CN112379034A - Identification model of rhubarb medicinal material primordium, construction method and identification method thereof - Google Patents

Identification model of rhubarb medicinal material primordium, construction method and identification method thereof Download PDF

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CN112379034A
CN112379034A CN202011474422.4A CN202011474422A CN112379034A CN 112379034 A CN112379034 A CN 112379034A CN 202011474422 A CN202011474422 A CN 202011474422A CN 112379034 A CN112379034 A CN 112379034A
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rhubarb
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王丽婷
董礼
胡莹莹
严敏嘉
胡敏
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Sanju Medical & Pharmaceutical Co ltd
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Abstract

The invention relates to the technical field of medicinal material identification, in particular to an identification model of rhubarb medicinal material primordium, a construction method and an identification method thereof. The common chromatographic peak is selected by detecting the fingerprint of the rhubarb drug, the peak area is taken as an independent variable and substituted into a typical discriminant function to obtain a corresponding coordinate point, and then the consistency of the coordinate point and the standard coordinate position is observed. The identification method is more intuitive and can quickly identify the type of the original rhubarb. And (4) adopting a classification function model, substituting the independent variable into a classification function for calculation, and comparing the discrimination values, wherein the maximum value is the primitive corresponding to the sample to be identified. The identification method is simple and rapid, and the identification result is more accurate. The identification method provided by the invention is simple in calculation, accurate in identification result, enriches identification means and is beneficial to establishment of internal control quality standards of enterprises.

Description

Identification model of rhubarb medicinal material primordium, construction method and identification method thereof
Technical Field
The invention relates to the technical field of medicinal material identification, in particular to an identification model of rhubarb medicinal material primordium, a construction method and an identification method thereof.
Background
Radix et rhizoma Rhei, which is dried root and rhizome of Rheum palmatum L., Rheum tanguticum Maxim. Ex Balf. or Rheum officinale Baill. of Polygonaceae. Collecting and digging stems and leaves at late autumn or before germination in spring, removing thin roots, scraping outer skin, cutting into sections or segments, stringing, and drying or directly drying.
At present, regarding the identification of the basic source of medicinal materials, the Chinese pharmacopoeia is identified by DNA barcode molecules. However, many varieties of medicinal materials do not have DNA barcodes. When controlling the medicinal material source, enterprises can only adopt an identification method in pharmacopeia standards, namely appearance shape identification, microscopic identification, thin-layer identification, chemical identification and the like. However, the above method still cannot identify different sources of the same variety of herbs. Therefore, the development of an economic and reliable identification method of the medicinal material source is of great significance.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a radix et rhizoma rhei medicinal material primitive identification model and a construction method thereof. Dimension reduction is carried out on 17 variables, and the problem of multiple linear collinearity is solved, so that the identification is more accurate. And moreover, the fingerprint spectrum analysis is carried out on the medicinal materials by utilizing the high performance liquid chromatography, so that the data is accurate and stable.
The second purpose of the invention is to provide a method for identifying the radix et rhizoma Rhei medicinal material primitive, which is characterized in that the discriminant score of a sample to be identified is calculated through a typical discriminant function, and the clustering condition of the sample is observed in a scatter diagram.
The third objective of the present invention is to provide another identification method for the radix et rhizoma Rhei medicinal material primitive, which calculates the discrimination value of the sample to be identified through a classification function, and takes the maximum value as the discrimination category of the sample. The identification method is simpler and quicker. And the identification result is more accurate. Enriches the identification means and is beneficial to the establishment of internal control quality standards of enterprises.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
a method for constructing a radix et rhizoma Rhei medicinal material primordial identification model comprises the following steps:
(1) respectively establishing three groups of standard fingerprints of medicinal rheum officinale, rheum tanguticum and rheum palmatum by adopting a liquid chromatography, independently calculating the proportion of 17 groups of characteristic peaks on the three groups of standard fingerprints to the total peak area, and sequentially obtaining three groups of independent variables X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12, X13, X14, X15, X16 and X17 from left to right;
(2) respectively substituting the three independent variables into a typical discriminant function for calculation to obtain three discriminant scores D1 and D2;
the typical discriminant function is:
function 1:
D1＝155.8×X1-39.9×X2-129.1×X3+117.5×X4+6.8×X5+489.9×X6+92.7×X7+786.4×X8+147.8×X9-46.6×X10+718.9×X11+167.5×X12+7.2×X13+333.8×X14-139.9×X15+1429.1×X16-53.8×X17-70.7；
function 2:
D2＝56.5×X1+1.7×X2+5.8×X3+4.4×X4-6.9×X5-54.5×X6+16×X7+70.4×X8+222×X9-22.6×X10-139×X11+18.4×X12+14.8×X13+76.4×X14+75.7×X15-137.1×X16+1.5×X17-8.8；
(3) taking D1 as abscissa and D2 as ordinate to obtain three groups of standard coordinate position scatter diagrams respectively corresponding to the medicinal rhubarb, the Tanggute rhubarb and the palmate rhubarb, namely a rhubarb medicinal material primitive identification model.
