Extraction process and quality detection method of Xinyu drink

Abstract

The invention discloses an extraction process and a quality detection method of Xinyu drink, and the extraction method comprises the following steps: taking the following medicinal materials of Xinyu drink prescription: coptis chinensis, dandelion, fructus forsythiae, radix aucklandiae, areca nut, lily, thunberg fritillary bulb and raw liquorice, adding water which is 8-12 times of the weight of the medicinal materials for soaking for 45 minutes for the first time, then decocting and extracting for 1-2 hours, adding water which is 6-8 times of the weight of the medicinal materials for the second time, decocting and extracting for 1-2 hours, merging the two decoction extracting solutions, and concentrating to obtain the Xinyu drink. And filtering to obtain the product. According to the method, the optimal chromatographic conditions such as the mobile phase composition, the gradient elution mode and the detection wavelength are screened out through a large number of experiments, the quality detection method of the Xinyu drink is established, the quality detection efficiency can be improved, meanwhile, the method is simple, convenient, high in accuracy, good in repeatability, good in stability and high in durability, and the method is suitable for popularization and application. And a scientific basis can be provided for quality evaluation and control of the Xinpu drink.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine detection, and particularly relates to a quality detection method for Xinpu Drink. Background Art

[0002] Xinpuyin is a modified Qingpuyin (composed of Qingmuxiang, dandelion, coptis root, white peony root, and licorice), which discards the nephrotoxic Qingmuxiang of the Aristolochiaceae family. The whole prescription is composed of Sichuan coptis root, dandelion, forsythia, costus root, areca nut, lily, Zhejiang Fritillaria, and raw licorice, and is mainly used to clear heat and dampness, strengthen the spleen and stomach, and regulate qi and activate blood circulation. In the Xinpuyin prescription, coptis root is the main medicine, which can remove dampness and heat, purge fire and detoxify. Dandelion and forsythia are the assistant medicines, which cooperate with the main medicine to clear heat and dampness, dissipate blood stasis and disperse nodules; costus root, areca nut, lily, Zhejiang Fritillaria work together to promote qi and remove stagnation, nourish yin and promote muscle growth; raw licorice harmonizes the medicinal properties. The combination of these medicines can clear heat and dampness, strengthen the spleen and stomach, and regulate qi and activate blood circulation. Clinical trials have shown that Xinpuyin combined with omeprazole has achieved good clinical efficacy in the treatment of Helicobacter pylori-related gastritis with spleen and stomach damp-heat syndrome, and can effectively improve patients' clinical symptoms. Compared with Western medicine, it has fewer side effects and better safety.

[0003] The content of effective ingredients obtained by different extraction methods of traditional Chinese medicine compound is quite different, which directly affects the clinical efficacy. There is no report on the optimization of the extraction process of Xinpuyin, and there are few quality detection methods for Xinpuyin. The present invention adopts high performance liquid chromatography to establish a quality detection method for Xinpuyin, which is of great significance to the quality evaluation of Xinpuyin and the formulation of quality standards. Summary of the invention

[0004] Purpose of the invention: The present invention uses orthogonal experimental design to establish a quality control system based on berberine, chicoric acid, forsythiaside, liquiritin and dry extract rate, determines the amount of water added, decoction time and decoction times as key process parameters, applies orthogonal design to optimize the extraction process of Xinpu Drink, and combines the AHP-entropy weight method to determine the comprehensive weight coefficient of each evaluation index, and performs comprehensive scoring to select the best extraction process.

[0005] Another object of the present invention is to establish a quality detection method with high precision and accuracy, good stability and repeatability by using high performance liquid chromatography.

[0006] Technical solution: In order to achieve the above purpose, the technical solution adopted by the present invention is:

[0007] A method for extracting Xinpuyin comprises the following steps:

[0008] Take the prescription medicinal materials of Xinpuyin, including 3-9 parts of coptis chinensis, 15-45 parts of dandelion, 10-30 parts of forsythia, 10-30 parts of costusroot, 10-30 parts of areca catechu, 30-90 parts of lily, 10-30 parts of thunbergii bulbs, and 9-27 parts of raw licorice. Add 8-12 times the volume of water of the medicinal materials for the first time, soak for 45 minutes, then decoct and extract for 1-2 hours. Add 6-8 times the amount of water for the second time, decoct and extract for 1-2 hours, combine the two decoction extracts, filter, and obtain.

