Construction method and application of characteristic chromatogram of rhizoma dryopteris crassirhizomae medicinal material, traditional Chinese medicine formula granules, decoction pieces or standard decoction

Abstract

The invention discloses a construction method of a characteristic chromatogram of a rhizoma dryopteris crassirhizomae medicinal material, formula granules, decoction pieces or standard decoction, and the construction method comprises the following steps: preparing a reference substance solution which comprises a reference substance solution of a reference medicinal material and a reference substance solution of a reference substance, the reference substance solution comprises a protocatechuic acid reference substance, a protocatechuic aldehyde reference substance, a neochlorogenic acid reference substance and a chlorogenic acid reference substance; taking a rhizoma dryopteris crassirhizomae medicinal material, formula granules, decoction pieces or a standard decoction to prepare a test solution; a preset amount of the reference substance solution and the test solution are taken and injected into a liquid chromatograph, the liquid chromatograph takes octadecylsilane chemically bonded silica as a filler, methanol as a mobile phase A and a phosphoric acid solution as a mobile phase B for gradient elution, and the characteristic chromatogram is established. The method can provide a data basis for quality control of rhizoma dryopteris crassirhizomae medicinal materials, formula granules, decoction pieces or standard decoction, and ensures the stability and controllability of product quality.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine quality analysis and detection, and in particular to a method for constructing a characteristic spectrum of a Chinese medicine, Chinese medicine formula granules, decoction pieces or standard decoctions and an application thereof. Background Art

[0002] Mianma Guanzhong is the dried rhizome and petiole residue of Dryopteris crassirhizoma Nakai, a plant of the Dryopteris family. It is included in the 2020 edition of the Chinese Pharmacopoeia. It has the effects of clearing away heat, detoxifying, and expelling parasites. It is used for abdominal pain caused by parasites and sores.

[0003] In the prior art, the research on Rhizoma Cynomorium is focused on the preparation method of water extract or formula granules, the multi-information thin-layer identification method of medicinal materials, and the use of phloroglucinol compounds. There are many studies on the phloroglucinol components of Rhizoma Cynomorium, such as Rhizoma ABB, Rhizoma Cynomorium ABBA and Rhizoma Cynomorium ABBP, which mainly play an anti-influenza virus role. At present, the 2020 edition of the Chinese Pharmacopoeia contains the standard of Rhizoma Cynomorium medicinal materials, using thin-layer identification and extracts as quality control methods, and there are no relevant provisions for characteristic spectra. "Study on HPLC Fingerprint of Rhizoma Cynomorium Formula Granules" uses high-performance liquid chromatography to establish the characteristic spectrum of Rhizoma Cynomorium Formula Granules, and identifies 12 common peaks, but there are problems such as rough baseline and poor separation of chromatographic peaks. From the application point of view, there are many counterfeit Rhizoma Cynomorium, such as Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, Rhizoma Cynomorium, etc. At present, the literature reports only distinguish the medicinal materials based on their properties, and lack objective and effective methods for identification. It can be seen that the quality of Mianma Guanzhong medicinal materials urgently needs to be standardized. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a method for constructing a characteristic spectrum of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions. The method has good reproducibility, accuracy and reliability, and can provide a data basis for mass production quality control of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions, thereby ensuring the stability and controllability of the product quality of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions.

[0005] In order to solve the above technical problems, the present invention provides a method for constructing a characteristic spectrum of a Mianma Guanzhong medicinal material, a formula granule, a decoction piece or a standard decoction, comprising the following steps: preparing a reference solution, the reference solution comprising a reference medicinal material reference solution and a reference substance reference solution, the reference substance reference solution comprising a protocatechuic acid reference substance, a protocatechuic aldehyde reference substance, a neochlorogenic acid reference substance, and a chlorogenic acid reference substance;

[0006] Take the medicinal material of Mianma Guanzhong, formula granules, decoction pieces or standard decoction to prepare the test solution;

[0007] Preset amounts of reference solution and test solution were taken and injected into a liquid chromatograph, which used octadecylsilane bonded silica gel as a filler, methanol as a mobile phase A, and phosphoric acid solution as a mobile phase B for gradient elution to establish a characteristic spectrum.

[0008] In one embodiment, the gradient elution is performed according to the following procedure:

[0009] 0-2min, mobile phase A is 3%, mobile phase B is 97%;

[0010] 2min~16min, mobile phase A from 3%→4%, mobile phase B from 97%→96%;

[0011] 16min-27min, mobile phase A from 4% to 10%, mobile phase B from 96% to 90%;

[0012] 27min~60min, mobile phase A from 10%→13%, mobile phase B from 90%→87%;

[0013] 60min~70min, mobile phase A changes from 13%→70%, and mobile phase B changes from 87%→30%.