The invention provides a method for constructing a radix et rhizoma Rhei medicinal material primordium identification model, and the model constructed by the method can accurately identify three radix et rhizoma Rhei medicinal material primordiums.
Preferably, the step of establishing three groups of standard fingerprint spectrums of the rhubarb horsetails, the rhubarb horsetails from Tanggu and the rhubarb horsetails from Zhang Ye includes: taking 0.5g of a rhubarb sample, sieving the rhubarb sample by a No. 2 sieve, placing the rhubarb sample in a conical flask with a plug, adding 25ml of water, heating and refluxing for 60 minutes, cooling the mixture, shaking up the mixture, filtering the mixture, and taking a subsequent filtrate for later use.
An identification model of rhubarb medicinal material primordium is obtained by the construction method.
The invention adopts principal component analysis and cluster analysis to establish a model capable of identifying three rhubarb medicinal material primordial genes, and the identification model is accurate and reliable through inspection, and the identification result can be accurately obtained through the model.
A method for identifying radix et rhizoma Rhei medicinal material primordium comprises making sample to be identified into sample solution, performing liquid chromatography to obtain sample map, calculating the ratio of 17 groups of characteristic peaks on the sample map to total peak area, sequentially obtaining independent variables X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12, X13, X14, X15, X16 and X17 from left to right; substituting into typical discriminant function to obtain discriminant scores D1 and D2 corresponding to the sample to be tested; obtaining a coordinate position corresponding to the to-be-tested sample by taking D1 as an abscissa and D2 as an ordinate; observing the consistency of the coordinate position of the sample to be tested and the standard coordinate position, thereby identifying the primordium of the sample to be identified; the standard coordinate positions are obtained by a classical discriminant function of the rhubarb, the rheum tanguticum Maxim and the rheum palmatum L.
Preferably, the standard coordinate position is obtained by the construction method of the radix et rhizoma Rhei medicinal material primitive identification model.
Preferably, the step of preparing the solution to be tested comprises: taking 0.5g of a rhubarb sample, sieving the rhubarb sample by a No. 2 sieve, putting the rhubarb sample into a conical flask with a plug, adding 25ml of water, heating and refluxing for 60 minutes, cooling the mixture, shaking up the mixture, filtering the mixture, and taking a subsequent filtrate to obtain the rhubarb extract.
In some embodiments, the rhubarb drug can be identified by the following method: firstly, a rheum officinale medicinal material basic identification model is constructed by the construction method to obtain three groups of standard coordinate position scatter diagrams respectively corresponding to medicinal rheum officinale, rheum tanguticum and rheum palmatum. And then obtaining the atlas and the independent variable of the sample to be detected, and substituting the atlas and the independent variable into the canonical discriminant function to obtain the coordinate position scatter diagram of the discriminant scores D1 and D2 corresponding to the sample to be detected. And finally, observing the consistency of the coordinate position of the sample to be identified and three groups of standard coordinate positions corresponding to the rhubarb, the radix et rhizoma rhei and the palmate rhubarb, thereby identifying the primordium of the sample to be identified.
Preferably, the identification model further comprises a standard coordinate position diagram of three basic crude drugs of medicinal rhubarb, palmate rhubarb and tanggu rhubarb.
The specific coordinates of the standard coordinate position map include:
medicinal rhubarb: abscissa 24.558, and ordinate 4.812.
Tanggute radix Et rhizoma Rhei: abscissa 11.264, and ordinate-6.171.
Rhubarb palmate leaf: abscissa-35.822, and ordinate 1.359.
In some embodiments, the rhubarb drug can be further identified by the following method: firstly, constructing a standard coordinate position diagram of three basic crude drugs of medicinal rhubarb, palmate rhubarb and Tanggute rhubarb according to the specific coordinates. And then obtaining the atlas and the independent variable of the sample to be detected, and substituting the atlas and the independent variable into the canonical discriminant function to obtain the coordinate points of the discriminant scores D1 and D2 corresponding to the sample to be detected. And finally, observing the consistency of the coordinate points and the standard coordinate position diagram, thereby identifying the primordial of the sample to be detected.
The identification method is simpler and more convenient, the identification is faster, and the identification result is clear at a glance.
The invention establishes a model capable of identifying three rhubarb medicinal material primordiums by detecting the fingerprint of the rhubarb medicinal material, selecting a common chromatographic peak, taking the peak area as the original data, and carrying out principal component analysis and clustering judgment, has simple calculation method, and compared with the common microscopic identification and thin-layer identification methods, the identification method provided by the invention can be deeper used for identifying different primordiums of the same variety, and enriches the identification means.