[0009] As a preferred embodiment, the above-mentioned method for extracting Xinpuyin comprises the following steps:

[0010] Take the prescription medicinal materials of Xinpuyin, including 3-9 parts of Coptis chinensis, 15-45 parts of Taraxacum, 10-30 parts of Forsythia suspensa, 10-30 parts of Costus root, 10-30 parts of Areca catechu, 30-90 parts of Lilium, 10-30 parts of Fritillaria thunbergii, and 9-27 parts of raw Licorice. Add 12 times the volume of water by weight of the medicinal materials for the first time, soak for 45 minutes, then decoct and extract for 1.5 hours. Add 8 times the amount of water for the second time, decoct and extract for 1.5 hours, combine the two decoction extracts, filter, and obtain.

[0011] A quality detection method for Xinpuyin comprises the following steps:

[0012] (1) Preparation of test solution:

[0013] Take the prescription medicinal materials of Xinpuyin, including coptis root, dandelion, forsythia, costusroot, areca nut, lily, fritillaria thunbergii, and raw licorice, soak them in water, then decoct and extract, combine the decoction extracts, filter, take the filtrate and put it in a volumetric flask, dilute to the mark with methanol, shake well, sonicate, filter, take the filtrate, centrifuge, take the supernatant, and obtain the test solution;

[0014] (2) Preparation of mixed reference solution:

[0015] Take appropriate amount of berberine, chicoric acid, forsythiaside, and cyperus feroxanthin reference substances, accurately weigh them, and dissolve them in methanol to prepare a mixed reference substance solution;

[0016] (3) Drawing of standard curve

[0017] Accurately pipette 2, 5, 10, 15, and 20 μL of the mixed reference solution of step (2) for injection and determination, respectively. Perform the determination twice in parallel for each portion, and draw a standard curve equation with the logarithm of the peak area as the ordinate Y and the logarithm of the reference content as the abscissa;

[0018] (4) Determination of test solution

[0019] Pipette 10 μL of the test solution from step (1), inject according to the chromatographic conditions, measure each sample twice in parallel, determine the peak area, and calculate the content of the index component in the sample according to the standard curve equation of the reference substance from step (3).

[0020] As a preferred embodiment, the quality detection method of Xinpuyin described above, the preparation method of the test solution is:

[0021] Take 3-9 parts of coptis chinensis, 15-45 parts of dandelion, 10-30 parts of forsythia, 10-30 parts of costusroot, 10-30 parts of areca catechu, 30-90 parts of lily, 10-30 parts of thunbergii, and 9-27 parts of raw licorice in the prescription of Xinpuyin. Add 12 times the volume of water by weight of the medicinal materials to soak for 45 minutes for the first time, then decoct and extract for 1.5 hours. Add 8 times the amount of water for the second time, decoct and extract for 1.5 hours, combine the two decoction extracts, filter, take 10mL of the filtrate in a 50mL volumetric flask, dilute to the scale with methanol, shake well, ultrasonicate for 30 minutes, filter, take the filtrate, centrifuge for 10 minutes, take the supernatant, and obtain the test solution.

[0022] As a preferred embodiment, in the above-mentioned quality detection method of Xinpuyin, the preparation method of the mixed reference solution in step (2) is:

[0023] Take appropriate amount of berberine, chicoric acid, forsythiaside and cyperus feroxanthin reference substances, accurately weigh them, dissolve them with methanol, and prepare mixed reference substance solutions with mass concentrations of 3.07 μg / mL, 10.63 μg / mL, 9.70 μg / mL and 5.10 μg / mL, respectively.

[0024] As a preferred embodiment, in the above-mentioned quality detection method of Xinpuyin, the chromatographic conditions of step (3) and step (4) are:

[0025] The chromatographic column was Hibar Purospher Star LP RP-18 endcapped, with specifications of 250 mm × 4.6 mm and 5 μm. The mobile phase was acetonitrile-0.05 mol / L potassium dihydrogen phosphate solution. The gradient elution was 0-7 min, 10%-17% acetonitrile; 7-14 min, 17% acetonitrile; 14-19 min, 17%-22% acetonitrile; 19-28 min, 22%-24% acetonitrile; 28-42 min, 24%-35% acetonitrile; 42-49 min, 35%-63% acetonitrile; 49-50 min, 63%-10% acetonitrile. The volume flow rate was 1 mL / min. The detection wavelengths were 237 nm, 277 nm, 330 nm and 345 nm. The column temperature was 25°C and the injection volume was 10 μL.

