UPLC (Ultra Performance Liquid Chromatography) map construction method and identification method for rhizoma menispermi medicinal material and formula granules

Abstract

The invention discloses a rhizoma menispermi medicinal material and formula granule UPLC (Ultra Performance Liquid Chromatography) identification method, which is characterized in that specified values of relative retention time of the rhizoma menispermi medicinal material and formula granule are determined through a high performance liquid chromatography, and the relative retention time of a corresponding test solution is respectively within + / -10% of the corresponding specified values of the rhizoma menispermi medicinal material and formula granule. The method can effectively identify the rhizoma menispermi medicinal material and the formula granules, can be used for identifying rhizoma menispermi and counterfeit berba aristolochiae mollissimae and subprostrate sophora medicinal materials and formula granules, and can also be used for identifying rhizoma coptidis, cortex phellodendri, caulis sinomenii and other medicinal materials containing alkaloid components and formula granules. The method overcomes the defects of the existing method, and has the characteristics of simplicity in operation, good stability, rich chromatographic peak information amount, good repeatability and high stability.

Description

Technical Field

[0001] The invention relates to the field of formula granule identification, in particular to a method for identifying northern bean root medicinal materials and formula granules. Background Art

[0002] The root of northern bean is the dried rhizome of the plant kudzuvine of the Menispermaceae family. The root of Sophora flavescens is the dried root and rhizome of the plant Sophora japonica of the Leguminosae family. The root of Aristolochia chinensis is a woody vine of the Aristolochiaceae family. The functions and efficacy of the root of northern bean and the root of Sophora flavescens are similar, and there is a possibility of mixing them. The appearance of the root of northern bean and the root of Sophora flavescens is similar, so it is easy to confuse them.

[0003] Chinese medicine formula granules are made from single-flavor pieces of medicine using water as the solvent, and are produced through physical methods of solid-liquid separation, concentration, drying, granulation and other processes. Chinese medicine formula granules retain multiple active ingredients in the medicinal materials, but the medicinal materials can no longer be distinguished by their appearance.

[0004] The root of northern bean contains a large number of alkaline compounds, such as magnolamine, batryticatine, batryticatine, and other alkaloid components. Since the alkaline compounds are easy to react with the free silanol on the silica gel, the peak shape is severely tailed. At present, the peak shape of the alkaline compounds is improved by adding a certain concentration of triethylamine (northern bean root) or 0.4mol / L ammonium chloride (Phellodendron), 4mmol / L potassium dihydrogen phosphate solution (Sophora flavescens), 25mmol / L ammonium acetate and 8mmol / L sodium dodecyl sulfate solution (ammonia water adjusted to pH 9.3) (Coptis chinensis) and other salt solutions in the mobile phase; since most chromatographic columns are used between pH: 2-8, adding 0.05% triethylamine (PH: 10.5) or salt solution in the mobile phase may exceed the tolerance range of the commonly used chromatographic columns, causing irreversible damage to the chromatographic column structure and reducing the service life. In addition, the repeatability is poor, and there are fewer separated chromatographic peaks. Summary of the invention

[0005] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a method for identifying northern bean root medicinal materials and formula granules, which can effectively identify northern bean root medicinal materials and formula granules, common counterfeit products such as Sophora flavescens and Rhizoma Cyperi, and Chinese medicine formula granules containing alkaloid components such as Caulis Sinomenii, Coptis chinensis, and Phellodendron chinense through high performance liquid chromatography.

