Quality control method of traditional Chinese medicine composition freeze-dried powder

Abstract

The invention discloses a quality control method of traditional Chinese medicine composition freeze-dried powder, which adopts an evaluation mode of combining reference index component content, fingerprint spectrum, paste yield and thin-layer chromatography to carry out quantity transfer analysis on the material reference of a Dabaixiang decoction. According to the method, comprehensive quality detection is carried out on the material reference of the Dazeheng lung-purging decoction through multiple technologies and multiple ways, and the quality standard of the material reference of the Dazeheng lung-purging decoction is drawn up, so that a scientific basis is provided for preparation development of the Dazeheng lung-purging decoction, and a thought is provided for development and research of similar prescriptions.

Description

Technical Field

[0001] The invention belongs to the technical field of quality detection of traditional Chinese medicine preparations, and in particular relates to a quality control method for freeze-dried powder of a traditional Chinese medicine composition. Background Art

[0002] The Daxiefei Decoction comes from the "Fuxing Jue Zangfu Use of Medicinal Methods", which consists of six herbs: tianlizi (boiled until black and mashed into a paste), rhubarb, white peony root, licorice (roasted), scutellaria and dried ginger. It is mainly used to treat phlegm in the chest, wheezing and unable to lie down, constipation, swelling of the body and face, and fullness, and those who want to have a smooth qi. Tianlizi can purge the lungs and calm wheezing, promote water circulation and reduce swelling, while rhubarb can purge accumulation, clear heat and purge fire, and is the main medicine; scutellaria can clear heat, dry dampness and purge fire, and assist rhubarb to clear and purge the evil of heat in the viscera; dried ginger warms the lungs and transforms fluid, and assists tianlizi, warming and expelling the evil of phlegm and fluid in the lungs, and is the minister medicine; white peony root can restrain yin and stop sweating, and is the adjuvant medicine; licorice can relieve urgency and relieve pain, harmonize the various medicines, assist peony, and transform yin with acid and sweet to nourish the lung deficiency, and is the guiding medicine. At present, the research on Daxiefei Decoction is mainly focused on clinical and application aspects. Modern pharmaceutical research shows that Daxiefei Decoction can treat chronic obstructive pulmonary disease (COPD) and prevent and treat pneumonia. It has significant efficacy, is safe to take, and has good development prospects.

[0003] At present, the reports on Daxiefei Decoction mainly focus on the evaluation of clinical efficacy, and there is no literature report on its quality control. The lack of quality control methods makes it difficult to effectively control the stability of the quality of the therapeutic preparation, thus affecting the production and quality assurance of the product. Summary of the invention

[0004] The purpose of the present invention is to provide a quality control method for a freeze-dried powder of a traditional Chinese medicine composition, namely, to conduct a comprehensive quality inspection on the material benchmark of Daxiefei Decoction and establish a quality standard for the material benchmark of Daxiefei Decoction, in order to provide a scientific basis for the preparation development of Daxiefei Decoction.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is: A quality control method for a freeze-dried powder of a traditional Chinese medicine composition, wherein the traditional Chinese medicine composition comprises scutellaria baicalensis, rhubarb, white peony root, roasted liquorice, scutellaria baicalensis and dried ginger, and a method for determining the content of a benchmark indicator component in the traditional Chinese medicine composition is established: S1. preparing a decoction of the Chinese medicine composition, freeze-drying it to obtain a lyophilized powder, and using the obtained lyophilized powder as a material reference; S2, prepare the test solution by taking the substance standard obtained in S1; S3, preparing a mixed reference solution consisting of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, paeoniflorin, liquiritigenin, baicalin, aloe-emodin, 6-gingerol, rhein, chrysophanol and physophanol methyl ether; S4, respectively preparing a negative sample solution that does not contain scutellaria baicalensis seeds, a negative sample solution that does not contain rhubarb, a negative sample solution that does not contain white peony root, a negative sample solution that does not contain licorice root, a negative sample solution that does not contain scutellaria baicalensis, and a negative sample solution that does not contain dried ginger in the traditional Chinese medicine composition; S5. Accurately weigh the test solution, mixed reference solution and negative sample solution respectively, and use high performance liquid chromatography to determine the content of the benchmark index components. The chromatographic conditions are set as follows: The chromatographic column was TC-C18 column, the detection wavelength was 254 nm, the column temperature was 30 °C, the flow rate was 1.0 mL / min, the injection volume was 10 μl, the mobile phase was A acetonitrile-methanol-B 0.1% formic acid aqueous solution, the volume ratio of acetonitrile-methanol was 4:6, and the gradient elution program was: 0-15 min, A was 5%→12%, B was 88%→95%; 15-40 min, A was 12%→20%, B was 80%→88%; 40-90 min, A was 20%→35%, B was 65%→80%; 90-110 min, A was 35%, B was 65%; 110-120 min, A was 35%→40%, B was 60%→65%; 120-160 min, A was 40%→72%, B was 28%→60%; 160-175 min, A is 72%→95%, B is 5%→28%; 175-180 min, A is 95%→100%, B is 0%→5%; 180-200 min, A is 100%, B is 0%; The liquid phase data is obtained by measurement and calculation to obtain the content of the benchmark index component in the traditional Chinese medicine composition.

[0006] Based on the above determination method, the content of each benchmark index component in the freeze-dried powder of the traditional Chinese medicine composition, i.e., the material benchmark, is further limited and required: the content of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside per 1g of material benchmark shall not be less than 0.033mg; the content of paeoniflorin per 1g of material benchmark shall not be less than 2.329mg; the content of liquiritin per 1g of material benchmark shall not be less than 0.100mg; the content of baicalin per 1g of material benchmark shall not be less than 5.130mg; the content of aloe-emodin per 1g of material benchmark shall not be less than 0.009mg; the content of 6-gingerol per 1g of material benchmark shall not be less than 0.200mg; the content of rhein per 1g of material benchmark shall not be less than 0.029mg; the content of chrysophanol per 1g of material benchmark shall not be less than 0.013mg; the content of physophanol methyl ether per 1g of material benchmark shall not be less than 0.006mg.

