Trichosanthes kirilowii Maxim and burdock soup contrast extract and preparation method thereof

Abstract

The invention discloses a trichosanthes kirilowii maxim and burdock soup contrast extract and a preparation method thereof, and belongs to the technical field of traditional Chinese medicine detection. The preparation method comprises the following steps: taking resin SEPABEADS SP825L, packing the resin SEPABEADS SP825L into a column, sequentially washing 6 column volumes with 95% ethanol and water, uniformly shaking until the water liquid level is flush with the resin surface, taking trichosanthes kirilowii maxim and burdock soup freeze-dried powder, adding water, carrying out ultrasonic treatment, loading a sample, washing 2 column volumes with water, standing and adsorbing overnight, washing 1 column volume with water next day, washing 3 column volumes with 80% ethanol, and carrying out freeze-drying to obtain the trichosanthes kirilowii maxim and burdock soup freeze-dried powder. And collecting the eluent, and freeze-drying to obtain the reference extract. The method for preparing the trichosanthes kirilowii Maxim and burdock soup reference extract provided by the invention passes a repetitive experiment, and the prepared reference extract can be used for controlling the quality of the trichosanthes kirilowii Maxim and burdock soup.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine detection, and in particular to a control extract of trichosanthes and burdock soup and a preparation method thereof. Background Art

[0002] Gualou Niubang Decoction is known for its functions of relieving liver depression, detoxifying and eliminating carbuncle. It is a classic prescription for treating breast diseases (mastitis, breast carbuncle, etc.). In modern Chinese medicine, it is mainly used to treat acute mastitis and other diseases, especially for the early inflammatory stage of acute mastitis. Gualou Niubang Decoction contains 12 herbs: dried tangerine peel, burdock seeds, gardenia, honeysuckle, licorice, trichosanthes seeds, scutellaria, radix trichosanthis, forsythia, thorns of Gleditsia sinensis, bupleurum and green peel. The classic prescription is a compound prescription composed of multiple herbs. Due to the complexity of its own ingredients and the diversity of influencing factors, how to objectively evaluate the quality control of the classic prescription has always been a difficult problem on the road to modernization of traditional Chinese medicine. In addition, chemical reference substances are difficult to develop, difficult to separate, unstable monomers, high production costs and technical requirements, and expensive testing costs. The supply of some difficult-to-obtain chemical reference substances is limited, and there are many restrictions in practical applications, making it difficult to promote.

[0003] The quality standard of prescription preparations is an important guarantee for the clinical efficacy of traditional Chinese medicine. At present, multi-component content determination is an effective quality control mode. However, this evaluation mode is often restricted by factors such as long experimental cycle, difficulty in preparing a single reference substance, and high cost. The use of control extracts instead of control medicinal materials or reference substances for quality control can truly and comprehensively evaluate the quality of the drug. At the same time, using the control extracts as a reference, the content of multiple components can be determined at the same time, which reflects the idea of ​​overall quality control of traditional Chinese medicine. Traditional Chinese medicine control extracts are a type of non-monomer component reference substances. Compared with single reference substances, control extracts are easy to prepare, low in price, good in stability, and relatively simple in preparation. As standard substances, they can greatly reduce the use of monomer reference substances, thereby saving rare resources of traditional Chinese medicine and reducing inspection costs; using the qualitative effect of control extracts, different origins of traditional Chinese medicines in classic prescriptions can be identified and the authenticity of the medicinal materials can be evaluated, so that the quality control of traditional Chinese medicines in classic prescriptions can be objectively evaluated. At present, there is no disclosure of the control extract of Gualou Niubang Decoction and the method for constructing its characteristic spectrum. Summary of the invention

[0004] The purpose of the present invention is to provide a reference extract of Gualou Niubang Decoction and a preparation method thereof, so as to solve the problems existing in the above-mentioned prior art. The method for preparing the reference extract of Gualou Niubang Decoction provided by the present invention has passed the repeatability experiment, and the prepared reference extract contains seven main components (gardenia glycoside, liquiritin, hesperidin, arctiin, baicalin, melaleuca A-7-O-β-glucuronide and wogonin), which can be used for quality control of Gualou Niubang Decoction.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] Technical solution 1: A control extract of trichosanthes and burdock soup, comprising the following ingredients: gardenia glycoside, liquiritigenin, hesperidin, arctiin, baicalin, melaleuca alternifolia A-7-O-β-glucuronide and wogonin.