[0006] The objective of the present invention is achieved through the following technical solutions:

[0007] A UPLC identification method for Radix Sophorae Flavescentis medicinal materials and formula granules, comprising the following steps:

[0008] (1) Preparation of reference solution: Take magnolia alkaloids, dauricine, and dauricine reference substances and dissolve them to obtain reference solution;

[0009] (2) The reference solution and the test solution were respectively injected into the high performance liquid chromatograph, and the mobile phase was passed through the chromatographic column by gradient elution. The chromatographic conditions were as follows:

[0010] The mobile phases are mobile phase A and mobile phase B; the mobile phase A is acetonitrile, and the mobile phase B is 0.1% phosphoric acid;

[0011] The gradient elution method is:

[0012] At 0 to 8 minutes, the mass fraction of the mobile phase A in the mobile phase is 3-8%, and the mass fraction of the mobile phase B in the mobile phase is 97-92%;

[0013] At 8 to 12 minutes, the mass fraction of the mobile phase A in the mobile phase is 8-15%, and the mass fraction of the mobile phase B in the mobile phase is 92-85%;

[0014] At 12 to 19 minutes, the mass fraction of the mobile phase A in the mobile phase is 15-18%, and the mass fraction of the mobile phase B in the mobile phase is 85-12%;

[0015] At 19-22 minutes, the mass fraction of the mobile phase A in the mobile phase is 18-20%, and the mass fraction of the mobile phase B in the mobile phase is 82-15%;

[0016] At 22-25 minutes, the mass fraction of the mobile phase A in the mobile phase is 20-23%, and the mass fraction of the mobile phase B in the mobile phase is 80-77%;

[0017] At 25-28 minutes, the mass fraction of the mobile phase A in the mobile phase is 23-26%, and the mass fraction of the mobile phase B in the mobile phase is 77-74%;

[0018] At 28 to 30 minutes, the mass fraction of the mobile phase A in the mobile phase is 26-35%, and the mass fraction of the mobile phase B in the mobile phase is 74-65%;

[0019] At 30 to 35 minutes, the mass fraction of the mobile phase A in the mobile phase is 35-85%, and the mass fraction of the mobile phase B in the mobile phase is 65-15%;

[0020] (3) Identification is performed by counting the number of characteristic peaks and relative retention time of characteristic spectra of multiple batches of samples and comparing the characteristic spectra of the samples to be tested with those of the reference substances.

[0021] Furthermore, the flow rate of the mobile phase through the chromatographic column is 0.2 ml per minute.

[0022] Furthermore, during the high performance liquid chromatography detection process, the column temperature of the chromatographic column is 25°C.

[0023] Furthermore, during the high performance liquid chromatography detection process, the filler is octadecylsilane bonded silica gel.

[0024] Furthermore, during the high performance liquid chromatography detection process, the detection wavelengths are 215nm and 284nm.

[0025] Furthermore, in the step (3), the prescribed value of the relative retention time of the Northern Root Radix and the formula granules is determined, and if the relative retention time of the corresponding sample solution to be tested falls within the range of ±10% of the corresponding prescribed value of the Northern Root Radix and the formula granules, it is judged to be the true Northern Root Radix and the formula granules.

[0026] The beneficial effects of the present invention are:

[0027] The invention can effectively identify the medicinal material and formula granules of Radix Sophorae Flavescentis by high performance liquid chromatography, and can be used to identify the medicinal materials and formula granules of Radix Sophorae Flavescentis and counterfeit Radix Sophorae Flavescentis, and can also be used to identify the medicinal materials and formula granules containing alkaloid components such as Rhizoma Coptidis, Cortex Phellodendri, Caulis Sinomenii, etc. The method overcomes the shortcomings of the existing methods, and has the characteristics of simple operation, good stability, rich chromatographic peak information, good repeatability, and high stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is the characteristic spectrum of the reference substance of magnolia alkaloid, dauricine and dauricine;

[0029] Figure 2 The characteristic spectrum of the Northern Root Formula Granules in Example 1;

[0030] Figure 3 The characteristic spectrum of the Radix Sophorae flavescentis medicinal material in Example 2;

[0031] Figure 4 The characteristic spectrum of the Qingfengteng formula granules in Example 3;

[0032] Figure 5 The characteristic spectrum of the Cortex Phellodendri formula particles in Example 4;