[0026] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0027] (1) The present invention weights the quality evaluation system based on the AHP-entropy weight method, clarifies the weight coefficients of the five indicators in the quality control system, and optimizes the modern extraction process of Xinpuyin to add 20 times of water, extract twice (the first time is 60% of the total water addition multiple, and the second time is 40% of the total water addition multiple), and boil for 1.5 hours each time. The verification test confirms that the selected optimal extraction process is stable and feasible, meets the requirements of large-scale production, and can provide a reference for the extraction application of Xinpuyin. The obtained process can ensure the feasibility of industrial production and the stability of product quality.

[0028] (2) The present invention screened out the best chromatographic conditions such as mobile phase composition, gradient elution mode, detection wavelength, etc. through a large number of experiments, and established a detection method for Xinpuyin. The method has the advantages of convenient operation, rapid detection, good method stability, high precision, good reproducibility, etc., and can accurately and reliably detect and control the quality of Xinpuyin. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 HPLC graphs of mixed reference substances at different wavelengths.

[0030] Figure 2 HPLC graphs of Xinpuyin test solution at different wavelengths. DETAILED DESCRIPTION

[0031] The present invention is further explained below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. After reading the present invention, various equivalent modifications of the present invention by those skilled in the art all fall within the scope defined by the claims attached to this application.

[0032] Example 1

[0033] 1. Instruments and Materials

[0034] 1.1 Instruments

[0035] Waters 2695 high performance liquid chromatograph, equipped with quaternary pump, column oven, autosampler, 2998 detector, Empower3 chromatography workstation, Waters Corporation, USA; Milli-Q ultrapure water machine, Millipore Corporation, USA; FA1104 electronic analytical balance, d = 0.1 mg, Shanghai Precision Scientific Instrument Co., Ltd.; MS105DU electronic analytical balance, d = 0.01 mg, Mettler-Toledo, Switzerland; KQ-500E ultrasonic cleaner, Kunshan Ultrasonic Instrument Co., Ltd.; DHG-9140A electric constant temperature blast drying oven, Shanghai Jinghong Experimental Equipment Co., Ltd.; AE-1106A multifunctional infrared oven, Zhongshan Aimi Life Appliance Co., Ltd.

[0036] 1.2. Drug testing

[0037] The reference substance berberine hydrochloride (batch number S01A10K943340, mass fraction 98%) was purchased from Shanghai Yuanye Biotechnology Co., Ltd. Chicoric acid (batch number 111752-202105, mass fraction 98.3%), forsythiaside (batch number 110821-202117, mass fraction 94.8%), and glycoside grass glycoside (batch number 111610-202209, mass fraction 95.2%) were all purchased from the China Food and Drug Inspection Institute. Potassium dihydrogen phosphate, chromatographic grade, Shanghai Aladdin Biochemical Technology Co., Ltd.; acetonitrile, chromatographic grade, Merck KGaA; methanol, chromatographic grade, Jiangsu Hanbang Technology Co., Ltd.; phosphoric acid, chromatographic grade, Shanghai Aladdin Biochemical Technology Co., Ltd.; other reagents were of analytical grade. Huanglian tablets (batch number 230201), lily (batch number 230201), and Fritillaria thunbergii (batch number 221201) were all purchased from Bozhou Wanzhong Chinese Medicine Pieces Factory; dandelion (batch number 220926) was purchased from Suzhou Boyuan Pharmaceutical Co., Ltd.; forsythia (batch number 230201) was purchased from Ma'anshan Jingquan Chinese Medicine Pieces Co., Ltd.; costusroot tablets (batch number 221201) were purchased from Chongqing Wanli Pharmaceutical Co., Ltd.; betel nut (batch number 220501) was purchased from Chongqing Wanli Pharmaceutical Co., Ltd.; licorice tablets (batch number 2204003) were purchased from Gansu Huaqingtang Pharmaceutical Pieces Co., Ltd. The identification reports of each piece of medicine show that they all meet the standards of the 2020 edition of the Chinese Pharmacopoeia.