[0007] Technical Solution 2: The preparation method of the control extract comprises the following steps: loading the column with resin SEPABEADSSP825L, washing 6 column volumes with 95% ethanol and water in sequence, shaking until the water level is flush with the resin surface, taking the freeze-dried powder of Trichosanthes and Burdock Decoction, ultrasonically treating it after adding water, loading it, washing it with water for 2 column volumes, allowing it to stand for adsorption overnight, washing it with water for 1 column volume the next day, washing it with 80% ethanol for 3 column volumes, collecting the eluate, and freeze-drying it to obtain the control extract.

[0008] Furthermore, the preparation method of the freeze-dried powder of trichosanthes and burdock soup comprises the following steps: taking the formula of trichosanthes and burdock soup, decocting with water and then freeze-drying to obtain the freeze-dried powder of trichosanthes and burdock soup.

[0009] Technical solution three: Application of the control extract in quality control of Trichosanthes and Burdock Decoction.

[0010] Technical solution 4: A method for constructing a characteristic spectrum of a trichosanthes and burdock soup extract, comprising the following steps:

[0011] a. Preparation of reference solution: Take gardenia glycoside, licorice glycoside, hesperidin, arctiin, baicalin, melaleucalyptus A-7-O-β-D glucuronide and wogonin reference substances, add solvent to obtain a reference solution;

[0012] b. Preparation of the test solution: dissolve the freeze-dried powder of Trichosanthes and Burdock Decoction to obtain the test solution;

[0013] c. injecting the reference solution and the test solution obtained in steps a and b into a high performance liquid chromatograph for detection, using the characteristic spectrum of the reference solution as a reference spectrum, selecting common peaks from the characteristic spectrum of the test solution, and constructing a characteristic spectrum of the reference extract of Trichosanthes and Burdock Decoction.

[0014] In step c, the HPLC detection conditions meet the following conditions:

[0015] Chromatographic column: Octadecylsilane bonded silica gel is used as filler;

[0016] Column temperature: 30°C;

[0017] Flow rate: 1.0 mL / min;

[0018] Injection volume: 10 μL;

[0019] Detection wavelength: 254nm, 280nm;

[0020] Mobile phase A was methanol-acetonitrile 10:1, and mobile phase B was 0.2% formic acid in water, gradient elution;

[0021] The gradient elution process is:

[0022] 0~10min, mobile phase A is 25~30%, mobile phase B is 75~70%,

[0023] 10-15min, mobile phase A is 30-35%, mobile phase B is 70-65%,

[0024] 15-20min, mobile phase A is 35-35%, mobile phase B is 65-65%,

[0025] 20-25min, mobile phase A is 35-40%, mobile phase B is 65-60%,

[0026] 25-30min, mobile phase A is 40-45%, mobile phase B is 60-55%,

[0027] 30-40min, mobile phase A is 45-50%, mobile phase B is 55-50%,

[0028] 40-50min, mobile phase A is 50-80%, mobile phase B is 50-20%,

[0029] 50~50.01min, mobile phase A is 80~25%, mobile phase B is 20~75%,

[0030] 50.01~70min, mobile phase A is 25~25%, mobile phase B is 75~75%;

[0031] Furthermore, the preparation method of the freeze-dried powder of trichosanthes and burdock soup comprises the following steps: taking the formula of trichosanthes and burdock soup, decocting with water and then freeze-drying to obtain the freeze-dried powder of trichosanthes and burdock soup.

