CN112546149A

CN112546149A - A Chinese medicinal composition for nourishing yin, clearing away lung-heat, relieving sore-throat, and removing toxic substance, and its preparation method and quality control method

Info

Publication number: CN112546149A
Application number: CN202011546393.8A
Authority: CN
Inventors: 郎伟君
Original assignee: Letai Pharmaceutical Co ltd
Current assignee: Letai Pharmaceutical Co ltd
Priority date: 2020-12-23
Filing date: 2020-12-23
Publication date: 2021-03-26

Abstract

The invention discloses a preparation method and a quality control method of a traditional Chinese medicine composition for nourishing yin, clearing away the lung-heat, relieving sore throat and detoxifying. According to the invention, the purpose of highest scutellaria baicalensis extract content is achieved by comparing the baicalin content of scutellaria baicalensis decoction pieces in different producing areas and determining that the scutellaria baicalensis from inner Mongolia is selected as one of the production raw materials; through the research on the column filling mode, the column filling sequence and the percolation temperature range of all medicinal materials in the production process, the optimal production process is selected to better control the product quality. By adopting HPLC to establish the fingerprint and comprehensively researching the attribution of each characteristic peak in the fingerprint, the main components in the prescription can be comprehensively controlled, so that the product quality is more stable and controllable.

Description

A Chinese medicinal composition for nourishing yin, clearing away lung-heat, relieving sore-throat, and removing toxic substance, and its preparation method and quality control method

Technical Field

The invention relates to a preparation method and a quality control method of a traditional Chinese medicine composition for nourishing yin, clearing away the lung-heat, relieving sore throat and detoxifying, and belongs to the technical field of medicines.

Background

The throat clearing and sore throat mixture is prepared from rehmannia root, dwarf lilyturf tuber, figwort root, weeping forsythia and baical skullcap root, has the effects of nourishing yin, clearing away the lung-heat, relieving sore throat and detoxifying, and is used for treating sore throat, dry throat, little saliva and swollen throat caused by yin deficiency, dryness-heat and fire toxin; acute tonsillitis and angina pectoris with the above syndrome. A large number of modern pharmacological studies show that the throat clearing and throat nourishing mixture has the effects of resisting bacteria, detoxifying, resisting radiation damage, improving T cell immune function, resisting tumors and the like; it is also used clinically for localized diphtheria, mild toxic diphtheria, acute tonsillitis, suppurative tonsillitis, angina, and radiation mucoreaction, with symptoms of yin deficiency and toxic fire flaming. In vitro tests show that the throat clearing and throat moistening mixture has inhibiting and killing effects on staphylococcus aureus, alpha-hemolytic streptococcus, beta-hemolytic streptococcus, pneumococcus, proteus and escherichia coli to different degrees.

The existing throat clearing and sore throat relieving mixture is prepared by pulverizing rehmanniae radix, radix Ophiopogonis, radix scrophulariae, fructus forsythiae and Scutellariae radix, soaking in ethanol as solvent, percolating, concentrating percolate, and concocting to obtain mixture. The method specifically comprises the following steps: pulverizing Scutellariae radix, radix Ophiopogonis, rehmanniae radix, fructus forsythiae, and radix scrophulariae, soaking in 57% ethanol as solvent for 24 hr, slowly percolating, collecting percolate, recovering ethanol under reduced pressure, concentrating, adding water, boiling for 30 min, standing for 48 hr, filtering, washing the residue with small amount of water, adding the washing solution into the filtrate, concentrating under reduced pressure, adding sodium benzoate, stirring, standing for 24 hr, filtering, adding water into the filtrate to desired amount, and stirring. The disadvantages of the existing preparation method are that: 1. the five medicinal materials of scutellaria, ophiopogon root, rehmannia, forsythia and figwort are packed in bags in a column filling mode in the percolation process, the number and the tightness of the bags in the bags have certain influence on the relative density of the finished product, and the tighter the medicinal bag is, the less the medicinal bag is, the lower the density of the finished product is. 2. The five medicinal materials of the scutellaria, the ophiopogon root, the rehmannia, the forsythia and the figwort are not fixed and packed in a column in a percolation process. Because the radix ophiopogonis and the rehmannia root have promotion effect on baicalin leaching, the fructus forsythiae has little influence on the baicalin leaching, and the radix scrophulariae has inhibition effect on the baicalin leaching, the existing column packing sequence has influence on the baicalin content, and is not the optimal scheme. 3. The environmental temperature of the percolation process and the temperature range of the solvent ethanol are not specified, and the content of the baicalin in the product can be influenced.

