Method for classifying characters of traditional Chinese medicine by orthogonal test and exploring relevance between characters and efficacy of traditional Chinese medicine

Abstract

The invention provides a method for classifying characters of traditional Chinese medicines and exploring relevance between the characters and medicine effects by using an orthogonal test. The method at least comprises the following steps: S1, multi-dimensional character measurement; s2, frequency analysis and correlation analysis; s3, carrying out orthogonal test classification; and S4, determining the content of the effective component spinosin. Through systematic character measurement, correlation analysis and orthogonal tests, the relevance between the traditional Chinese medicine appearance characters and the content of the effective component spinosin is established, the problems of incomplete analysis, low efficiency and insufficient accuracy in the prior art are solved, and a scientific basis is provided for traditional Chinese medicine quality evaluation and high-quality germplasm screening.

Description

Technical Field

[0001] This invention relates to the field of exploration technology of spinosin content, and in particular to the classification of Chinese herbal medicine properties using orthogonal experiments and the exploration of their correlation with efficacy. Background Technology

[0002] Existing methods for exploring the content of effective components in traditional Chinese medicine (TCM) often lack a systematic analysis of the correlation between the appearance characteristics and intrinsic quality (such as the content of effective components). Traditional methods may rely solely on single-dimensional trait measurements or simple content determinations, making it difficult to comprehensively reveal the influence of multiple factors on the content of effective components. Furthermore, experimental designs may suffer from inaccurate variable control, insufficient data, or unscientific analytical methods, resulting in the inability to efficiently screen germplasm resources with high content of target components. Summary of the Invention

[0003] In view of this, the present invention provides a method for classifying the characteristics of traditional Chinese medicine and exploring their correlation with efficacy using orthogonal experiments. Through systematic trait measurement, correlation analysis, and orthogonal experimental design, the correlation between appearance characteristics and the content of the active ingredient Spinosin is established, solving the problems of incomplete analysis, low efficiency, and insufficient accuracy in the prior art, and providing a scientific basis for the quality evaluation of traditional Chinese medicine and the screening of high-quality germplasm.

[0004] The technical solution of this invention is implemented as follows: First, the characteristics of the source of traditional Chinese medicine are systematically measured, and frequency and correlation analyses are performed. Based on this experience, orthogonal experiments are used to classify the characteristics of traditional Chinese medicine and explore their correlation with efficacy. That is, according to the differences in the characteristics of traditional Chinese medicine, the characteristics are divided into several different factors. Then, based on the factors, each factor is divided into several levels according to the magnitude of the factor index. Finally, according to the factor level table of orthogonal experiments in mathematical statistics, statistically significant groups are screened out. The content of the corresponding characteristics of traditional Chinese medicine and the active ingredient spinosin is also explored. The method includes at least the following steps:

[0005] S1 Multidimensional Trait Measurement

[0006] Data on the source of the Chinese medicinal herb Ziziphus jujuba var. spinosa were measured, the Ziziphus jujuba var. spinosa were numbered, and their transverse and longitudinal diameters were accurately measured and photographed using vernier calipers.

[0007] Peel the jujube pulp and measure the jujube pit. Use pliers to peel the pulp, wash and dry the pit, and measure the transverse diameter, longitudinal diameter, xylem thickness, symmetry, and number of kernels of the pit;

[0008] Data on jujube seed kernels were measured using a stereomicroscope.

[0009] The fruit, fruit core, and kernel transverse diameter and longitudinal diameter are measured by vernier caliper and stereomicroscope, and the kernel is placed on one side and recorded. The basic trait data of the fruit core and kernel are obtained, and the measured kernel is packed.

[0010] S2 Frequency analysis and correlation analysis

[0011] The Pearson correlation coefficients between various factors are analyzed to determine the correlation between the number of kernels, concave-convex degree, size, fruit core, and traditional Chinese medicine shape.

[0012] According to the differences in kernel spinon content and climate factors in different production areas, and the influence of trait factors on spinon content, the concave-convex degree, longitudinal diameter length, and longitudinal diameter transverse diameter ratio are selected as factors, and the kernel germplasm with high spinon content is screened.