[0032] Furthermore, the formula of the Trichosanthes and Burdock Soup is: 3.73g tangerine peel, 3.73g burdock fruit, 3.73g gardenia, 3.73g honeysuckle, 3.73g liquorice, 3.73g trichosanthes seed, 3.73g scutellaria, 3.73g trichosanthes root, 3.73g forsythia, 3.73g gleditsia spine, 1.85g bupleurum and 1.85g green peel.

[0033] Furthermore, in step a, the solvent is methanol.

[0034] Furthermore, in step c, the detection conditions of the high performance liquid chromatography include: using SEPABEADS SP825L macroporous resin as column packing.

[0035] Technical solution 5: A method for identifying the extract of Trichosanthes and Burdock Decoction, comprising the following steps:

[0036] The freeze-dried powder of the Gualou Niubang decoction extract to be identified is weighed to prepare a test solution of the Gualou Niubang decoction extract, and the test solution is injected into a high performance liquid chromatograph for detection to obtain a corresponding characteristic chromatogram 1 to be identified; the characteristic chromatogram to be identified is compared with a corresponding characteristic spectrum 2 constructed by the characteristic spectrum construction method of the Gualou Niubang decoction extract, and if the characteristic chromatogram 1 to be identified is consistent with the characteristic chromatogram 2, it is identified as qualified in quality.

[0037] The present invention discloses the following technical effects:

[0038] The invention establishes a reference extract of trichosanthes and burdock soup and a preparation method thereof, as well as a method for constructing a characteristic spectrum of the reference extract. Through repeated experiments, it is confirmed that the reference extract prepared by the invention contains seven main components (gardenia glycoside, liquiritin, hesperidin, arctiin, baicalin, melaleuca A-7-O-β-glucuronide and wogonin), and can be used for quality control of trichosanthes and burdock soup. The preparation method of the control extract of Gualou Niubang Decoction is as follows: take 200mL of resin SEPABEADS SP825L and load it into a column, rinse 6 column volumes (1200mL) with 95% ethanol, then rinse 6 column volumes with water, shake well, wait for the resin to naturally settle to a constant volume, wait for the water liquid level to be flush with the resin surface, take 3g of Gualou Niubang Decoction freeze-dried powder (hereinafter referred to as GNDGF), add 20mL of water, ultrasonicate for 20min until completely dissolved, load the sample, rinse 2 column volumes (400mL) with water, let it stand for adsorption overnight, rinse 1 column volume with water on the second day, rinse 3 column volumes with 80% ethanol, collect the eluate, and freeze-dry. In addition, the present invention also provides an identification method for the extract of trichosanthes and burdock soup. The present invention takes the traditional Chinese medicine classic recipe trichosanthes and burdock soup as a benchmark, prepares the corresponding trichosanthes and burdock soup control extract on the basis of the traditional decoction method, and conducts a characteristic spectrum study on the extract, which can be used for qualitative identification of the corresponding decoction.

[0039] The preparation of the control extract is of great significance: first, it provides technical support for the quality control of control extracts and compound preparations applied by the China Food and Drug Inspection Institute, which is highly creative; second, it responds to the call for multi-index overall quality control of traditional Chinese medicine, while avoiding the repetition of pre-treatment, prescription, extraction, ingredient identification and other processes, making the experimental operation simpler and reducing resource consumption. It can provide guarantee for the quality control of classic Chinese medicine decoctions and lay a certain foundation for their international promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0041] Figure 1 Picture of loading sample for column;

[0042] Figure 2 It is the eluent for each group of water parts;

[0043] Figure 3 20% ethanol eluent for each group;

[0044] Figure 4 The eluent was 50% ethanol for each group;

[0045] Figure 5 The eluent was 80% ethanol for each group;

[0046] Figure 6 The eluent was 95% ethanol for each group;

[0047] Figure 7 Load pictures for reproducible experiments;