Disclosure of Invention

In order to overcome the defects of the existing preparation method of the throat clearing and throat moistening mixture, the invention aims to provide a preparation method and a quality control method of a traditional Chinese medicine composition for nourishing yin, clearing away the lung-heat, relieving sore throat and detoxifying.

In order to achieve the purpose, the invention adopts the following technical means:

the invention relates to a preparation method of a traditional Chinese medicine composition for nourishing yin, clearing away the lung-heat, relieving sore-throat and detoxifying, which comprises the following steps:

(1) pulverizing five traditional Chinese medicinal materials of rehmannia root, radix ophiopogonis, figwort root, weeping forsythia and baical skullcap root into coarse powder, wherein the baical skullcap root is bulk-packed, and the four traditional Chinese medicinal materials of the radix ophiopogonis, the rehmannia root, the weeping forsythia and the figwort root are respectively packaged and decocted in bags;

(2) loading the Chinese medicinal materials obtained in the step (1) into a column according to the sequence of scutellaria baicalensis-ophiopogon root-rehmannia root-forsythia suspense-radix scrophulariae from bottom to top, using ethanol as a solvent, soaking for 24 hours, slowly percolating, collecting percolate, recovering ethanol under reduced pressure, concentrating, taking out, adding water, boiling for 30 minutes, standing for 48 hours, filtering, washing filter residues with a small amount of water, adding washing liquor into the filtrate, concentrating under reduced pressure, adding sodium benzoate, stirring, standing for 24 hours, filtering, taking the filtrate, adding water to a certain amount, and stirring uniformly to obtain the Chinese medicinal composition.

Wherein, preferably, the weight percentages of the traditional Chinese medicinal materials are as follows: 14.6 percent of rehmannia root, 13.0 percent of dwarf lilyturf tuber, 21.2 percent of figwort root, 25.6 percent of weeping forsythia capsule and 25.6 percent of baical skullcap root.

Wherein, preferably, 50-60% v/v ethanol is used as solvent, slowly percolating at a speed of 1ml per minute after soaking for 24 hr, and collecting percolate, preferably, 57% v/v ethanol is used as solvent.

Wherein, the environmental temperature and the solvent ethanol temperature during the percolation are preferably controlled at 20-30 ℃.

Wherein, preferably, the scutellaria baicalensis in the inner Mongolia producing area is selected to be used as a production raw material so as to achieve the purpose of the highest content of scutellaria baicalensis extracts.

Preferably, the rehmannia root, the dwarf lilyturf tuber, the figwort root and the baical skullcap root are prepared into traditional Chinese medicine decoction pieces, and the weeping forsythia is selected and then is used as the medicine.

Processing rehmannia decoction pieces: removing impurities, cleaning, moistening, slicing, and drying.

Processing radix ophiopogonis decoction pieces: removing impurities, cleaning, moistening, rolling, and drying.

Processing radix scrophulariae decoction pieces: removing residual rhizome and impurities, cleaning, moistening, slicing, and drying; or micro bubble, steaming, slightly drying, slicing, and drying.

Processing of scutellaria baicalensis decoction pieces: removing impurities, decocting in boiling water for 10 min, taking out, sealing, slicing, and drying; or steaming for half an hour, taking out, slicing, and drying (avoiding exposure to sunlight).

Wherein, the preferable method also comprises the step of carrying out quality control on the main components in the prescription by adopting an HPLC to establish a quality control method.

Preferably, the quality control method comprises the following steps:

(1) preparation of control solutions:

a: precisely weighing 10.01mg of verbascoside reference substance, placing in a 100mL measuring flask, adding methanol to scale to obtain verbascoside reference substance solution;

b: accurately weighing 10.09mg of a cinnamic acid reference substance, placing the cinnamic acid reference substance into a 100mL measuring flask, and adding methanol to the scale to obtain a cinnamic acid reference substance solution;

c: accurately weighing 10.03mg of ferulic acid reference substance, placing into a 100mL measuring flask, adding methanol to scale to obtain ferulic acid reference substance solution;

d: accurately weighing 10.05mg of baicalin reference substance, placing into a 100mL measuring flask, adding methanol to scale to obtain baicalin reference substance solution;

e: accurately weighing baicalein reference substance 10.03mg, placing into a 100mL measuring flask, adding methanol to scale to obtain baicalein reference substance solution;

f: accurately weighing wogonin reference substance 10.11mg, placing into 100mL measuring flask, adding methanol to scale to obtain wogonin reference substance solution;

