients are obtained, so that patients need to take larger dosage to ensure the curative effect when taking the traditional Chinese medicine decoction pieces. In addition, because the extract powder has stronger hygroscopicity, excessive instability can be shown in the actual production process of the preparation, and although the hygroscopicity of the extract can be reduced to a certain extent by adding auxiliary materials, the effect is often poor, and the addition of the auxiliary materials can also increase the volume of the preparation, so that the dosage of patients is increased. Therefore, a preparation which not only ensures curative effect, but also is convenient to carry and use and controllable and stable in quality needs to be developed.

Disclosure of Invention

In order to solve the technical problems, the invention aims to provide a preparation method of an anti-rheumatism traditional Chinese medicine preparation, the traditional Chinese medicine preparation and application.

In order to achieve the above object, the present invention provides the following technical solutions:

the preparation method of the anti-rheumatism traditional Chinese medicine preparation comprises the following raw materials in parts by weight:

14-16 parts of geranium, 5-7 parts of liquorice, 11-13 parts of radix angelicae pubescentis, 11-13 parts of gentiana macrophylla, 9-11 parts of elder, 9-11 parts of rhizoma corydalis, 9-11 parts of angelica sinensis and 7-9 parts of chamomile;

the preparation method comprises the following steps:

taking geranium, liquorice and chamomile according to parts by weight, extracting with water under reflux, and refining to obtain an aqueous extract;

reflux-extracting radix Gentianae Marcrophyllae, radix Angelicae sinensis, ramulus Sambuci Williamsii, rhizoma corydalis and radix Angelicae Pubescentis with ethanol solution to obtain ethanol extract;

concentrating the water extract and the alcohol extract under reduced pressure respectively, drying to obtain extract fine powder, adding binder and part of raw material fine powder, mixing and granulating.

Preferably, the refining comprises: concentrating the water extract under reduced pressure until the ratio of the mass of the medicinal materials to the volume of the liquid medicine is 1 (1-1.2), standing, and filtering to obtain supernatant.

Preferably, the amount of the part of raw material fine powder is 5-10% of the total weight of the extract fine powder.

Preferably, the partial raw material is selected from one or two of angelica and radix angelicae pubescentis.

Preferably, the binder is an ethanol solution with a concentration of 92-93%.

Preferably, the concentration of the ethanol solution is 65-75%, the volume of the added ethanol solution is 9-11 times of the weight of the raw materials, the reflux extraction is carried out for 1-3 times, and each time of extraction is 80-120 min.

Preferably, the temperature of the reduced pressure concentration is 60-80 ℃.

Preferably, the drying temperature is 60-80 ℃.

The invention also provides a traditional Chinese medicine preparation prepared by the preparation method.

Preferably, the traditional Chinese medicine preparation is in the form of granules.

The invention also provides application of the traditional Chinese medicine preparation in preparing a medicine for treating rheumatoid arthritis.

Compared with the prior art, the invention has the following beneficial effects:

(1) According to the preparation method, the natural sedimentation method is used for removing impurities and refining the water extract, so that the cost is low, compared with other impurity removing methods such as the water extraction and alcohol sedimentation method, the high-speed centrifugation method, the clarifier adsorption method and the like, the impurity removing effect is better, the effective components can be reserved to the greatest extent, and meanwhile, the production cost is lower.

(2) The preparation method only uses part of raw material fine powder as a filler, can reduce the moisture absorption rate of the extract and increase the stability of the preparation production process. In addition, as only part of raw material fine powder is used as the filling agent, the finally prepared preparation does not contain other auxiliary materials irrelevant to the drug effect, and the dosage of patients is reduced.

(3) The solvent dosage, the extraction time, the extraction temperature and the drying temperature in the preparation method can furthest extract the effective components in the raw materials, improve the drug effect of the preparation and reduce the dosage of patients.

(4) The traditional Chinese medicine preparation has the effects of dispelling wind-cold-dampness, clearing and activating the channels and collaterals and relieving pain, has obvious curative effect and has unique advantages for treating rheumatoid arthritis.

Detailed Description

The invention provides a preparation method of an antirheumatic traditional Chinese medicine preparation, the traditional Chinese medicine preparation and application, and all raw material components are commercial products well known to those skilled in the art unless specified.