[0048] Figure 8 The results of the investigation when resin A was used as column filler; among them, 1. Gardenia glycoside, 2. Licorice glycoside, 3. Hesperidin, 4. Arctiin, 5. Baicalin, 6. Melaleuca alternifolia A-7-O-β-D-glucuronide, 7. Wogonin (from top to bottom: mixed standard 254nm, mixed standard 280nm, water part, 20% ethanol, 50% ethanol, 80% ethanol, 95% ethanol);

[0049] Fig. 9 The results of the investigation when resin H was used as column filler; among them, 1. Gardenia glycoside, 2. Licorice glycoside, 3. Hesperidin, 4. Arctiin, 5. Baicalin, 6. Melaleuca alternifolia A-7-O-β-D-glucuronide, 7. Wogonin (from top to bottom: mixed standard 254nm, mixed standard 280nm, water part, 20% ethanol, 50% ethanol, 80% ethanol, 95% ethanol);

[0050] Fig.10 The results of the investigation when resin S was used as column filler; among them, 1. Gardenia glycoside, 2. Licorice glycoside, 3. Hesperidin, 4. Arctiin, 5. Baicalin, 6. Melaleuca alternifolia A-7-O-β-D-glucuronide, 7. Wogonin (from top to bottom: mixed standard 254nm, mixed standard 280nm, water part, 20% ethanol, 50% ethanol, 80% ethanol, 95% ethanol);

[0051] Fig.11 The effect of 2g sample loading on the retained position of each component in the elution result; among them, 1. Gardenia glycoside, 2. Licorice glycoside, 3. Hesperidin, 4. Arctiin, 5. Baicalin, 6. Melaleuca alternifolia A-7-O-β-D-glucuronide, 7. Wogonin (from top to bottom: mixed standard 254nm, mixed standard 280nm, water position, 20% ethanol, 50% ethanol, 80% ethanol, 95% ethanol);

[0052] Fig.12 The effect of 3g sample loading on the retained position of each component in the elution result; among them, 1. Gardenia glycoside, 2. Licorice glycoside, 3. Hesperidin, 4. Arctiin, 5. Baicalin, 6. Melaleuca alternifolia A-7-O-β-D-glucuronide, 7. Wogonin (from top to bottom: mixed standard 254nm, mixed standard 280nm, water position, 20% ethanol, 50% ethanol, 80% ethanol, 95% ethanol);

[0053] Fig.13 The effect of 4g sample loading on the retained position of each component in the elution result; among them, 1. Gardenia glycoside, 2. Licorice glycoside, 3. Hesperidin, 4. Arctiin, 5. Baicalin, 6. Melaleuca alternifolia A-7-O-β-D-glucuronide, 7. Wogonin (from top to bottom: mixed standard 254nm, mixed standard 280nm, water position, 20% ethanol, 50% ethanol, 80% ethanol, 95% ethanol);

[0054] Fig.14 This is the situation of importing the repetitive sample injection results into the traditional Chinese medicine fingerprint similarity software, among which, 1: Gardenia jasminoides; 2: Licorice inoside; 3: Hesperidin; 4: Arctiin; 5: Baicalin; 6. Melaleuca alternifolia A-7-O-β-D-glucuronide, 7. Wogonin S1:1-1; S2:2-1; S3:3-1; S4:4-1; S5:5-1; S6: mixed standard. DETAILED DESCRIPTION

[0055] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0056] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0057] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0058] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0059] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0060] Example 1

[0061] 1. Instruments and test drugs

[0062] Analytical glass column (inner diameter 4.9 cm, column length 50 cm). Resin S: SEPABEADS SP825L (Mitsubishi), purchased from Beijing Green Baicao Technology Development Co., Ltd. Resin H: HPD-600, Resin A: ADS-17; all purchased from Cangzhou Baoen Adsorption Material Technology Co., Ltd.