(2) preparation of test solution

Taking 1mL of a test sample, adding water to dilute the test sample to 50mL, shaking up, and taking a subsequent filtrate as a test sample solution;

(3) HPLC analysis

Chromatographic conditions are as follows: liquid chromatograph: agilent 1260 high performance liquid chromatograph;

a chromatographic column: an Agilent ZORBAXSB-C18 column with specification of 4.6mm × 250mm, 5 μm; the mobile phase was acetonitrile (A) -0.3% v/v glacial acetic acid in water (B) and the gradient elution procedure was as follows:

detection wavelength: 270 nm; column temperature: 30 ℃; flow rate: 1.0 ml/min; recording chromatogram for 90 minutes, and the sample injection amount is 10 mu l;

(4) quality evaluation of to-be-detected product based on fingerprint

And after the detection is finished, obtaining a chromatogram, evaluating by adopting software of a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2012 edition), and analyzing with a contrast map, wherein the product is qualified if the similarity is more than 0.9.

The traditional Chinese medicine composition prepared by the preparation method is also within the protection scope of the invention.

Furthermore, the invention also provides application of the traditional Chinese medicine composition in preparing medicines for nourishing yin, clearing away the lung-heat, relieving sore throat and detoxifying.

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

1. the extract content of the medicinal materials is reduced compared with the extract content of the decoction pieces prepared by the medicinal materials, so that the medicinal materials of the invention, namely the scutellaria baicalensis, the rehmannia glutinosa, the ophiopogon root and the figwort root, are selected and prepared after being prepared, and the forsythia is selected and prepared.

2. The invention determines to select the inner Mongolia radix scutellariae as the production raw material by comparing the baicalin content data of radix scutellariae decoction pieces in different producing areas, so as to achieve the purpose of the highest content of radix scutellariae extract.

3. The method determines and selects the following Chinese medicinal materials by comparing different column filling modes and column filling sequences of Chinese medicinal materials in a percolation process and comparing different percolation environment temperatures and solvent ethanol temperatures: 1. the scutellaria baicalensis is scattered and infiltrated, and the radix ophiopogonis, the rehmannia, the fructus forsythiae and the radix scrophulariae are all packed into a column in a wrapping and decocting bag mode; 2. selecting a column filling sequence of scutellaria baicalensis, ophiopogon root, rehmannia root, forsythia suspense and figwort from bottom to top;

4. the environmental temperature of the percolation process and the temperature of the solvent ethanol are controlled at 20-30 ℃. The optimal production process scheme is determined by selecting the column filling mode and the column filling sequence so as to better control the product quality.

5. By adopting HPLC to establish the fingerprint and comprehensively researching the attribution of each characteristic peak in the fingerprint, the main components in the prescription can be comprehensively controlled, so that the product quality is more stable and controllable.

Drawings

FIG. 1 shows the similarity of the fingerprints of 4 batches of samples.

Detailed Description

The present invention will be further described with reference to the following examples, but the present invention is not limited to the following examples. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.

Example 1 selection of Scutellaria baicalensis Georgi production area

The traditional Chinese medicine Scutellaria is a dried root of Scutellaria (Scutellaria baicalensis Georgi) of Labiatae, is originally recorded in Shen nong Ben Cao Jing, has the functions of clearing heat and removing toxicity, and is one of the traditional Chinese medicines commonly used in clinic. The chemical components of the scutellaria baicalensis comprise flavonoids, phenolic acids, phenethyl alcohol, amino acids, sterols, essential oil, trace elements and the like. The active component of the flavone is baicalin, and the baicalin has inhibitory effect on various viruses. Due to the geographical factors and the growth environment, the chemical composition and the content of active ingredients of the scutellaria baicalensis medicinal materials in different areas are different. According to the invention, the production place selection of scutellaria baicalensis is determined by comparing the content of baicalin in the scutellaria baicalensis decoction pieces in different production places. The detection result shows that the baicalin of the inner Mongolia baical skullcap root decoction pieces is easier to leach, so that the inner Mongolia baical skullcap root is selected as a production raw material to ensure the content level of the baicalin.