Preferably, the chromatographic conditions of the liquid chromatographic analysis are as follows: using octadecylsilane chemically bonded silica as filler, acetonitrile as mobile phase A, and 0.05-0.5% phosphoric acid solution as mobile phase B, and performing gradient elution as follows:
Figure BDA0002834642910000041
Figure BDA0002834642910000051
the chromatographic conditions of the liquid chromatography analysis adopted when the typical discriminant function and the classification function are established are as described above, so that the identification result obtained when the sample to be detected is analyzed by adopting the standard is more accurate.
Preferably, octadecylsilane bonded silica is used as a filler, and the column length is 150mm, the inner diameter is 2.1mm, and the particle diameter is 1.6 μm.
Preferably, the number of theoretical plates is not less than 3000 calculated by emodin peak.
Preferably, the chromatographic conditions are: the flow rate was 0.30ml per minute.
Preferably, the column temperature is 25 ℃.
Preferably, the detection wavelength is 260 nm.
The identification result obtained when the sample to be detected is analyzed by adopting the standard is more accurate.
A method for identifying a radix et rhizoma Rhei medicinal material primordium comprises the following steps:
(1) establishing a fingerprint of a sample to be detected by adopting a liquid chromatography, calculating the proportion of 17 groups of characteristic peaks on the fingerprint to the total peak area, and sequentially obtaining independent variables A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, A12, A13, A14, A15, A16 and A17 from left to right;
(2) substituting the independent variables into classification functions corresponding to the medicinal rhubarb, the Tanggute rhubarb and the palmate rhubarb to calculate to obtain three groups of discrimination values Y1, Y2 and Y3;
the classification function is:
medicinal rhubarb:
Y1＝4.1×A1-0.97×A2-3.14×A3+2.91×A4+0.13×A5+11.77×A6+2.35×A7+19.65×A8+4.7×A9-1.25×A10+16.98×A11+4.2×A12+0.25×A13+8.56×A14-3.07×A15+34.44×A16-1.32×A17-2.09；
tanggute radix Et rhizoma Rhei:
Y2＝1.41×A1-0.46×A2-1.49×A3+1.3×A4+0.12×A5+5.85×A6+0.94×A7+8.42×A8+0.29×A9-0.39×A10+8.96×A11+1.77×A12-0.01×A13+3.29×A14-2.04×A15+16.94×A16-0.62×A17-8.25；
rhubarb palmate leaf:
Y3＝-5.5×A1+1.43×A2+4.63×A3-4.2×A4-0.25×A5-17.62×A6-3.3×A7-28.08×A8-4.99×A9+1.64×A10-25.94×A11-5.98×A12-0.24×A13-11.85×A14+5.11×A15-51.38×A16+1.93×A17+1.88；
(3) and comparing the discrimination values Y1, Y2 and Y3, wherein the maximum value is the primordium corresponding to the sample to be detected.
The invention also provides another identification method of the radix et rhizoma Rhei medicinal material primitive, which adopts a classification function model, only requires substituting independent variables into a classification function for calculation during application, and then compares the discrimination values Y1, Y2 and Y3. The identification method is simpler and quicker. And the identification result is more accurate. The identification method of rhubarb provided by the invention enriches identification means and is beneficial to establishing internal control quality standards for enterprises.
Preferably, the step of establishing the fingerprint of the sample to be detected includes: taking 0.5g of a rhubarb sample, sieving the rhubarb sample by a No. 2 sieve, placing the rhubarb sample in a conical flask with a plug, adding 25ml of water, heating and refluxing for 60 minutes, cooling the mixture, shaking the mixture evenly, filtering the mixture, taking a subsequent filtrate for later use, and detecting the filtrate by a high performance liquid chromatography to obtain the fingerprint spectrum of the sample to be detected.
Preferably, the analysis conditions of the liquid chromatography are as follows: using octadecylsilane chemically bonded silica as filler, acetonitrile as mobile phase A, and 0.05-0.5% phosphoric acid solution as mobile phase B, and performing gradient elution as follows:
Figure BDA0002834642910000061
Figure BDA0002834642910000071
the method collects the original data through the high performance liquid chromatography, and the data collected by the method more accurately lays a foundation for identifying the radix et rhizoma Rhei medicinal material primordium.
The chromatographic conditions of the liquid chromatography analysis adopted when the typical discriminant function and the classification function are established are as described above, so that the identification result obtained when the sample to be detected is analyzed by adopting the standard is more accurate.
Preferably, octadecylsilane bonded silica is used as a filler, and the column length is 150mm, the inner diameter is 2.1mm, and the particle diameter is 1.6 μm.
Preferably, the number of theoretical plates is not less than 3000 calculated by emodin peak.
Preferably, the chromatographic conditions are: the flow rate was 0.30ml per minute;
preferably, the column temperature is 25 ℃;
preferably, the detection wavelength is 260 nm.
The chromatographic conditions of the sample to be detected are more accurate when the standard with the flow rate of 0.30ml per minute, the column temperature of 25 ℃ and the detection wavelength of 260nm is adopted for analysis.