[0063] Gardeniain (110749-202320, 98.1%), glycyrrhizin (111610-202209, 95.2%), hesperidin (110721-202220, 97.2%), arctiin (110819-202414, 96.6%) and baicalin (110715-202223, 97.2%) were purchased from China Food and Drug Inspection Institutes; melaleuca alternifolia A-7-O-β-D-glucuronide (23111396, 98%) and baicalin (23080601, 98%) were purchased from Beijing BetterKang Biotechnology Co., Ltd.

[0064] 2. Experimental Methods

[0065] 2.1 Preparation of reference solution

[0066] Gardeniain, glycyrrhizin, hesperidin, arctiin, baicalin, melaleuca alternifolia A-7-O-β-D-glucuronide and wogonin were dissolved in methanol to prepare reference stock solutions with concentrations of 1120 μg / mL, 534 μg / mL, 438 μg / mL, 166 μg / mL, 1702 μg / mL, 342 μg / mL and 624 μg / mL, respectively.

[0067] Accurately pipette 1 mL of each of the above-mentioned reference substance mother solutions, dilute to a 10 mL volumetric flask with methanol, and mix by ultrasonication to obtain a mixed standard solution with concentrations of 112.0 μg / mL, 53.4 μg / mL, 43.8 μg / mL, 16.6 μg / mL, 170.2 μg / mL, 34.2 μg / mL, and 62.4 μg / mL, respectively.

[0068] 2.2 Preparation of freeze-dried powder of Trichosanthes and Burdock Decoction

[0069] Take 3.73g of dried tangerine peel, 3.73g of burdock fruit, 3.73g of gardenia, 3.73g of honeysuckle, 3.73g of liquorice, 3.73g of trichosanthes seed, 3.73g of scutellaria, 3.73g of trichosanthes root, 3.73g of forsythia, 3.73g of gleditsia spine; 1.85g of bupleurum and 1.85g of green tangerine peel, add 400mL of water, boil until the temperature reaches 160mL, and freeze-dry to obtain freeze-dried powder of trichosanthes and burdock decoction (GNDGF).

[0070] 2.3 Comparison of elution solutions

[0071] 200mL of resin (soaked in 95% ethanol until it naturally settles and the resin volume remains unchanged). Load the column, rinse 6 column volumes (1200mL) with 95% ethanol, then rinse 6 column volumes with water, shake well, wait until the resin naturally settles and the volume remains unchanged, wait until the water level is flush with the resin surface, accurately weigh the GNDGF-01# sample, add 20mL of water, ultrasonicate for 20min until it is completely dissolved, load the sample, rinse 2 column volumes (400mL) with water, collect the water eluate, let it stand for adsorption overnight, then rinse 1 column volume with water, combine the water eluate, discard it, rinse 3 column volumes (600mL) each with 20% ethanol, 50% ethanol, and 80% ethanol in turn, rinse 2 column volumes (400mL) with 95% ethanol, respectively, combine the 4 ethanol eluates, recover the reagents until there is no ethanol smell, freeze-dry, and obtain each elution part. Load the column and load the sample (2 column volumes have been rinsed in the beaker) see Figure 1 , water fraction eluent (3 column volumes) see Figure 2 ; 20% ethanol eluent (3 column volumes) see Figure 3 ; 50% ethanol eluent (3 column volumes) see Figure 4 ; 80% ethanol eluent (3 column volumes) see Figure 5 ; 95% ethanol eluent (2 column volumes) see Figure 6 .

[0072] 2.4 Investigation of different loading amounts

[0073] According to the loading method of 2.3, resin S was selected, and different loading amounts (2g, 3g, 4g) were investigated to determine the loading amount of the control extract of Trichosanthes and Arctium Decoction.

[0074] 2.5 Investigation of different resins

[0075] According to the loading method in 2.3, different column packing materials (resin A, resin H, and resin S) were investigated respectively, and the loading amount was selected as 3g to determine the resin type of the control extract of Gualou Niubang Decoction. The investigation of different loading amounts and column packing materials is shown in Table 1.