1. Purchasing radix Scutellariae from different producing areas

The Scutellariae radix is mainly distributed in provinces and autonomous regions such as Heilongjiang, Jilin, Liaoning, Hebei, Henan, Shandong, Shanxi, inner Mongolia, Gansu, Shaanxi, Ningxia, etc. Wherein the yield of inner Mongolia, Shanxi and Hebei is larger. The invention respectively purchases a batch of scutellaria baicalensis medicinal materials with medicinal material producing areas of Hebei, inner Mongolia and Shanxi, and the medicinal materials are labiate plants through inspection and identification. Detecting the relative density and baicalin content of three batches of radix Scutellariae materials respectively purchased from Hebei, inner Mongolia and Shanxi, and recording data. The medicinal material supplier is Daqing Tongshi Chinese medicinal decoction piece Co.

2. The relative density and baicalin content items of the scutellaria baicalensis medicinal materials in different producing areas are detected, and the detection results are shown in the following table 1.

TABLE 1 relative Density and baicalin content of Scutellariae radix in different production areas

Serial number	Item	Relative density	Baicalin content (mg/ml)
				1	Inner Mongolia radix scutellariae (after processing)	1.17	14.28
2	Hebei radix Scutellariae (processed)	1.17	11.98
				3	Shanxi Baikal skullcap root medicine (after processing)	1.16	12.36

And (4) conclusion: according to the detection data of the relative density and the baicalin content of the scutellaria baicalensis in different producing areas, the baicalin content of purchased inner Mongolia scutellaria baicalensis is higher than that of the scutellaria baicalensis in Hebei and Shanxi producing areas, so that the scutellaria baicalensis in the inner Mongolia producing areas is selected as one of the raw materials.

Example 2 study of the percolation Process

1. Research on column filling mode and sequence of medicinal materials in percolation process

Through the comparison of different column filling modes and column filling sequences of the Chinese medicinal materials in the percolation process, an optimal production process scheme is selected to better control the product quality.

1.1 study of column packing mode of each medicinal material in percolation process

The first experimental example:

the formula (weight ratio of each raw material component): 14.6 percent of rehmannia root, 13.0 percent of dwarf lilyturf tuber, 21.2 percent of figwort root, 25.6 percent of weeping forsythia capsule and 25.6 percent of baical skullcap root.

The production process comprises the following steps: pulverizing Scutellariae radix, radix Ophiopogonis, rehmanniae radix, fructus forsythiae, and radix scrophulariae, soaking in 57% v/v ethanol as solvent for 24 hr, slowly percolating at a speed of about 1ml per minute, collecting percolate of about 6000ml, recovering ethanol under reduced pressure, concentrating to about 1400ml, adding water 800ml, boiling for 30 min, standing for 48 hr, filtering, washing residue with small amount of water, adding the washing solution into the filtrate, concentrating under reduced pressure to about 1000ml, adding sodium benzoate 3g, stirring, standing for 24 hr, filtering, collecting filtrate, adding water to 1000ml, and stirring. Wherein, the scutellaria baicalensis, the ophiopogon root, the rehmannia, the forsythia and the figwort root are prepared into a medicine in a diffusing and permeating mode, the relative density and the baicalin content of a finished product produced by the medicine are detected, and data are recorded. (see Table 2)

Experiment example two:

the formula (weight ratio of each raw material component) is the same as that of the experimental example one.

The production process is the same as the first experimental example.

Wherein, the five medicinal materials of the scutellaria, the ophiopogon root, the rehmannia, the forsythia and the figwort are packaged and decocted in bags, each medicinal material is respectively packaged in one bag, and the five bags are totally used for detecting the relative density and the baicalin content of a finished product produced by the medicinal materials and recording data. (see Table 2)

Experiment example three:

The production process is the same as the first experimental example.

Wherein, the five medicinal materials of the scutellaria, the ophiopogon root, the rehmannia, the forsythia and the figwort are packed and decocted in a bag mode, the scutellaria, the ophiopogon root and the rehmannia are packed one bag respectively, the forsythia and the figwort are packed two bags respectively, and the seven bags are totally used, the relative density and the baicalin content of the finished products are detected, and the data are recorded. (see Table 2)

Experimental example four:

The production process is the same as the first experimental example.

Wherein, the five medicinal materials of the scutellaria, the ophiopogon root, the rehmannia, the forsythia and the figwort are packed and decocted in bags, the scutellaria and the ophiopogon root are packed one bag each, the rehmannia, the forsythia and the figwort are evenly divided into two bags, eight bags are used, the relative density and the baicalin content of the finished products are detected, and the data is recorded. (see Table 2)

Experimental example five:

The production process is the same as the first experimental example.