[0013] S3 Classification of orthogonal test

[0014] The orthogonal test method is used, the concave-convex degree, longitudinal diameter transverse diameter ratio, and longitudinal diameter length are taken as influencing factors, two levels are selected, and the kernel produced in Shandong with the highest content is divided into four groups according to the orthogonal test method, which lays a foundation for subsequent exploration of the relationship between traits and quality.

[0015] S4 Determination of effective component content

[0016] The kernel in S3 is ground, weighed, ultrasonically extracted with methanol, centrifuged, and the standard sample is prepared. The spinon content is determined by high performance liquid chromatograph to establish the correlation between trait data and spinon content.

[0017] Further, the experimental instruments used for appearance trait identification include camera, vernier caliper, stereomicroscope, beaker, and 6-inch tweezers.

[0018] Further, the traditional Chinese medicine sample transverse diameter and longitudinal diameter are measured to two decimal places, and the unit is mm.

[0019] Further, in the S3 orthogonal test method, the two levels of concave-convex degree are full and not full, the two levels of longitudinal diameter transverse diameter ratio are 1.0 and 1.2, and the two levels of longitudinal diameter length are 7mm and 5.5mm. The experimental variable setting is clear.

[0020] Further, the experimental instruments used for content determination include one-millionth analytical balance, ultrasonic instrument, centrifuge, high performance liquid chromatograph, organic phase chromatography filtration device, and water phase chromatography filtration device.

[0021] Further, the reagents used for content determination include methanol, acetonitrile, and water.

[0022] Further, in the S4 content determination step, the test sample powder is weighed accurately to 0.001 g, and three portions of each test sample are weighed for parallelism; 2 mL of methanol is taken by a 5 mL pipette into an EP tube containing the test sample, and ultrasonic extraction is performed for 20 min, twice; the centrifugal speed is 9800 r / min, and the time is 10 min.

[0023] Further, an advantage and disadvantage identification model is established based on the obtained fruit kernel trait data and chemical composition characteristics after testing.

[0024] The present application has the following advantages due to the above technical solutions:

[0025] Systematicness of multi-dimensional trait measurement: the source of traditional Chinese medicine Zizyphi Semen, Zizyphi Endocarp and Zizyphi Sement, and the transverse diameter, longitudinal diameter, concave-convex degree, and xylem thickness of the above are accurately measured by using a vernier caliper, a body mirror and the like, a large amount of data is obtained, the sufficiency and reliability of the data are ensured, and the appearance features affecting the quality are comprehensively covered.

[0026] Precision of correlation analysis and screening: the correlation between the number of fruit kernels, concave-convex degree, size and the shape of fruit kernels and fruit is determined through Pearson correlation coefficient analysis and significance test.

[0027] High efficiency of orthogonal test design: the orthogonal test method is used, reasonable levels of key factors such as concave-convex degree, aspect ratio and longitudinal length are combined, multi-factor experiments are scientifically grouped into four groups, the influence of each factor on the content of effective components is effectively analyzed while the number of experiments is reduced, and the experimental efficiency and variable control accuracy are improved.

[0028] The content of effective components is determined by using high performance liquid chromatography, the direct correlation between trait data and quality indicators is established, the germplasm with high content of target components can be accurately screened, and quantitative basis is provided for traditional Chinese medicine quality evaluation.

[0029] Universality and reproducibility of the method: the experimental instruments (vernier caliper, body mirror, high performance liquid chromatograph and the like) and reagents (methanol, acetonitrile and the like) are conventional analysis tools, the steps are clear and standard, the method is suitable for the exploration of the content of effective components of various traditional Chinese medicines or fruit kernels, and has wide technical popularization value.

[0030] Construction of advantage and disadvantage identification model: based on the trait data, trait classification and chemical composition characteristics obtained by testing, an objective advantage and disadvantage identification model can be established, new technical means are provided for traditional Chinese medicine variety identification and quality control, and the subjectivity of traditional experience identification is made up.