[0033] Figure 6 The characteristic spectrum of the Rhizoma Coptidis formula granules in Example 5;

[0034] Figure 7 The characteristic spectrum of Xungufeng medicinal materials in Example 6;

[0035] Figure 8 The characteristic spectrum of the Xungufeng formula granules in Example 7;

[0036] Fig. 9This is the characteristic spectrum of the medicinal material of Sophora flavescens in Example 8;

[0037] Fig.10 This is the characteristic spectrum of the safflower root formula granules in Example 9;

[0038] Fig.11 This is the characteristic spectrum of the northern bean root formula granules in comparative example 1;

[0039] Fig.12 This is the characteristic spectrum of the northern bean root medicinal material in comparative example 2. DETAILED DESCRIPTION

[0040] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0041] It should be noted that the illustrations provided in the following embodiments are only schematic illustrations of the basic concept of the present invention, and thus the drawings only show components related to the present invention rather than being drawn according to the number, shape and size of components in actual implementation. In actual implementation, the type, quantity and proportion of each component may be changed arbitrarily, and the component layout may also be more complicated.

[0042] The following specific embodiments all use high performance liquid chromatography to detect the test products to obtain corresponding characteristic spectra:

[0043] The mobile phases in the high performance liquid chromatography detection process are mobile phase A and mobile phase B; the mobile phase A is acetonitrile, and the mobile phase B is 0.1% phosphoric acid;

[0044] In the high performance liquid chromatography detection process, the mobile phase is passed through the chromatographic column by gradient elution, and the gradient elution method is:

[0045] At 0 to 8 minutes, the mass fraction of the mobile phase A in the mobile phase is 3-8%, and the mass fraction of the mobile phase B in the mobile phase is 97-92%;

[0046] At 8 to 12 minutes, the mass fraction of the mobile phase A in the mobile phase is 8-15%, and the mass fraction of the mobile phase B in the mobile phase is 92-85%;

[0047] At 12 to 19 minutes, the mass fraction of the mobile phase A in the mobile phase is 15-18%, and the mass fraction of the mobile phase B in the mobile phase is 85-12%;

[0048] At 19-22 minutes, the mass fraction of the mobile phase A in the mobile phase is 18-20%, and the mass fraction of the mobile phase B in the mobile phase is 82-15%;

[0049] At 22-25 minutes, the mass fraction of the mobile phase A in the mobile phase is 20-23%, and the mass fraction of the mobile phase B in the mobile phase is 80-77%;

[0050] At 25-28 minutes, the mass fraction of the mobile phase A in the mobile phase is 23-26%, and the mass fraction of the mobile phase B in the mobile phase is 77-74%;

[0051] At 28 to 30 minutes, the mass fraction of the mobile phase A in the mobile phase is 26-35%, and the mass fraction of the mobile phase B in the mobile phase is 74-65%;

[0052] At 30 to 35 minutes, the mass fraction of the mobile phase A in the mobile phase is 35-85%, and the mass fraction of the mobile phase B in the mobile phase is 65-15%;

[0053] During the HPLC detection process, the flow rate of the mobile phase through the chromatographic column is 0.2 ml per minute;

[0054] The column temperature of the chromatographic column is 25°C; the filler is octadecylsilane bonded silica gel, with a specification of 2.1×100mm, 1.7μm;

[0055] During the high performance liquid chromatography detection process, the detection wavelengths are 215 nm and 284 nm; the theoretical plate number calculated based on the bean curd acid A peak should be no less than 10,000;

[0056] For high performance liquid chromatography, please refer to General Chapter 0512 of the 2020 Edition of the Chinese Pharmacopoeia.

[0057] Prepare the medicinal materials or formula granules to be identified into test solution:

[0058] Take an appropriate amount of the medicinal material or formula granules to be identified, grind them into powder, take about 0.1g, put it in a stoppered conical flask, add 25ml of 50% methanol, ultrasonically treat (power 500W, frequency 35kHz) for 20 minutes, take it out, cool, shake well, filter, and take the filtrate to obtain the test solution.