Wherein, the scutellaria baicalensis is bulk-loaded, the four medicinal materials of the ophiopogon root, the rehmannia root, the forsythia and the figwort root are packaged and decocted in a bag mode, each medicinal material is averagely divided into two bags, eight bags in total are adopted, the relative density and the baicalin content of a finished product produced by the method are detected, and data are recorded. (see Table 2)

TABLE 2 test results of column packing method for medicinal materials

1.2 conclusion

The five medicinal materials are respectively tested in different column filling modes of loose infiltration and bag packing, and the test result table shows that the number and the tightness of the bag packing bags have certain influence on the relative density of the finished product, except the scutellaria medicinal material, the tighter the other medicinal material bags are, the less the medicine is leached, and the smaller the density of the finished product is. Therefore, the column mounting method of the fifth embodiment is selected.

2. Research on column packing sequence of medicinal materials in percolation process

2.1 study of the column packing sequence of the medicinal materials in the percolation Process

The first experimental example:

The production process comprises the following steps: pulverizing Scutellariae radix, radix Ophiopogonis, rehmanniae radix, fructus forsythiae, and radix scrophulariae, soaking in 57% v/v ethanol as solvent for 24 hr, slowly percolating at a speed of about 1ml per minute, collecting percolate of about 6000ml, recovering ethanol under reduced pressure, concentrating to about 1400ml, adding water 800ml, boiling for 30 min, standing for 48 hr, filtering, washing residue with small amount of water, adding the washing solution into the filtrate, concentrating under reduced pressure to about 1000ml, adding sodium benzoate 3g, stirring, standing for 24 hr, filtering, collecting filtrate, adding water to 1000ml, and stirring.

Wherein, the five medicinal materials of the baical skullcap root, the dwarf lilyturf tuber, the rehmannia root, the weeping forsythia and the figwort root adopt a disordered diffusing and permeating mode, the relative density and the baicalin content of a finished product produced by the five medicinal materials are detected, and data are recorded. (see Table 3)

Experiment example two:

The production process is the same as the first experimental example.

Wherein, the five medicinal materials of scutellaria baicalensis, ophiopogon root, rehmannia glutinosa, forsythia suspensa and figwort are respectively filled in columns from bottom to top, and the five medicinal materials are all packed and decocted in bags, the relative density and the baicalin content of the finished products produced by the five medicinal materials are detected, and the data is recorded. (see Table 3)

Experiment example three:

The production process is the same as the first experimental example.

The five medicinal materials of figwort, forsythia, scutellaria, ophiopogon root and rehmannia are respectively filled in columns from bottom to top, and the five medicinal materials are all packaged and decocted in bags, and the relative density and the baicalin content of the finished products produced by the five medicinal materials are detected and the data are recorded. (see Table 3)

Experimental example four:

The production process is the same as the first experimental example.

Wherein, the baical skullcap root is bulked, the other four medicinal materials of the dwarf lilyturf tuber, the rehmannia root, the weeping forsythia and the figwort root are all packed and decocted in a bag mode, columns are arranged from bottom to top according to the sequence of the baical skullcap root, the dwarf lilyturf tuber, the rehmannia root, the weeping forsythia and the figwort root, the relative density and the baicalin content of a finished product produced by the columns are detected, and data are recorded. (see Table 3)

TABLE 3 test results of the column loading sequence of herbs

2.2 conclusion

Because the radix ophiopogonis and the rehmannia root have promotion effect on baicalin leaching, the forsythia has little influence on the baicalin leaching, the figwort has inhibition effect on the baicalin leaching, and the relative density of the product is influenced most, the bottom-to-top column packing sequence of the scutellaria baicalensis, the radix ophiopogonis, the rehmannia root, the forsythia and the figwort is most suitable.

And (3) knotting: by comprehensively studying the column filling mode and the column filling sequence of all medicinal materials in the percolation process, the method finally determines that the scutellaria baicalensis is bulk-loaded, the ophiopogon root, the rehmannia root, the forsythia and the figwort are all packaged and decocted in bags, and percolation is carried out according to the column filling sequence of the scutellaria baicalensis-ophiopogon root-rehmannia root-forsythia-figwort from bottom to top.

3. Temperature study of percolation process

3.1 the relative density and baicalin content of the products produced by different percolation environmental temperatures and solvent ethanol temperature ranges are detected, data are compared, and the most suitable percolation environmental temperature and solvent ethanol temperature range are selected to better control the product quality.