[0038] Example 1

[0039] 1. A quality detection method for Xinpuyin, comprising the following steps:

[0040] (1) Preparation of test solution:

[0041] Take the prescription medicinal materials of Xinpuyin, including 3g of Coptis chinensis, 15g of Taraxacum officinale, 10g of Forsythia suspensa, 10g of Aucklandia lappa, 10g of Areca catechu, 30g of Lilium, 10g of Fritillaria thunbergii, and 9g of Licorice root. Add 15 times the volume of water by weight of the medicinal materials and soak for 45 minutes for the first time, then decoct and extract for 1.5 hours. Add 10 times the volume of water for the second time, decoct and extract for 1.5 hours, combine the extracts of the two decoctions, filter, take 10mL of the filtrate in a 50mL volumetric flask, dilute to the mark with methanol, shake well, sonicate for 30 minutes, filter, take the filtrate, centrifuge for 10 minutes, take the supernatant, and obtain the test solution;

[0042] (2) Preparation of mixed reference solution:

[0043] Take appropriate amount of berberine, chicoric acid, forsythiaside and cyperus feroxanthin reference substances, accurately weigh them, dissolve them with methanol, and prepare mixed reference substance solutions with mass concentrations of 3.07 μg / mL, 10.63 μg / mL, 9.70 μg / mL and 5.10 μg / mL, respectively.

[0044] (3) Drawing of standard curve

[0045] 2, 5, 10, 15, 20 μL of the mixed reference solution of step (2) were accurately drawn and injected for determination, and the chromatographic conditions were (chromatographic column Hibar Purospher Star LP RP-18 endcapped, specification 250 mm×4.6 mm, 5 μm; mobile phase was acetonitrile-0.05 mol / L potassium dihydrogen phosphate solution; gradient elution 0-7 min, 10%-17% acetonitrile; 7-14 min, 17% acetonitrile; 14-19 min, 17%-22% acetonitrile; 19-28 min, 22%-24% acetonitrile; 28-42 min, 24%-35% acetonitrile; 42-49 min, 35%-63% acetonitrile; 49-50 min, 63%-10% acetonitrile; volume flow rate 1 mL / min; detection wavelengths 237 nm, 277 nm, 330 nm, 345 nm; column temperature 25°C, injection volume 10 μL). Each sample was measured twice in parallel, and the logarithm of the peak area was used as the ordinate Y, and the logarithm of the reference substance content was used as the abscissa to draw the standard curve equation; the regression equations of liquiritin, forsythiaside, chicoric acid, and berberine were obtained as Y=2048074.5319X-469.0732(R 2 =0.9999), Y=813991X-500.71(R 2 =0.9999), Y=2471712.7709X+1200.6098(R 2 =0.9998),Y=3561132.1406X+1398.4878(R 2 =0.9998); each component showed a good linear relationship in the range of 0.0102-0.1020 μg, 0.0194-0.1941 μg, 0.0213-0.2126 μg, and 0.0171-0.1708 μg, respectively.

[0046] (4) Determination of test solution

[0047] Take 10 μL of the test solution in step (1) and inject it under the same chromatographic conditions as in step (3) above. Perform two parallel measurements for each sample, determine the peak area, and calculate the content of the index component in the test solution according to the standard curve equation of the reference substance in step (3). See the following table.

[0048] Detection ingredients Licorice glycoside Forsythiaside Chicoric acid Berberine <![CDATA[Mass fraction / (mg·g -1 )]]> 0.62 0.67 0.44 2.65 .

[0049] 2. Methodological investigation

[0050] (1) Precision test,

[0051] Accurately pipette 5 μL of the above mixed reference solution, inject 6 times according to the above chromatographic conditions, and record the peak area. The results show that the RSDs of the peak areas of liquiritin, forsythiaside, chicoric acid, and berberine are 2.82%, 2.54%, 0.18%, and 2.13%, respectively, indicating that the instrument has good precision.

[0052] (2) Repeatability test

[0053] Six test sample solutions were prepared in parallel according to the above method and injected according to the above chromatographic conditions. Each test sample was injected twice in parallel, and the peak area was recorded and the content was calculated. The results showed that the RSDs of the peak areas of liquiritin, forsythiaside, chicoric acid, and berberine were 0.65%, 2.72%, 0.49%, and 0.20%, respectively, indicating that the method had good repeatability.

[0054] (3) Stability test

[0055] Accurately pipette 10 μL of the same test solution, and determine the peak area of ​​each index component according to the above chromatographic conditions at 0, 2, 4, 6, 12, 20, and 24 hours after sample preparation. The results showed that the RSD values ​​of the peak areas of liquiritin, forsythiaside, chicoric acid, and berberine were 0.74%, 2.54%, 1.13%, and 0.75%, respectively, indicating that the test solution had good stability within 24 hours.