[0059] Take appropriate amount of magnolia alkaloid, daurosuline and daurosuline reference substance, weigh them accurately, add methanol to make a mixed solution containing 40 μg of daurosuline, daurosuline and magnolia alkaloid per 1 ml, which is used as the reference substance solution.

[0060] Accurately pipette 0.5 μl of the reference substance solution, inject it into the liquid chromatograph, and use the high performance liquid chromatography method of Example 3 to detect the reference substance solution to obtain the reference substance characteristic spectrum, such as Figure 1 shown.

[0061] Accurately pipette 0.5 μl of the test solution and reference solution respectively, inject into the liquid chromatograph for determination.

[0062] Embodiment 1:

[0063] This embodiment provides a method for identifying the Northern Bean Root Formula Granules. The HPLC detection method is as described in the above steps. The characteristic spectrum of the Northern Bean Root Formula Granules is as follows: Figure 2 As shown. The characteristic spectrum of the Northern Bean Root Formula Granules presents 15 characteristic peaks, and the retention times of the 15 characteristic peaks in the characteristic spectrum of the Northern Bean Root reference medicinal material reference correspond to each other. Among the 15 characteristic peaks in the characteristic spectrum of the Northern Bean Root Formula Granules, 3 peaks correspond to the retention times of the corresponding reference substance peaks in the characteristic spectrum of magnolamine, dauricine, and dauricine reference substance reference substance, respectively. The peak corresponding to the magnolia pine reference peak is the S1 peak. The relative retention times of peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 7, peak 9 and S1 peak are calculated. The peak corresponding to the dauricine reference peak is the S2 peak. The relative retention times of peak 10, peak 12, peak 14, peak 15 and S2 peak are calculated. The specified values ​​of relative retention times are determined to be 0.19 (peak 1), 0.31 (peak 2), 0.50 (peak 3), 0.52 (peak 4), 0.64 (peak 5), 0.66 (peak 6), 0.78 (peak 7), 1.08 (peak 9), 1.14 (peak 10), 0.98 (peak 12), 1.03 (peak 14), and 1.10 (peak 15).

[0064] If the relative retention time of the test solution falls within the range of ±10% of the above standard value, the test product is judged to be the northern bean root formula granules. The relative retention time data of the characteristic spectra of 6 batches of northern bean root formula granules and the samples to be tested are shown in Table 1. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of northern bean root formula granules.

[0065] The sample to be tested has 15 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, so it is determined to be the northern bean root formula granules.

[0066] Embodiment 2:

[0067] This embodiment provides a method for identifying a medicinal material of Radix sibiricum. The high performance liquid chromatography detection is the same as in Example 1. The characteristic spectrum of the medicinal material of Radix sibiricum is as follows: Figure 3There are 17 characteristic peaks in the characteristic spectrum of the northern bean root medicinal material, and 3 of the 17 characteristic peaks in the characteristic spectrum of the northern bean root medicinal material correspond to the retention times of the corresponding reference substance peaks in the characteristic spectrum of magnolia alkaloids, dauricine, and dauricine reference substance. The peak corresponding to the magnolia pine reference peak is the S1 peak. The relative retention times of peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 7, peak 8, peak 10, peak 11 and the S1 peak are calculated. The peak corresponding to the dauricine reference peak is the S2 peak. The relative retention times of peak 13, peak 15, peak 16, peak 17 and the S2 peak are calculated. The specified values ​​of the relative retention times are determined to be 0.50 (peak 1), 0.52 (peak 2), 0.54 (peak 3), 0.60 (peak 4), 0.64 (peak 5), 0.66 (peak 6), 0.76 (peak 7), 0.78 (peak 8), 1.08 (peak 10), 1.14 (peak 11), 0.98 (peak 13), 1.03 (peak 15), 1.05 (peak 16), and 1.10 (peak 17).