In addition, the method for obtaining the typical discriminant function and the classification function comprises the following steps:
(1) three batches of medicinal rhubarb, palmate rhubarb and Tanggute rhubarb are taken respectively to establish the fingerprint spectra of three rhubarb medicinal material primordia.
Specifically, the step of establishing the fingerprint of the rhubarb medicinal material comprises the following steps: preparing a test solution and a reference solution, precisely sucking 1 mu l of each of the test solution and the reference solution, injecting the solutions into a high performance liquid chromatograph, and recording a chromatogram.
Wherein the step of preparing the test solution comprises: taking 0.5g of rhubarb medicine powder, sieving the rhubarb medicine powder by a No. 2 sieve, placing the rhubarb medicine powder in a conical flask with a plug, adding 25ml of water, heating and refluxing for 60 minutes, cooling the mixture, shaking the mixture evenly, filtering the mixture, and taking the subsequent filtrate as a test solution.
The step of preparing a reference solution comprises: taking an emodin reference substance, precisely weighing, and adding methanol to obtain a solution with emodin content of 50 μ g/mL as a reference substance solution.
The high performance liquid chromatograph uses octadecylsilane chemically bonded silica as a filler, the column length is 150mm, the inner diameter is 2.1mm, the particle size is 1.6 mu m, acetonitrile is used as a mobile phase A, 0.1% phosphoric acid solution is used as a mobile phase B for gradient elution, the flow rate is 0.30ml per minute, the column temperature is 25 ℃, the detection wavelength is 260nm, and the number of theoretical plates is not less than 3000 according to the emodin peak.
The gradient elution was performed according to the following procedure: 0-1 min, wherein the content of mobile phase A is 2% → 11%, and the content of mobile phase B is 98% → 89%; 1-3 min, wherein the mobile phase A is 11% and the mobile phase B is 89%; 3-6 min, wherein the mobile phase A is 11% → 15%, and the mobile phase B is 89% → 85%; 6-8 min, the mobile phase A is 15%, and the mobile phase B is 85%.
Furthermore, the inventors also performed precision tests, stability tests, and repeatability tests. The precision test comprises the following steps: and taking the same sample, carrying out continuous sample introduction for 6 times, recording chromatograms, and calculating the peak area RSD of 17 groups of 6 chromatographic peaks, wherein the RSD is less than or equal to 1.4%. The stability test comprises the following steps: sampling the same sample at 0, 1, 2, 4, 8 and 24 hours for detection, recording chromatogram, and calculating peak area RSD of 17 groups of 6 chromatographic peaks, wherein RSD is less than or equal to 3.2%. The step of repeatability tests comprises: taking 6 parts of the same rhubarb medicinal material, adopting the same preparation method and chromatographic detection, and calculating the peak areas RSD of 17 groups of 6 chromatographic peaks, wherein the RSD is less than or equal to 4.1 percent.
(2) Collecting common chromatographic peaks of the fingerprint, and calculating the proportion of each peak in the common chromatographic peaks in the total peak area to obtain modeling data. The modeling data is shown in fig. 1.
(3) And carrying out principal component analysis on the modeling data, and then carrying out k-means classification to obtain a typical discriminant function model and a classification function model.
Specifically, the modeling data is used to obtain normalized conversion coefficient data of each variable, as shown in fig. 2. Extracting the normalized conversion coefficient data of each variable to obtain three principal components, wherein the three principal components are shown as a principal component coefficient table in fig. 3 and a principal component score table in fig. 4. Wherein the first principal component cumulative interpretation variable is 51.9%; the second principal component cumulative interpretation variable is 83.6%; the third principal component cumulative interpretation variable is 91.5%. It is stated that the results from the principal component analysis can explain most of the original variable information. Since the second principal component interpretation cumulative variable obtained by principal component analysis reached 83.6%, classification was performed with the first and second principal components (K-means classification), and the classification was defined as 3 classes, as shown in fig. 5. And then, classifying and distinguishing three main components of the main component analysis by using independent variables and classifying numbers obtained by k mean value classification to obtain a typical discriminant function and a classification function. The coefficients of the canonical discriminant function are shown in fig. 6, the scores of the canonical discriminant function class centroids are shown in fig. 7, and the coefficients of the class discriminant function are shown in fig. 8. And finally obtaining a typical discriminant function and a classification function through analysis.
The typical discriminant function and the classification function provided by the invention adopt a principal component analysis method, 3 principal components are extracted from the original data, and the chromatographic peaks of the principal components are not clearly influenced, so that the quality of medicinal materials can be prevented from being counterfeited. And moreover, a typical discriminant function model and a classification function model are obtained by adopting clustering judgment analysis, and the two function models are linear models, so that the calculation is simpler, and the application of a user is facilitated.
Compared with the prior art, the invention has the beneficial effects that:
(1) the high performance liquid chromatography is adopted to collect the original data, and the data acquisition is accurate.