[0056] (4) Sample recovery test

[0057] Weigh 6 portions of the test solution with known content accurately, add the reference solution with known concentration accurately in turn, prepare the test solution according to the above method, and inject and measure according to the above chromatographic conditions. Calculate the recovery rate of each index component. The results showed that the average recovery rates of liquiritin, forsythiaside, chicoric acid, and berberine were 102.13%, 104.34%, 99.67%, and 100.58%, respectively, and the RSD values ​​were 0.23%, 1.45%, 0.89%, and 2.13%, respectively, indicating that this method has good accuracy.

[0058] The present invention screens out the best chromatographic conditions such as mobile phase composition, gradient elution mode, detection wavelength, etc. through a large number of experiments, and establishes a detection method for Xinpuyin. The method has high accuracy, good repeatability and good stability.

[0059] Example 2 Investigation of the extraction process of Xinpuyin

[0060] 1. Investigation on water absorption rate of Xinpuyin decoction pieces

[0061] Weigh the slices according to the ratio of the prescription flavor: 3g of Sichuan coptis root, 15g of dandelion, 10g of forsythia, 10g of costus root, 10g of betel nut, 30g of lily, 10g of fritillaria thunbergii, and 9g of raw licorice. Make 5 parallel portions, record the weight, add 10 times the amount (970mL) of water, place at room temperature (25°C), filter out the water at 15min, 30min, 45min, 60min, and 75min, weigh the slices after soaking, and calculate the water absorption rate of the slices at each time. The calculation formula is as follows, and the test results are shown in Table 1. The results show that after the slices are soaked for 45min, the water absorption rate of the slices has basically reached saturation, and 45min is finally determined as the soaking time.

[0062] Table 1. Results of immersion time investigation

[0063] Soaking time (min) 15 30 45 60 75 Water absorption (%) 75.57 88.55 108.47 118.57 121.21 .

[0064] 2. Determination of dry paste rate

[0065] Take the water extract of Xinpuyin, shake it well and accurately measure 10mL, place it in an evaporating dish that has been dried to constant weight, evaporate it in a water bath and dry it in an oven at 105℃ for 3h, move it to a dryer and cool it for 30min, quickly weigh it accurately, and calculate the dry paste rate. Calculation formula: Dry paste rate (%) = (m×v) / (M×V)×100%, where m represents the mass of dry paste (g), v represents the total volume of water extract (mL), M represents the mass of the raw material (g), and V represents the sampling volume (mL).

[0066] 3. Extraction orthogonal test

[0067] (1) Determination of critical quality attributes (CQAs) and critical process parameters (CPPs): Based on the theory of monarch, minister, assistant and envoy, five index components, namely berberine in monarch herb Coptis chinensis, chicoric acid in minister herb Taraxacum mongolicum, forsythiaside in Forsythia suspensa, liquiritin in envoy herb Glycyrrhiza uralensis and dry extract rate, were selected as CQAs of the extraction process of Xinpuyin. Three factors, namely solid-liquid ratio (A), extraction time (B) and extraction times (C), were selected as CPPs of the preparation process of Xinpuyin.

[0068] (2) Orthogonal experimental design: Weigh 9 pieces of medicinal materials from the same batch with the same dosage ratio of the prescribed medicinal ingredients. The three factors of solid-liquid ratio (A), extraction time (B), and number of extractions (C) were investigated. The contents of berberine, chicoric acid, forsythiaside, and glycyrrhizin in the extracts and the dry extract rate were used as evaluation indicators. The extraction was carried out according to the L9 (34) orthogonal table. The test results are shown in Table 2.

[0069] Table 2 Orthogonal experimental design and response values

[0070]

[0071]