[0076] Table 1 Investigation of different sample loading amounts and column packings

[0077]

[0078]

[0079] 2.6 Repeat the experiment

[0080] Take 200mL of resin SEPABEADS SP825L (soak in 95% ethanol until it naturally settles and the volume of the resin remains unchanged). Load the column, rinse with 95% ethanol for 6 column volumes (1200mL), then rinse with water for 6 column volumes, shake well, wait until the resin naturally settles and the volume remains unchanged, wait until the water level is flush with the resin surface, take an appropriate amount of GNDGF according to Table 2, add 20mL of water, ultrasonicate for 20min until it is completely dissolved, load the sample, rinse with water for 2 column volumes (400mL), collect the water eluate, let it stand for adsorption overnight, rinse with water for 1 column volume on the second day, combine the water eluate, rinse with 80% ethanol for 3 column volumes (X-1), 1 column volume (X-2), 1 column volume (X-3), 1 column volume (X-4), and finally rinse with 95% ethanol for 2 column volumes, and store the obtained eluates by number. Figure 7 Samples were loaded for repeatability experiments, and the repeats of different samples are shown in Table 2.

[0081] Table 2 Repetition of different samples

[0082] Column number sample Sample loading / g 1# GNDGF-01# 3 2# GNDGF-01# 3 3# GNDGF-01# 3 4# GNDGF-08# 3 5# GNDGF-09# 3

[0083] 3. Experimental results

[0084] 3.1 Results of investigation on different column packings

[0085] The results of the investigation of different column packings are shown in Figure 8 , Resin A, since Gardenia glycoside was detected in the water eluent, 20% ethanol can elute most of the main components, and 50% ethanol can basically elute; Fig. 9 It can be seen that when resin H is used as column filler, geniposide is mainly eluted at 20% ethanol, and other components are concentrated at 50% ethanol. Fig.10 It can be seen that when using resin S, the target component is mainly concentrated in the 50% ethanol part. Therefore, SEPABEADS SP825L macroporous resin with good adsorption performance and more concentrated elution components was selected as the column filler.

[0086] 3.2 Results of different loading amounts

[0087] Fig.11 The effect of 2g sample loading on the location of each component retained in the elution results; Fig.12 The effect of 3g sample loading on the location of each component retained in the elution result; Fig.13 The results show that different loading amounts (2, 3, 4 g) have no effect on the locations of the components retained in the elution results, and 3 g was selected as the loading amount.

[0088] 3.3 Repeatability experimental results

[0089] The repetitive sample injection results ( Fig.14 ) was imported into the Chinese medicine fingerprint similarity software (2012 version) to calculate the similarity. The results are shown in Table 3. The similarity results of the same batch of samples repeated 3 times (S1-S3) were above 0.998, and the similarity results of 3 different batches of samples were repeated was greater than 0.985, indicating that the method has good repeatability.

[0090] Table 3 Similarity results of repeatability test

[0091] S1 S2 S3 S4 S5 S1 1.000 1.000 0.998 0.986 0.994 S2 1.000 1.000 0.998 0.985 0.994 S3 0.998 0.998 1.000 0.989 0.997 S4 0.986 0.985 0.989 1.000 0.994 S5 0.994 0.994 0.997 0.994 1.000

[0092] 3.4 Repeatability test eluent concentration results

[0093] The content of components in each elution part was calculated according to the standard curve. The elution rate of gardenia glycoside, glycyrrhizin, hesperidin and melaleucalyptus A glycoside in part X-1 was 100%. The contents of arctiin, baicalin and wogonin in part X-1 accounted for 99.47%-99.86%, 98.74%-99.24%, and 96.14%-97.90% of the total content, respectively. The results of the content of target components in part X-1 are shown in Table 4, and the concentration unit is μg / mL.