[0031] The above summary is intended to illustrate only and is not intended to serve as an limitation in any manner. In addition to the illustrative aspects, embodiments and features described above, further aspects, embodiments and features will become apparent to those skilled in the art from the following detailed description, which, taken in conjunction with the drawing, discloses a preferred embodiment. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 A flow chart of the steps of the present application;

[0033] Figure 2 A longitudinal diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application;

[0034] Figure 3 A transverse diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application;

[0035] Figure 4 A longitudinal diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application;

[0036] Figure 5 A transverse diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application;

[0037] Figure 6 A longitudinal diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application;

[0038] Figure 7 A transverse diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application;

[0039] Figure 8 A longitudinal diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application;

[0040] Figure 9 A longitudinal diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application;

[0041] Figure 10 A longitudinal diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application; Figure 1

[0042] Figure 11 A longitudinal diameter distribution graph of the Chinese wingnut for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application; Figure 2

[0043] Figure 12 A standard curve graph for exploring the content of spinozine of the Chinese wingnut in an embodiment of the present application. DETAILED DESCRIPTION

[0044] ​​It should be apparent that the following description illustrates by way of example only a number of embodiments of the present disclosure. Those skilled in the art will readily understand other advantages and / or uses of the present disclosure from the description of the example embodiments without departing from the spirit of the disclosure. It is to be clearly understood that the embodiments described are merely a few of the embodiments of the present disclosure, and are not all-inclusive of all embodiments. The present disclosure can also be implemented or applied in other different specific embodiments, and the details in the specification can be modified or changed based on different views and applications without departing from the spirit of the present disclosure. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict. Based on the embodiments in the present disclosure, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present disclosure.

[0045] It should be noted that the various aspects of the embodiments described below are within the scope of the appended claims. It should be apparent that the aspects described herein can be embodied in a wide variety of forms and that any specific structure and / or function described herein is merely illustrative. Based on the teachings provided herein one skilled in the art will appreciate that one aspect described herein can be implemented independently of any other aspects and that the various aspects described herein can be combined in various ways. For example, an apparatus can be implemented or a method can be practiced using any number of the aspects set forth herein. In addition, such an apparatus can be implemented or such a method can be practiced using other structure and / or functionality in addition to or other than one or more of the aspects set forth herein.

[0046] It should also be noted that the drawings included in the following embodiments are only to illustrate the basic concept of the present disclosure in a schematic manner, and only the components related to the present disclosure are shown in the drawings, not drawn according to the number, shape and size of the components when actually implemented, the actual implementation of each component can be a random change in shape, number and proportion, and the layout of the components can be more complex.

[0047] The present application will be described in detail below in conjunction with the drawings.

[0048] The present application provides a method for classifying traditional Chinese medicine properties by orthogonal test and exploring the correlation between the properties and the efficacy, i.e. according to the differences in the properties of traditional Chinese medicine, the properties of traditional Chinese medicine are divided into several different factors, then on the basis of the factors, according to the size of the factor index, each factor is divided into several levels, and finally according to the factor level table of orthogonal test in mathematical statistics, the group with statistical significance is screened out. And explore its (the corresponding properties of traditional Chinese medicine) and the content of the effective component spinozine, the method at least includes the following steps:

[0049] S1 Multi-dimensional property measurement

[0050] The data of the source of the Chinese medicine Suanzao is measured, the Suanzao is numbered, the horizontal diameter and the vertical diameter are accurately measured by using a vernier caliper, and a photograph is taken for recording;

[0051] The Suanzao pulp is peeled, and the data of the Suanzao kernel is measured. The pulp is peeled with forceps, the kernel is washed and dried, and the horizontal diameter, the vertical diameter, the thickness of the wood part, the symmetry condition and the number of kernels are measured;

[0052] The data of the Suanzao kernel is measured by using a body mirror;

[0053] The fruit, the kernel and the horizontal diameter and the vertical diameter of the kernel are measured by using a vernier caliper, and the one side placement condition is observed and recorded. The basic trait data of the kernel and the kernel are obtained by the system measurement, and the measured kernel is packed.