[0007] In order to further realize the present invention, the preparation method of Daxiefei Decoction recorded in the original prescription is "Tinglizi (boiled black and mashed into mud), rhubarb, peony root (3 liang each), licorice (roasted), scutellaria baicalensis, dried ginger (1 liang each), the above six ingredients, add 5 liters of water, boil to obtain 2 liters, divide into warm portions and take again, take twice a day", and according to literature research, 1 liang in the Eastern Han Dynasty is equal to 15.4 g now, and 1 liter is equal to 200 mL. The preparation method of the water decoction of the replica Chinese medicine composition is as follows: take 46.2 g of stir-fried tinglizi, 46.2 g of rhubarb, 46.2 g of white peony root, 15.4 g of roasted licorice, 15.4 g of scutellaria baicalensis and 15.4 g of dried ginger, add 1000 mL of pure water, soak for 1 h, boil it with high heat and then turn to low heat to decoct for 1 h until the medicinal liquid is 400 ml. The obtained medicinal liquid is the water decoction of the traditional Chinese medicine composition. Combined with modern technology, the water decoction of the traditional Chinese medicine composition is filtered and the filtrate is concentrated to 185 mL under reduced pressure to obtain the concentrated water decoction of the traditional Chinese medicine composition. The concentrated water decoction of the traditional Chinese medicine composition is placed in a -80°C refrigerator for 24 h, and then freeze-dried for 48 h. The obtained freeze-dried powder is the material benchmark.

[0008] Since the stability of the preparation process and the influence of the origin and batch of the medicinal pieces are important links in quality control, in order to further realize the present invention, it is necessary to control the paste yield (powder yield) of the material benchmark to 15.48%-22.40%.

[0009] The paste yield of the present invention fluctuates within the range of ±30% of its average value, thereby ensuring the stability of the preparation process.

[0010] The inventors found that quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside in the benchmark indicative component of the material benchmark of the present invention will only peak when 70% methanol is used as the solvent, and because quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside is the indicative component of the monarch drug Lepidium melongena, 70% methanol is determined as the sample solvent. In order to further realize the present invention, a method for preparing a test solution based on the material benchmark is designed: accurately weigh 0.1000 g of the material benchmark, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, make up the lost weight with 70% methanol, cool, pass through a 0.45 μm microporous filter membrane, and take the filtrate to obtain.

[0011] The RSDs of the specificity, precision, repeatability and stability test results of the content determination method for 9 benchmark indicator components, including quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, paeoniflorin, liquiritigenin, baicalin, aloe-emodin, 6-gingerol, rhein, chrysophanol and physophanol methyl ether, established by the present invention are all less than 3.0%, indicating that the above test results are all good; the mass concentrations of the benchmark indicator components have a good linear relationship within their respective ranges; and the RSDs of the average sample recovery rates are all less than 3.0%, indicating that the method has good accuracy.

[0012] The present invention establishes an HPLC fingerprint spectrum, performs attribution of characteristic peaks and similarity evaluation, that is, by analyzing liquid phase data, calculates the similarity between the fingerprint spectrum of the test sample and the fingerprint spectrum of the reference substance, and limits the similarity to no less than 0.7 as a basis for quality control.

[0013] In order to further control the quality of the Chinese medicine composition of the present invention, the contents of total polysaccharides, total flavonoids and total polyphenols in the material benchmark are also limited, that is, the contents of total polysaccharides, total flavonoids and total polyphenols in the material benchmark shall not be less than 16.89%, 28.40% and 0.49%, respectively.

[0014] The present invention also establishes a method for determining the content of total polysaccharides, total flavonoids and total polyphenols in the material standard, including a method for determining the content of total polysaccharides as follows: S1. Prepare the test solution by taking the material standard and prepare the glucose reference solution; S2. Use ultraviolet spectrophotometry, use phenol solution and concentrated sulfuric acid as color developing agents, and the detection wavelength is 490nm. Determine and calculate to obtain; The total flavonoids content was determined as follows: S1. Prepare the test solution by taking the material standard and prepare the rutin reference solution; S2. Using ultraviolet spectrophotometry, with aluminum nitrate solution and sodium hydroxide solution as color developing agents, and a detection wavelength of 500 nm, the results were determined and calculated; The total polyphenol content was determined by: S1. Prepare the test solution by taking the material standard and prepare the gallic acid reference solution; S2. Use ultraviolet spectrophotometry, use Folin phenol solution and sodium carbonate solution as color developing agents, and the detection wavelength is 760nm. Determine and calculate to obtain the result.

[0015] The present invention also carries out qualitative identification on rhubarb, liquorice, scutellaria baicalensis and dried ginger in the material standard.

[0016] In particular, the thin layer chromatography identification method is used to qualitatively identify the scutellaria baicalensis in the material standard, including the following steps: S1. Prepare the test solution by taking the material standard; S2. preparing a scutellaria baicalensis medicinal material reference solution, a baicalin reference solution, a baicalein reference solution, and a negative sample solution not containing scutellaria baicalensis; S3. Absorb the solutions obtained from S1 and S2 and spot them on the same silica gel G thin layer plate respectively. Use ethyl acetate-butanone-formic acid-water-methanol with a volume ratio of 5:3:0.6:1:0.4 as the developing agent. After development, take out, dry, spray with 5% ferric chloride ethanol test solution, and examine under sunlight.

[0017] Compared with the prior art, the present invention has the following beneficial effects: The present invention adopts an evaluation model that combines the content of benchmark indicator components, fingerprints, paste yield and thin-layer chromatography, and conducts comprehensive quality testing on the freeze-dried powder of the Chinese medicine composition of the present invention, i.e., the material benchmark, through multiple technologies and multiple approaches, and formulates the quality standard of the material benchmark, in order to provide a scientific basis for the preparation development of Daxiefei Decoction, and provide ideas for the development and research of similar prescriptions. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is the fingerprint superposition diagram in Experimental Example 4 of the present invention, wherein S1-S15 are HPLC fingerprints of 15 batches of material standards, and R is the reference spectrum; Figure 2 HPLC chromatograms of the test solution, mixed reference solution and each single-flavor decoction piece solution of the material benchmark of S1 in Experimental Example 4 of the present invention, wherein 3 is quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside; 4 is paeoniflorin; 6 is liquiritigenin; 8 is baicalin; 12 is aloe-emodin; 13 is 6-gingerol; 14 is rhein; 16 is chrysophanol; 17 is physophanol methyl ether; Figure 3 HPLC chromatograms of the test solution, mixed reference solution and each negative sample solution of the substance reference of S1 in Experimental Example 4 of the present invention, wherein 3 is quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside; 4 is paeoniflorin; 6 is liquiritigenin; 8 is baicalin; 12 is aloe-emodin; 13 is 6-gingerol; 14 is rhein; 16 is chrysophanol; 17 is physophanol methyl ether; Figure 4 is the standard curve of total polysaccharides in Experimental Example 5 of the present invention; Figure 5 is the standard curve of total flavonoids in Experimental Example 5 of the present invention; Figure 6 is the standard curve of total polyphenols in Experimental Example 5 of the present invention; Figure 7It is the thin layer identification diagram in Experimental Example 6 of the present invention, wherein: A is the thin layer identification diagram of rhubarb, 1-3 are the test solution, 4 are the rhubarb medicinal material reference solution, 5 are the rhein reference solution, 6 are the rhein acid reference solution, and 7 are the negative sample solution; B is the thin layer identification diagram of licorice, 1-3 are the test solution, 4 are the licorice medicinal material reference solution, 5 are the glycyrrhizic acid reference solution, and 6 are the negative sample solution; C is the thin layer identification diagram of scutellaria baicalensis, 1-3 are the test solution, 4 are the scutellaria baicalensis medicinal material reference solution, 5 are the baicalin reference solution, 6 are the baicalein reference solution, and 7 are the negative sample solution; D is the thin layer identification diagram of dried ginger, 1-3 are the test solution, 4 are the dried ginger medicinal material reference solution, 5 are the gingerol reference solution, and 6 are the negative sample solution. DETAILED DESCRIPTION

[0019] The quality control method of a freeze-dried powder of a traditional Chinese medicine composition provided by the present invention is further described below in conjunction with experimental examples and accompanying drawings.

