Traditional Chinese medicine powder boiling process and dosage determination method

Abstract

The invention discloses a traditional Chinese medicine powder boiling process and a dosage determination method. The method comprises the following steps: preparing the traditional Chinese medicine decoction powder; determining evaluation index components; establishing a fingerprint spectrum or a characteristic fingerprint spectrum; evaluation index component content determination: establishing an evaluation index component content determination method, and determining the content; determining a powder boiling process; determining the dosage of the decoction powder: preparing decoction of the decoction powder with different multiples by using the determined process of the decoction powder, comparing the similarity of fingerprints or characteristic fingerprints of the decoction of the decoction powder with different doses and the traditional decoction of the decoction piece by taking the traditional decoction piece as a reference, and determining the dosage of the decoction powder according to the weighted comprehensive score of the content of the evaluation index component. The selected fingerprint spectrum or characteristic fingerprint spectrum similarity and the weighted comprehensive score of the evaluation index component content are closest to the dosage of a traditional decoction piece decoction, the dosage is determined as the boiling powder dosage, and the pharmacodynamic test verifies the dosage. The problems that traditional Chinese medicine decoction pieces are not easy to distinguish, prone to gelatinization during decoction, difficult to adjust, difficult to convert dosage and the like are effectively solved.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine, and particularly relates to a decocting process and dosage determination method for traditional Chinese medicine powder. Background Art

[0002] Traditional Chinese medicine powder is a form of medication in which traditional Chinese medicine is made into coarse powder or granules within a certain range, decocted with water, and taken with or without filtering the residue. This form is one of the traditional medication methods in clinical traditional Chinese medicine, starting from the pre-Qin period and being most prevalent during the Tang and Song dynasties. The decocting of traditional Chinese medicine powder simplifies the leaching conditions of active ingredients. Whether the active ingredients in the medicine can be completely leached or retained directly affects the clinical efficacy. For traditional Chinese medicine powder, after the medicinal materials are pulverized, the diffusion area increases, the dissolved substances increase, and the dissolution rate accelerates. As the degree of pulverization increases, the chance of cell destruction increases, and the substances inside the cells directly contact water and dissolve, thus increasing the leaching amount, and the decocting time and dosage can be reduced. Compared with the traditional decoction of sliced traditional Chinese medicine, traditional Chinese medicine powder has great advantages in saving medicinal materials, reducing costs, reducing decocting costs, and increasing the extraction rate of active ingredients of drugs. However, there are also problems such as difficulty in identifying traditional Chinese medicine powder slices, easy gelatinization during decocting, difficult dispensing, and difficult dosage conversion. Preparing suitable traditional Chinese medicine powder, optimizing the decocting process, and scientifically converting the decocting dosage are problems that need to be solved urgently at present. Summary of the Invention

[0003] In view of the defects of the prior art, the present invention provides a decocting process and dosage determination method for traditional Chinese medicine powder. Based on the "chemical composition - biological effect" model, the present invention determines the process and dosage of traditional Chinese medicine powder, effectively solving the problems such as difficulty in identifying traditional Chinese medicine powder slices, easy gelatinization during decocting, difficult dispensing, and difficult dosage conversion.

[0004] The technical solution of the present invention is as follows.

[0005] In view of the problems such as difficulty in identifying traditional Chinese medicine powder slices, easy gelatinization during decocting, and difficult dispensing, the traditional Chinese medicine is prepared into granules that can pass through 4 mesh but not through 10 mesh. The traditional Chinese medicine granules of this size have good uniformity, are easy to identify, are not easy to gelatinize during decocting, can shorten the decocting time, and are convenient for dispensing. In view of the problem of difficult dosage conversion, the present invention uses the quality marker of traditional Chinese medicine or other active pharmaceutical ingredients as the evaluation index component, establishes a fingerprint or characteristic fingerprint including the evaluation index component; establishes a high-performance liquid chromatography detection method for the content of the evaluation index component; optimizes the decocting process with the weighted comprehensive score of the content of the evaluation index component as the index; determines the dosage of traditional Chinese medicine powder with the similarity of the fingerprint or characteristic fingerprint and the weighted comprehensive score of the content of the evaluation index component as the index, and verifies it through pharmacodynamic experiments.