[0068] If the relative retention time of the test solution falls within the range of ±10% of the above specified value, the test sample is judged to be the northern bean root medicinal material. The relative retention time data of the characteristic spectra of 6 batches of northern bean root medicinal materials and the samples to be tested are shown in Table 2. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of northern bean root medicinal materials.

[0069] The sample to be tested has 17 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, so it is determined to be the medicinal material of Northern Bean Root.

[0070] Embodiment 3:

[0071] This embodiment provides a method for identifying the Sinomenium sinomenium formula granules. The high performance liquid chromatography detection is the same as in Example 1. The characteristic spectrum of the Sinomenium sinomenium formula granules is as follows: Figure 4 As shown. There are 10 characteristic peaks in the characteristic spectrum of Qingfengteng formula granules. The peak corresponding to the peak 7 reference peak is the S peak. The relative retention time of peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 8, peak 9, peak 10 and S peak is calculated, and the specified values ​​of relative retention time are determined to be: 0.29 (peak 1), 0.32 (peak 2), 0.60 (peak 3), 0.64 (peak 4), 0.66 (peak 5), 0.89 (peak 6), 1.08 (peak 8), 1.14 (peak 9), 1.36 (peak 10).

[0072] If the relative retention time of the test solution falls within the range of ±10% of the above specified value, the test product is judged to be Caulis Sinensis Formula Granules. The relative retention time data of the characteristic spectra of 6 batches of Caulis Sinensis Formula Granules and the samples to be tested are shown in Table 3. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of Caulis Sinensis Formula Granules.

[0073] The sample to be tested has 10 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, and is determined to be Sinomene Atractylodes formula granules.

[0074] Embodiment 4:

[0075] This embodiment provides a method for identifying the Phellodendron chinense formula granules. The high performance liquid chromatography detection is the same as in Example 1. The characteristic spectrum of the Phellodendron chinense formula granules is as follows: Figure 5 As shown. There are 9 characteristic peaks in the characteristic spectrum of Phellodendron chinense formula granules, and the peak corresponding to the peak 6 reference peak is the S peak. The relative retention time of peak 1, peak 2, peak 3, peak 4, peak 5, peak 7, peak 8, peak 9 and S peak was calculated, and the specified values ​​of relative retention time were determined to be 0.64 (peak 1), 0.68 (peak 2), 0.84 (peak 3), 0.86 (peak 4), 0.87 (peak 5), 1.08 (peak 7), 1.18 (peak 8), and 1.31 (peak 9).

[0076] If the relative retention time of the test solution falls within the range of ±10% of the above specified value, the test sample is judged to be Phellodendron chinense formula granules. The relative retention time data of the characteristic spectra of 6 batches of Phellodendron chinense formula granules and the samples to be tested are shown in Table 4. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of Phellodendron chinense formula granules.

[0077] The sample to be tested has 9 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, so it is determined to be Phellodendron chinense formula granules.

[0078] Embodiment 5:

[0079] This embodiment provides a method for identifying the Rhizoma Coptidis Formula Granules. The HPLC detection is the same as in Example 1. The characteristic spectrum of the Rhizoma Coptidis Formula Granules is as follows: Figure 6 As shown. There are 7 characteristic peaks in the characteristic spectrum of Huanglian formula granules. The peak corresponding to the peak 4 reference peak is the S peak. The relative retention time of peak 1, peak 2, peak 3, peak 5, peak 6, peak 7 and S peak is calculated, and the specified values ​​of relative retention time are determined to be: 0.41 (peak 1), 0.75 (peak 2), 0.86 (peak 3), 1.08 (peak 5), 1.13 (peak 6), 1.31 (peak 7);

[0080] If the relative retention time of the test solution falls within the range of ±10% of the above specified value, the test sample is judged to be Coptis chinensis formula granules. The relative retention time data of the characteristic spectra of 6 batches of Coptis chinensis formula granules and the samples to be tested are shown in Table 5. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of Coptis chinensis formula granules.

