CN113466361A - Method for evaluating quality of gastrointestinal powder - Google Patents
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
The invention discloses a method for evaluating the quality of gastrointestinal powder, which comprises the following steps: preparing a reference solution; preparing a test solution; HPLC chromatograms of the reference substance and the test substance are obtained by adopting a high performance liquid chromatograph; measuring the relative peak area of the common peak of different samples; calculating the similarity between the HPLC (high performance liquid chromatography) spectrum and the control spectrum of different test samples; performing cluster analysis on different test articles; carrying out principal component analysis on common peaks obtained by the fingerprint spectrums of different test articles; and calculating the score of the principal component and the comprehensive score according to the score matrix of the principal component. The invention provides reference for the quality control method of the gastrointestinal powder; various similarity evaluation modes and a cluster analysis method based on the application of SPSS statistical software are established, and the consistency of the overall similarity and improvement degree similarity evaluation method and the cluster analysis result is verified in a two-way mode; the method is based on the analysis method of SPSS statistical software, and the relevance of the index effective components in the monarch drug and the quality is found.
Description
Technical Field
The invention relates to the technical field of pharmacodynamic evaluation, in particular to a quality evaluation method of gastrointestinal powder.
Background
The gastrointestinal powder is an OTC medicament developed by Guangxi Yuan Antang pharmaceutical industry Co Ltd, is prepared by compatibility of a plurality of traditional Chinese medicines such as cinnamon leaf, fructus evodiae, folium artemisiae argyi, fructus amomi, clove, dried orange peel, tuckahoe, baeckea frutescens, swamp mahogany and the like, is an external patch for treating diarrhea, abdominal pain and borborborygmus caused by cold-dampness accumulation, and has obvious effect in a combined treatment scheme. The chemical components of the gastrointestinal powder are complex, and a plurality of components directly influence the functional indications. The gastrointestinal powder currently has only a few researches on content measurement, clinical combination treatment effect observation and the like. The fingerprint spectrum technology is suitable for multi-component complex samples, can comprehensively evaluate the quality of a preparation by an undetermined compound, and is an effective method for ensuring the consistency and the stability of the preparation.
At present, a traditional Chinese medicine fingerprint similarity evaluation system is a common evaluation method, but the software has defects, and the significance is difficult to reflect on samples with small differences and preparations with stable processes. The medium fingerprint atlas has the common characteristics of large data volume and difficult processing, and the clustering analysis and the principal component analysis can reduce the dimension of data, retain main information and improve the processing process.
China is wide in territory, has complex natural geographic environments, and has different environments such as sunshine, temperature, soil texture and the like, which can cause batch difference of medicinal materials. The Chinese patent medicine can generate chemical reaction in the processing process, the complexity of chemical component analysis in the Chinese patent medicine is further increased, and the difficulty of batch research is increased; and because the processing technology of a pharmaceutical factory is basically shaped, the component types and the content of the finished medicines in each batch tend to be similar, thereby increasing the difficulty of batch quality evaluation.
The demand of traditional Chinese medicines and traditional Chinese medicine preparations is rapidly increased all over the world, so the evaluation and control of the quality of medicinal materials are particularly critical, and because the traditional Chinese medicines are difficult to completely characterize all compounds, but the compounds usually have synergistic effect in the aspect of treatment, and a proper analysis method is necessary to find. The fingerprint is a spectrum or chromatogram obtained by measuring various chemical components of the traditional Chinese medicine, has a plurality of corresponding analysis techniques, is one of common methods by combining HPLC, can research gene segments of the traditional Chinese medicine, and is an effective method for evaluating the quality of the traditional Chinese medicine, identifying the truth and falseness, distinguishing species and ensuring the consistency and stability.
Therefore, it is an urgent technical problem to be solved by those skilled in the art to provide a simple, rapid and reproducible quality evaluation method for gastrointestinal powder.