In the invention, the preparation method comprises the following raw materials in parts by weight: 14-16 parts of geranium, 5-7 parts of liquorice, 11-13 parts of radix angelicae pubescentis, 11-13 parts of gentiana macrophylla, 9-11 parts of sambucus chinensis, 9-11 parts of rhizoma corydalis, 9-11 parts of angelica sinensis and 7-9 parts of chamomile, and the preparation method is that: 15 parts of geranium, 6 parts of liquorice, 12 parts of radix angelicae pubescentis, 12 parts of gentiana macrophylla, 10 parts of elder, 10 parts of rhizoma corydalis, 10 parts of angelica sinensis and 8 parts of chamomile.

In the invention, the medicines are used in proportion to have the effects of dispelling wind, dredging collaterals and strengthening tendons and bones. Specifically, the geranium wilfordii is bitter and flat, has the effects of dispelling wind and dredging collaterals, strengthening tendons and bones, and tonifying middle-jiao and xies, and is a monarch drug. Qin Marcrophylla is good at dispelling wind-dampness, relaxing tendons and activating collaterals, relieving joint pain, and penetrating skin to exogenous pathogenic factors; radix Angelicae Pubescentis, with pungent and warm natured drugs, is good at dispelling pathogenic wind, cold and dampness from joints; chamomile has the effects of diminishing inflammation and relieving pain, and the chamomile, the chamomile and the chamomile are all ministerial drugs. Rhizoma corydalis has effects of promoting blood circulation, removing blood stasis and relieving pain; bone fracture setting, tendon reunion, blood circulation promoting, pain relieving, wind dispelling and dampness eliminating effects are taken as adjuvant drugs; dang Gui is a drug for regulating blood. Licorice root, radix Glycyrrhizae has the effects of clearing heat and detoxicating, relieving spasm and pain, and harmonizing other drugs, and has the actions of tonifying and strengthening qi, and is a side-effect drug. The seven medicines together play the roles of dispelling wind-cold dampness, dredging the meridian passage and relieving pain.

The preparation method of the invention comprises the following steps:

concentrating and drying the water extract and the alcohol extract respectively to obtain extract fine powder, adding the adhesive and part of raw material fine powder, mixing and granulating.

In the invention, the geranium, the chamomile and the liquorice are extracted by water reflux, so that the main components of gallic acid in the geranium, polysaccharide in the liquorice and flavonoid active substances rich in the chamomile are obtained, and the components can regulate inflammatory factors and play an anti-inflammatory role.

Specifically, the water reflux extraction comprises the following steps: soaking geranium, liquorice and chamomile in water with the volume of 8-12 times of the weight of the raw materials for 90-150 min, and reflux-extracting for 1-3 times, wherein each time is 60-120 min. In the water reflux extraction, the water addition ratio is preferably 10 to 12 times, more preferably 11 times. The soaking time is preferably 120 to 150min, more preferably 130min. The number of times of the reflux extraction is preferably 2 to 3 times, more preferably 2 times. The time for each extraction is preferably 60 to 100min, more preferably 90min.

In the invention, the geranium, licorice and jersey Gan Jushui extract needs to be refined, so as to remove impurities and macromolecular components irrelevant to the drug effect, reduce the extract yield, improve the product quality and reduce the dosage of patients. At present, the impurity removal refining is carried out by adopting methods such as a water extraction and alcohol precipitation method, a macroporous resin adsorption method, a high-speed centrifugation method, an adsorption clarification method and the like in the industry, but researches show that compared with other methods such as a water extraction and alcohol precipitation method, a high-speed centrifugation method, a clarifier method and the like, the method can furthest retain the index components in the traditional Chinese medicine extract and can furthest reduce the extract yield by using a natural sedimentation method.

Specifically, the natural sedimentation method comprises the following steps: concentrating the water extract under reduced pressure to obtain medicinal materials: 1 to 1.2 percent of liquid medicine, and filtering and taking supernatant after standing. The medicinal materials are as follows: the liquid medicine refers to the ratio of the weight of solute in the water extract to the volume of the liquid medicine, and is preferably 1:1.1. Preferably, the pressure of the reduced pressure concentration is-0.08 to 0.07Mpa. Preferably, the temperature of the reduced pressure concentration is 60 to 80 ℃, more preferably 65 to 75 ℃, and even more preferably 70 ℃. As an alternative embodiment, the temperature of the standing may be 2 to 8 ℃, preferably 4 ℃, and the time of the standing may be 20 to 30 hours, preferably 24 hours.