[0081] The sample to be tested has 7 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, so it is determined to be Coptis chinensis formula granules.

[0082] Embodiment 6:

[0083] This embodiment provides a method for identifying the medicinal material of Xungufeng. The high performance liquid chromatography detection is the same as in Example 1. The characteristic spectrum of the medicinal material of Xungufeng is as follows Figure 7 As shown. There are 13 characteristic peaks in the characteristic spectrum of Xungufeng medicinal materials. The peak corresponding to the peak 6 reference peak is the S peak. The relative retention time of peak 1, peak 2, peak 3, peak 4, peak 5, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12, peak 13 and S peak was calculated, and the specified values ​​of relative retention time were determined as follows: 0.41 (peak 1), 0.53 (peak 2), 0.59 (peak 3), 0.68 (peak 4), 0.79 (peak 5), 1.14 (peak 7), 1.35 (peak 8), 1.42 (peak 9), 1.44 (peak 10), 1.46 (peak 11), 1.51 (peak 12), 1.52 (peak 13);

[0084] If the relative retention time of the test solution falls within the range of ±10% of the above specified value, the test sample is judged to be a Xun Gu Feng medicinal material. The relative retention time data of the characteristic spectra of 6 batches of Xun Gu Feng medicinal materials and the samples to be tested are shown in Table 6. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of Xun Gu Feng medicinal materials.

[0085] The sample to be tested has 13 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, so it is determined to be a Xungufeng medicinal material.

[0086] Embodiment 7:

[0087] This embodiment provides a method for identifying Xungufeng granules. The HPLC detection is the same as in Example 1. The characteristic spectrum of Xungufeng is as follows: Figure 8 As shown. There are 12 characteristic peaks in the characteristic spectrum of Xungufeng granules. The peak corresponding to the peak 6 reference peak is the S peak. The relative retention time of peak 1, peak 2, peak 3, peak 4, peak 5, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12 and S peak was calculated, and the specified values ​​of relative retention time were determined to be: 0.41 (peak 1), 0.53 (peak 2), 0.59 (peak 3), 0.68 (peak 4), 0.79 (peak 5), 1.14 (peak 7), 1.35 (peak 8), 1.42 (peak 9), 1.44 (peak 10), 1.46 (peak 11), 1.51 (peak 12);

[0088] If the relative retention time of the test solution falls within the range of ±10% of the above specified value, the test sample is judged to be Xungufeng granules. The relative retention time data of the characteristic spectra of 6 batches of Xungufeng granules and the samples to be tested are shown in Table 7. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of Xungufeng granules.

[0089] The sample to be tested has 12 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, and is determined to be Xungufeng formula granules.

[0090] Embodiment 8:

[0091] This embodiment provides a method for identifying Sophora flavescens medicinal materials. The high performance liquid chromatography detection is the same as in Example 1. The characteristic spectrum of Sophora flavescens medicinal materials is as follows: Fig. 9 As shown. There are 9 characteristic peaks in the characteristic spectrum of Sophora flavescens medicinal material. The peak corresponding to the peak 5 reference peak is the S peak. The relative retention time of peak 1, peak 2, peak 3, peak 4, peak 6, peak 7, peak 8, peak 9 and S peak was calculated, and the specified values ​​of relative retention time were determined to be: 0.14 (peak 1), 0.15 (peak 2), 0.35 (peak 3), 0.42 (peak 4), 1.35 (peak 6), 1.50 (peak 7), 1.55 (peak 8), 1.59 (peak 9);

[0092] If the relative retention time of the test solution falls within the range of ±10% of the above specified value, the test sample is judged to be Sophora flavescens medicinal material. The relative retention time data of the characteristic spectra of 6 batches of Sophora flavescens medicinal materials and the samples to be tested are shown in Table 8. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of Sophora flavescens medicinal material granules.

[0093] The sample to be tested has 9 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, so it is determined to be Sophora flavescens.