Disclosure of Invention
In view of the above, the invention provides a method for evaluating the quality of gastrointestinal powder, which is used for establishing an HPLC fingerprint of the gastrointestinal powder, evaluating the similarity and performing cluster analysis on multiple batches of gastrointestinal powder, wherein the principal component analysis can study the weight of chemical components in the gastrointestinal powder, and provides a basis for establishing the intra-batch and inter-batch quality control standards of products and promoting more studies.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for evaluating the quality of gastrointestinal powder comprises the following steps:
(1) respectively weighing hesperidin, cinnamic acid, cinnamaldehyde, eugenol, evodiamine and rutaecarpine, dissolving with methanol to obtain a mixed solution containing the above 6 components, and filtering the mixed solution to obtain a reference solution;
(2) weighing intestine and stomach powder, dissolving in methanol, performing ultrasonic treatment, cooling to room temperature, shaking, filtering, and collecting filtrate to obtain test solution;
(3) detecting the reference solution and the test solution by adopting a high performance liquid chromatograph to obtain HPLC chromatograms of the reference and the test solution;
(4) introducing the chromatogram into traditional Chinese medicine chromatogram fingerprint similarity evaluation system software, taking the chromatogram of any test sample as a reference spectrum, performing chromatographic peak matching by adopting a multi-point correction method, generating a reference spectrum by a median method, identifying the chromatographic peak by using a reference substance, determining peaks belonging to hesperidin, cinnamic acid, cinnamaldehyde, eugenol, evodiamine and rutaecarpine, setting the peak belonging to eugenol as the reference peak, and determining the relative peak area of the common peak of different test samples;
(5) calculating the similarity between the HPLC (high performance liquid chromatography) spectrum of the test sample and the reference spectrum by respectively adopting traditional Chinese medicine chromatogram fingerprint similarity evaluation system software, an overall similarity formula and an improvement degree similarity formula, calculating the overall similarity Q between two fingerprint spectrums by using a formula (1), and calculating the improvement degree similarity Q' between the two fingerprint spectrums by using a formula (2);
wherein, ai、bjPeak areas of the same chromatographic peak in the chromatograms A and B are respectively, and n is the number of chromatographic peaks;
(6) taking the relative peak area of the common peak as original data, measuring the squared Euclidean distance by an interclass connection method by using SPSS software, and carrying out cluster analysis on different samples;
(7) carrying out standardization processing on peak areas of the common peaks by using SPSS software, and carrying out principal component analysis on the common peaks obtained by the finger prints of different samples to obtain the characteristics and the variance of a correlation matrix;
(8) and calculating the score of the principal component and the comprehensive score according to the score matrix of the principal component by using SPSS software.
Furthermore, in the step (1), the reference substance solution contains hesperidin 97.52 μ g/mL, cinnamic acid 1.14 μ g/mL, cinnamaldehyde 8.68 μ g/mL, eugenol 7.93 μ g/mL, evodiamine 10.04 μ g/mL and rutaecarpine 4.26 μ g/mL.
The beneficial effect of adopting the further scheme is that: the qualitative analysis of the effective component groups of some monarch drugs in the prescription is helpful for establishing the fingerprint spectrum of the gastrointestinal powder, and defining chemical components influencing the quality evaluation, and the information quantity provided by the main component analysis method for the experiment is increased.
Further, the ratio of the mass of the powder for intestine and stomach to the volume of the methanol in the step (2) is 1: 50.
Furthermore, in the step (2), the ultrasonic power is 500W, the ultrasonic frequency is 40KHz, and the ultrasonic time is 30 min.
The beneficial effect of adopting the further scheme is that: the method for preparing the test solution can simply, quickly and efficiently obtain the test solution with required impurities, and is beneficial to subsequent experimental research.