In the invention, gentiana macrophylla, angelica sinensis, elderberry, rhizoma corydalis and radix angelicae pubescentis are extracted by an alcohol extraction method, so that the active ingredients of ferulic acid in angelica sinensis, osthole in radix angelicae pubescentis, gentiopicroside in gentiana macrophylla, iridoid compounds in elderberry and alkaloids in rhizoma corydalis are obtained, and the above ingredients have the effects of anti-inflammatory, analgesic, rheumatoid arthritis treatment and the like.

Specifically, the ethanol reflux extraction comprises the following steps: mixing radix gentianae macrophyllae, chinese angelica, elder, rhizoma corydalis and radix angelicae pubescentis, adding ethanol solution with the volume of 9-11 times of the weight of the raw materials and the concentration of 65-75%, and carrying out reflux extraction for 1-3 times, wherein each time of extraction is 80-100 min. The multiple of the addition of the ethanol solution is preferably 10 times. The concentration of the ethanol solution is preferably 70%. The number of reflux extractions is preferably 2 to 3, more preferably 2. The time for each extraction is preferably 90min.

Preferably, the alcohol extract and the refined water extract are respectively concentrated under reduced pressure at 60-80 ℃ and-0.08-0.07 Mpa until the relative density is 1.20-1.25 and 1.15-1.20, then are dried under vacuum at 60-80 ℃ to respectively obtain alcohol extract and water extract dry extract, the dry extract is crushed and sieved to obtain fine powder, the fine powder is uniformly mixed with auxiliary materials, and the mixture is added with an adhesive for granulating. Preferably, the fine powder of the dry extract is required to pass through a 80-100 mesh sieve.

In the present invention, the temperature at which the alcohol extract and the aqueous extract are concentrated under reduced pressure is 60 to 80 ℃, preferably 65 to 75 ℃, more preferably 70 ℃. The temperature of the vacuum drying is 60 to 80 ℃, preferably 65 to 75 ℃, more preferably 70 ℃. According to the invention, the contents of ferulic acid, osthole and gentiopicroside in the alcohol extract and the content of gallic acid in the water extract are high in the temperature range, the loss is minimum, and the content is reduced when the temperature range is exceeded.

In the invention, only part of fine powder of medicinal raw materials is used as a filler. Firstly, the content of active ingredients in the preparation can be increased by using the medicine fine powder as a filling agent, so that the addition of ingredients irrelevant to the medicine effect is reduced; and secondly, researches show that the hygroscopicity of the extract is too strong, and only part of the fine powder of the raw material medicaments in the formula is added as a filler, so that the problem of the strong hygroscopicity of the fine powder of the extract can be effectively solved, the forming rate and the mobile phase are improved, and the stability of the preparation is improved compared with other types of fillers such as sucrose and CMC-Na.

Specifically, the fine powder of part of the medicinal materials is selected from one or more of herba Erodii seu Geranii, glycyrrhrizae radix, radix Angelicae Pubescentis, radix Gentianae Marcrophyllae, ramulus Sambuci Williamsii, rhizoma corydalis, and radix Angelicae sinensis, preferably radix Angelicae sinensis and/or radix Angelicae Pubescentis, and more preferably radix Angelicae Pubescentis. Preferably, the fine powder of the medicinal materials is sieved by a sieve of 80-100 meshes. The addition amount of the medicinal fine powder is 5-10% of the total dry extract, preferably 6-9%, more preferably 8% by weight.

In the present invention, an ethanol solution having a concentration of 92 to 93% is preferably used as the binder, and an ethanol solution having a concentration of 93% is more preferably used. The preparation method provided by the invention only uses part of medicinal material raw material fine powder as a filler, does not use other types of fillers such as starch, dextrin, sucrose and the like, and does not add other auxiliary materials except the medicinal material fine powder and the ethanol solution, so when the ethanol solution is used as a binder, the concentration of the ethanol is very critical, the difficulty degree of granulation, the appearance of particles and the like can be influenced due to too high or too low concentration, the problems of over-adhesion, difficult particle sizing, difficult sieving and the like occur due to too low concentration of the ethanol solution, the soft material is too dry due to too high concentration of the ethanol, the particles are loose, too much fine powder, and the granulation cannot be completed due to too high or too low concentration.

The invention provides an antirheumatic traditional Chinese medicine preparation, which is prepared by the preparation method. As an alternative embodiment, the formulation contains the particles obtained by the above preparation method and a pharmaceutically acceptable carrier. Preferably, the dosage form can be an oral preparation, can be granules, capsules, emplastrum, tablets, sustained-release preparations or the like, and more preferably, the dosage form is granules.