[0020] Experimental Example 1: Preparation of freeze-dried powder of Chinese medicine decoction as material reference: 1. Instruments and test drugs 1. Instruments: 1260Ⅱ high performance liquid chromatograph (Agilent Technologies, USA); N1210B rotary evaporator (Shanghai Airang Instrument Co., Ltd.); ultra-low temperature refrigerator (Panasonic Corporation, Japan); FDU-2110 freeze dryer (Shanghai Airang Instrument Co., Ltd.); ABJ80-4M electronic analytical balance (1 / 10,000); ABT100-5M electronic analytical balance (1 / 100,000) (Cohen, Germany); DT1028H ultrasonic cleaner; DHG-9070AE electric heating blast drying oven (Guangdong Taihongjun Scientific Instrument Co., Ltd.).

[0021] 2. Drug testing (1) Reference substances: quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside (batch number zc2306001), liquiritin (batch number zc06001), baicalin (batch number zc2307001), 6-gingerol (batch number zc2308001), Lanzhou Zhichun Biotechnology Co., Ltd., purity ≥ 98.0%; rhein (batch number 19010521), rutin (batch number 1609225), gallic acid (batch number 18102509), D-anhydrous-glucose (batch number 22070596), Lanzhou Plant Biotechnology Co., Ltd., purity ≥ 98.0 %; Rhein (batch number 110756-200110), Rhein methyl ether (batch number 110758-2000509), Aloe-emodin (batch number 110795-200504), Rhein (batch number 110796-200513), Paeoniflorin (batch number 110736-200630), National Institute for the Control of Food and Biological Products, purity ≥98.0%.

[0022] (2) Reagents: Petroleum ether (batch number: 20231101), anhydrous ethanol (batch number: 20221005), ethyl formate (batch number: 20230201), and glacial acetic acid (batch number: 20230504) were all analytical grade, from Tianjin Damao Chemical Reagent Factory; ferric chloride (batch number: 20210301) and n-butanol (batch number: 20200301) were all analytical grade, from Yantai Shuangshuang Chemical Co., Ltd.; ethyl acetate (batch number: 20180103) was analytical grade, from Lianlong Bohua Tianjin Pharmaceutical Chemical Co., Ltd.; vanillin (batch number: 20201113) was from Tianjin Komiou Chemical Reagent Co., Ltd.; formic acid (batch number: 20200825) was chromatographic grade, from Tianjin Damao Chemical Reagent Factory; methanol (batch number: 1143407112) and acetonitrile (batch number: JA100930) were all chromatographic grade, from Merck, Germany.

[0023] (3) Medicinal materials: All batches of stir-fried scutellaria baicalensis seeds, rhubarb, white peony root, dried ginger, roasted licorice, and scutellaria baicalensis slices were provided by the company. The specific information is shown in Table 1. They were identified as Artemisia sophora japonica, a plant of the cruciferous family. Description sophia (L.) Webb.ex Prantl. Rheum palmatum Rheum palmatum L . Dried root of Paeonia lactiflora, Ranunculaceae Paeonia lactiflora Pall. Zingiber officinale Zingiber officinale Rosc. Dried rhizomes of Licorice Glycyrrhiza uralensis Fisch. Scutellaria baicalensis Scutellaria baicalensis Georgi The dried roots all meet the requirements of Part I of the 2020 edition of the Chinese Pharmacopoeia.

[0024] 2. Preparation of water decoction and freeze-dried powder of Chinese medicine composition The six medicinal pieces were randomly combined and sorted according to the random number table method. The 15 batches of water decoctions of the Chinese medicine composition corresponded to the batches of medicinal pieces (as shown in Table 2). The preparation method of the Chinese medicine composition recorded in the original prescription, namely, Daxiefei Decoction, is "Tinglizi (boiled black and mashed into a paste), Rhubarb, Peony Root (3 liang each), Licorice (roasted), Scutellaria Baicalensis, Dry Ginger (1 liang each). Add the above six ingredients to 5 liters of water, boil and extract 2 liters, divide into portions and take warm, twice a day". According to literature research, 1 liang in the Eastern Han Dynasty is equal to 15.4 g today, and 1 liter is equal to 200 mL. After conversion, 46.2 g of stir-fried scutellaria baicalensis seeds, 46.2 g of rhubarb, 46.2 g of white peony root, 15.4 g of roasted liquorice root, 15.4 g of scutellaria baicalensis and 15.4 g of dried ginger were taken, soaked in 1000 mL of pure water for 1 h, boiled over high heat and then simmered on low heat for 1 h until the liquid reached 400 ml. The obtained liquid was the decoction of the traditional Chinese medicine composition. The filtrate after filtration was concentrated under reduced pressure to 185 mL to obtain a concentrated decoction of the traditional Chinese medicine composition. The concentrated decoction was placed in a -80°C refrigerator for 24 h, and then freeze-dried for 48 h. The obtained freeze-dried powder of the traditional Chinese medicine composition was used as the material benchmark.