[0054] S2 Frequency analysis and correlation analysis

[0055] The Pearson correlation coefficients of the fruit, the kernel and the kernel are analyzed to determine the correlation of the number of kernels, the concave-convex degree, the size, the kernel and the appearance of traditional Chinese medicine;

[0056] The influence of climate and production place factors on the content of the effective component spinon of Suanzao kernel is explored, on the basis of which, the influence of trait factors on the content of spinon is combined, the concave-convex degree, the length of the vertical diameter, the ratio of the vertical diameter to the horizontal diameter are taken as indexes, and the kernel germplasm with high content of spinon is screened out.

[0057] S3 Classification of orthogonal test

[0058] The orthogonal test method is adopted, the concave-convex degree, the ratio of the vertical diameter to the horizontal diameter and the length of the vertical diameter are taken as influencing factors, two levels are taken for each, the influence of climate and production place factors on the content of the effective component spinon of Suanzao kernel is explored, it is concluded that the quality of Suanzao kernel in Shandong is optimal, on the basis of which, the Suanzao kernel in Shandong is divided into four groups according to the orthogonal test method, which lays a foundation for subsequent exploration of the relationship between traits and quality.

[0059] S4 Determination of the content of the effective component spinon

[0060] After the kernel in S3 is grouped, ground and weighed, the content of spinon is determined by using methanol ultrasonic extraction, centrifugation, preparation of a standard sample and high performance liquid chromatography, so as to establish the correlation between trait data and the content of spinon.

[0061] The optimal and poor identification model is established by the obtained kernel trait data and the characteristics of chemical composition after the test is completed.

[0062] Further, the experimental instruments used for appearance trait identification include a camera, a vernier caliper, a body mirror, a beaker and 6-inch forceps.

[0063] Furthermore, the experimental instruments used for content determination include a 0.01% analytical balance, an ultrasonic instrument, a centrifuge, a high-performance liquid chromatograph, an organic phase chromatography filtration device, and an aqueous phase chromatography filtration device.

[0064] Furthermore, the reagents used for content determination include methanol, acetonitrile, and water.

[0065] This invention also provides an example of exploring the spinoxin content in jujube using the method of this invention:

[0066] Extensive basic data were measured on jujube, jujube kernel, and jujube seed, and frequency analysis was performed using SPSS software. Based on this, orthogonal experiments were conducted for classification.

[0067] Experimental instruments: camera, vernier calipers, stereoscope, beaker, 6-inch needle-nose pliers.

[0068] Reagent: Purified water.

[0069] Experimental samples: procured from Shandong province.

[0070] Measurement of jujubes: The jujube samples were numbered and marked. The jujube samples were taken one by one, and the transverse and longitudinal diameters of the jujubes were measured with vernier calipers. The photos were taken and the data were recorded in mm, accurate to two decimal places. A total of 142 sets of data were obtained.

[0071] Measurement of jujube pits: Use needle-nose pliers to peel off the pulp, wash the pulp off the surface of the pit with water, wipe it dry with a damp cloth, measure the transverse and longitudinal diameters of the jujube pit, observe the thickness of the xylem in the pit, and whether the pit is symmetrical, take photos, and record the data.

[0072] Use needle-nose pliers to break open the jujube pits, record the number of jujube kernels, and put the kernels into a plastic bag with a designated number. A total of 113 sets of data on the transverse diameter and longitudinal diameter of the jujube pits were obtained.

[0073] Measurement of jujube kernels: Take out the numbered jujube kernels from the bag in sequence, place them under a stereomicroscope, and use a plotting tool with a scale of 100μm and a vertical line to measure the transverse and longitudinal diameters of the jujube kernels. Observe whether one side of the jujube kernel can be laid flat on a horizontal surface and record it as "can" (A) or "cannot" (B). Record the data in the experimental record book.

[0074] The 142 sets of recorded data were entered into SPSS software for frequency analysis, and histograms were obtained.