[0006] A decocting process and dosage determination method for traditional Chinese medicine powder, characterized by comprising the following steps:

[0007] (1) Preparation of decoction pieces in powder form: The traditional Chinese medicine is crushed using a coarse crusher;

[0008] (2) Determination of evaluation index components: Select quality markers and other pharmacodynamic components as evaluation index components; Determination of quality markers: Predict the quality markers of traditional Chinese medicine based on five aspects: quality transfer and traceability, component uniqueness, effectiveness, measurability, and the compatibility environment of compound prescriptions;

[0009] (3) Establishment of fingerprint or characteristic fingerprint: Establish a fingerprint or characteristic fingerprint of traditional Chinese medicine decoction containing evaluation index components;

[0010] (4) Determination of the content of evaluation index components: Using the chromatographic conditions of the fingerprint or characteristic fingerprint of traditional Chinese medicine decoction obtained in step (3), establish a method for determining the content of evaluation index components and determine their content;

[0011] (5) Determination of the decoction pieces in powder form process: Use single-factor investigation, orthogonal experiment, and weighted scoring method to determine the final decoction pieces in powder form process;

[0012] (6) Determination of the dosage of decoction pieces in powder form: Using the determined decoction pieces in powder form process, prepare decoction pieces in powder form of 0.1 to 1 times and more than five different multiples. Taking the traditional decoction pieces as a reference, compare the similarity of the fingerprint or characteristic fingerprint of the decoction pieces in powder form with different dosages and the traditional decoction pieces, as well as the weighted comprehensive score of the content of evaluation index components. Select the dosage with the fingerprint or characteristic fingerprint similarity and the weighted comprehensive score of the content of evaluation index components closest to the traditional decoction pieces as the dosage of decoction pieces in powder form, and verify it through pharmacodynamic experiments.

[0013] In the above method, in step (1), if the decoction pieces in powder form of traditional Chinese medicine are taken after removing the residue, the traditional Chinese medicine is crushed into particles with a size of 4 to 10 meshes.

[0014] In the above method, in step (1), if the decoction pieces in powder form of traditional Chinese medicine are taken together, the traditional Chinese medicine is crushed into fine powder with a size of 60 to 80 meshes.

[0015] In the above method, in step (3), the fingerprint or characteristic fingerprint is a high-performance liquid chromatography fingerprint or characteristic fingerprint; The specific steps are as follows: 1. Selection of chromatographic conditions; 2. Determination of the preparation methods of reference substances and test samples; 3. Methodological investigation; 4. Establishment of fingerprint or characteristic fingerprint; 5. Evaluation of the similarity of fingerprint or characteristic fingerprint.

[0016] In the above method, in step (4), the method for determining the content of evaluation index components is a high-performance liquid chromatography detection method. The specific steps are as follows: 1. Selection of chromatographic conditions; 2. Determination of the preparation methods of reference substances and test samples; 3. Methodological investigation; 4. Establishment of a high-performance liquid chromatography detection method for determining the content of evaluation index components; 5. Detection of the content of evaluation index components.

[0017] Compared with the prior art, the advantages of the present invention are as follows:

[0018] Currently, the decocted powder of traditional Chinese medicine is difficult to distinguish, easy to gelatinize during decoction, difficult to filter, difficult to dispense, and difficult to convert the dosage. The decocted powder of traditional Chinese medicine is difficult to distinguish, easy to gelatinize during decoction, and difficult to filter and dispense, mainly because the particle size of the decocted powder is too fine. According to the characteristics of the taking method of the decocted powder, the present invention determines the particle size for taking the decocted powder after removing the residue as being able to pass through a 4-mesh sieve but not able to pass through a 10-mesh sieve, which can partially preserve the characteristics of the decocted pieces, facilitating identification, effectively avoiding gelatinization during decoction, being easy to filter, and convenient for dispensing, and preferably solving the problems such as the difficulty in distinguishing, easy gelatinization during decoction, difficult filtration, and difficult dispensing of the current decocted powder of traditional Chinese medicine. The difficulty in converting the dosage of the decocted powder of traditional Chinese medicine is mainly due to reasons such as the evaluation indexes selected in the prior art being not scientific and reasonable enough. The present invention selects the quality markers of traditional Chinese medicine and other pharmacodynamic components as the evaluation index components, establishes a fingerprint or characteristic fingerprint containing the evaluation index components, establishes a detection method for the content of the evaluation index components, and uses methods such as single-factor investigation, orthogonal test, and weighted scoring to optimize the decoction process. Using the determined decoction process, decocted powder decoctions with different multiples (5 or more doses can be selected from 0.1 to 1 times according to needs) are prepared. Taking the traditional decoction of decocted pieces as a reference, the similarity of the fingerprint or characteristic fingerprint of the decocted powder decoction with different doses and the traditional decoction of decocted pieces, as well as the weighted comprehensive score of the content of the evaluation index components, are compared. The dose with the fingerprint or characteristic fingerprint similarity and the weighted comprehensive score of the content of the evaluation index components closest to the traditional decoction of decocted pieces is selected and determined as the decocted powder dose, and it is verified through pharmacodynamic experiments. Determining the dose of the decocted powder of traditional Chinese medicine based on the "chemical composition - biological effect" model is more scientific and reasonable. Description of the Drawings

[0019] Figure 1 is the fingerprint of Danggui Buxue Decoction;

[0020] Figure 2 is the UHPLC chromatogram of the reference substance;

[0021] Figure 3 is the UHPLC chromatogram of the decoction of Danggui Buxue Decoction with decocted pieces;

[0022] Figure 4 is the UHPLC chromatogram of the decocted powder of Danggui Buxue Decoction;

[0023] Figure 5 is the result of the investigation on the soaking time;

[0024] Figure 6 is the result of the investigation on the water addition amount;

[0025] Figure 7 is the result of the investigation on the decoction time.

[0026] Figure 8 Orthogonal test results.

[0027] Figure 9 To generate overlay maps and reference maps. DETAILED DESCRIPTION

[0028] Example embodiments will now be described more fully with reference to the accompanying drawings. However, example embodiments can be implemented in a variety of forms and should not be construed as limited to the examples set forth herein; rather, these example embodiments are provided so that the description of the present disclosure will be more comprehensive and complete, and the concept of the example embodiments will be fully conveyed to those skilled in the art.

[0029] Example 1

[0030] Study on the cooking process and dosage of Danggui Buxue Decoction. In this embodiment, Danggui Buxue Decoction includes two Chinese medicines, Radix Astragali and Radix Angelicae Sinensis.

[0031] The method comprises the following steps:

[0032] (1) Preparation by boiling

[0033] First, the astragalus is processed into 2-5 mm segments, preferably 2-3 mm segments; the angelica is processed into 2-10 mm thick slices, preferably 3-5 mm thick slices. Then it is crushed by a coarse crusher equipped with a 4-mesh screen, and the medicinal materials are added in small amounts and crushed. After crushing, the finished product rate of astragalus powder is 83.5%, and the finished product rate of angelica powder is 85%.

[0034] (2) Determination of evaluation index components

[0035] The quality markers of Danggui Buxue Decoction, ferulic acid, verbascosin, verbascosin glucoside, formononetin, formononetin and the active ingredient ligusticolide I were selected as evaluation index components, and the weights were assigned using the hierarchical analysis method: ferulic acid (22.99%), verbascosin (22.99%), verbascosin glucoside (36.50%), formononetin (4.33%), formononetin (6.50%), ligusticolide I (6.69%).