The technical solutions of the present invention will be clearly and completely described in the following in connection with the embodiments of the present invention. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Example 1

15g of geranium, 6g of liquorice and 8g of chamomile are taken according to parts by weight, 11 times of water is added, the mixture is soaked for 130min, reflux extraction is carried out for 2 times, each time of extraction is carried out for 90min, and the water extract is decompressed and concentrated to the medicinal materials at 70 ℃: the liquid medicine is 1:1.1, and the supernatant is filtered after being placed for 24 hours under the low temperature environment of 4 ℃.

Taking 12g of gentiana macrophylla, 10g of Chinese angelica, 10g of elder, 10g of rhizoma corydalis and 12g of radix angelicae pubescentis according to parts by weight, crushing part of radix angelicae pubescentis raw materials, sieving with a 90-mesh sieve to obtain radix angelicae pubescentis fine powder, mixing the rest raw materials, adding 10 times of 70% ethanol solution, carrying out reflux extraction for 2 times, and extracting for 90min each time.

Concentrating the water extract and the alcohol extract at 70 ℃ under reduced pressure respectively until the relative density is 1.15-1.20 and 1.20-1.25, then drying in vacuum at 70 ℃, and crushing to obtain extract fine powder. Taking 93% ethanol solution as binder, adding extract fine powder and radix Angelicae Pubescentis fine powder of 8% of the weight of the extract powder, mixing, and granulating to obtain granule.

Example 2

The preparation method was the same as in example 1, except that after the completion of granulation, the granules were filled into capsules to prepare capsules.

Example 3

Compared with the example 1, the gentiana macrophylla, the angelica sinensis, the elder, the rhizoma corydalis and the radix angelicae pubescentis are mixed and then added with 10 times of 70% ethanol solution, and the mixture is extracted under reflux for 3 times, each time for 90min, and the rest are the same.

Example 4

Compared with the example 1, the water extract is decompressed and concentrated to the medicinal materials at 70 ℃ during the refining: the liquid medicine is 1:1, the liquid medicine is placed for 24 hours under the low temperature environment of 4 ℃ and then filtered, the supernatant is taken, and the rest are the same.

Example 5

Compared with the example 1, the water extract is decompressed and concentrated to the medicinal materials at 70 ℃ during the refining: the liquid medicine is 1:2, the liquid medicine is placed for 24 hours under the low temperature environment of 4 ℃ and then filtered, the supernatant is taken, and the rest are the same.

Example 6

In comparison with example 1, the aqueous extract and the alcoholic extract were concentrated under reduced pressure at 60℃respectively, the remainder being the same.

Example 7

In comparison with example 1, the concentrated aqueous and alcoholic extracts were dried at 60℃under vacuum, the remainder being the same.

Example 8

In comparison with example 1, an ethanol solution having a concentration of 92% was used as a binder, and the remainder were the same.

Example 9

The amount of the fine powder of radix Angelicae Pubescentis added was 10% as compared with example 1, and the rest was the same.

Example 10

Compared with the example 1, the rest part of the Chinese angelica raw material is crushed and passes through a 90-mesh screen, the dosage is 8% of the weight of the extract powder, and the rest is the same.

Example 11

Compared with example 1, the rest part of the Chinese angelica and the radix angelicae pubescentis fine powder are crushed and pass through a 90-mesh screen, and the Chinese angelica fine powder accounting for 5% of the weight of the extract powder and the radix angelicae pubescentis fine powder accounting for 5% of the weight of the extract powder are used as filling agents, and the rest parts are the same.

Example 12

The preparation method was the same as in example 1 except that after the completion of granulation, the granules were compressed into tablets and prepared into tablets.

Example 13

Concentrating the water extract and the alcohol extract at 70 ℃ under reduced pressure respectively until the relative density is 1.15-1.20 and 1.20-1.25, then drying in vacuum at 70 ℃, and crushing to obtain extract fine powder. Adding 580g vaseline into the extract fine powder and radix Angelicae Pubescentis fine powder with weight of 8% of the extract powder, stirring to obtain Chinese medicinal paste, spreading the Chinese medicinal paste (thickness 1.5cm, diameter 8 cm) on plaster cloth, and making into Chinese medicinal patch.

Comparative example 1

Compared with the example 1, the gentiana macrophylla, the angelica sinensis, the elder, the rhizoma corydalis and the radix angelicae pubescentis are mixed and then added with 8 times of 80% ethanol solution, and the mixture is extracted for 2 times under reflux, each time for 90min, and the rest are the same.