The first experimental example:

the formula (the weight percentages of the raw material components) is as follows: 14.6 percent of rehmannia root, 13.0 percent of dwarf lilyturf tuber, 21.2 percent of figwort root, 25.6 percent of weeping forsythia capsule and 25.6 percent of baical skullcap root.

Wherein, the process production environment temperature and the solvent ethanol temperature are adjusted to be below 15 ℃, the crushed five medicinal materials are added with ethanol solvent for dipping, the percolation process is adopted for production, the percolation liquid is concentrated and prepared into mixture, the relative density and the baicalin content are detected, and the data is recorded. (see Table 4)

Experiment example two:

The production process is the same as the first experimental example.

Wherein, the process production environment temperature and the solvent ethanol temperature are adjusted to be more than 20 ℃ but less than 25 ℃, the crushed five medicinal materials are added with ethanol solvent for dipping, the percolation process is adopted for production, the percolation liquid is concentrated and prepared into mixture, the relative density and the baicalin content are detected, and the data is recorded. (see Table 4)

Experiment example three:

The production process is the same as the first experimental example.

Wherein, the environmental temperature of the process production and the temperature of the solvent ethanol are adjusted to be more than 25 ℃ and less than 30 ℃, the crushed five medicinal materials are added with the ethanol solvent for soaking, the percolation process is adopted for production, the percolation liquid is concentrated and prepared into mixture, the relative density and the baicalin content are detected, and the data is recorded. (see Table 4)

TABLE 4 test results in different temperature ranges

3.2 conclusion

The detection value of the baicalin content of the product produced in the temperature range of the experimental example I is lower than that of the products produced in the experimental examples II and III, which shows that the percolation temperature has certain influence on the product quality. In the experimental process, when the percolation environment and the temperature of the solvent ethanol are lower than 20 ℃, the viscosity of the ethanol is increased to influence the fluidity of the ethanol, and further the percolation effect is possibly influenced. In order to better ensure the product quality, the environmental temperature of the percolation process and the temperature of the solvent ethanol are preferably controlled within the range of 20-30 ℃ by integrating the test data of the second experimental example and the third experimental example.

Example 3 preparation of a Chinese medicinal composition for nourishing yin, clearing lung-heat, relieving sore-throat and removing toxic substance

According to the experimental results of example 2, the finally determined preparation method of the traditional Chinese medicine composition for nourishing yin, clearing away the lung-heat, relieving sore-throat and detoxifying comprises the following steps:

(1) weighing the following Chinese medicinal materials in percentage by weight:

14.6 percent of rehmannia root, 13.0 percent of dwarf lilyturf tuber, 21.2 percent of figwort root, 25.6 percent of weeping forsythia capsule and 25.6 percent of baical skullcap root.

(2) The rehmannia root, the dwarf lilyturf tuber, the figwort root and the baical skullcap root are prepared into traditional Chinese medicine decoction pieces, and the weeping forsythia is selected and then added into the medicine.

Processing radix ophiopogonis decoction pieces: -removing impurities, washing, moistening, flattening, and drying.

Processing radix scutellariae decoction pieces (inner Mongolia producing area): removing impurities, decocting in boiling water for 10 min, taking out, sealing, slicing, and drying; or steaming for half an hour, taking out, slicing, and drying (avoiding exposure to sunlight).

(3) Pulverizing the five traditional Chinese medicinal materials of rehmannia root, radix ophiopogonis, figwort root, weeping forsythia and baical skullcap root into coarse powder, wherein the baical skullcap root is bulk-packaged, and the four traditional Chinese medicinal materials of the radix ophiopogonis, the rehmannia root, the weeping forsythia and the figwort root are respectively packaged and decocted in bags;

(4) and (3) loading the Chinese medicinal materials obtained in the step (2) into columns in the order of scutellaria baicalensis-ophiopogon root-rehmannia root-forsythia suspense-radix scrophulariae from bottom to top, adjusting the ambient temperature and the temperature of solvent ethanol to 20-30 ℃, using 57% v/v ethanol as a solvent, soaking for 24 hours, slowly percolating at the speed of about 1ml per minute, collecting about 6000ml of percolate, recovering ethanol under reduced pressure, concentrating to about 1400ml, adding 800ml of water, boiling for 30 minutes, standing for 48 hours, filtering, washing filter residues with a small amount of water, adding a washing solution into the filtrate, concentrating under reduced pressure to about 1000ml, adding 3g of sodium benzoate, stirring uniformly, standing for 24 hours, filtering, taking the filtrate, adding water to 1000ml, and stirring uniformly to obtain the Chinese medicinal composition.