[0036] (3) Fingerprint establishment

[0037] Taking the relative retention time, separation degree and peak shape of chromatographic peaks of ferulic acid, calycosin, calycosin glucoside, formononetin, formononetin and ligusticum lactone I as the investigation factors, the appropriate mobile phase was selected to establish the fingerprint of Danggui Buxue Decoction. The details are as follows:

[0038] 1. Chromatographic conditions

[0039] Chromatographic conditions: Agilent 1290 ultra-high performance liquid chromatograph (model: Agilent 1290 Infinity II), chromatographic column (InfinityLab Poroshell 120 Aq-C18, 2.1×100 mm 2.7 μm), pre-column (UHPLC Guard 3PK, InfinityLab Poroshell 120 Aq-C18 2.1×5 mm 2.7 μm). The mobile phase was acetonitrile (A)-0.1% glacial acetic acid (B) gradient elution (0-10 min, 5% A-18% A; 10-15 min, 18% A; 15-25 min, 18% A-19% A; 25-37 min, 19% A-22% A; 37-40 min, 22% A-34% A; 40-45 min, 34% A), detection wavelength 260 nm, flow rate 0.3 mL·min -1 , column temperature 30 °C, injection volume 5 μL.

[0040] 2. Preparation of reference substances and test samples

[0041] 2.1 Preparation method of reference substance solution

[0042] Accurately weigh the reference substances of ferulic acid, calycosin, calycosin-7-O-β-D-glucoside, ononin, formononetin, and senkyunolide I, prepare reference substance solutions respectively, and then accurately measure each reference substance solution to prepare a mixed reference substance solution with a certain concentration (ferulic acid 63.82 μg / mL, calycosin 37.26 μg / mL, calycosin-7-O-β-D-glucoside 130.5732 μg / mL, ononin 32.99 μg / mL, formononetin 17.94 μg / mL, senkyunolide I 19.79 μg / mL), and set aside.

[0043] 2.2 Preparation method of test sample solution

[0044] Prepare the decoction powder solution of Danggui Buxue Decoction. Accurately measure 10 mL of the decoction powder solution, place it in a 25 mL volumetric flask, add methanol to volume, shake well, transfer to a centrifuge tube and centrifuge (10000 r·min -1 ) for 5 min, take the supernatant, and filter through a 0.22 μm microporous filter membrane.

[0045] 3. Establishment of fingerprint

[0046] Take the decoction of traditional Chinese herbal pieces and the decocted powder solutions of Danggui Buxue Decoction at 0.5 times, 0.6 times, 0.7 times, 0.8 times, 0.9 times and 1 time doses (marked as S1 - S7, where the decoction of traditional Chinese herbal pieces is S1, 0.5 times is S2, 0.6 times is S3, 0.7 times is S4, 0.8 times is S5, 0.9 times is S6, 1 time is S7). Prepare the test solutions according to the method under "2.2", inject samples for determination under the chromatographic conditions in item "1.", and record the chromatograms. See details in Figure 1 It is the fingerprint of Danggui Buxue Decoction, where 3 represents ferulic acid, 4 represents calycosin - 7 - O - β - D - glucoside, 5 represents senkyunolide I, 7 represents ononin, 8 represents calycosin, and 10 represents formononetin.

[0047] (4) Determination of the content of the evaluation index components: Regarding the relative retention time, resolution and peak shape of the chromatographic peaks of ferulic acid, calycosin, calycosin - 7 - O - β - D - glucoside, formononetin, ononin, and senkyunolide I as the investigation factors, select the appropriate mobile phase, establish an HPLC method for the determination of the content of the evaluation index components of Danggui Buxue Decoction, and determine their contents. Figure 2 It is the UHPLC chromatogram of the reference substance; Figure 3 It is the UHPLC chromatogram of the traditional Chinese herbal pieces of Danggui Buxue Decoction; Figure 4 It is the UHPLC chromatogram of the decocted powder of Danggui Buxue Decoction; where 3 is ferulic acid, 4 is calycosin - 7 - O - β - D - glucoside, 5 is senkyunolide I, 7 is ononin, 8 is calycosin, and 10 is formononetin.

[0048] Specifically as follows:

[0049] 1. Chromatographic conditions

[0050] Same as the chromatographic conditions of the fingerprint.

[0051] 2. Preparation of reference substance and test solution

[0052] The preparation methods of the reference substance and test solution are the same as those of the reference substance and test solution in the fingerprint.

[0053] (5) Determination of the decocted powder process.