[0025] Experimental Example 2: Determination of the content of 9 benchmark index components in the material standard: 1. Content determination method: S1. Prepare a decoction of the Chinese medicine composition by the method of Experimental Example 1, freeze-dry, and obtain a lyophilized powder, and use the obtained lyophilized powder as a material reference; S2. Prepare the test solution by taking the substance standard: accurately weigh 0.1000 g of the substance standard obtained in S1, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, make up the weight loss with 70% methanol, cool, filter through a 0.45 μm microporous membrane, and take the filtrate to obtain the test solution; S3. Prepare mixed reference solution: (1) Accurately weigh appropriate amounts of quercetin-3-O-β-D-glucoside-7-O-β-D-gentiobioside, paeoniflorin, liquiritin, baicalin, aloe-emodin, 6-gingerol, rhein, chrysophanol and physophanol methyl ether reference substances, mix them, and dissolve them in methanol by ultrasonication to prepare mixed reference stock solutions with mass concentrations of 0.112, 0.432, 0.134, 0.324, 0.092, 0.30, 0.21, 0.092 and 0.144 mg / ml, respectively; (2) Using the stepwise dilution method, accurately pipette 1 ml of the mixed reference stock solution and dilute it with methanol into mixed reference solutions of different concentrations: 2x, 4x, 6x, 8x, and 10x; S4. Prepare respectively a negative sample solution containing no scutellariae seeds, a negative sample solution containing no rhubarb, a negative sample solution containing no white peony root, a negative sample solution containing no licorice root, a negative sample solution containing no scutellariae baicalensis, and a negative sample solution containing no dried ginger in the traditional Chinese medicine composition: (1) Preparation of negative sample solution without scutellaria baicalensis seeds Take 46.2 g of rhubarb, 46.2 g of white peony root, 15.4 g of roasted liquorice, 15.4 g of scutellaria baicalensis and 15.4 g of dried ginger, soak in 1000 mL of pure water for 1 h, boil over high heat, then turn to low heat and decoct for 1 h until the liquid is 400 ml, to obtain a decoction of the Chinese medicine composition without scutellaria baicalensis seeds, and concentrate the filtered filtrate under reduced pressure to 185 mL to obtain a decoction concentrate of the Chinese medicine composition without scutellaria baicalensis seeds. Place the obtained decoction concentrate of the Chinese medicine composition in a -80 ° C refrigerator for 24 h, and freeze-dry for 48 h. The obtained freeze-dried powder is a negative sample lacking scutellaria baicalensis seeds. Accurately weigh 0.1000 g of the negative sample, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, and lyophilize with 70 Add % methanol to make up for the weight loss, cool, filter through a 0.45 μm microporous membrane, and take the filtrate to obtain the negative sample solution without scutellaria baicalensis seeds; (2) Preparation of negative sample solution without rhubarb Take 46.2 g of stir-fried scutellaria baicalensis seeds, 46.2 g of white peony root, 15.4 g of roasted liquorice root, 15.4 g of scutellaria baicalensis and 15.4 g of dried ginger, add 1000 mL of pure water and soak for 1 h, boil with high heat and then turn to low heat to decoct for 1 h until the liquid is 400 ml, to obtain a water decoction of the Chinese medicine composition without rhubarb, and the filtered filtrate is concentrated under reduced pressure to 185 mL to obtain a water decoction concentrate of the Chinese medicine composition without rhubarb, place the obtained water decoction concentrate of the Chinese medicine composition in a -80°C refrigerator for 24 h, and freeze-dry for 48 h. The obtained lyophilized powder is a negative sample without rhubarb, accurately weigh 0.1000 g of the negative sample, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, make up the weight loss with 70% methanol, cool, pass through a 0.45 μm microporous filter membrane, and take the subsequent filtrate to obtain a negative sample solution without rhubarb; (3) Preparation of negative sample solution without white peony root Take 46.2 g of stir-fried scutellaria baicalensis seeds, 46.2 g of rhubarb, 15.4 g of roasted liquorice, 15.4 g of scutellaria baicalensis and 15.4 g of dried ginger, add 1000 mL of pure water and soak for 1 h, boil with high heat and then turn to low heat to decoct for 1 h until the liquid is 400 ml, to obtain a decoction of the Chinese medicine composition without white peony, and the filtered filtrate is concentrated under reduced pressure to 185 mL to obtain a decoction concentrate of the Chinese medicine composition without white peony, place the obtained decoction concentrate of the Chinese medicine composition in a -80 ° C refrigerator for 24 h, and freeze-dry for 48 h. The obtained lyophilized powder is a negative sample without white peony, accurately weigh 0.1000 g of the negative sample, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, make up the weight loss with 70% methanol, cool, pass through a 0.45 μm microporous filter membrane, and take the filtrate to obtain a negative sample solution without white peony; (4) Preparation of negative sample solution without licorice root Take 46.2 g of stir-fried scutellaria baicalensis seeds, 46.2 g of rhubarb, 46.2 g of white peony root, 15.4 g of scutellaria baicalensis and 15.4 g of dried ginger, add 1000 mL of pure water and soak for 1 h, boil with high heat and then turn to low heat to decoct for 1 h until the liquid is 400 ml, to obtain a decoction of the Chinese medicine composition without licorice, and the filtered filtrate is concentrated under reduced pressure to 185 mL to obtain a decoction concentrate of the Chinese medicine composition without licorice, place the obtained decoction concentrate of the Chinese medicine composition in a -80°C refrigerator for 24 h, and freeze-dry for 48 h. The obtained lyophilized powder is a negative sample without licorice, accurately weigh 0.1000 g of the negative sample, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, make up the weight loss with 70% methanol, cool, pass through a 0.45 μm microporous filter membrane, and take the filtrate to obtain a negative sample solution without licorice; (5) Preparation of negative sample solution without Scutellaria baicalensis Take 46.2 g of stir-fried scutellaria baicalensis seeds, 46.2 g of rhubarb, 46.2 g of white peony root, 15.4 g of roasted licorice and 15.4 g of dried ginger, add 1000 mL of pure water and soak for 1 h, boil over high heat and then turn to low heat to decoct for 1 h until the liquid is 400 ml, to obtain a decoction of the Chinese medicine composition without scutellaria baicalensis, and the filtrate after filtration is concentrated under reduced pressure to 185 mL to obtain a decoction concentrate of the Chinese medicine composition without scutellaria baicalensis. Place the obtained decoction concentrate of the Chinese medicine composition in a -80 ° C refrigerator for 24 h, and freeze-dry for 48 h. The obtained freeze-dried powder is a negative sample without scutellaria baicalensis. Accurately weigh 0.1000 g of the negative sample, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, and lyophilize with 70 % methanol to make up for the loss in weight, cool, filter through a 0.45 μm microporous membrane, and take the filtrate to obtain a negative sample solution that does not contain Scutellaria baicalensis; (6) Preparation of negative sample solution without dried ginger Take 46.2 g of stir-fried scutellaria baicalensis seeds, 46.2 g of rhubarb, 46.2 g of white peony root, 15.4 g of roasted licorice and 15.4 g of scutellaria baicalensis, add 1000 mL of pure water and soak for 1 h, boil with high heat and then turn to low heat to decoct for 1 h until the liquid is 400 ml, to obtain a decoction of the Chinese medicine composition without dried ginger, and the filtered filtrate is concentrated under reduced pressure to 185 mL to obtain a concentrated decoction of the Chinese medicine composition without dried ginger, and place the obtained concentrated decoction of the Chinese medicine composition in a -80 ° C refrigerator for 24 h, and freeze-dry for 48 h. The obtained lyophilized powder is a negative sample without dried ginger, accurately weigh 0.1000 g of the negative sample, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, make up the weight loss with 70% methanol, cool, pass through a 0.45 μm microporous filter membrane, and take the filtrate to obtain a negative sample solution without dried ginger; S5. Accurately weigh the test solution, mixed reference solution and negative sample solution respectively, and use high performance liquid chromatography to determine the content of the benchmark index components. The chromatographic conditions are set as follows: The chromatographic column was TC-C18 column, the detection wavelength was 254 nm, the column temperature was 30 °C, the flow rate was 1.0 mL / min, the injection volume was 10 μl, the mobile phase was A acetonitrile-methanol-B 0.1% formic acid aqueous solution, the volume ratio of acetonitrile-methanol was 4:6, and the gradient elution program was: 0-15 min, A was 5%→12%, B was 88%→95%; 15-40 min, A was 12%→20%, B was 80%→88%; 40-90 min, A was 20%→35%, B was 65%→80%; 90-110 min, A was 35%, B was 65%; 110-120 min, A was 35%→40%, B was 60%→65%; 120-160 min, A was 40%→72%, B was 28%→60%; 160-175 min, A is 72%→95%, B is 5%→28%; 175-180 min, A is 95%→100%, B is 0%→5%; 180-200 min, A is 100%, B is 0%; The liquid phase data is obtained by measurement and calculation to obtain the content of the benchmark index component in the material benchmark.