[0072] 4. Calculation of combined weights of AHP-entropy weight method

[0073] 4.1 AHP ​​subjective weighting

[0074] (1) Construction of judgment matrix: Modern pharmacological studies have shown that Coptis chinensis has anti-pathogenic, anti-inflammatory, anti-tumor, anti-diarrhea, platelet aggregation inhibition, gastric juice secretion inhibition and other effects. Due to its significant antibacterial activity, it is known as the "Chinese herbal antibiotic". Among them, the main component berberine plays an important role in antibacterial activity. As a broad-spectrum antibacterial drug, it has strong antibacterial efficacy. The most abundant active ingredient in dandelion is phenolic compounds, and the phenolic compound chicoric acid is also considered to be the main component of plant extracts that exert anti-inflammatory and antioxidant activities. Forsythin is one of the main active ingredients of the Chinese herbal medicine Forsythia suspensa, which has antioxidant, anti-inflammatory, and anti-aging effects. Combined with the concept of Q-Marker, the relevant literature analysis on the screening of licorice Q-Marker was carried out, and it is recommended that the flavonoid compound liquiritin can be used as the Q-Marker of licorice. The subjective AHP method was used to determine the priority of each indicator as follows: berberine content > chicoric acid content > forsythin content > liquiritin content = dry paste rate. Then, according to the AHP theory judgment matrix 1 to 9 scale, the relative importance of n indicators in the same level is scored, and the results are shown in Table 3. Berberine is the main active ingredient of the monarch drug Coptis chinensis. It is more important than chicoric acid, so it is marked as 4. It is more important than forsythiaside, so it is marked as 5. It is more important than licorice glycoside dry cream rate, so it is marked as 6. And so on, according to formula (1), the judgment matrix A is constructed. The data is shown in Table 4. ef Indicates the comparison result of factor e relative to factor f.

[0075] Table 3. Criteria for constructing judgment matrix using AHP

[0076]

[0077]

[0078] Table 4 Judgment priority matrix for pairwise comparison of indicators

[0079] Berberine Chicoric acid Forsythiaside Licorice glycoside Dry paste rate Berberine 1 4 5 6 6 Chicoric acid 1 / 4 1 3 4 4 Forsythiaside 1 / 5 1 / 3 1 2 2 Licorice glycoside 1 / 6 1 / 4 1 / 2 1 1 Dry paste rate 1 / 6 1 / 4 1 / 2 1 1 .

[0080] (2) Calculate the subjective weight coefficient (W j S ): First, perform geometric mean (square root method) on the judgment matrix A and calculate the initial weight coefficient (Wj′) according to formula (2), a jnrepresents the comparison result of the jth (j=1, 2, ..., n) index factor to the nth (n=1, 2, ..., j) index factor, and then calculates the normalized W according to formula (3): j S , and get the W of 5 indicators j S , the results are shown in Table 5.

[0081] W′ j =(a j1 a j2 …a jn ) 1 / n (2)

[0082]

[0083] (3) Consistency check: Perform a consistency check on the matrix. First, calculate the maximum characteristic root (λ) according to formula (4). max ), and then the consistency index (CI) was calculated according to formula (5), and the CI was 0.04, and the consistency ratio (CR) (CR = CI / RI, RI is the degree of freedom index) was 0.03, both of which were less than 0.1, indicating that the matrix was consistent.

[0084]

[0085] CI=(λ max -n) / (n-1) (5)

[0086] 4.2 Objective weighting of information entropy

[0087] (1) Establishing the original data matrix of orthogonal experiment: There are m samples, n evaluation indicators, and the original data matrix R = (Xi, j) m × n. The original data matrix (R) is established with berberine, chicoric acid, forsythia glycosides, glycyrrhizin content and dry extract rate as evaluation indicators.

[0088] (2) Convert the original data into a probability matrix: According to formula (6), the original data matrix R is converted into a probability matrix (P), P ij It represents the probability of the i-th sample under the j-th index.

[0089]

[0090] (3) Calculate the information entropy of each indicator (H j ) and objective weight coefficient (W j O ):H jThe smaller it is, the higher the degree of data dispersion under the j index is, and the greater the amount of information it provides. According to formula (7), the H of each index is calculated. j They are 0.9806, 0.9946, 0.9833, 0.9898, and 0.9934, respectively. The W of each index is calculated according to formula (8): j O .

[0091]

[0092]

[0093] 4.3 Determination of comprehensive weight coefficient (Wj)

[0094] Using AHP to get W j S , entropy weight method to obtain W j O , calculate the Wj of each index according to formula (9), and the results are shown in Table 5.

[0095]

[0096] Table 5 Combination weights

[0097] Evaluation indicators <![CDATA[W j S ]]> <![CDATA[W j O ]]> <![CDATA[W j ]]> Berberine 0.53 0.33 0.70 Chicoric acid 0.23 0.23 0.21 Forsythiaside 0.11 0.11 0.05 Licorice glycoside 0.07 0.07 0.02 Dry paste rate 0.07 0.07 0.02 .

[0098] 5. Comprehensive rating

[0099] The Wj obtained by the AHP-entropy weight method shows that berberine accounts for 70%, chicoric acid accounts for 21%, forsythin accounts for 5%, liquiritin accounts for 2%, and the dry cream rate accounts for 2%. The obtained content data are standardized and the C of each index is calculated according to formula (10): ij The results are shown in Table 6. max(x j )、min(x j ) are the maximum and minimum values ​​of the same indicator in the jth indicator of the ith experiment.