[0075] Depend on Figure 1 It can be seen that the longitudinal axis sequence of jujube is continuous and exhibits obvious normal distribution characteristics.

[0076] Depend on Figure 2It can be seen that the horizontal diameter series of the Chinese date has continuity, but the normal distribution is not obvious. The analysis is due to the different degree of dehydration of the pulp, so the value fluctuates greatly.

[0077] From Figure 3 It can be seen that the longitudinal diameter of the Chinese date kernel has the characteristics of normal distribution, but a data fault appears at 9.5 mm.

[0078] From Figure 4 It can be seen that the frequency distribution of the horizontal diameter of the Chinese date kernel has two normal distribution peaks, and a fault also appears at 6.50 mm, and a second peak appears after the fault. Therefore, it can be preliminarily concluded that there are at least two categories of Chinese date kernels in this batch of samples.

[0079] From Figure 5 It can be seen that the longitudinal diameter of the Chinese date kernel has the characteristics of normal distribution.

[0080] From Figure 6 It can be seen that the horizontal diameter of the Chinese date kernel has a fault at 4.50-5.00 mm, and then a normal distribution peak appears, and there are two peaks in the normal distribution graph of the horizontal diameter of the Chinese date kernel and the Chinese date kernel, that is, the Chinese date can be roughly divided into two groups, and through records and observations, it can be seen that the thickness of the xylem, the symmetry of the date kernel, and the number of date kernels are different between the two groups.

[0081] From Figure 7 , Figure 8 It can be seen that the ratio of the longitudinal diameter to the horizontal diameter of the Chinese date and the ratio of the longitudinal diameter to the horizontal diameter of the Chinese date kernel both have two normal distribution trends.

[0082] From Figure 9 , 10 It can be seen that the ratio of the longitudinal diameter to the horizontal diameter of the two batches of Chinese date kernels is different.

[0083] Frequency analysis - all characteristic data have the characteristics of normal distribution.

[0084] Standard analysis

[0085] The measurement parameter statistical table (Table 1) is as follows

[0086]

[0087] a. There are multiple modes. The smallest value is shown.

[0088] Through the above table (measurement parameter statistical table), a large amount of data is formulated as follows:

[0089] 1. Symmetry of date kernel: observe along the middle line of the date kernel, if both sides are symmetrical, it is considered that the date kernel is symmetrical, and is recorded as "1.00";

[0090] 2. If one side is thicker and the other side is thinner, it is considered as asymmetric, and is recorded as "2.00";

[0091] 3. Wood thickness: using a pair of tweezers to squeeze the jujube kernel, if it is not easy to break and the thickness is observed to be thicker, it is considered as thicker, and is recorded as "1.00"; if it is easy to break and the thickness is observed to be thinner, it is considered as thinner, and is recorded as "2.00";

[0092] 4. Concave-convex degree: place the jujube kernel on a horizontal surface with the side with obvious fine lines facing down, contact the horizontal surface, and give a rotating force. Among them, the one with larger rotating speed is considered as full, and is recorded as "2.00"; the one with rotating circumference <2r is considered as one side flat, and is recorded as "1.00".

[0093] Counting rule table (Table 2)

[0094]

[0095] In summary, the concave-convex degree, the ratio of longitudinal diameter to transverse diameter, and the longitudinal diameter length indeed have an impact on the shape of jujube kernel, and complete fractal screening can be carried out accordingly.

[0096] Correlation analysis (correlation between concave-convex degree, jujube kernel symmetry, wood thickness, and jujube kernel transverse diameter)

[0097] Correlation coefficient table (Table 3) is as follows

[0098]

[0099]

[0100] *. At the 0.05 level (two-tailed), the correlation is significant.

[0101] **. At the 0.01 level (two-tailed), the correlation is significant.

[0102] From the correlation coefficient, the Pearson correlation coefficient r between the number of jujube kernels and the symmetry of jujube kernels is 0.949, P=0.000<0.001, which means that the number of jujube kernels and the symmetry of jujube kernels are positively linearly correlated, and the linearity is significant.