[0026] 2. Methodological Investigation: 1. Specificity experiment: Accurately weigh the test solution, mixed reference solution and negative sample solution, and analyze them under the chromatographic conditions of S5 in "I. Content determination method". The result of negative sample solution is as follows: Figure 3 As shown, there is no interference at the peak time of the determined components, indicating that the method has good specificity.

[0027] 2. Linear relationship investigation Take the mixed reference stock solution prepared in S3 (1) in "I. Content determination method" and the five mixed reference substance solutions of different concentrations prepared in S3 (2), and perform the sample injection and determination under the chromatographic conditions of S5 in "I. Content determination method". （X） is the mass concentration of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, paeoniflorin, liquiritin, baicalin, aloe-emodin, 6-gingerol, rhein, chrysophanol and physophanol methyl ether, and the peak area is the ordinate. （Y） , linear regression was performed, and the regression equation obtained is shown in Table 3. It can be seen that the linear relationship between each component is good within its respective range.

[0028] 3. Precision test: Take one portion of the test solution and inject it continuously for 6 times according to the chromatographic conditions of S5 in "I. Content Determination Method". The RSD values ​​of the peak areas of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, paeoniflorin, liquiritigenin, baicalin, aloe-emodin, 6-gingerol, rhein, chrysophanol and physophanol methyl ether were 1.26%, 0.46%, 1.09%, 0.60%, 1.53%, 0.37%, 0.19%, 0.36% and 0.56%, respectively. The RSD values ​​of the above components were all less than 2.0%, indicating that the instrument had good precision.

[0029] 4. Repeatability investigation: Six portions of the same batch of test solution were taken and injected separately under the chromatographic conditions of S5 in "I. Content determination method". The RSDs of the mass fractions of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, paeoniflorin, liquiritigenin, baicalin, aloe rhubarb, 6-gingerol, rhein, chrysophanol and physophanol methyl ether were 1.64%, 1.07%, 1.10%, 1.16%, 1.38%, 1.74%, 1.07%, 0.91% and 1.30%, respectively. The RSD values ​​of the above components were all less than 2.0%, indicating that the method had good repeatability.

[0030] 5. Stability investigation: Take one portion of the test solution and inject it at 0, 4, 10, 17 and 24 h under the chromatographic condition of S5 in "I. Content Determination Method". The RSDs of the peak areas of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, paeoniflorin, liquiritigenin, baicalin, aloe-emodin, 6-gingerol, rhein, chrysophanol and physophanol methyl ether were 2.11%, 2.66%, 1.05%, 0.26%, 1.73%, 1.69%, 2.50%, 0.97% and 1.40%, respectively. The RSD values ​​of the above components were all less than 3.0%, indicating that the sample had good stability.

[0031] 6. Sample recovery investigation Take 6 portions of the substance with the determined content of each benchmark index component (0.050 g) and place them in 1 ml volumetric flasks. Add 50%, 100%, and 150% of the benchmark index component to the mixed reference solution, respectively. Prepare the test solution according to the method of S2 in "I. Content determination method". Prepare 3 portions in parallel for each mass concentration. Under the chromatographic conditions of S5 in "I. Content Determination Method", the samples were injected and determined respectively, and the recoveries of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, paeoniflorin, liquiritigenin, baicalin, aloe-emodin, 6-gingerol, rhein, chrysophanol and physophanol methyl ether were calculated respectively. The results showed that the average recoveries of the nine components were 94.97%, 95.83%, 95.54%, 96.09%, 92.01%, 95.23%, 97.17%, 94.06% and 96.80%, respectively, and the RSDs were 1.68%, 2.70%, 1.90%, 2.18%, 2.65%, 2.59%, 2.71%, 2.71% and 1.73%, respectively. The average recoveries of the benchmark index components were between 92% and 98%, and the RSDs were all less than 3.0%, indicating that the method had good accuracy.

[0032] III. Determination of the content limits of the nine benchmark indicators in the material benchmark By determining the contents of 9 benchmark components in 15 batches of material bases, the minimum content in each group was determined as the content limit, that is, the content of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside per 1g of material base shall not be less than 0.033mg; the content of paeoniflorin per 1g of material base shall not be less than 2.329mg; the content of liquiritin per 1g of material base shall not be less than 0.100mg; the content of baicalin per 1g of material base shall not be less than 5.130mg; the content of aloe-emodin per 1g of material base shall not be less than 0.009mg; the content of 6-gingerol per 1g of material base shall not be less than 0.200mg; the content of rhein per 1g of material base shall not be less than 0.029mg; the content of chrysophanol per 1g of material base shall not be less than 0.013mg; the content of physophanol methyl ether per 1g of material base shall not be less than 0.006mg, as shown in Table 4 and Table 4.

[0033] Experimental Example 3: Benchmark cream yield test: The concentrated decoction of the Chinese medicine composition obtained in Experimental Example 1 was converted to the material standard in proportion to obtain the dry paste mass of the material standard, and the actual paste yield of the material standard was calculated. The corresponding single-flavor decoction piece solutions and negative sample solutions were prepared in the same way to obtain their respective dry paste masses and determine the single-flavor decoction piece paste yield. The theoretical paste yield of the material standard was calculated according to the formula theoretical paste yield = ∑ single-flavor decoction yield × single-flavor decoction piece mass / total amount of the whole prescription.