Further, the chromatographic conditions of the high performance liquid chromatograph in the step (3) are as follows:
a chromatographic column: AgilentXDB-C18(250 mm. times.4.6 mm. times.5 μm);
mobile phase: acetonitrile (a) -0.2% phosphoric acid solution (B);
gradient elution: 0-10 min, 22% A, 10-60 min, 22% -45% A, 60-70 min and 45% A; flow rate: 1.0 mL/min;
detection wavelength: 226 nm;
column temperature: 25 ℃;
sample introduction amount: 10 μ L.
The beneficial effect of adopting the further scheme is that: high performance liquid chromatography is the mainstream chemical component analysis means at present, and the experimental result with high acceptance can be obtained by the method. The obtained chromatographic condition is one of the most key factors for establishing the fingerprint spectrum, so that the reliability of the experiment can be tested, and the spectrum and data information can be provided for the next analysis.
The invention has the beneficial effects that:
the invention establishes the gastrointestinal powder HPLC fingerprint, compares the results of the gastrointestinal powder HPLC fingerprint in different evaluation modes of a traditional Chinese medicine fingerprint similarity evaluation system, overall similarity, improvement degree similarity and the like, performs clustering and principal component analysis by using SPSS software, obtains 18 common peaks from the HPLC fingerprint, and determines 6 compounds by using a reference substance. The cluster analysis result is consistent with the overall similarity and improvement degree similarity analysis result, and gastrointestinal dispersions of different batches can be distinguished, and in the main component analysis, the 6 compounds are not only index effective components of monarch drugs in the prescription, but also are larger weight indexes and are key factors for evaluating the quality of the gastrointestinal dispersions. The method is simple, convenient, rapid and good in reproducibility, is an effective means for performing batch-to-batch analysis on the quality of the multi-batch gastrointestinal powder, and has high application value.
Compared with different evaluation modes such as a traditional Chinese medicine fingerprint similarity evaluation system, overall similarity and improvement degree similarity, the invention can better find the inter-batch difference of the overall similarity and the improvement degree similarity. The cluster analysis is to collect data of similar objects with the same properties, and to perform cluster analysis on related data obtained from the fingerprint by using SPSS software, and the cluster analysis can be used as preliminary exploratory analysis and can intuitively and simply obtain some conclusions. The conclusion obtained by the analysis method is consistent with the analysis result of the overall similarity and the improved degree similarity, and the practicability of the overall similarity and the improved degree similarity is verified in a two-way mode.
The principal component analysis adopted by the method is a bilinear model method, a maximum variance principle is utilized, a plurality of independent variables contained in original data are subjected to linear fitting, new low-dimensional variables replace original high-dimensional variables, namely principal components, all principal components are not related to each other, so that the principal components can reflect most of information of the original variables, the contained information is not overlapped with each other, and the dimension reduction of the data is further realized. The method is provided for searching the correlation of common components in gastrointestinal powder, the SPSS software is used for carrying out principal component analysis on the related data obtained from the fingerprint, and the scores of the principal components are converted from the variance and weight results, so that the method has important reference value for evaluating the quality of medicinal materials.
By combining the research means and the analysis results, the quality control method of the intestines and stomach in bulk and between batches can be expanded and improved compared with the prior method, and the method keeps pace with the modernization of the traditional Chinese medicine, and is beneficial to the supervision of products.
Compared with the prior art, the invention establishes a fingerprint spectrum based on HPLC (high performance liquid chromatography) for the gastrointestinal powder; the invention provides reference for a quality control method of the gastrointestinal powder which mainly adopts hesperidin, cinnamic acid, cinnamaldehyde, eugenol, evodiamine, rutaecarpine and the like; the invention establishes a plurality of similarity evaluation modes for the gastrointestinal powder and a cluster analysis method based on the application of SPSS statistical software, and bidirectionally verifies the consistency of the overall similarity and improvement degree similarity evaluation method and the cluster analysis result; the invention establishes a principal component analysis method based on SPSS statistical software for the gastrointestinal powder, and discovers the correlation between index effective components in monarch drugs and the quality through the principal component analysis result.