Comparative example 2

Compared with the example 1, the gentiana macrophylla, the angelica sinensis, the elder, the rhizoma corydalis and the radix angelicae pubescentis are mixed and then added with 10 times of 80% ethanol solution, and the mixture is subjected to reflux extraction for 2 times, each time for 120min, and the rest are the same.

Comparative example 3

Compared with the example 1, the gentiana macrophylla, the angelica sinensis, the elder, the rhizoma corydalis and the radix angelicae pubescentis are mixed and then added with 12 times of 70% ethanol solution, and the mixture is subjected to reflux extraction for 2 times, each time for 120min, and the rest are the same.

Comparative example 4

Compared with example 1, the aqueous extract was purified by centrifugation at 3000rpm for 20min to obtain a supernatant filtrate, and the remainder were the same.

Comparative example 5

Compared with example 1, the water extract was purified by water extraction and alcohol precipitation, absolute ethanol was added to the water extract to a concentration of 60%, and the mixture was left to stand at 4℃for 24 hours and filtered, and the rest was the same.

Comparative example 6

Compared with the example 1, the water extract is refined by using a chitosan clarification method, the chitosan solution with the volume of 4% of the water extract is slowly added into the water extract in a trickle way, the water extract is uniformly mixed and then is put into a water bath kettle with the temperature of 60 ℃ for heat preservation for 1h, then is kept stand for 24h at the low temperature of 4 ℃ and then is filtered, and the rest is the same.

Comparative example 7

Compared with the example 1, the ZTC1+1-II natural clarifying method is used for refining the water extract, the natural clarifying agent A component accounting for 8% of the volume of the water extract is slowly added firstly, then the water extract is put into a water bath at 60 ℃ for 1h and then is taken out, the natural clarifying agent B component accounting for 4% of the volume of the water extract is slowly added, then the water extract is put into the water bath at 60 ℃ for 1h and then is taken out, the water extract is kept stand for 24h in a low-temperature environment at 4 ℃ and is filtered, and the rest is the same.

Comparative example 8

In comparison with example 1, the aqueous extract and the alcoholic extract were concentrated under reduced pressure at 50℃respectively, the remainder being the same.

Comparative example 9

In comparison with example 1, the aqueous extract and the alcoholic extract were concentrated under reduced pressure at 90℃respectively, the remainder being the same.

Comparative example 10

In comparison with example 1, the concentrated aqueous extract and the alcoholic extract were dried at 50℃under vacuum, the remainder being the same.

Comparative example 11

In comparison with example 1, the concentrated aqueous extract and the alcoholic extract were dried at 90℃under vacuum, the remainder being the same.

Comparative example 12

In comparison with example 1, ethanol with a concentration of 88% was used as binder, the remainder being the same.

Comparative example 13

In comparison with example 1, ethanol with a concentration of 95% was used as binder, the remainder being the same.

Comparative example 14

Compared with example 1, 10% of sucrose was used as filler based on the total weight of the dry extract, and the rest was the same.

Comparative example 15

In comparison with example 1, lactose was used as filler in an amount of 10% by weight based on the total weight of the dry extract, the remainder being the same.

Comparative example 16

As compared with example 1, CMC-Na was used as filler in an amount of 10% by weight based on the total dry extract weight, the remainder being the same.

Test example 1

1. Detection method

(1) Determination of extractum yield

Placing 50mL of sample solution into a dried constant-weight evaporation dish, evaporating to dryness, transferring into a 105 ℃ oven for drying for 3 hours, placing into a dryer for cooling and drying for 1 hour, precisely weighing, and calculating the extract yield according to the following formula:

(2) Method for measuring gentiopicroside content in extract

The measurement was performed by high performance liquid chromatography, and the chromatographic conditions and standard information were as follows:

chromatographic column: agilent C18, 250mm x 4.6mm,5 μm; mobile phase: methanol-water= (22:78); sample injection amount: 5. Mu.L; volume flow rate: 1.0mL/min; column temperature: 30 ℃; detection wavelength: 270nm; the gentiopicroside reference substance is provided by Chinese food and drug verification institute, and has batch number of 110770-201918.

(3) Method for measuring content of ferulic acid and osthole in extract

chromatographic column: agilent C18 column, 250mm 4.6mm,5 μm; the mobile phases A and B are acetonitrile and 0.1% phosphoric acid solution mobile phases respectively, and gradient elution is carried out; sample injection amount: 10. Mu.L; volume flow rate: 1.0mL/min; detection wavelength: 320nm; column temperature: 30 ℃; the reference substance is provided by Chinese food and drug verification institute, with ferulic acid batch number 110773-201614, osthole batch number 110822-201710.