[0034] As shown in Table 5, the average value of the actual paste yield of 15 batches of material standards is 18.11%, and the range of ±30% is 15.48%-22.40%. The actual paste yield fluctuates within the range of ±30% of its average value, indicating that the preparation process of the material standard is relatively stable, and the origin and batch of the medicinal pieces have little effect on it.

[0035] Experimental Example 4: Material-based fingerprint study: 1. Chromatographic conditions, preparation of test solution and preparation of mixed reference solution: see Experimental Example 2.

[0036] 2. See Experimental Example 2 for precision test.

[0037] 3. Repeatability study see Experimental Example 2.

[0038] 3. Stability investigation see Experimental Example 2.

[0039] 4. Fingerprint establishment and similarity evaluation According to the method in Experimental Example 2, 15 batches of sample solutions were prepared. The chromatographic conditions in Experimental Example 2 were used for sample injection and measurement. The chromatograms of each batch were recorded and analyzed using the "Chinese Herbal Medicine Chromatographic Fingerprint Similarity Evaluation System (2004.A Edition)" software (refer to the chromatogram: S6; time window width: 0.1 min; median method; similarity calculation: multi-point correction and full spectrum peak matching), the fingerprints of the test solutions of 15 batches of substance benchmarks were calculated by software, and the reference fingerprints were generated. The results showed that the similarities between the fingerprints of the test solutions of 15 batches of substance benchmarks (S1-S15) and the reference fingerprints (R) were 0.789, 0.808, 0.854, 0.877, 0.815, 0.822, 0.937, 0.944, 0.967, 0.883, 0.903, 0.889, 0.939, 0.925, and 0.933, respectively, all greater than 0.7, indicating that the overall similarity of the 15 batches of substance benchmarks was good. The fingerprint overlay diagram is shown in Figure 1 .

[0040] 5. Identification and attribution of chromatographic peaks The decoction of the Chinese medicine composition numbered S1 in Table 2 was taken and prepared as a material reference according to the method of Experimental Example 1. The test solution of the material reference of S1 was prepared by the method in Experimental Example 2. The fingerprints of the test solution and the single-flavor decoction solution, the negative sample solution and the mixed reference solution prepared in Experimental Example 3 were compared. It was found that the test solution of the material reference of S1 had 17 common peaks at a wavelength of 254 nm, such as Figure 2-3 As shown, peaks 3 (quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside) and 5 belong to the Tenglizi decoction piece, peaks 1 and 4 (peonidin) belong to the white peony decoction piece, peaks 2, 6 (liquiritin) and 15 belong to the liquorice decoction piece, peaks 8 (baicalin), 9, 10 and 11 belong to the scutellaria decoction piece, peak 13 (6-gingerol) belongs to the dried ginger decoction piece, and peaks 7, 12 (aloe-emodin), 14 (rhein), 16 (chrysophanol) and 17 (physophanol methyl ether) belong to the rhubarb decoction piece. It can be seen that the material standard prepared by the process under Experimental Example 1 has a relatively complete transfer of its main components from the decoction piece to the material standard, and the attribution relationship is relatively clear.

[0041] Experimental Example 5: Determination of the content of total polysaccharides, total flavonoids and total polyphenols in the material standard: 1. Determination of total polysaccharides: Accurately weigh 0.01091g of glucose reference substance, place it in a 25ml volumetric flask, add purified water to dissolve and dilute to the scale, shake well, and obtain a glucose reference solution with a concentration of 0.4364mg / ml. Accurately pipette 0.8, 1.0, 1.2, 1.4, and 1.6ml of the above glucose reference solution into a 10ml volumetric flask, add purified water to the scale, and shake well. Accurately pipette 2ml of each of the above solutions into a stoppered measuring cylinder, add 1.0ml of 4% phenol solution respectively, shake well, quickly add 7ml of concentrated sulfuric acid, shake well, keep warm in a 40℃ water bath for 30min, take out, cool in an ice water bath for 5min, take out, use the corresponding reagent as a blank, and measure the absorbance at a wavelength of 490nm using a UV-visible photometer. See the standard curve of total polysaccharides. Figure 4 .

[0042] Prepare the test solution: accurately weigh 0.1 g of 15 batches of material standards, dilute to a 25 ml volumetric flask with purified water, shake well, then take 5 ml and dilute to a 25 ml volumetric flask, shake well again, then take 1 ml and dilute to a 10 ml volumetric flask, accurately pipette 2 ml of the sample solution into a stoppered measuring cylinder, add 1.0 ml of 4% phenol solution respectively, shake well, quickly add 7 ml of concentrated sulfuric acid, shake well, keep warm in a 40 ° C water bath for 30 min, take out, cool in an ice water bath for 5 min, take out, use the corresponding reagent as a blank, and measure the absorbance at a wavelength of 490 nm according to the UV-Vis photometer. Measure its content according to the standard curve. As shown in Table 6, the content of total polysaccharides in the 15 batches of material standards is 16.89-27.77%.

[0043] 2. Determination of total flavonoids: Accurately weigh 0.00507g of rutin reference substance, place it in a 25ml volumetric flask, add 70% alcohol to the scale, shake well, and obtain a rutin reference substance solution with a concentration of 0.2028mg / ml. Accurately aspirate 2.5ml, 3.5ml, 4.5ml, 5.5ml, and 6.5ml of rutin reference substance solution, place them in 25ml volumetric flasks, add 0.4ml of 5% sodium nitrite solution, shake well, and let it stand for 6min, add 0.4ml of 10% aluminum nitrate solution, let it stand for 6min, add 4ml of 4% sodium hydroxide solution, add 70% ethanol to the scale, let it stand for 15min, use 70% ethanol as blank, measure the absorbance at a wavelength of 500nm, and draw a standard curve, see Figure 5 .

[0044] Preparation of test solution: accurately weigh 0.1g of freeze-dried powder of each batch of 15 batches of the Chinese medicine composition of the present invention into a 25ml volumetric flask, add 70% ethanol, ultrasonic for 40min, make up the weight, fix the volume, and then take 5ml from it into a 25ml volumetric flask, add 70% ethanol to fix the volume to the scale line, and the test solution is obtained. Accurately draw 3ml of the sample solution, put it in a stoppered measuring cylinder, add 0.4ml of 5% sodium nitrite solution, shake well and place for 6min, add 0.4ml of 10% aluminum nitrate solution, place for 6min, add 4ml of 4% sodium hydroxide solution, use 70% ethanol as a blank, measure the absorbance at a wavelength of 500mn, substitute it into the standard curve equation, and obtain the total flavonoid content in the sample. As shown in Table 6, the total flavonoid content in the 15 batches of material standards is 28.40-189.87%.