Drawings
FIG. 1 is an HPLC comparison fingerprint of gastrointestinal powder in an embodiment of the present invention;
FIG. 2 is an HPLC chromatogram of a mixed control in an example of the invention;
in the figure, 1-hesperidin; 7-cinnamic acid; 9-cinnamaldehyde; 11-eugenol; 14-evodiamine; 16-rutaecarpine;
FIG. 3 is an HPLC fingerprint of 10 batches of samples in the example of the present invention;
FIG. 4 is a schematic diagram of a clustering result of 10 batches of samples in the embodiment of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Instrument and reagent
Waterse2695 series Quaternary gradient Pump high Performance liquid chromatograph (Watts, USA); an XS205 electronic balance (mettler-toledo, switzerland); KS-500DE ultrasonic cleaner (Kunshan Jielimei ultrasonic Instrument Co., Ltd.)
Hesperidin (batch No. 110721-; eugenol (batch number: MUST-16040204), evodiamine (batch number: MUST-16040313), rutaecarpine (batch number: MUST-16040314), and reference substances were purchased from Dowmatte Biotech, Inc., and the purity was 96.2%, 98.8%, 98.7%, 99.00%, 99.99%, and 99.97% in this order.
The number of 10 batches of gastrointestinal powder is S1-S10, the batch numbers are 201812051, 201812052, 201812061, 201812062, 201812063, 201812071, 201812072, 201812073, 201812081 and 201812082 in sequence, and the gastrointestinal powder is provided by Guangxi Source Antang pharmaceutical industry Co.
Acetonitrile is chromatographically pure; other reagents are analytically pure; the water is purified water.
Examples
A method for evaluating the quality of gastrointestinal powder comprises the following steps:
(1) respectively weighing hesperidin, cinnamic acid, cinnamaldehyde, eugenol, evodiamine and rutaecarpine, adding methanol to dissolve to obtain a mixed solution containing the 6 components, and filtering the mixed solution to obtain a reference solution, wherein the hesperidin mass concentration in the reference solution is 97.52 mu g/mL, the cinnamic acid mass concentration is 1.14 mu g/mL, the cinnamaldehyde mass concentration is 8.68 mu g/mL, the eugenol mass concentration is 7.93 mu g/mL, the evodiamine mass concentration is 10.04 mu g/mL, and the rutaecarpine mass concentration is 4.26 mu g/mL;
(2) weighing 0.5g of intestine and stomach powder, dissolving in 25ml of methanol, performing ultrasonic treatment at 500W and 40KHz for 30min, cooling to room temperature, shaking, filtering, and collecting filtrate to obtain 10 batches of test solution;
(3) detecting the reference solution and 10 batches of test solution by using a high performance liquid chromatograph to obtain HPLC chromatograms of the reference and test solutions;
the chromatographic conditions of the high performance liquid chromatograph are as follows:
a chromatographic column: AgilentXDB-C18(250 mm. times.4.6 mm. times.5 μm);
mobile phase: acetonitrile (a) -0.2% phosphoric acid solution (B);
gradient elution: 0-10 min, 22% A, 10-60 min, 22% -45% A, 60-70 min and 45% A; flow rate: 1.0 mL/min;
detection wavelength: 226 nm;
column temperature: 25 ℃;
sample introduction amount: 10 μ L.
(4) Introducing the chromatogram into traditional Chinese medicine chromatogram fingerprint similarity evaluation system software, taking the chromatogram of S1 as a reference chromatogram, performing chromatogram peak matching by adopting a multipoint correction method, generating a reference chromatogram R by a median method, see figure 1, determining 18 common peaks, identifying the chromatogram peaks by using a reference substance, wherein the peaks of 1, 7, 9, 11, 14 and 16 are hesperidin, cinnamic acid, cinnamaldehyde, eugenol, evodiamine and rutaecarpine respectively, the HPLC chromatogram of the mixed reference substance is shown in figure 2, and the HPLC superposed fingerprint of 10 batches of samples is shown in figure 3. The peak area of the 11 th peak is large, and the separation degree and the symmetry factor both meet the requirements, so the reference peak (S) is set, the relative peak area of the common peak of 10 batches of samples is measured, and the result is shown in Table 1.