[0103] The Pearson correlation coefficient r between the number of jujube kernels and the concave-convex degree is 0.933, P=0.000<0.001, which means that the number of jujube kernels and the concave-convex degree are positively linearly correlated, and the linearity is significant.

[0104] The Pearson correlation coefficient r between the symmetry of jujube kernel and the concave-convex degree is 0.912, P=0.000<0.001, which means that the number of jujube kernels and the concave-convex degree are positively linearly correlated, and the linearity is significant.

[0105] Pearson correlation coefficient r = 0.681, P = 0.000 < 0.001 between the jujube kernel number and the concave-convex degree, that is, the jujube kernel number and the concave-convex degree are positively linearly correlated, and the correlation is relatively large.

[0106] In summary, when the jujube kernel number is 1, the jujube kernel is relatively full, and the jujube kernel is asymmetric; when the jujube kernel number is 2, the jujube kernel is relatively flat, and the jujube kernel is symmetric; under the same ratio of longitudinal diameter to transverse diameter, the longer the transverse diameter of the jujube kernel, the thicker the xylem.

[0107] The size correlation coefficient table between jujube, jujube kernel and jujube kernel is as follows (Table 4)

[0108]

[0109] **. At the 0.01 level (two-tailed), the correlation is significant.

[0110] Pearson correlation coefficient r = 0.867, P = 0.000 < 0.001, the transverse diameter of jujube and the transverse diameter of jujube kernel are positively linearly correlated, but since r = 0.867 < 0.905, the correlation is not stable enough, and the analysis reason is that the drying degree of jujube pulp is not the same, resulting in linear fluctuation.

[0111] Pearson correlation coefficient r = 0.875, P = 0.000 < 0.001, the transverse diameter of jujube and the transverse diameter of jujube kernel are positively linearly correlated, but since r = 0.875 < 0.905, the correlation is not stable enough, and the analysis reason is that the drying degree of jujube pulp is not the same, resulting in linear fluctuation.

[0112] Pearson correlation coefficient r = 0.918, P = 0.000 < 0.001, the transverse diameter of jujube kernel and the transverse diameter of jujube kernel are positively linearly correlated, and after removing the pulp, there is no influence of the corresponding pulp drying degree, and the linearity is significant.

[0113] Linear regression:

[0114] The variance analysis table of regression significance test of jujube kernel number and xylem thickness (Table 5) is as follows

[0115]

[0116] a. Dependent variable: jujube kernel number.

[0117] b. Predictive variable: (constant), xylem thickness.

[0118] F = 24.920, P = 0.000 < 0.001, the regression equation has high statistical significance.

[0119] The coefficient table (Table 6) is as follows

[0120]

[0121]

[0122] According to the coefficient table, the regression equation is y = 0.805 + 0.456.

[0123] According to Table 1-Table 6, the relationship between the jujube and the jujube core in the shape is: the jujube with longer transverse diameter and longitudinal diameter, and the longitudinal to transverse ratio close to 1, is round and full, the peeled jujube core also has longer transverse diameter and longitudinal diameter, and the longitudinal to transverse ratio close to 1, the xylem thickness is thicker, and the xylem is asymmetric on both sides, and the jujube kernel number is 1 (common) or 3 (rare).

[0124] The jujube with shorter transverse diameter and longer longitudinal diameter, and the longitudinal to transverse ratio greater than 1, is thin. The peeled jujube core also has shorter transverse diameter and longer longitudinal diameter, and the longitudinal to transverse ratio greater than 1. The xylem is thin, and the xylem is symmetric on both sides, and the jujube kernel number is 2.

[0125] The relationship between the jujube core and the jujube kernel is: the xylem thickness is thicker, and the xylem is asymmetric on both sides, and the jujube kernel number is 1 or 3, and the jujube kernel on one side is protruding, and the longitudinal to transverse ratio is close to 1.

[0126] The xylem is thin, and the xylem is symmetric on both sides, and the jujube kernel number is 2, and the jujube kernel on one side is flat, and the longitudinal to transverse ratio is greater than 1.