[0045] 3. Determination of total polyphenols: Accurately weigh 0.00251 g of gallic acid reference substance, place it in a 25 ml volumetric flask, and dilute to the mark with purified water to obtain a gallic acid reference substance solution with a concentration of 0.1004 mg / ml. Accurately pipette 1.5, 2.0, 2.5, 3.0, and 3.5 ml of the above reference substance solution into a 25 ml brown volumetric flask, add 1.5 ml of Folin phenol reagent in turn, react for 8 min, then add 1.5 ml of 10% sodium carbonate solution, dilute to the mark with purified water, and stand at 25°C for 60 min (protected from light). Use the corresponding reagent as blank, measure the absorbance at a wavelength of 760 nm, and make a standard curve, see Figure 6 , and we get the regression equation.

[0046] Preparation of test solution: accurately weigh 0.1g of freeze-dried powder of each batch of 15 batches of the Chinese medicine composition of the present invention, put it in a 25ml volumetric flask, dilute to the mark with purified water, then take 5ml from it to a 25ml volumetric flask, add purified water to dilute to the mark, and the test solution is obtained. Accurately draw 2ml of the sample solution into a 25ml test tube, add 1.5ml of Folin phenol reagent, react for 8 min, then add 1.5ml of 10% sodium carbonate solution, add purified water to dilute to the mark, stand at 25°C for 60 min (protected from light), use the corresponding reagent as a blank, measure the absorbance at a wavelength of 760nm, substitute it into the regression equation, and calculate the total polyphenol content. As shown in Table 6, the total polyphenol content in the 15 batches of material basis is 0.49-2.24%.

[0047] IV. Determination of the content limits of total polysaccharides, total flavonoids and total polyphenols in the material standards: By determining the contents of total polysaccharides, total flavonoids and total polyphenols in 15 batches of material benchmarks, the minimum content in each group was determined as the content limit, that is, the content of total polysaccharides shall not be less than 16.89%, the content of total flavonoids shall not be less than 28.40%, and the content of total polyphenols shall not be less than 0.49%.

[0048] Experimental Example 6: Qualitative identification of rhubarb, liquorice, scutellaria and dried ginger in the material standard: 1. Thin layer chromatography identification of rhubarb Weigh the decoctions of the Chinese medicine compositions numbered S1, S2, and S3 in Table 2 respectively, and prepare them as material standards according to the method of Experimental Example 1. Accurately weigh 0.0782g, 0.0713g, and 0.0695g of the material standards respectively, and according to the thin layer chromatography method of the medicinal materials and decoction pieces of rhubarb in the 2020 edition of the "Chinese Pharmacopoeia (Part I)", obtain the material standard test solution, rhubarb medicinal material reference solution, aloe-rhubarb reference solution, rhein reference solution, chrysophanol reference solution, physophanol methyl ether reference solution, and negative sample solution without rhubarb. Thin layer chromatography identification of rhubarb was performed, and the results showed that the same orange-yellow fluorescent spots appeared in the material standard test solution at the corresponding position of the chromatogram of the rhubarb medicinal material reference solution, and the negative sample solution without rhubarb showed no interference. See details. Figure 7 A.

[0049] 2. TLC Identification of Licorice Weigh the decoctions of the Chinese medicine compositions numbered S1, S2, and S3 in Table 2 respectively, and prepare them as material standards according to the method of Experimental Example 1. Accurately weigh 2.3172g, 2.1291g, and 2.0780g of the material standards respectively, and obtain the material standard test solution, licorice medicinal material reference solution, liquiritin reference solution, and negative sample solution without licorice according to the thin layer chromatography method of the 2020 edition of the "Chinese Pharmacopoeia (Part I)". Thin layer chromatography identification of licorice was performed, and the material standard test solution showed the same orange-yellow fluorescent spot at the corresponding position of the chromatogram of the licorice medicinal material reference solution, and the negative sample solution without licorice showed no interference. See details. Figure 7 B.

[0050] 3. TLC Identification of Scutellaria baicalensis Weigh the decoctions of the Chinese medicine compositions numbered S1, S2, and S3 in Table 2 respectively, and prepare them as material standards according to the method of Experimental Example 1, respectively. Accurately weigh 2.0009 g, 2.1302 g, and 2.0778 g of the material standards, respectively, to prepare the test solutions. According to the thin layer chromatography method of the medicinal materials and decoction pieces of Scutellaria baicalensis in the 2020 edition of the "Chinese Pharmacopoeia (Part I)", the material standard test solution, Scutellaria baicalensis medicinal material reference solution, baicalin reference solution, baicalein reference solution, and a negative sample solution without Scutellaria baicalensis were obtained.

[0051] According to the thin layer chromatography method (General Rule 0502), the above solutions were respectively spotted on the same silica gel G thin layer plate, and ethyl acetate-butanone-formic acid-water-methanol (volume ratio of 5:3:0.6:1:0.4) was used as the developing agent. After development, the plate was taken out, dried, sprayed with 5% ferric chloride ethanol test solution, and placed under sunlight for inspection. The results showed that at the corresponding position of the chromatogram of the scutellaria baicalensis medicinal material reference solution, the chromatogram of the material reference test solution showed spots of the same color, and there was no interference with the negative sample solution control without scutellaria baicalensis. See details. Figure 7 C.

[0052] 4. Thin layer chromatography identification of dried ginger Weigh the decoctions of the Chinese medicine compositions numbered S1, S2, and S3 in Table 2 respectively, and prepare them as material standards according to the method of Experimental Example 1. Accurately weigh 2.3247g, 2.1295g, and 2.0784g of the material standards respectively, and according to the thin layer chromatography method of dried ginger for medicinal materials and decoction pieces in the 2020 edition of the "Chinese Pharmacopoeia (Part I)", obtain the material standard test solution, dried ginger medicinal material reference solution, 6-gingerol reference solution, and negative sample solution without dried ginger. Thin layer chromatography identification of dried ginger was performed, and the results showed that at the corresponding position of the chromatogram of the dried ginger medicinal material reference solution, the material standard test solution showed spots of the same color in the chromatogram, and there was no interference with the negative sample solution control without dried ginger. See details. Figure 7 D.