(4) Method for measuring gallic acid content in water extract

chromatographic column: agilent C18, 250mm x 4.6mm,5 μm; mobile phase: methanol-0.2% phosphoric acid (5:95); sample injection amount: 5L; volume flow rate: 1.0mL/min; detection wavelength: 266nm; gallic acid reference substance is provided by Chinese food and drug inspection institute, and has batch number of 110831-201605.

(5) Determination of the content of Total polysaccharide

The total polysaccharide content was determined using uv-vis spectrophotometry. Taking a proper amount of D-anhydrous glucose reference substance, precisely weighing, placing into a 10mL volumetric flask, and fixing the volume to a scale with pure water to obtain a reference substance solution.

And precisely transferring 0.3mL of the sample solution, 0.3mL of the D-anhydrous glucose control solution by a pipette, placing the sample solution in a 10mL glass test tube, adding 1mL of 5% phenol solution and 5mL of concentrated sulfuric acid solution, mixing, standing for 10min, then placing the test tube in boiling water, heating for 20min, taking out the test tube, rapidly cooling the mixture to room temperature by cold water, and detecting absorbance at 483.2 nm. The content was calculated by standard curve method.

(6) Measurement of Forming Rate

And weighing a proper amount of prepared compound geranium granules, and calculating the forming rate by taking the qualified compound geranium granules which pass through a No. 1 sieve and cannot pass through a No. 5 sieve.

Molding rate (%) =acceptable pellet weight/total pellet weight×100%.

(7) Determination of moisture absorption Rate

And (3) taking a saturated sodium chloride solution dryer, placing the dryer in a constant temperature incubator at 25 ℃ for 24 hours, and placing a temperature and humidity recorder in the dryer to keep the internal humidity of the vessel constant at 75%. Precisely weighing 2g of uniformly mixed particles, placing the uniformly mixed particles in a constant-weight flat weighing bottle, precisely weighing the total weight, placing the obtained product in the dryer (25 ℃, opening the bottle cap), fully absorbing moisture by the particles after 2 days, and calculating the moisture absorption rate.

Moisture absorption (%) = (weight after moisture absorption-weight before moisture absorption)/weight before moisture absorption×100%.

(8) Measurement of the angle of repose

2. Detection result

(1) Alcohol extraction process performance detection result

The composite score is calculated according to the following formula:

composite score = gentiopicroside/maximum content x 0.4+ ferulic acid/maximum content x 0.2+ osthole/maximum content x 0.2+ extract yield/maximum extract amount x 0.2

TABLE 1 results of alcohol extraction Process Performance test

As can be seen from Table 1, the three active ingredients of ferulic acid, osthole and gentiopicroside in the extract obtained by the alcohol extraction process of examples 1 and 3 have high contents, and the comprehensive score is above 95. Comparative examples 1 to 3 were lower in the content of three active ingredients in the extract than examples 1 and 3 due to unsuitable ethanol concentration, extraction time and fold of the ethanol solution. It can be seen that the effect of the alcohol extraction process of the present invention is superior to that of comparative examples 1 to 3.

(2) Performance test results of refining method

The composite score is calculated according to the following formula:

comprehensive score = gallic acid retention/maximum retention value x 0.4+ total polysaccharide retention/maximum retention value x 0.3+ extract yield reduction rate/maximum reduction rate x 0.3

Table 2 results of refining process performance test

As can be seen from Table 2, the refining methods of examples 1, 4 and 5 can effectively reduce the extract yield, and retain gallic acid and polysaccharide in the aqueous extract to the maximum extent, and the comprehensive scores are above 90. Comparative examples 4 to 7 used centrifugation, water extraction and alcohol precipitation, chitosan clarification and ZTC1+1-II natural clarification, respectively, and the effects of lowering the extract yields of comparative examples 4, 6 and 7 were not satisfactory, and the retention of gallic acid and total polysaccharides were not as good as in examples 1, 4 and 5. Comparative example 5 uses the water extraction and alcohol precipitation method, and the extract is reduced similar to that of example 1, but the loss of gallic acid and polysaccharide as active ingredients is too high. It can be seen that the technical effects of the present invention are superior to those of comparative examples 4 to 7.