[0094] Embodiment 9:

[0095] This embodiment provides a method for identifying Sophora flavescens granules. The high performance liquid chromatography detection is the same as in Example 1. The characteristic spectrum of Sophora flavescens formula granules is as follows: Fig.10 As shown. There are 8 characteristic peaks in the characteristic spectrum of Sophora flavescens formula granules. The peak corresponding to the peak 4 reference peak is the S peak. The relative retention time of peak 1, peak 2, peak 3, peak 5, peak 6, peak 7, peak 8 and S peak was calculated, and the specified values ​​of relative retention time were determined to be: 0.14 (peak 1), 0.15 (peak 2), 0.42 (peak 3), 1.35 (peak 5), 1.51 (peak 6), 1.54 (peak 7), 1.59 (peak 8);

[0096] If the relative retention time of the test solution falls within the range of ±10% of the above specified value, the test sample is judged to be Sophora flavescens granules. The relative retention time data of the characteristic spectra of 6 batches of Sophora flavescens formula granules and the samples to be tested are shown in Table 9. The standard range is established based on the average relative retention time of the characteristic spectra of 6 batches of Sophora flavescens formula granules.

[0097] The sample to be tested has 8 characteristic peaks, and the relative retention times are all within the range of ±10% of the standard value, so it is determined to be Sophora flavescens formula granules.

[0098]

[0099]

[0100]

[0101]

[0102]

[0103] Comparative Example 1:

[0104] This comparative example refers to the quality detection method of a standard decoction of Radix Sophorae Flavescentis disclosed in CN 112858497 A to detect the Radix Sophorae Flavescentis formula granules in Example 1, and the steps are as follows:

[0105] S1. Preparation of the test solution: Take the product, place it in a stoppered conical flask, add 50% methanol, ultrasonically treat for 20 minutes, power 600W, frequency 40kHz, cool, shake well, filter, and take the filtrate to obtain;

[0106] S2 detection: the test solution obtained in step S1 was injected into a high performance liquid chromatograph, and the obtained high performance liquid chromatogram was compared with the obtained HPLC characteristic spectrum to detect the quality and authenticity of the northern bean root formula granules;

[0107] The chromatographic conditions in the HPLC instrument meet the following requirements:

[0108] Chromatographic column: Octadecylsilane bonded silica gel is used as filler, column length is 250 mm, inner diameter is 4.6 mm, particle size is 5 μm;

[0109] Chromatographic column temperature: 30°C;

[0110] Mobile phase: acetonitrile as mobile phase A, 0.05% triethylamine solution as mobile phase B;

[0111] Mobile phase flow rate: 1.0 mL / min;

[0112] Injection volume: 10 μL;

[0113] Detection wavelength: 284nm;

[0114] The number of theoretical plates calculated based on the dauricine peak should be no less than 6000;

[0115] The mobile phase gradient elution program is:

[0116]

[0117] The test results are as follows Fig.11As shown, the method disclosed in CN112858497 A can only identify 5 characteristic peaks, while the present invention can identify 15 characteristic peaks of the northern bean root formula granules.

[0118] The characteristic spectrum chromatographic conditions of the present invention are acetonitrile-0.1% phosphoric acid solution, and the method has good repeatability. However, the method disclosed in CN112858497A uses acetonitrile-0.05% triethylamine solution, requires an alkaline-resistant chromatographic column, causes greater damage to the chromatographic column, has poor repeatability, and the number of characteristic peaks is unstable. After repeated use, the separation degree of characteristic peaks deteriorates.

[0119] In addition, the ultra-high performance liquid chromatography analysis time of the present invention is relatively short, thus saving costs, whereas the ordinary liquid phase analysis time used in the method disclosed in CN112858497 A is relatively long.