TABLE 1 relative peak area of common peaks
| Peak number | S1 | S2 | S3 | S4 | S5 | S6 | S7 | | S9 | S10 | |
| 1 | 1.099 | 1.173 | 0.991 | 1.017 | 1.235 | 0.934 | 1.015 | 1.030 | 1.106 | 0.921 | |
| 2 | 0.055 | 0.067 | 0.073 | 0.079 | 0.067 | 0.081 | 0.093 | 0.057 | 0.064 | 0.050 | |
| 3 | 0.018 | 0.023 | 0.023 | 0.023 | 0.027 | 0.023 | 0.028 | 0.016 | 0.018 | 0.014 | |
| 4 | 0.031 | 0.035 | 0.032 | 0.031 | 0.039 | 0.033 | 0.034 | 0.030 | 0.033 | 0.027 | |
| 5 | 0.107 | 0.112 | 0.108 | 0.101 | 0.094 | 0.104 | 0.114 | 0.108 | 0.115 | 0.105 | |
| 6 | 0.032 | 0.035 | 0.033 | 0.031 | 0.038 | 0.030 | 0.037 | 0.038 | 0.040 | 0.032 | |
| 7 | 0.028 | 0.031 | 0.032 | 0.030 | 0.052 | 0.032 | 0.033 | 0.027 | 0.028 | 0.023 | |
| 8 | 0.020 | 0.019 | 0.020 | 0.020 | 0.032 | 0.020 | 0.021 | 0.018 | 0.018 | 0.016 | |
| 9 | 0.363 | 0.351 | 0.385 | 0.369 | 0.388 | 0.368 | 0.413 | 0.370 | 0.395 | 0.384 | |
| 10 | 0.234 | 0.247 | 0.243 | 0.234 | 0.235 | 0.257 | 0.285 | 0.291 | 0.284 | 0.276 | |
| 11 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | |
| 12 | 0.044 | 0.051 | 0.046 | 0.048 | 0.055 | 0.048 | 0.052 | 0.042 | 0.045 | 0.026 | |
| 13 | 0.199 | 0.212 | 0.184 | 0.184 | 0.236 | 0.185 | 0.202 | 0.187 | 0.196 | 0.168 | |
| 14 | 0.212 | 0.264 | 0.232 | 0.230 | 0.245 | 0.247 | 0.250 | 0.200 | 0.256 | 0.217 | |
| 15 | 0.131 | 0.137 | 0.121 | 0.121 | 0.155 | 0.120 | 0.134 | 0.124 | 0.129 | 0.109 | |
| 16 | 0.125 | 0.162 | 0.147 | 0.144 | 0.156 | 0.143 | 0.157 | 0.125 | 0.158 | 0.130 | |
| 17 | 0.066 | 0.026 | 0.022 | 0.022 | 0.029 | 0.022 | 0.026 | 0.024 | 0.025 | 0.021 | |
| 18 | 0.051 | 0.057 | 0.051 | 0.046 | 0.059 | 0.050 | 0.050 | 0.047 | 0.046 | 0.026 |
(5) Similarity between HPLC (high performance liquid chromatography) spectra of different test samples and a reference spectrum is calculated by adopting traditional Chinese medicine chromatogram fingerprint similarity evaluation system software, an overall similarity formula and an improvement degree similarity formula;
wherein, the overall similarity can quantitatively reflect the relative difference of the samples, and the similarity Q of the fingerprint A and the fingerprint B is calculated by a formula (1); when the difference of the peak areas does not exceed 100%, the improvement degree similarity can accurately reflect the relative difference between the sample and the reference, and the similarity Q' of the finger prints A and B is calculated by a formula (2);
ai，bjthe peak areas of the same chromatographic peak in the chromatograms A and B are shown, and n is the number of chromatographic peaks;
the results of HPLC (high performance liquid chromatography) fingerprint spectrums and comparison spectrums of 10 batches of samples under different similarity calculation methods are shown in Table 2, and the results show that the similarity results of the traditional Chinese medicine chromatographic fingerprint similarity evaluation system are all larger than 0.996, which indicates that the chemical components of the gastrointestinal powder in each batch are basically consistent, but the difference between the 10 batches of samples and the reference cannot be reflected. The similarity results of 10 samples calculated by adopting the overall similarity and improvement degree similarity formula are all between 0.88 and 0.96, wherein 8 samples exceed 0.91, 3, 4, 6, 7 and 9 samples have higher similarity with a reference, and 5 and 10 samples have the largest difference with the reference. The results of the overall similarity and the improvement degree similarity of each batch are basically consistent, which indicates that the two evaluation methods are applicable to gastrointestinal diseases.