(2) The invention provides a typical discriminant function model and a method for identifying the radix et rhizoma Rhei medicinal material primitive thereof through principal component analysis and clustering judgment, and the primitive is identified through observing the consistency of the coordinate position of a sample to be identified and a standard coordinate position. The identification method is simple and clear at a glance for the identification result.
(3) The invention provides a method for identifying a radix et rhizoma Rhei medicinal material primitive by using a classification function model, which is implemented by only substituting independent variables into a classification function for calculation and then comparing discrimination values Y1, Y2 and Y3. The identification method is simpler and quicker. And the identification result is more accurate.
(4) The invention provides two identification models and identification methods thereof, the model identification calculation is simple, the identification results of the two identification models are accurate and reliable, the identification means is enriched, and the establishment of the internal control quality standard of enterprises is facilitated.
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In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is modeling data for obtaining canonical discriminant functions and classification functions;
FIG. 2 is normalized conversion coefficient data for each variable obtained from modeling data;
FIG. 3 is a table of principal component coefficients obtained by extracting normalized conversion coefficient data for each variable;
FIG. 4 is a table of principal component scores obtained by extracting normalized conversion coefficient data for each variable;
FIG. 5 is a K-means classification number results table;
FIG. 6 is a table of typical discriminant function coefficients;
FIG. 7 is a typical discriminant function class centroid score table;
FIG. 8 is a table of classification discrimination function coefficients;
fig. 9 is a position diagram of 17 groups of characteristic peaks of a fingerprint of a rhubarb drug provided by an embodiment of the invention;
FIG. 10 is a three-group standard coordinate position scattergram of Rheum officinale Baill, Rheum tanguticum Maxim and Rheum palmatum obtained by the construction method described in example 1;
FIG. 11 is a standard coordinate position diagram of the three basic crude drugs of rhubarb horsetails for medicine, Tanggute rhubarb horsetails and Zuoyehuang as described in example 4.
Detailed Description
The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and the detailed description, but those skilled in the art will understand that the following described embodiments are some, not all, of the embodiments of the present invention, and are only used for illustrating the present invention, and should not be construed as limiting the scope of the present invention. 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. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1
The method for constructing the radix et rhizoma Rhei medicinal material primitive identification model comprises the following steps:
(1) establishing fingerprint spectrums of medicinal rhubarb, Tanggute rhubarb and palmate rhubarb: respectively taking 0.5g of medicinal rhubarb, Tanggute rhubarb and palmate rhubarb, sieving the medicinal rhubarb, the Tanggute rhubarb and the palmate rhubarb respectively, putting the medicinal rhubarb, the Tanggute rhubarb and the palmate rhubarb into a conical flask with a plug, adding 25ml of water, heating and refluxing the mixture for 60 minutes, cooling the mixture, shaking the mixture evenly, filtering the mixture, taking subsequent filtrate for later use, and then detecting the subsequent filtrate by using a high performance liquid chromatography to respectively obtain the fingerprint spectrums of the three groups of rhubarb.
The conditions of the high performance liquid chromatography are as follows: octadecylsilane chemically bonded silica is used as a filler, the column length is 150mm, the inner diameter is 2.1mm, the particle size is 1.6 mu m, acetonitrile is used as a mobile phase A, 0.1% phosphoric acid solution is used as a mobile phase B for gradient elution, the flow rate is 0.30ml per minute, the column temperature is 25 ℃, the detection wavelength is 260nm, and the number of theoretical plates is not less than 3000 according to the emodin peak.
The gradient elution procedure was as follows:
time (min) Mobile phase A (%) Mobile phase B (%)
0～1 2→11 98→89
1～3 11 89
3～6 11→15 89→85
6～8 15 85
。
(2) Collecting common chromatographic peaks of the fingerprint, wherein the common chromatographic peaks comprise 17 groups of characteristic peaks, and the positions of the 17 groups of characteristic peaks are shown in figure 9. Then, the proportion of 17 groups of characteristic peaks on the three groups of rhubarb fingerprint to the total peak area is independently calculated, and three groups of independent variables are sequentially obtained from left to right, which is shown in the following table 1.
Table 117 group characteristic peak to total peak area ratio statistical table
Figure BDA0002834642910000121
(3) And respectively substituting the three groups of independent variables into typical discriminant functions to obtain three groups of discriminant scores D1 and D2, which are shown in Table 2. And respectively taking the three groups of discrimination scores D1 as abscissa and D2 as ordinate to obtain three groups of standard coordinate position scatter diagrams respectively corresponding to the rhubarb horsetails, the rhubarb horsetails of Tanggute and the rhubarb palmate leaf.