[0120] Comparative Example 2:

[0121] The difference between this comparative example and comparative example 1 is that the northern bean root medicinal material in Example 2 is tested with reference to the quality test method of a northern bean root standard decoction disclosed in CN 112858497 A. The test results are as follows: Fig.12 As shown, the method disclosed in CN112858497 A can only identify 5 characteristic peaks, while the Radix Sophorae flavescentis medicinal material of the present invention can identify 17 characteristic peaks, and more components can be identified.

[0122] The method disclosed in CN112858497 A can only identify 5 characteristic peaks in the northern bean root medicinal material and the formula granules, and cannot distinguish the northern bean root medicinal material and the formula granules. The northern bean root medicinal material of the present invention can identify 17 characteristic peaks, and the northern bean root formula granules can identify 15 characteristic peaks, which can effectively distinguish the northern bean root formula granules and the medicinal material.

[0123] In summary, the above embodiments are merely examples of the preferred embodiments of the present invention and do not include all implementation scopes of the invention.

Claims

1. A UPLC identification method for Radix Sophorae Flavescentis medicinal materials and formula granules, characterized in that: The following steps are involved: (1) Preparation of reference solution: Take magnolia alkaloids, dauricine, and dauricine reference substances and dissolve them to obtain reference solution; (2) The reference solution and the test solution were respectively injected into the high performance liquid chromatograph, and the mobile phase was passed through the chromatographic column by gradient elution. The chromatographic conditions were as follows: The mobile phases are mobile phase A and mobile phase B; the mobile phase A is acetonitrile, and the mobile phase B is 0.1% phosphoric acid; The gradient elution method is: At 0 to 8 minutes, the mass fraction of the mobile phase A in the mobile phase is 3-8%, and the mass fraction of the mobile phase B in the mobile phase is 97-92%; At 8 to 12 minutes, the mass fraction of the mobile phase A in the mobile phase is 8-15%, and the mass fraction of the mobile phase B in the mobile phase is 92-85%; At 12 to 19 minutes, the mass fraction of the mobile phase A in the mobile phase is 15-18%, and the mass fraction of the mobile phase B in the mobile phase is 85-12%; At 19-22 minutes, the mass fraction of the mobile phase A in the mobile phase is 18-20%, and the mass fraction of the mobile phase B in the mobile phase is 82-15%; At 22-25 minutes, the mass fraction of the mobile phase A in the mobile phase is 20-23%, and the mass fraction of the mobile phase B in the mobile phase is 80-77%; At 25-28 minutes, the mass fraction of the mobile phase A in the mobile phase is 23-26%, and the mass fraction of the mobile phase B in the mobile phase is 77-74%; At 28-30 minutes, the mass fraction of the mobile phase A in the mobile phase is 26-35%, and the mass fraction of the mobile phase B in the mobile phase is 74-65%; At 30 to 35 minutes, the mass fraction of the mobile phase A in the mobile phase is 35-85%, and the mass fraction of the mobile phase B in the mobile phase is 65-15%; (3) Identification is performed by counting the number of characteristic peaks and relative retention time of characteristic spectra of multiple batches of samples and comparing the characteristic spectra of the samples to be tested with those of the reference substances.

2. The method according to claim 1, characterized in that The flow rate of the mobile phase through the chromatographic column was 0.2 ml per minute.

3. The method according to claim 1, characterized in that During the high performance liquid chromatography detection process, the column temperature of the chromatographic column is 25°C.

4. The method according to claim 1, characterized in that: In the high performance liquid chromatography detection process, the filler is octadecylsilane bonded silica gel.

5. The method according to claim 1, characterized in that During the high performance liquid chromatography detection process, the detection wavelengths are 215 nm and 284 nm.

6. The method according to claim 1, characterized in that In the step (3), the prescribed value of the relative retention time of the Northern Root Radix and the formula granules is determined. If the relative retention time of the corresponding sample solution to be tested falls within the range of ±10% of the corresponding prescribed value of the Northern Root Radix and the formula granules, it is judged to be the true Northern Root Radix and the formula granules.

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