[0127] Classification of jujube kernels: using orthogonal test method to classify jujube kernels produced in Shandong area.

[0128] From the previous sampling of jujube and frequency analysis, correlation analysis, it is known that the factors affecting the shape of jujube kernel mainly include three factors, which are concave-convex degree, longitudinal to transverse ratio, and longitudinal diameter length.

[0129] Among them, the longitudinal to transverse ratio and the longitudinal diameter length both have the characteristics of continuity and normal distribution.

[0130] Taking two levels of each of the three factors (concave-convex degree, longitudinal to transverse ratio, and longitudinal diameter length), an orthogonal test scheme is obtained, which has a total of 4 groups.

[0131] Factor level table

[0132]

[0133] Orthogonal test scheme

[0134]

[0135] Select the above by orthogonal experiment of 4 groups of jujube kernel content (spinosin) determination, the specific steps are as follows:

[0136] Step 1: with mortar 4 jujube kernel grinding into powder, respectively, placed in 5 mL EP tube, do a good job of marking, for standby;

[0137] Step 2: using a millionth balance, accurately weigh 0.1 g (precision to 0.001 g) of test powder, each test sample three times, a total of 12, used for parallel experiment, standby;

[0138] Step 3: using 5 mL pipette 2 mL methanol in 5 mL EP tube with test sample, ultrasonic 20 min, 2 times, centrifugation 9800 r / min, 10 min. Absorb the supernatant in 1.5 mL EP tube, standby;

[0139] Step 4: accurately weigh 5 mg of jujube saponin A and spinosin, placed in 5 mL volumetric flask, namely (1 mg / mL) standard.

[0140] In which, the chromatographic conditions: Eclipse XDB-C18 (4.6 mm x 250 mm, 5 μm); mobile phase: acetonitrile (B) - water (A), gradient elution (0-3 min, 20% B→29% B; 3-8 min, 29% B→30% B; 8-18 min, 30% B→80% B, 18-24 min, 80% B→20% B); detection wavelength: 335 nm; flow rate: 1 mL·min-1; column temperature: 30 ℃; injection volume: 10 μL.

[0141] Step 5: the standard curve concentration gradient table is as follows (Table 7) and standard curve graph ( Figure 11 ).

[0142] Instrument includes FA2004 electronic balance, VGT-1990QTD, H1650-w, SPD-20A, organic phase chromatography filter, water phase chromatography filter.

[0143] Reagent using methanol (analytical pure), acetonitrile (chromatographic pure), pure water.

[0144] Standard curve concentration gradient table (Table 7)

[0145]

[0146]

[0147] (Shandong) different characteristics of jujube kernel spinosin content table (Table 8)

[0148]

[0149] Orthogonal test scheme and direct analysis table (table 9)

[0150]

[0151] From the above table 7-table 9, the maximum range of concave-convex degree (A), the second maximum range of longitudinal diameter length, and the minimum range of aspect ratio.

[0152] The content of spinozine is taken as an index, and the range is taken as a standard, so it can be known that factor A (concave-convex degree) has the greatest influence on the content of spinozine, factor C (longitudinal diameter length) is the secondary factor influencing the content of spinozine, and factor B (aspect ratio) has no influence on the content of spinozine basically, that is, the more flat the one side of the Semen Ziziphi Spinosae is, the thinner the xylem of the Semen Ziziphi Spinosae is, and the more symmetrical the Semen Ziziphi Spinosae is, and the higher the content of spinozine is.

[0153] The above is only a specific embodiment of the present application, but the protection scope of the present application is not limited to this, and any person skilled in the art can easily think of various changes or replacements within the technical range disclosed by the present application, and these should be covered in the protection scope of the present application.