TABLE 2 discrimination scores of Rheum officinale, Rheum tanguticum Maxim and Rheum palmatum
Discriminant score D1 D2
Rhubarb root and rhizome 1 26.9 9.6
Radix Et rhizoma Rhei Palmati 2 -39.6 -1.9
Tanggute radix Et rhizoma Rhei 3 6.5 -2.9
Example 2
The radix et rhizoma Rhei identification model of this embodiment is a scattergram of three sets of standard coordinate positions respectively corresponding to Rheum officinale Baill, Rheum tanguticum Maxim and Rheum palmatum obtained by the construction method of embodiment 1, as shown in FIG. 10. In fig. 10, (1) is the coordinate position point of rhubarb horsetails, 2) is the coordinate position point of radix Et rhizoma Rhei, and 3) is the coordinate position point of Rheum palmatum.
Example 3
The identification method of the rhubarb medicinal material primitive comprises the following steps:
firstly, the fingerprint of the rhubarb sample to be identified is established, and all the parameters are consistent with the parameters of establishing the fingerprints of the rhubarb horsetails, the rhubarb horsetails from Tang ancient times and the rhubarb horsetails in the example 1. The independent variables of the rhubarb samples to be identified were then obtained, see table 3 below. And substituting the standard discriminant function into a typical discriminant function to obtain discriminant scores D1 and D2 (see the following table 4) corresponding to the rheum officinale sample to be identified and a coordinate point thereof.
TABLE 3 statistical table of ratio of characteristic peak to total peak area of rhubarb sample to be identified
Figure BDA0002834642910000131
TABLE 4 discrimination score of Rheum officinale sample to be identified
Discriminant score D1 D2
Rhubarb sample to be identified 14.7 12.4
And finally, observing the consistency of the coordinate point positions of the rhubarb sample to be identified and three groups of standard coordinate position scatter diagrams respectively corresponding to the rhubarb for medical use, the rhubarb for Tang Gu and the rhubarb for palm leaf obtained in the embodiment 2, thereby identifying the primordium of the sample to be detected.
As can be seen from fig. 10, the coordinate position of the rheum officinale sample to be identified (4) is consistent with the coordinate position of the rheum officinale (1), and therefore, the origin of the rheum officinale sample to be identified is rheum officinale.
Example 4
The method for identifying three rhubarb medicinal material primordial factors, which is described in the embodiment, comprises the following steps:
(1) establishing a fingerprint of a rhubarb sample to be identified: taking 0.5g of a rhubarb sample to be identified, sieving the rhubarb sample by a No. 2 sieve, placing the rhubarb sample in a conical flask with a plug, adding 25ml of water, heating and refluxing for 60 minutes, cooling, shaking up, filtering, taking a subsequent filtrate for standby, and detecting by using a high performance liquid chromatography to obtain the fingerprint spectrum of the sample to be detected.
The conditions of the high performance liquid chromatography are as follows: octadecylsilane chemically bonded silica is used as a filler, the column length is 150mm, the inner diameter is 2.1mm, the particle size is 1.6 mu m, acetonitrile is used as a mobile phase A, 0.1% phosphoric acid solution is used as a mobile phase B for gradient elution, the flow rate is 0.30ml per minute, the column temperature is 25 ℃, the detection wavelength is 260nm, and the number of theoretical plates is not less than 3000 according to the emodin peak.
The gradient elution procedure was as follows:
time (min) Mobile phase A (%) Mobile phase B (%)
0～1 2→11 98→89
1～3 11 89
3～6 11→15 89→85
6～8 15 85
。
(2) The ratio of 17 groups of characteristic peaks on the fingerprint of the rhubarb sample to be identified to the total peak area is calculated, and independent variables are obtained sequentially from left to right, as shown in the following table 5.
TABLE 5 statistical table of ratio of characteristic peak to total peak area of rhubarb sample to be identified
Figure BDA0002834642910000141
(3) Making a standard coordinate position diagram of three basic crude drugs including medicinal rhubarb, Tanggute rhubarb and palmate rhubarb, wherein the specific coordinates of the standard coordinate position diagram are as follows:
medicinal rhubarb: abscissa 24.558, ordinate 4.812;
tanggute radix Et rhizoma Rhei: abscissa 11.264, ordinate-6.171;
rhubarb palmate leaf: abscissa-35.822, and ordinate 1.359.
The standard coordinate position diagram is shown in fig. 11, wherein (a) in fig. 11 is a standard coordinate position point of rhubarb for medicinal use, (b) is a standard coordinate position point of rheum tanguticum, and (c) is a standard coordinate position point of rheum palmatum.
(4) Substituting the independent variables into a classical discriminant function to obtain discriminant scores D1 and D2 of the rhubarb sample to be identified, and drawing a coordinate point of the rhubarb sample to be identified by taking D1 as an abscissa and D2 as an ordinate, namely (5) in FIG. 11.
(5) As is apparent from the observation of fig. 11, the coordinate point (5) of the rhubarb sample to be identified coincides with the (a) -standard coordinate position point of rhubarb horsetails, and therefore, the origin of the rhubarb sample to be identified is rhubarb horsetails.