[14] .

[0100]

[0101] Table 6 C of each index component ij value

[0102] Berberine Chicoric acid Forsythiaside Licorice glycoside Dry paste rate <![CDATA[C1]]> 0.00 0.47 0.00 0.00 0.00 <![CDATA[C2]]> 0.73 0.21 0.56 0.45 0.61 <![CDATA[C3]]> 0.20 0.74 0.74 0.03 0.99 <![CDATA[C4]]> 0.15 0.16 0.26 0.34 0.59 <![CDATA[C5]]> 0.90 1.00 1.00 1.00 1.00 <![CDATA[C6]]> 0.26 0.47 0.28 0.38 0.31 <![CDATA[C7]]> 0.24 0.05 0.49 0.59 0.84 <![CDATA[C8]]> 0.27 0.05 0.54 0.62 0.34 <![CDATA[C9]]> 1.00 0.00 0.62 0.24 1.00 .

[0103] The comprehensive score was calculated according to formula (11), and the results are shown in Table 6. As shown in Table 6, the comprehensive score of experiment 5 was the highest. The orthogonal results showed that the amount of water added, extraction time, and extraction times had no significant effect on berberine and chicoric acid, but had a significant effect on liquiritin. The decoction time and extraction times had a significant effect on forsythin. Through intuitive analysis, considering the production efficiency and energy consumption cost, the optimal extraction process was determined to be adding 20 times the amount of water of the slices, extracting twice (the first time was 60% of the total water addition multiple, and the second time was 40% of the total water addition multiple), and decoction for 1.5 hours each time.

[0104] Overall score = C ij小檗碱 ×70%+C ij菊苣酸 ×21%+C ij连翘苷 ×5%+C ij苷草苷 ×2%+C ij干浸膏 ×2%(11).

[0105] 6. Verification test

[0106] The optimized extraction process parameters were verified, and the test was repeated 3 times. The contents of berberine, chicoric acid, forsythiaside, and liquiritin were determined and the dry extract rate was calculated according to the detection method of Example 1. The results are shown in Table 7. The verification test results show that the measured index component contents and dry extract rates are similar to the water extraction process values, and the RSD values ​​of the index components are all less than 3.0%, indicating that the extraction process determined by this method is stable, feasible, and has good repeatability.

[0107] Table 7 Verification test results

[0108]

[0109] Xinpuyin is used clinically to treat Helicobacter pylori. As a clinical experience prescription for clearing away heat and dampness, strengthening the spleen and stomach, and regulating qi and activating blood circulation, it has achieved good clinical efficacy in clinical treatment and can effectively improve the clinical symptoms of patients; compared with Western medicine, it has fewer side effects and better safety. Animal experiments have shown that the mechanism of Xinpuyin in treating Helicobacter pylori is to regulate immunity, resist inflammation, promote the repair of gastric mucosal damage, and improve pathological changes.

[0110] The present invention weights the quality evaluation system based on the AHP-entropy weight method, clarifies the weight coefficients of 5 indicators in the quality control system, and optimizes the modern extraction process of Xinpuyin to add 20 times of water, extract twice (the first time is 60% of the total water addition multiple, and the second time is 40% of the total water addition multiple), and boil for 1.5 hours each time. The verification test confirms that the selected optimal extraction process is stable and feasible, meets the requirements of large-scale production, and can provide a reference for the extraction application of Xinpuyin. The obtained process can ensure the feasibility of industrial production and the stability of product quality.

[0111] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for extracting Xinpuyin, characterized in that: The steps include: Take the prescription medicinal materials of Xinpuyin, including 3-9 parts of coptis chinensis, 15-45 parts of dandelion, 10-30 parts of forsythia, 10-30 parts of costusroot, 10-30 parts of areca catechu, 30-90 parts of lily, 10-30 parts of thunbergii bulbs, and 9-27 parts of raw licorice. Add 8-12 times the volume of water of the medicinal materials for the first time, soak for 45 minutes, then decoct and extract for 1-2 hours. Add 6-8 times the amount of water for the second time, decoct and extract for 1-2 hours, combine the two decoction extracts, filter, and obtain.