[0054] 1. Single - factor investigation

[0055] According to the requirements of the "Specification for Chinese Medicine Decoction in Medical Institutions", the soaking time, the amount of water added, and the decocting time were investigated. Six levels were set for each factor. Using ferulic acid, calycosin, calycosin-7-O-β-D-glucoside, formononetin, ononin, and senkyunolide I as evaluation indicators for weighted comprehensive scoring, three of these levels were selected for the orthogonal experiment. The soaking time was investigated at 10 min, 15 min, 20 min, 25 min, 30 min, and 35 min; the amount of water added was investigated at 8 times, 10 times, 12 times, 14 times, 16 times, and 18 times; and the decocting time was selected at 20 min, 25 min, 30 min, 35 min, 40 min, and 45 min. The investigation results showed that for the soaking time of 20 min, 25 min, and 30 min (as shown in Figure 5 ), for the amount of water added of 14 times, 16 times, and 18 times (as shown in Figure 6 ), and for the decocting time of 35 min, 40 min, and 45 min (as shown in Figure 7 ), the comprehensive scores were relatively high. Therefore, the soaking time was selected as 20 min, 25 min, and 30 min; the amount of water added was selected as 14 times, 16 times, and 18 times; and the decocting time was selected as 35 min, 40 min, and 45 min for the orthogonal experiment.

[0056] 2. Orthogonal experiment: Soaking times of 20 min, 25 min, and 30 min, amounts of water added of 14 times, 16 times, and 18 times, and decocting times of 35 min, 40 min, and 45 min were selected for the orthogonal experiment. Using the contents of ferulic acid, calycosin, calycosin-7-O-β-D-glucoside, formononetin, ononin, and senkyunolide I as indicators for weighted comprehensive scoring, the decoction-dispersion process was optimized. The results showed (as shown in Figure 8 ) that the highest comprehensive score was obtained with 18 times the amount of water added, soaking for 20 min, and decocting for 45 min. Therefore, 18 times the amount of water added, soaking for 20 min, and decocting for 45 min were selected as the decoction-dispersion process for Danggui Buxue Decoction. After verification, this process was stable and feasible, so it was determined as the decoction-dispersion process for Danggui Buxue Decoction.

[0057] Table 1 Factors and Levels

[0058]

[0059] Table 2 Orthogonal Design

[0060]

[0061] Table 3 Orthogonal Intuitive Analysis

[0062]

[0063] Table 4 Variance Analysis

[0064]

[0065] (6) Determination of the decoction pieces dosage for decocting in dispersion form

[0066] 1. Determination based on the similarity of fingerprint chromatograms

[0067] Using the determined decocting in dispersion form process, prepare the decoction pieces of Danggui Buxue Decoction in dispersion form at 0.5 times, 0.6 times, 0.7 times, 0.8 times, 0.9 times, and 1.0 times the amount of the decoction pieces respectively. Prepare the decoction pieces of Danggui Buxue Decoction according to the requirements of the "Regulations on the Decoction of Traditional Chinese Medicines in Medical Institutions" (marked as S1 - S7, where the decoction of traditional decoction pieces is S1, 0.5 times is S2, 0.6 times is S3, 0.7 times is S4, 0.8 times is S5, 0.9 times is S6, and 1 time is S7). Prepare the test solution according to the above method, inject the sample under the above chromatographic conditions for determination, record the chromatogram, import the data into the "Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints (2012 Edition)", use the average method and multi-point calibration method, select a time width of 0.1 min, generate the superimposed chromatogram and the reference chromatogram, as shown in Figure 9 A in it. A total of 10 common peaks are calibrated. By comparing with the reference substance ( Figure 9 B in it), 6 components are identified, namely ferulic acid, calycosin - 7 - O - glucoside, senkyunolide I, ononin, calycosin, and formononetin. The similarity between the fingerprint chromatograms of the decoction pieces of Danggui Buxue Decoction at different dosage multiples and the reference chromatogram is > 0.99, and the similarity of the decoction pieces of Danggui Buxue Decoction at 0.9 times the dosage reaches 0.999.

[0068] 2. Determination based on the weighted comprehensive score of the content of the evaluation index components

[0069] Taking the contents of ferulic acid, calycosin, calycosin - 7 - O - glucoside, formononetin, ononin, and senkyunolide I as the indexes for weighted comprehensive scoring, select the dosage that is closest to the decoction pieces of Danggui Buxue Decoction in terms of the comprehensive score. The result shows that the decocting in dispersion form at 0.9 times the amount of the decoction pieces is the closest.