Example 4 quality control method

In the embodiment, the fingerprint is established by HPLC, the specificity is strong, the attribution of each characteristic peak in the fingerprint is comprehensively researched, each peak is respectively attributed to each medicinal material, the main components in the product can be comprehensively controlled, and the product quality is more stable and controllable.

1. Operation step (1) preparation of control solution:

(2) preparation of test solution

1mL of a sample (4 batches of the sample prepared in example 3) was taken, diluted to 50mL with water, shaken up, and the subsequent filtrate was taken as a sample solution.

(3) HPLC analysis

Chromatographic conditions are as follows: liquid chromatograph: agilent 1260 high performance liquid chromatograph (DAD-detector, autosampler);

a chromatographic column: agilent ZORBAXSB-C18 column (4.6 mm. times.250 mm, 5 μm); the mobile phase was acetonitrile (a) -0.3% aqueous glacial acetic acid (B) and the gradient elution procedure is as in table 5:

TABLE 5 gradient elution procedure

(4) quality evaluation of to-be-detected product based on fingerprint

2. Establishment of fingerprint

Preparing a test solution and a reference solution according to the method, determining according to chromatographic conditions, recording chromatograms, introducing 4 batches of chromatograms into a traditional Chinese medicine chromatogram fingerprint similarity evaluation system 2012 edition for evaluation, matching chromatographic peaks by taking 01 batches of sample chromatograms as reference chromatograms, and generating the reference fingerprint by a median method.

Similarity calculation adopts similarity software, namely 'traditional Chinese medicine chromatogram fingerprint similarity evaluation system 2012 edition' and takes comparison fingerprints as reference fingerprints, and calculates the similarity of the fingerprints of 4 batches of test articles, and the result is shown in table 6 and figure 1.

TABLE 64 test sample solution similarity calculation results

The results show that the similarity of the fingerprints obtained by 4 batches of samples under the 270nm detection condition is greater than 0.99, and the fingerprints have higher similarity (figure 1).

3. Determination of characteristic peaks

According to the principle that the retention time is stable, the peak area is relatively high and the detection rate reaches 100%, 10 chromatographic peaks with good repeatability and strong specificity are selected as characteristic peaks, baicalin 5, which is an effective component with moderate retention time and large peak area, is taken as a reference peak, and the relative retention time of each characteristic peak is calculated, which is shown in table 7.

TABLE 74 relative retention time of characteristic peaks of test solutions

The determination of the specified value of the relative retention time of the characteristic peak takes the average value of the relative retention time of 10 characteristic peaks in 4 batches of test sample solutions as the specified value, and determines the fluctuation range of the specified value by combining the influence of each variation factor on the relative retention time of the characteristic peak in methodology inspection. Finally, the following steps are provided: the test sample has 10 characteristic peaks in the characteristic spectrum, the chromatographic peak corresponding to baicalin reference substance is S peak, and the relative retention time of each characteristic peak and peak (S) is calculated to be within + -5% of the specified value. The values of the characteristic peaks relative to the retention time were 0.719 (peak 1), 0.741 (peak 2), 0.774 (peak 3), 0.857 (peak 4), 1.000 (peak 5, S), 1.094 (peak 6), 1.151 (peak 7), 1.207 (peak 8), 1.436 (peak 9), 1.740 (peak 10).

4. Identification of chromatographic Peak

Comparing the main peaks with each control under 270nm wavelength detection, wherein the 5 peak is baicalin, the 1 peak is ferulic acid, the 2 peak is verbascoside, the 8 peak is cinnamic acid, the 9 peak is baicalein, and the 10 peak is wogonin.

5. Methodology investigation

(1) Precision test

Precisely sucking the same sample solution, continuously feeding samples for 6 times in the same day, and recording peak area. The relative retention time and relative peak area of the common peak are both less than 2.0% by taking the retention time and peak area of baicalin as reference. The similarity is all within 0.99, which accords with the technical requirement of fingerprint spectrum and shows that the precision of the instrument is good.

(2) Repeatability test

Taking 6 samples, respectively preparing test solution, detecting according to the chromatographic conditions, recording chromatogram, and taking the retention time and peak area of baicalin as reference, wherein the RSD is less than 2.0% and the relative retention time and relative peak area of the common peak are both used as reference. The similarity is all 0.99, and the technical requirement of the fingerprint spectrum is met, so that the repeatability of the method is good.