(3) Performance results of reduced pressure concentration temperature

TABLE 3 detection results of reduced pressure concentration temperature

As can be seen from Table 3, the alcohol extracts of examples 1 and 6 were high in ferulic acid, osthole, gentiopicroside and gallic acid in the water body, wherein the content of the above components was the highest in example 1. The temperatures of comparative examples 8 and 9 were too low and too high, respectively, and the contents of the active ingredients in the extracts were lower than those of examples 1 and 6. It can be seen that the technical effect of the concentration process of the present invention is superior to that of comparative examples 8 and 9.

(4) Performance results at vacuum drying temperature

TABLE 4 detection results of vacuum drying temperatures

As can be seen from Table 4, the alcohol extract and the aqueous extract of examples 1 and 7 were high in ferulic acid, osthole, gentiopicroside and gallic acid, and the content of the above components was the highest in example 1. The drying temperatures of comparative examples 10 and 11 were too low and too high, respectively, and the contents of the active ingredients in the extracts were lower than those of examples 1 and 7. It can be seen that the technical effect of the vacuum drying process of the present invention is superior to that of comparative examples 10 and 11.

(5) Performance results of the adhesive

TABLE 5 detection results of adhesives

Group of	Particle morphology
		Example 1	The soft material has moderate viscosity, is easy to granulate and sift, and has uniform granules
Example 8	The soft material has moderate viscosity, is easy to granulate and screen, and has slightly bad granule color
		Comparative example 12	Adding small amount to quickly agglomerate, become sticky, not easy to mix uniformly and difficult to granulate
Comparative example 13	The soft material is dry, is easy to granulate, has loose granules and too much fine powder

As can be seen from table 5, the binders of examples 1 and 8 were able to be successfully granulated, with example 1 being the most effective. The ethanol concentrations of comparative examples 12 and 13 as binders were too low and too high, respectively, resulting in unsuccessful granulation. It can be seen that the formulation of the present invention is sensitive to ethanol concentration, either too high or too low to be successfully granulated.

(6) Performance results of the adjuvant

The composite score is calculated as follows:

composite score = molding rate/maximum molding rate x 0.5+ minimum moisture absorption rate/moisture absorption rate x 0.3+ minimum angle of repose/angle of repose x 0.2

Table 6 detection results of adjuvants

Group of	Molding rate (%)	Moisture absorption Rate (%)	Angle of repose (°)	Comprehensive scoring
					Example 1	94.38	8.73	20.82	98.66
Example 9	91.42	9.29	21.34	94.85
					Example 10	92.60	8.99	20.79	96.90
Example 11	89.65	8.53	21.61	96.13
					Comparative example 14	—	—	—	—
Comparative example 15	—	—	—	—
					Comparative example 16	—	—	—	—

The auxiliary materials and the extract powder of comparative examples 14 to 16 were granulated to form a black thick paste, and were agglomerated, and were not granulated. As can be seen from Table 6, the granules obtained by the preparation methods of examples 1 and 9 to 11 were high in molding rate, low in moisture absorption rate and good in fluidity, and the effect of example 1 was the best and the composite score was the highest. Therefore, only part of medicinal fine powder is added as the filler, so that the problems of high hygroscopicity and low forming rate of the extract fine powder can be effectively solved.

Test example 2

Clinical data

1. Western diagnostic criteria

Diagnosis was made according to the relevant diagnosis criteria for rheumatoid diseases, established by the American society of rheumatology, 1987, and diagnosis of rheumatoid arthritis, therapeutic guidelines:

(1) Morning stiffness for at least 1h;

(2) 3 or more joints are involved;

(3) The joint swelling between the wrist, the metacarpophalangeal or the proximal interphalangeal is more than or equal to 6 weeks;

(4) The involvement of the symmetrical joint is more than or equal to 6 weeks;

(5) There are rheumatoid subcutaneous nodules;

(6) A hand X-ray film change;

(7) Serum rheumatoid factor positive.

The rheumatoid arthritis can be diagnosed by providing 4 or more than 4 of the 7.

2. Diagnostic criteria for traditional Chinese medicine

The diagnosis of the traditional Chinese medicine accords with cold arthralgia of the traditional Chinese medicine science and the clinical research guidelines of the new traditional Chinese medicine, and clinically manifests as cold pain and swelling of joints, increased pain when encountering cold, reduced pain, fixed pain, unfavorable flexion and extension of joints, cold limbs, white coating, wiry and tight pulse and the like.

3. Inclusion criteria: meets the Western diagnosis standard; meets the diagnostic standard of traditional Chinese medicine; the sex is unlimited, and the age is 20-70 years old; continuously taking the Chinese medicine orally and checking regularly; and signing an informed consent form.