[0070] Table 5 Results of sample determination (μg / mL, n = 3)

[0071]

[0072] 3. Verification by pharmacodynamic experiments

[0073] Using rats with qi - blood deficiency model, taking blood routine, organ index, and swimming time as indexes, verify through pharmacodynamic experiments whether the dosage determined by the fingerprint chromatogram and the weighted comprehensive score of the content of the evaluation index components is reasonable and feasible. The results show that the decoction pieces of Danggui Buxue Decoction in dispersion form at 0.9 times the dosage has the same pharmacodynamic effect as the decoction pieces of Danggui Buxue Decoction.

[0074] Table 6 Effects of Danggui Buxue Decoction on the blood routine of rats with qi - blood deficiency model

[0075]

[0076] Table 7 Effects of Danggui Buxue Decoction on Organ Index and Swimming Time of Rats with Qi and Blood Deficiency Model

[0077]

[0078] Although the embodiments of the present invention have been shown and described above, it can be understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those of ordinary skill in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present invention.

Claims

1. A Chinese medicine decoction process and dosage determination method, characterized in that: The steps include: (1) Preparation of Chinese medicine powder: crush the Chinese medicine using a coarse crusher; (2) Determination of evaluation index components: Select quality markers as evaluation index components; Determination of quality markers: Predict the quality markers of traditional Chinese medicine based on five aspects: mass transfer and traceability, ingredient specificity, effectiveness, measurability, and compound compatibility environment; (3) Establishment of fingerprint or characteristic fingerprint: Establishment of fingerprint or characteristic fingerprint of Chinese herbal medicine decoction including evaluation index components; (4) Determination of the content of the evaluation index component: using the chromatographic conditions of the fingerprint or characteristic fingerprint of the traditional Chinese medicine decoction obtained in step (3), establishing a method for determining the content of the evaluation index component, and determining its content; (5) Determination of the cooking process: The final cooking process was determined by using single factor investigation, orthogonal test and weighted comprehensive scoring method; (6) Determination of the decoction dosage: Use a determined decoction process to prepare decoctions of 0.1 to 1 times or more than 5 different multiples. Take the traditional decoction as a reference, compare the fingerprint or characteristic fingerprint similarity between the decoctions of different doses and the traditional decoction, and the weighted comprehensive score of the evaluation index component content. Select the dose with the fingerprint or characteristic fingerprint similarity and the weighted comprehensive score of the evaluation index component content closest to the traditional decoction, and determine it as the decoction dosage.

2. The Chinese medicine decoction process and dosage determination method as claimed in claim 1, characterized in that: In step (1), if the Chinese medicine is boiled and the residue is removed for consumption, the Chinese medicine is crushed into particles of 4-10 mesh sizes.

3. The Chinese medicine decoction process and dosage determination method as claimed in claim 1, characterized in that: In step (1), if the Chinese medicine is boiled and taken together, the Chinese medicine is crushed into fine powder of 60-80 mesh.

4. The Chinese medicine decoction process and dosage determination method as claimed in claim 1, characterized in that: In step (3), the fingerprint or characteristic fingerprint is a high performance liquid chromatography fingerprint or characteristic fingerprint; the specific steps are as follows:

1. Selection of chromatographic conditions; 2. Determination of the preparation method of reference and test samples; 3. Methodological investigation; 4. Establishment of fingerprint or characteristic fingerprint; 5. Evaluation of the similarity of fingerprint or characteristic fingerprint.

5. The Chinese medicine decoction process and dosage determination method as claimed in claim 1, characterized in that: In step (4), the method for determining the content of the evaluation index component is a high performance liquid chromatography detection method. The specific steps are as follows:

1. Selection of chromatographic conditions; 2. Determination of the preparation method of the reference substance and the test substance; 3. Methodological investigation; 4. Establishment of a high performance liquid chromatography detection method for the content of the evaluation index component; 5. Detection of the content of the evaluation index component.

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