Example 5
The method for identifying three rhubarb medicinal material primordial factors, which is described in the embodiment, comprises the following steps:
(1) establishing a fingerprint of a rhubarb sample to be identified: taking 4 groups of samples, and establishing a fingerprint spectrum according to the following parameters:
taking 0.5g of a rhubarb sample to be identified, sieving the rhubarb sample by a No. 2 sieve, placing the rhubarb sample in a conical flask with a plug, adding 25ml of water, heating and refluxing for 60 minutes, cooling, shaking up, filtering, taking a subsequent filtrate for standby, and detecting by using a high performance liquid chromatography to obtain the fingerprint spectrum of the sample to be detected.
The conditions of the high performance liquid chromatography are as follows: octadecylsilane chemically bonded silica is used as a filler, the column length is 150mm, the inner diameter is 2.1mm, the particle size is 1.6 mu m, acetonitrile is used as a mobile phase A, 0.1% phosphoric acid solution is used as a mobile phase B for gradient elution, the flow rate is 0.30ml per minute, the column temperature is 25 ℃, the detection wavelength is 260nm, and the number of theoretical plates is not less than 3000 according to the emodin peak.
The gradient elution procedure was as follows:
Figure BDA0002834642910000151
Figure BDA0002834642910000161
(2) the ratio of 17 groups of characteristic peaks on the fingerprint of 4 groups of rheum officinale samples to be identified to the total peak area is calculated, and independent variables are obtained sequentially from left to right as shown in the following table 6.
TABLE 6 statistical table of ratio of characteristic peak to total peak area of rhubarb sample to be identified
Figure BDA0002834642910000162
(3) Substituting the independent variables into classification functions corresponding to the medicinal rhubarb, the Tanggute rhubarb and the palmate rhubarb to calculate to obtain three groups of discrimination values Y1, Y2 and Y3, as shown in the following table 7.
TABLE 7 discrimination values of Rheum officinale samples to be identified
The discrimination value Y1-medicinal rhubarb Y2-Tanggute radix Et rhizoma Rhei Y3-Zuoyehuang
Rhubarb
6 to be identified 0.004 -0.005 -1.041
Rhubarb 7 to be identified 0.505 0.261 -1.808
Rhubarb 8 to be identified 0.065 0.151 -1.257
Rhubarb 9 to be identified -1.296 -0.518 0.772
(4) And comparing the discrimination values Y1, Y2 and Y3, wherein the maximum value is the primordium corresponding to the sample to be detected.
As shown in Table 7, for the sample to be identified of rhubarb No. 6, the value of Y1 is the largest, and the corresponding primitive is rhubarb horsetails, therefore, the primitive of the sample to be identified No. 6 is rhubarb horsetails.
For the rhubarb sample No. 7 to be identified, the value of Y1 is the largest, and the corresponding primitive is rhubarb horsetails, therefore, the primitive of the sample No. 7 to be identified is rhubarb horsetails.
For the rheum officinale sample to be identified No. 8, the Y2 value is the largest, and the corresponding primitive is Tanggute radix Et rhizoma Rhei, so the primitive of the rheum officinale sample to be identified No. 8 is Tanggute radix Et rhizoma Rhei.
For the rheum officinale sample to be identified No. 9, the value of Y3 is the largest, and the corresponding primitive is rheum palmatum, so the primitive of the rheum palmatum sample to be identified No. 9 is rheum palmatum.
While particular embodiments of the present invention have been illustrated and described, it will be appreciated that the above embodiments are merely illustrative of the technical solution of the present invention and are not restrictive; those of ordinary skill in the art will understand that: modifications may be made to the above-described embodiments, or equivalents may be substituted for some or all of the features thereof without departing from the spirit and scope of the present invention; the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention; it is therefore intended to cover in the appended claims all such alternatives and modifications that are within the scope of the invention.

Claims (10)

1. A method for constructing a radix et rhizoma Rhei medicinal material primitive identification model is characterized by comprising the following steps:
(1) respectively establishing three groups of standard fingerprints of medicinal rheum officinale, rheum tanguticum and rheum palmatum by adopting a liquid chromatography, independently calculating the proportion of 17 groups of characteristic peaks on the three groups of standard fingerprints to the total peak area, and sequentially obtaining three groups of independent variables X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12, X13, X14, X15, X16 and X17 from left to right;
(2) respectively substituting the three independent variables into a typical discriminant function for calculation to obtain three discriminant scores D1 and D2;
the typical discriminant function is:
function 1:
D1＝155.8×X1-39.9×X2-129.1×X3+117.5×X4+6.8×X5+489.9×X6+92.7×X7+786.4×X8+147.8×X9-46.6×X10+718.9×X11+167.5×X12+7.2×X13+333.8×X14-139.9×X15+1429.1×X16-53.8×X17-70.7；
function 2:
D2＝56.5×X1+1.7×X2+5.8×X3+4.4×X4-6.9×X5-54.5×X6+16×X7+70.4×X8+222×X9-22.6×X10-139×X11+18.4×X12+14.8×X13+76.4×X14+75.7×X15-137.1×X16+1.5×X17-8.8；
(3) taking D1 as abscissa and D2 as ordinate to obtain three groups of standard coordinate position scatter diagrams respectively corresponding to the medicinal rhubarb, the Tanggute rhubarb and the palmate rhubarb, namely a rhubarb medicinal material primitive identification model.