4. Exclusion criteria: severe advanced rheumatoid arthritis, joint deformation, and inability to self-care in life; other rheumatic diseases and severe heart-lung, liver-kidney dysfunction; pregnant or lactating women; while receiving other related treatments; those treated with glucocorticoids and antirheumatic drugs (DMARDs) within 6 months; allergic to oral administration.

5. General data

Clinical observations 1081 cases were confirmed in rheumatoid arthritis patients, of which men 648 and women 433.

6. Therapeutic method

The granule prepared in example 1 was taken three times daily, 10 to 20g each time, with warm water after meals, for 3 months.

7. Efficacy assessment index

And (3) observing the indexes:

(1) Clinical symptom observation: the swelling degree, tenderness and pain point number of the joints before and after treatment are divided into 'none, mild, moderate and severe' according to the degree, and 0, 2, 4 and 6 are respectively recorded; the pain number is calculated according to the actual pain joint number; morning stiffness time is the time from morning to the time when stiffness disappears.

(2) The Chinese medicine symptoms before and after treatment are scored according to the symptoms of morning stiffness, tenderness joint number, swelling joint number, pain score and joint swelling, namely, symptoms of no, mild, moderate and severe, respectively scored for 0, 2, 4 and 6.

(3) Observing the serum inflammatory factors before and after treatment to detect the level of rheumatoid factors, C-reactive proteins and blood sedimentation;

8. curative effect judging index

And (3) healing: the clinical symptoms are obviously improved, and the laboratory indexes are recovered to be normal;

the method is effective: the clinical symptoms are partially improved, and laboratory indexes are improved;

invalidation: clinical symptoms, laboratory indexes and the like are not obviously improved, and even aggravated.

9. Therapeutic results

TABLE 7 Effect of the granules of example 1 on treating rheumatoid arthritis

Healing of the wound	Effective and effective	Invalidation of	Total effective rate
				325(30.06％)	610(56.43％)	146(13.51％)	86.49％

As shown in Table 7, the granule of the present invention cured 30.06% after treating rheumatoid arthritis, and was 56.43% effective, and the total effective rate was 86.49%. Therefore, the granule of the present invention can effectively treat rheumatoid arthritis.

The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims

1. The preparation method of the anti-rheumatism traditional Chinese medicine preparation is characterized in that the traditional Chinese medicine preparation comprises the following raw materials:

herba Erodii seu Geranii, glycyrrhrizae radix, radix Angelicae Pubescentis, radix Gentianae Marcrophyllae, ramulus Sambuci Williamsii, rhizoma corydalis, radix Angelicae sinensis and flos Matricariae Chamomillae;

the preparation method comprises the following steps:

taking 14-16 parts of geranium, 5-7 parts of liquorice and 7-9 parts of chamomile, extracting with water under reflux, and refining to obtain an aqueous extract;

taking, by weight, 11-13 parts of gentiana macrophylla, 9-11 parts of angelica sinensis, 9-11 parts of elder, 9-11 parts of corydalis tuber and 11-13 parts of radix angelicae pubescentis, and carrying out reflux extraction with an ethanol solution to obtain an ethanol extract;

concentrating the water extract and the alcohol extract under reduced pressure respectively, drying to obtain extract fine powder, adding ethanol solution with the concentration of 92-93% and fine powder of angelica and/or radix angelicae pubescentis raw materials with the total weight of 5-10% of the total weight of the extract fine powder, mixing and granulating.

2. The method of manufacturing according to claim 1, wherein the refining comprises: concentrating the water extract under reduced pressure until the ratio of the mass of the medicinal materials to the volume of the liquid medicine is 1 (1-1.2), standing, and filtering to obtain supernatant.

3. The preparation method according to claim 1, wherein the concentration of the ethanol solution is 65-75%, the volume of the added ethanol solution is 9-11 times of the weight of the raw material, and the reflux extraction is performed for 1-3 times, and each time the extraction is performed for 80-120 min.

4. The method according to claim 1, wherein the reduced pressure concentration is carried out at a temperature of 60 to 80 ℃ and the drying is carried out at a temperature of 60 to 80 ℃.

5. A Chinese medicinal preparation prepared by the preparation method of any one of claims 1 to 4.

6. The traditional Chinese medicine preparation according to claim 5, wherein the dosage form of the traditional Chinese medicine preparation comprises a emplastrum, a granule, a tablet or a capsule.

7. The use of a Chinese medicinal preparation according to claim 5 or 6 in the preparation of a medicament for the treatment of rheumatoid arthritis.