Claims

1. A method for controlling the quality of a freeze-dried powder of a traditional Chinese medicine composition, wherein the traditional Chinese medicine composition comprises scutellaria baicalensis, rhubarb, white peony root, roasted liquorice, scutellaria baicalensis and dried ginger, characterized in that: Establish a method for determining the content of the benchmark index components in the traditional Chinese medicine composition: S1. preparing a decoction of the Chinese medicine composition, freeze-drying it to obtain a lyophilized powder, and using the obtained lyophilized powder as a material reference; S2, prepare the test solution by taking the substance standard obtained in S1; S3, preparing a mixed reference solution consisting of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, paeoniflorin, liquiritigenin, baicalin, aloe-emodin, 6-gingerol, rhein, chrysophanol and physophanol methyl ether; S4, respectively preparing a negative sample solution that does not contain scutellaria baicalensis seeds, a negative sample solution that does not contain rhubarb, a negative sample solution that does not contain white peony root, a negative sample solution that does not contain licorice root, a negative sample solution that does not contain scutellaria baicalensis, and a negative sample solution that does not contain dried ginger in the traditional Chinese medicine composition; S5. Accurately weigh the test solution, mixed reference solution and negative sample solution respectively, and use high performance liquid chromatography to determine the content of the benchmark index components. The chromatographic conditions are set as follows: The chromatographic column was TC-C18, the detection wavelength was 254 nm, the column temperature was 30 °C, the flow rate was 1.0 mL / min, the injection volume was 10 μl, the mobile phase was A acetonitrile-methanol-B 0.1% formic acid aqueous solution, the volume ratio of acetonitrile-methanol was 4:6, and the gradient elution program was: 0-15 min, A is 5%→12%, B is 88%→95%; 15-40 min, A is 12%→20%, B is 80%→88%; 40-90 min, A is 20%→35%, B is 65%→80%; 90-110 min, A is 35%, B is 65%; 110-120 min, A is 35%→40%, B is 60%→65%; 120-160 min, A: 40%→72%, B: 28%→60%; 160-175 min, A: 72%→95%, B: 5%→28%; 175-180 min, A is 95%→100%, B is 0%→5%; 180-200 min, A is 100%, B is 0%; The liquid phase data is obtained by measurement and calculation to obtain the content of the benchmark index component in the traditional Chinese medicine composition.

2. The quality control method of the lyophilized powder of the Chinese medicine composition according to claim 1, characterized in that: The content of each benchmark indicator component in the material benchmark: The content of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside per 1g of the substance shall not be less than 0.033mg; The content of paeoniflorin per 1g of substance shall not be less than 2.329mg; The content of liquiritin per 1g of substance shall not be less than 0.100mg; The content of baicalin per 1g of substance shall not be less than 5.130mg; The content of aloe-emodin per 1g of substance shall not be less than 0.009mg; The content of 6-gingerol in each 1g of the substance shall not be less than 0.200mg; The content of rhein per 1g of substance shall not be less than 0.029mg; The content of chrysophanol per 1g of substance shall not be less than 0.013mg; The content of physaccharin methyl ether per 1g of substance shall not be less than 0.006mg.

3. The quality control method of the lyophilized powder of the Chinese medicine composition according to claim 2, characterized in that: The preparation method of the material standard described in S1 is as follows: take 46.2 g of stir-fried scutellaria baicalensis seeds, 46.2 g of rhubarb, 46.2 g of white peony root, 15.4 g of roasted liquorice root, 15.4 g of scutellaria baicalensis and 15.4 g of dried ginger, add 1000 mL of pure water and soak for 1 hour, boil over high heat and then simmer over low heat for 1 hour until the medicinal liquid is 400 ml, the obtained medicinal liquid is the water decoction of the traditional Chinese medicine composition, filter, and concentrate the filtrate under reduced pressure to 185 mL to obtain a concentrated water decoction of the traditional Chinese medicine composition, place the obtained concentrated water decoction of the traditional Chinese medicine composition in a -80°C refrigerator for 24 hours, and then freeze-dry for 48 hours, and the obtained freeze-dried powder is the material standard.

4. The quality control method of the freeze-dried powder of the Chinese medicine composition as claimed in claim 3, characterized in that: The cream yield of the substance benchmark is 15.48%-22.40%.

5. The quality control method of the freeze-dried powder of the Chinese medicine composition as claimed in claim 4, characterized in that: The method for preparing the test solution in S2 is as follows: accurately weigh 0.1000 g of the substance standard, add 70% methanol to a 1 ml volumetric flask, ultrasonicate at a frequency of 50 Hz for 30 min, make up the loss in weight with 70% methanol, cool, filter through a 0.45 μm microporous filter membrane, and obtain the filtrate.

6. The quality control method of the freeze-dried powder of the Chinese medicine composition according to claim 5, characterized in that: Analyze the liquid phase data obtained in S5, calculate the fingerprint of the test solution by software to generate a reference fingerprint, and then calculate the similarity between each test solution fingerprint and the reference fingerprint, and the similarity shall not be less than 0.

7.

7. The quality control method of the freeze-dried powder of the Chinese medicine composition according to claim 1, characterized in that: The quality control method also includes the determination of the total polysaccharide, total flavonoid and total polyphenol content in the material benchmark. The total polysaccharide content in the material benchmark shall not be less than 16.89%, the total flavonoid content shall not be less than 28.40%, and the total polyphenol content shall not be less than 0.49%.

8. The quality control method of the freeze-dried powder of the Chinese medicine composition according to claim 7, characterized in that: The total polysaccharide content determination method is: S1. Prepare the test solution by taking the material standard and prepare the glucose reference solution; S2. Use ultraviolet spectrophotometry, use phenol solution and concentrated sulfuric acid as color developing agents, and the detection wavelength is 490nm. Determine and calculate to obtain; The total flavonoids content determination method is: S1. Prepare the test solution by taking the material standard and prepare the rutin reference solution; S2. Using ultraviolet spectrophotometry, with aluminum nitrate solution and sodium hydroxide solution as color developing agents, and a detection wavelength of 500 nm, the results were determined and calculated; The total polyphenol content determination method is: S1. Prepare the test solution by taking the material standard and prepare the gallic acid reference solution; S2. Use ultraviolet spectrophotometry, use Folin phenol solution and sodium carbonate solution as color developing agents, and the detection wavelength is 760nm. Determine and calculate to obtain the result.

9. The quality control method of the freeze-dried powder of the Chinese medicine composition according to claim 8, characterized in that: The quality control method also includes qualitative identification of rhubarb, liquorice, scutellaria baicalensis and dried ginger in the freeze-dried powder of the traditional Chinese medicine composition.

10. The quality control method of the freeze-dried powder of the Chinese medicine composition according to claim 9, characterized in that: The qualitative identification of the radix scutellariae is carried out by thin layer chromatography, comprising the following steps: S1. Prepare the test solution by taking the material standard; S2. preparing a scutellaria baicalensis medicinal material reference solution, a baicalin reference solution, a baicalein reference solution, and a negative sample solution not containing scutellaria baicalensis; S3. Absorb the solutions obtained from S1 and S2 and spot them on the same silica gel G thin layer plate respectively. Use ethyl acetate-butanone-formic acid-water-methanol with a volume ratio of 5:3:0.6:1:0.4 as the developing agent. After development, take out, dry, spray with 5% ferric chloride ethanol test solution, and examine under sunlight.

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