[0094] Table 4 Elution site sample concentration

[0095]

[0096] The final preparation method of the control extract of Trichosanthes and Burdock Decoction is as follows: take 200mL of resin SEPABEADS SP825L (soaked in 95% ethanol until it naturally settles to a constant volume of the resin) and load it into a column, rinse 6 column volumes (1200mL) with 95% ethanol, then rinse 6 column volumes with water, shake well, wait for the resin to naturally settle to a constant volume, wait for the water level to be flush with the resin surface, take 3g of GNDGF, add 20mL of water, ultrasonicate for 20min until completely dissolved, load the sample, rinse 2 column volumes (400mL) with water, let stand for adsorption overnight, rinse 1 column volume with water on the second day, rinse 3 column volumes with 80% ethanol, collect the eluate, recover the reagent under reduced pressure until there is no ethanol smell, and lyophilize to obtain.

[0097] Example 2 A method for constructing a characteristic spectrum of a trichosanthes and burdock soup extract

[0098] a. Preparation of reference solution: Take gardenia glycoside, licorice glycoside, hesperidin, arctiin, baicalin, melaleucalyptus A-7-O-β-D glucuronide and wogonin reference substances, add methanol to obtain a reference solution;

[0099] b. Preparation of test solution: add water to the freeze-dried powder of Gualou Niubang Decoction and ultrasonically dissolve it to obtain the test solution (the preparation method of freeze-dried powder of Gualou Niubang Decoction comprises the following steps: take the formula of Gualou Niubang Decoction, add water, boil it, and then freeze-dry it to obtain freeze-dried powder of Gualou Niubang Decoction. The formula of Gualou Niubang Decoction is: 3.73g of dried tangerine peel, 3.73g of burdock seeds, 3.73g of gardenia, 3.73g of honeysuckle, 3.73g of liquorice, 3.73g of trichosanthes seeds, 3.73g of scutellaria, 3.73g of trichosanthes root, 3.73g of forsythia, 3.73g of sapodilla spines, 1.85g of bupleurum and 1.85g of green peel);

[0100] c. Inject the reference solution and the test solution obtained in steps a and b into a high performance liquid chromatograph for detection, use the characteristic spectrum of the reference solution as a reference spectrum, select common peaks from the characteristic spectrum of the test solution, and construct the characteristic spectrum of the reference extract of Trichosanthes and Burdock Decoction. The high performance liquid chromatography detection conditions meet the following requirements: chromatographic column: octadecylsilane bonded silica gel as filler; column temperature: 30°C; flow rate: 1.0mL / min; injection volume: 10μL; detection wavelength: 254nm, 280nm; mobile phase A is methanol-acetonitrile 10:1, mobile phase B is 0.2% formic acid aqueous solution, gradient elution; gradient elution process: 0-10min, mobile phase A is 25-30%, mobile phase B is 75-70%, 10-15min, mobile phase A is 30-35%, mobile phase B is 70-65%, 15-20min, mobile phase A is 35-35%, mobile phase B is The detection conditions of the high performance liquid chromatography include: using SEPABEADS SP825L macroporous resin as column filler.

[0101] The embodiments described above are only descriptions of the preferred embodiments of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A control extract of Gualou Niubang decoction, characterized in that: Includes the following ingredients: gardenia glycoside, liquiritigenin, hesperidin, arctiin, baicalin, melaleuca alternifolia A-7-O-β-glucuronide and wogonin.

2. The method for preparing the control extract according to claim 1, characterized in that: The following steps are involved: Take the resin SEPABEADS SP825L column, wash it with 95% ethanol and water for 6 column volumes in sequence, shake it until the water level is flush with the resin surface, take the freeze-dried powder of Trichosanthes and Burdock Decoction, add water and then ultrasonicate it, load it, wash it with water for 2 column volumes, let it stand for adsorption overnight, wash it with water for 1 column volume the next day, wash it with 80% ethanol for 3 column volumes, collect the eluate, and freeze-dry it to obtain the control extract.

3. The preparation method according to claim 2, characterized in that: The preparation method of the freeze-dried powder of trichosanthes and burdock soup comprises the following steps: taking the formula of trichosanthes and burdock soup, boiling it with water, and then freeze-drying it to obtain the freeze-dried powder of trichosanthes and burdock soup.