2. The method for extracting Xinpuyin according to claim 1, characterized in that: The steps include: Take the prescription medicinal materials of Xinpuyin, including 3-9 parts of Coptis chinensis, 15-45 parts of Taraxacum, 10-30 parts of Forsythia suspensa, 10-30 parts of Costus root, 10-30 parts of Areca catechu, 30-90 parts of Lilium, 10-30 parts of Fritillaria thunbergii, and 9-27 parts of raw Licorice. Add 12 times the volume of water by weight of the medicinal materials for the first time, soak for 45 minutes, then decoct and extract for 1.5 hours. Add 8 times the amount of water for the second time, decoct and extract for 1.5 hours, combine the two decoction extracts, filter, and obtain.

3. A quality detection method for Xinpuyin, characterized in that: The steps include: (1) Preparation of test solution: Take the prescription medicinal materials of Xinpuyin, including coptis root, dandelion, forsythia, costusroot, areca nut, lily, fritillaria thunbergii, and raw licorice, soak them in water, then decoct and extract, combine the decoction extracts, filter, take the filtrate and put it in a volumetric flask, dilute to the mark with methanol, shake well, sonicate, filter, take the filtrate, centrifuge, take the supernatant, and obtain the test solution; (2) Preparation of mixed reference solution: Take appropriate amount of berberine, chicoric acid, forsythiaside, and liquiritin reference substances, weigh accurately, and dissolve them in methanol to prepare a mixed reference substance solution; (3) Drawing of standard curve Accurately pipette 2, 5, 10, 15, and 20 μL of the mixed reference solution of step (2) for injection and determination, respectively. Perform the determination twice in parallel for each portion, and draw a standard curve equation with the logarithm of the peak area as the ordinate Y and the logarithm of the reference content as the abscissa; (4) Determination of test solution Pipette 10 μL of the test solution from step (1), inject according to the chromatographic conditions, measure each sample twice in parallel, determine the peak area, and calculate the content of the index component in the sample according to the standard curve equation of the reference substance from step (3).

4. The quality detection method of Xinpuyin according to claim 3, characterized in that: The preparation method of the test solution is: Take 3-9 parts of coptis chinensis, 15-45 parts of dandelion, 10-30 parts of forsythia, 10-30 parts of costusroot, 10-30 parts of areca catechu, 30-90 parts of lily, 10-30 parts of thunbergii, and 9-27 parts of raw licorice in the prescription of Xinpuyin. Add 12 times the volume of water by weight of the medicinal materials to soak for 45 minutes for the first time, then decoct and extract for 1.5 hours. Add 8 times the amount of water for the second time, decoct and extract for 1.5 hours, combine the two decoction extracts, filter, take 10mL of the filtrate in a 50mL volumetric flask, dilute to the scale with methanol, shake well, ultrasonicate for 30 minutes, filter, take the filtrate, centrifuge for 10 minutes, take the supernatant, and obtain the test solution.

5. The quality detection method of Xinpuyin according to claim 3, characterized in that: The preparation method of the mixed reference substance solution in step (2) is: Take appropriate amount of berberine, chicoric acid, forsythiaside and cyperus feroxanthin reference substances, accurately weigh them, dissolve them with methanol, and prepare mixed reference substance solutions with mass concentrations of 3.07 μg / mL, 10.63 μg / mL, 9.70 μg / mL and 5.10 μg / mL, respectively.

6. The quality detection method of Xinpuyin according to claim 3 is characterized in that: The chromatographic conditions of step (3) and step (4) are: The chromatographic column was Hibar Purospher Star LP RP-18 endcapped, with specifications of 250 mm × 4.6 mm, 5 μm; the mobile phase was acetonitrile-0.05 mol / L potassium dihydrogen phosphate solution; gradient elution; volume flow rate was 1 mL / min; detection wavelengths were 237 nm, 277 nm, 330 nm, and 345 nm; and the column temperature was 25 °C.

7. The quality detection method of Xinpuyin according to claim 6, characterized in that: The gradient elution conditions are as follows: 0-7 min, 10%-17% acetonitrile; 7-14 min, 17% acetonitrile; 14-19 min, 17%-22% acetonitrile; 19-28 min, 22%-24% acetonitrile; 28-42 min, 24%-35% acetonitrile; 42-49 min, 35%-63% acetonitrile; 49-50 min, 63%-10% acetonitrile.

8. The quality detection method of Xinpuyin according to claim 3 is characterized in that: The regression equations of liquiritin, forsythiaside, chicoric acid and berberine were Y=2048074.5319X-469.0732, Y=813991X-500.71, Y=2471712.7709X+1200.6098, and Y=3561132.1406X+1398.4878.

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