[0154] Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A method for classifying the properties of traditional Chinese medicines and exploring the relevance of the properties to the efficacy of the medicines using orthogonal tests, characterized in that, The method comprises at least the following steps: S1 multi-dimensional trait measurement The data of the source Zizyphus jujuba Mill. of the traditional Chinese medicine Zizyphi Semen Praeparatum is measured, the Zizyphus jujuba Mill. is numbered, the horizontal diameter and the vertical diameter are accurately measured by using a vernier caliper, and a photograph is taken for recording; The Zizyphus jujuba Mill. pulp is peeled, and the data of the Zizyphus jujuba Mill. kernel is measured. The pulp is peeled by using a pincer, the kernel is washed and dried, and the horizontal diameter, the vertical diameter, the thickness of the xylem, the symmetry condition and the number of kernels are measured; The data of the Zizyphi Semen Praeparatum is measured by using a body mirror. The fruit, the kernel and the kernel horizontal diameter and the vertical diameter are measured by using a vernier caliper, the kernel is observed by placing one side and recorded, the basic trait data of the kernel and the kernel are obtained, and the measured kernel is packed. S2 frequency analysis and correlation analysis The Pearson correlation coefficients of the corresponding Zizyphus jujuba Mill., Zizyphus jujuba Mill. kernel and Zizyphi Semen Praeparatum are analyzed, and the correlation of the number of kernels, the concave-convex degree, the size, the Zizyphus jujuba Mill. kernel and the traditional Chinese medicine appearance is determined; The influence of climate and production place factors on the content of the effective component spinozine of Zizyphi Semen Praeparatum is explored, the influence of the trait factors on the content of the effective component spinozine is combined on the basis, the concave-convex degree, the vertical diameter length and the like are taken as indexes, and the kernel germplasm with high content of spinozine is screened. S3 classification of orthogonal test The orthogonal test method is adopted, the concave-convex degree, the ratio of the vertical diameter to the horizontal diameter and the vertical diameter length are taken as influence factors, two levels are taken for each, the Zizyphi Semen Praeparatum produced in Shandong with the highest content in the climate and production place factors is divided into four groups, and the foundation is laid for subsequent exploration of the relationship between the traits and the quality. S4 determination of the content of the effective component The kernel after grouping in S3 is ground, weighed, ultrasonically extracted by using methanol, centrifuged, the standard sample is prepared, the content of the effective component spinozine is determined by using a high performance liquid chromatograph, and the correlation between the trait data and the content of spinozine is established.

2. The orthogonal test for classifying the properties of traditional Chinese medicine and exploring the correlation between the properties and the efficacy according to claim 1, characterized in that, The horizontal diameter and the vertical diameter of the traditional Chinese medicine sample in S1 are accurately measured to two decimal places, and the unit is mm. 3.The method of classifying the properties of traditional Chinese medicines and exploring the correlation between the properties and the pharmacodynamic effects of the traditional Chinese medicines using orthogonal tests according to claim 1, characterized in that, In the orthogonal test method in S3, there are three factors, which are the concave-convex degree, the ratio of the vertical diameter to the horizontal diameter and the vertical diameter length. The two levels of the concave-convex degree are full and not full, the two levels of the ratio of the vertical diameter to the horizontal diameter are 1.0 and 1.2, the two levels of the vertical diameter length are 7 mm and 5.5 mm, and the experimental variable setting is determined.

4. The orthogonal test for classifying the properties of traditional Chinese medicine and exploring the correlation between the properties and the efficacy according to claim 1, characterized in that, In the content determination step in S4, the powder of the test sample is accurately weighed to 0.001 g, three parts of the test sample are taken for parallel measurement; 2 mL of methanol is taken by using a 5 mL pipette in an EP tube containing the test sample, ultrasonic extraction is performed for 20 min, 2 times, the centrifugal speed is 9800 r / min, and the time is 10 min.

5. The orthogonal test for classifying the properties of traditional Chinese medicine and exploring the correlation between the properties and the efficacy according to claim 1, characterized in that, The traditional Chinese medicine good and bad identification model is established by using the obtained Zizyphus jujuba Mill., Zizyphus jujuba Mill. kernel and Zizyphi Semen Praeparatum trait data and the characteristics of the chemical composition.

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