4. Use of the control extract according to claim 1 in quality control of Trichosanthes and Burdock Decoction.

5. A method for constructing a characteristic spectrum of a Gualou Niu Bang decoction extract, characterized in that: The following steps are involved: a. Preparation of reference solution: Take gardenia glycoside, licorice glycoside, hesperidin, arctiin, baicalin, melaleucalyptus A-7-O-β-D glucuronide and wogonin reference substances, add solvent to obtain a reference solution; b. Preparation of the test solution: dissolve the freeze-dried powder of Trichosanthes and Burdock Decoction to obtain the test solution; c. injecting the reference solution and the test solution obtained in steps a and b into a high performance liquid chromatograph for detection, using the characteristic spectrum of the reference solution as a reference spectrum, selecting common peaks from the characteristic spectrum of the test solution, and constructing a characteristic spectrum of the reference extract of Trichosanthes and Burdock Decoction. In step c, the HPLC detection conditions meet the following conditions: Chromatographic column: Octadecylsilane bonded silica gel is used as filler; Column temperature: 30°C; Flow rate: 1.0 mL / min; Injection volume: 10 μL; Detection wavelength: 254nm, 280nm; Mobile phase A was methanol-acetonitrile 10:1, and mobile phase B was 0.2% formic acid in water, gradient elution; The gradient elution process is: 0~10min, mobile phase A is 25~30%, mobile phase B is 75~70%, 10-15min, mobile phase A is 30-35%, mobile phase B is 70-65%, 15-20min, mobile phase A is 35-35%, mobile phase B is 65-65%, 20-25min, mobile phase A is 35-40%, mobile phase B is 65-60%, 25-30min, mobile phase A is 40-45%, mobile phase B is 60-55%, 30-40min, mobile phase A is 45-50%, mobile phase B is 55-50%, 40-50min, mobile phase A is 50-80%, mobile phase B is 50-20%, 50~50.01min, mobile phase A is 80~25%, mobile phase B is 20~75%, 50.01~70min, mobile phase A is 25~25%, mobile phase B is 75~75%.

6. The method for constructing a characteristic map according to claim 5, characterized in that: The preparation method of the freeze-dried powder of trichosanthes and burdock soup comprises the following steps: taking the formula of trichosanthes and burdock soup, boiling it with water, and then freeze-drying it to obtain the freeze-dried powder of trichosanthes and burdock soup.

7. The method for constructing a characteristic map according to claim 6, characterized in that: The formula of the trichosanthes and burdock soup is: 3.73g of dried tangerine peel, 3.73g of burdock fruit, 3.73g of gardenia, 3.73g of honeysuckle, 3.73g of liquorice, 3.73g of trichosanthes seed, 3.73g of scutellaria, 3.73g of trichosanthes root, 3.73g of forsythia, 3.73g of gleditsia spine, 1.85g of bupleurum and 1.85g of green peel.

8. The method for constructing a characteristic spectrum according to claim 6, characterized in that: In step a, the solvent is methanol.

9. The method for constructing a characteristic spectrum according to claim 6, characterized in that: In step c, the detection conditions of the high performance liquid chromatography include: using SEPABEADS SP825L macroporous resin as column filler.

10. A method for identifying an extract of Gualou Niubang decoction, characterized in that: The following steps are involved: Weigh the freeze-dried powder of the Gualou Niubang decoction extract to be identified, prepare a test solution of the Gualou Niubang decoction extract, inject the test solution into a high performance liquid chromatograph for detection, and obtain a corresponding characteristic chromatogram 1 to be identified; compare the characteristic chromatogram to be identified with the corresponding characteristic spectrum 2 constructed by the characteristic spectrum construction method of the Gualou Niubang decoction extract according to claim 5, if the characteristic chromatogram 1 to be identified is consistent with the characteristic chromatogram 2, it is identified as qualified in quality.

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