2. The identification model of radix et rhizoma Rhei medicinal material primordium is obtained by the construction method of claim 1.
3. A method for identifying a radix et rhizoma Rhei medicinal material primordium is characterized in that a sample to be identified is prepared into a sample solution to be tested, liquid chromatography analysis is carried out to obtain a sample map to be tested, the proportion of 17 groups of characteristic peaks on the sample map to the total peak area is calculated, and independent variables X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12, X13, X14, X15, X16 and X17 are sequentially obtained from left to right; substituting into typical discriminant function to obtain discriminant scores D1 and D2 corresponding to the sample to be tested;
obtaining a coordinate position corresponding to the to-be-tested sample by taking D1 as an abscissa and D2 as an ordinate;
observing the consistency of the coordinate position of the sample to be tested and the standard coordinate position, thereby identifying the primordium of the sample to be identified;
the standard coordinate positions are obtained by a classical discriminant function of the rhubarb, the rheum tanguticum Maxim and the rheum palmatum L.
4. An identification method according to claim 3, characterized in that the chromatographic conditions of the liquid chromatographic analysis are: using octadecylsilane chemically bonded silica as filler, acetonitrile as mobile phase A, and 0.05-0.5% phosphoric acid solution as mobile phase B, and performing gradient elution as follows:
time (min) Mobile phase A (%) Mobile phase B (%) 0～1 2→11 98→89 1～3 11 89 3～6 11→15 89→85 6～8 15 85
。
5. An identification method according to claim 4, characterized in that said chromatographic conditions are: the flow rate was 0.30ml per minute;
preferably, the column temperature is 25 ℃.
6. An identification method according to claim 4, characterized in that said chromatographic conditions are: the detection wavelength was 260 nm.
7. The identification method of the radix et rhizoma Rhei medicinal material primordium is characterized by comprising the following steps:
(1) establishing a fingerprint of a sample to be detected by adopting a liquid chromatography, calculating the proportion of 17 groups of characteristic peaks on the fingerprint to the total peak area, and sequentially obtaining independent variables A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, A12, A13, A14, A15, A16 and A17 from left to right;
(2) substituting the independent variables into classification functions corresponding to the medicinal rhubarb, the Tanggute rhubarb and the palmate rhubarb to calculate to obtain three groups of discrimination values Y1, Y2 and Y3;
the classification function is:
medicinal rhubarb:
Y1＝4.1×A1-0.97×A2-3.14×A3+2.91×A4+0.13×A5+11.77×A6+2.35×A7+19.65×A8+4.7×A9-1.25×A10+16.98×A11+4.2×A12+0.25×A13+8.56×A14-3.07×A15+34.44×A16-1.32×A17-2.09；
tanggute radix Et rhizoma Rhei:
Y2＝1.41×A1-0.46×A2-1.49×A3+1.3×A4+0.12×A5+5.85×A6+0.94×A7+8.42×A8+0.29×A9-0.39×A10+8.96×A11+1.77×A12-0.01×A13+3.29×A14-2.04×A15+16.94×A16-0.62×A17-8.25；
rhubarb palmate leaf:
Y3＝-5.5×A1+1.43×A2+4.63×A3-4.2×A4-0.25×A5-17.62×A6-3.3×A7-28.08×A8-4.99×A9+1.64×A10-25.94×A11-5.98×A12-0.24×A13-11.85×A14+5.11×A15-51.38×A16+1.93×A17+1.88；
(3) and comparing the discrimination values Y1, Y2 and Y3, wherein the maximum value is the primordium corresponding to the sample to be detected.
8. The identification method according to claim 7, wherein the analysis conditions of the liquid chromatography are: using octadecylsilane chemically bonded silica as filler, acetonitrile as mobile phase A, and 0.05-0.5% phosphoric acid solution as mobile phase B, and performing gradient elution as follows:
Figure FDA0002834642900000031
Figure FDA0002834642900000041
9. an identification method according to claim 8, characterized in that the chromatographic conditions are: the flow rate was 0.30ml per minute;
preferably, the column temperature is 25 ℃.
10. An identification method according to claim 8, characterized in that the chromatographic conditions are: the detection wavelength was 260 nm.
CN202011474422.4A 2020-12-14 2020-12-14 Identification model of rhubarb medicinal material primordium, construction method and identification method thereof Pending CN112379034A (en)

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