(3) Stability test

Injecting the same sample solution into liquid chromatograph at 0, 4, 8, 12, 24, and 48h after preparation, respectively, and recording peak area of each component with retention time and peak area of baicalein as reference. The relative retention time of all the common peaks is basically consistent with the relative peak area, the RSD is less than 2.0%, the similarity is 0.99, the technical requirements of the fingerprint spectrum are met, and the stability of the test solution in 48 hours is shown.

(4) Chromatographic peak assignment

Taking other medicinal materials except certain medicinal materials in the prescription respectively, preparing into negative control samples according to the process, taking appropriate amount of each negative control sample, and preparing into negative control solution according to the preparation method of the test solution. Taking the corresponding decoction pieces of each unit prescription medicine respectively, preparing corresponding single medicinal material test samples according to the prescription extraction process, taking a proper amount of each test sample, and preparing corresponding single medicinal material test sample solution according to the test sample solution preparation method.

Respectively taking 10 mu l of test solution, each negative control solution and each single medicinal material test solution, recording chromatograms according to the chromatographic conditions under the item 'two', and determining the attribution of each characteristic peak according to the ultraviolet absorption and relative retention time of each chromatographic peak. The fingerprint spectrum of the product has 10 characteristic peaks, and the characteristic peaks belonging to Scutellariae radix are No. 5, No. 9, and No. 10 peaks; the characteristic peaks attributed to radix scrophulariae are peaks No. 1 and No. 8, and the characteristic peak attributed to rehmanniae radix is peak No. 2. The three medicines of radix ophiopogonis, fructus forsythiae, radix scutellariae, rehmannia and radix scrophulariae in the prescription can be reflected in the fingerprint spectrum.

6. Conclusion

(1) And (4) measuring samples of the 4 batches, recording chromatograms, and determining a common peak, wherein the total peak area of the common peak is more than 90%.

(2) And calculating the retention time, the relative retention time, the peak area and the peak area percentage of each common peak by taking the baicalin as a reference peak. The average of the relative peak areas, and the standard deviation thereof.

(3) Similarity analysis is carried out on the fingerprints of 4 batches of samples by adopting a pharmacopoeia committee 'Chinese medicine chromatogram fingerprint similarity evaluation system 2012 edition'.

(4) The comparison fingerprints of 4 batches of samples are generated by a median method, the similarity between the 4 batches of samples and the comparison fingerprints is respectively more than 0.99, and the result shows that the similarity between the batches is better and the quality is stable.

Claims

1. A preparation method of a traditional Chinese medicine composition for nourishing yin, clearing away the lung-heat, relieving sore throat and detoxifying is characterized by comprising the following steps:

2. The preparation method of claim 1, wherein the weight percentages of the traditional Chinese medicinal materials are as follows: 14.6 percent of rehmannia root, 13.0 percent of dwarf lilyturf tuber, 21.2 percent of figwort root, 25.6 percent of weeping forsythia capsule and 25.6 percent of baical skullcap root.

3. The method according to claim 1, wherein 50-60% v/v ethanol is used as a solvent, and after 24 hours of immersion, the solution is slowly percolated at a rate of 1ml per minute, and the percolate is collected, preferably 57% v/v ethanol is used as a solvent.

4. The method according to claim 1, wherein the temperature of the solvent ethanol and the ambient temperature during the percolation are controlled to 20 to 30 ℃.

5. The method of claim 1, wherein the scutellaria baicalensis from the inner Mongolia origin is selected as a raw material for production, so as to achieve the purpose of the highest content of scutellaria baicalensis extract.

6. The preparation method of claim 1, wherein the rehmannia root, the dwarf lilyturf tuber, the figwort root and the baical skullcap root are prepared into decoction pieces of Chinese herbal medicines, and the weeping forsythia is selected and used as the medicine.

7. The method of claim 1, further comprising quality control of the main ingredient in the prescription by establishing a quality control method using HPLC.

8. The method of claim 7, wherein the quality control method comprises the steps of:

(1) preparation of control solutions:

(2) preparation of test solution

(3) HPLC analysis

(4) quality evaluation of to-be-detected product based on fingerprint

9. The Chinese medicinal composition prepared by the preparation method according to any one of claims 1 to 8.

10. The use of the composition of claim 9 in the preparation of a medicament for nourishing yin to clear lung heat, relieving sore throat and removing toxic substances.