TABLE 210 similarity of HPLC finger print and control profiles for lots of samples
(6) Taking the relative peak area of the common peak as original data, measuring the squared Euclidean distance by an inter-group connection method by using SPSS software, and carrying out cluster analysis on 10 batches of samples; the results are shown in FIG. 4. The 10 samples can be clustered into three categories: s3, S4, S6, S7 and S9 are in the same category; s8 and S10 are the same; s1, S2 and S5 are in the same category. The classification result is consistent with the overall similarity and the improvement degree similarity result.
(7) Carrying out standardization processing on peak areas of the common peaks by using SPSS software, and carrying out principal component analysis on the common peaks obtained by the finger prints of different samples to obtain the characteristics and the variance of a correlation matrix; some results are shown in Table 3. As a result, the characteristic value lambda of the first 4 components is larger than 1, and the cumulative variance contribution rate of the characteristic values reaches 90.662%, so that the first 4 components are taken as main components. As can be seen from the main factor load matrix in table 4, the main component 1 mainly reflects information of hesperidin, cinnamaldehyde, eugenol, evodiamine, rutaecarpine and peaks 5, 6 and 10, the main component 2 mainly reflects information of cinnamic acid and peaks 2, 3, 4 and 12, the main component 3 mainly reflects information of peaks 8, 13, 15 and 17, and the main component 4 mainly reflects information of peak 18.
TABLE 3 principal Components eigenvalues and contribution ratios
TABLE 4 principal component load matrix
(8) And (3) calculating the score and the comprehensive score of the principal component by using SPSS software according to the score matrix of the principal component, wherein the score corresponds to the overall quality of the gastrointestinal powder of different batches, and ranking is carried out according to the score, and the result is shown in a table 5. The quality of 10 batches of gastrointestinal powder is S8, S9, S6, S3, S7, S10, S4, S5, S1 and S2 from high to low in sequence.
Table 510 sample principal component scores, composite scores, and rankings
The traditional Chinese medicine chromatogram fingerprint similarity evaluation system recommended by the State pharmacopoeia Committee is a common analysis software, and the cosine of an included angle is used as a similarity evaluation index. The similarity calculated by using the method is between 0.996 and 1.000, which shows that the similarity of each spectrum peak in proportion is higher, but the linear fluctuation of the peak area is not detected, and the types and the contents of the components of each batch of preparation are basically stable due to the process, so the method has defects in the evaluation of the preparation, and is more suitable for the variety identification of medicinal materials. Therefore, the overall similarity and the improvement degree similarity are introduced for comparative evaluation, the overall similarity result is between 0.886 and 0.955, the improvement degree similarity result is between 0.878 and 0.951, and the results are consistent with the clustering result, which indicates that the two methods are both suitable for evaluation of the gastrointestinal scattering fingerprint similarity.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (5)
1. The method for evaluating the quality of the gastrointestinal powder is characterized by comprising the following steps of:
(1) respectively weighing hesperidin, cinnamic acid, cinnamaldehyde, eugenol, evodiamine and rutaecarpine, dissolving with methanol to obtain a mixed solution containing the above 6 components, and filtering the mixed solution to obtain a reference solution;
(2) weighing intestine and stomach powder, dissolving in methanol, performing ultrasonic treatment, cooling to room temperature, shaking, filtering, and collecting filtrate to obtain test solution;
(3) detecting the reference solution and the test solution by adopting a high performance liquid chromatograph to obtain HPLC chromatograms of the reference and the test solution;
(4) introducing the chromatogram into traditional Chinese medicine chromatogram fingerprint similarity evaluation system software, taking the chromatogram of any test sample as a reference spectrum, performing chromatographic peak matching by adopting a multi-point correction method, generating a reference spectrum by a median method, identifying the chromatographic peak by using a reference substance, determining peaks belonging to hesperidin, cinnamic acid, cinnamaldehyde, eugenol, evodiamine and rutaecarpine, setting the peak belonging to eugenol as the reference peak, and determining the relative peak area of the common peak of different test samples;
(5) calculating the similarity between the HPLC (high performance liquid chromatography) spectrum of the test sample and the reference spectrum by respectively adopting traditional Chinese medicine chromatogram fingerprint similarity evaluation system software, an overall similarity formula and an improvement degree similarity formula, calculating the overall similarity Q between two fingerprint spectrums by using a formula (1), and calculating the improvement degree similarity Q' between the two fingerprint spectrums by using a formula (2);
wherein, ai、bjAre respectively colorPeak areas of the same chromatographic peak in the spectrograms A and B, wherein n is the number of chromatographic peaks;
(6) taking the relative peak area of the common peak as original data, measuring the squared Euclidean distance by an interclass connection method by using SPSS software, and carrying out cluster analysis on different samples;
(7) carrying out standardization processing on peak areas of the common peaks by using SPSS software, and carrying out principal component analysis on the common peaks obtained by the finger prints of different samples to obtain the characteristics and the variance of a correlation matrix;
(8) and calculating the score of the principal component and the comprehensive score according to the score matrix of the principal component by using SPSS software.
2. The quality evaluation method of gastrointestinal powder according to claim 1, wherein in the step (1), the reference solution contains hesperidin in a mass concentration of 97.52 μ g/mL, cinnamic acid in a mass concentration of 1.14 μ g/mL, cinnamaldehyde in a mass concentration of 8.68 μ g/mL, eugenol in a mass concentration of 7.93 μ g/mL, evodiamine in a mass concentration of 10.04 μ g/mL, and rutaecarpine in a mass concentration of 4.26 μ g/mL.
3. The method for evaluating the quality of the gastrointestinal powder according to claim 1, wherein the volume ratio of the mass of the gastrointestinal powder to the methanol in the step (2) is 1: 50.
4. The method for evaluating the quality of the gastrointestinal powder according to claim 1, wherein the ultrasonic power in the step (2) is 500W, the ultrasonic frequency is 40KHz, and the ultrasonic time is 30 min.
5. The method for evaluating the quality of the gastrointestinal powder according to claim 1, wherein the chromatographic conditions of the high performance liquid chromatograph in the step (3) are as follows:
a chromatographic column: agilent XDB-C18(250 mm. times.4.6 mm. times.5 μm);
mobile phase: acetonitrile (a) -0.2% phosphoric acid solution (B);
gradient elution: 0-10 min, 22% A, 10-60 min, 22% -45% A, 60-70 min and 45% A; flow rate: 1.0 mL/min;
detection wavelength: 226 nm;
column temperature: 25 ℃;
sample introduction amount: 10 μ L.
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