[0014] In one embodiment, the concentration of the phosphoric acid solution is 0.05% to 0.15%.

[0015] In one embodiment, the preset amount of the reference solution and the test solution is 0.5 μL to 1.5 μL;

[0016] The liquid chromatograph uses octadecylsilane bonded silica gel as a filler, with a column length of 150 mm, an inner diameter of 3 mm, a particle size of 2.7 μm, and a column temperature of 23° C. to 27° C.;

[0017] When performing gradient elution, the flow rate of the mobile phase is 0.38-0.42 mL / min;

[0018] The ultraviolet detection wavelength of the liquid chromatograph is 300nm-310nm.

[0019] In one embodiment, the test solution is prepared by the following method:

[0020] Take the medicinal materials, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii, grind them into powder, weigh 1g-2g, put them in a stoppered conical flask, add 15mL-25mL of 65%-75% methanol, ultrasonically treat for 25min-35min, cool, shake well, filter, and take the filtrate to obtain the product.

[0021] In one embodiment, the reference substance solution is prepared according to the following method:

[0022] Take appropriate amount of protocatechuic acid reference substance, protocatechuic aldehyde reference substance, neochlorogenic acid reference substance, and chlorogenic acid reference substance, accurately weigh them, and add 70% methanol to make a mixed solution containing 50 μg of each per 1 ml, which serves as the reference substance solution.

[0023] In one embodiment, the control medicinal material reference solution is prepared by the following method:

[0024] Take about 1.5g to 2g of the control medicinal material of Rhizoma Cynomorii, place it in a stoppered conical flask, add 25mL to 35mL of water, heat and reflux for 25min to 35min, filter, evaporate to dryness, add 15mL to 25mL of 65% to 75% methanol to the residue, ultrasonically treat for 25min to 35min, filter, and take the filtrate as the control medicinal material reference solution.

[0025] In one embodiment, the characteristic spectrum includes 12 common peaks, among which peak No. 2 corresponds to protocatechuic acid, peak No. 4 corresponds to protocatechuic aldehyde, peak No. 5 corresponds to neochlorogenic acid, and peak No. 8 corresponds to chlorogenic acid.

[0026] In one embodiment, in the characteristic spectrum of the Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction, the protocatechuic aldehyde peak is used as the reference peak S1, and the relative retention times of peaks 1 to 3, 5 to 7 and the reference peak S1 are consistent with: peak 1 0.275-0.32, peak 2 0.585-0.597, peak 3 0.796-0.894, peak 5 1.253-1.292, peak 6 1.431-1.528, peak 7 1.516-1.609;

[0027] Taking the chlorogenic acid peak as the reference peak S2, the relative retention times of peaks 9 to 12 and the reference peak S2 are consistent with: peak 9 1.022-1.040, peak 10 1.142-1.168, peak 11 1.177-1.223, peak 12 1.265-1.328.

[0028] Correspondingly, the present invention also provides a characteristic spectrum of the Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction, and the characteristic spectrum of the Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction is obtained by the above-mentioned method for constructing the characteristic spectrum of the Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction.

[0029] And, the use of the characteristic spectrum of the Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction in any one of (1) to (3):

[0030] (1) Authenticity identification and ingredient testing of the medicinal materials, formula granules, decoction pieces or standard decoctions of Rhizoma Cynomorii;

[0031] (2) Distinguishing between the medicinal materials, formula granules, decoction pieces or standard decoctions of Rhizoma Cyrthospermi and the counterfeit Rhizoma Cyrthospermi;

[0032] (3) Differentiate between the medicinal materials, formula granules, decoction pieces or standard decoctions of Rhizoma Cynomorium and counterfeit Rhizoma Cynomorium.

[0033] The implementation of the present invention has the following beneficial effects:

[0034] The present invention establishes a characteristic spectrum of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions through the construction method of the characteristic spectrum, and the characteristic spectrum can fully display the chemical composition characteristics of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions, and the characteristic peak information is rich, the baseline is stable, and the peak separation is good, and the quality information of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions can be fully reflected, so as to achieve the purpose of comprehensively and effectively controlling the quality of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions. And it can realize the identification and distinction between Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions and counterfeit Ziqi Guanzhong and counterfeit Guanzhong. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 It is a chromatogram of the durability of the characteristic spectrum of the Mianma Guanzhong formula granules in Example 1 of the present invention-different column temperatures;

[0036] Figure 2 It is a chromatogram of the durability of the characteristic spectrum of the Mianma Guanzhong formula granules in Example 1 of the present invention-different flow rate inspection;

[0037] Figure 3 This is a superimposed chromatogram of the Mianma Guanzhong formula granules in Example 1 of the present invention;

[0038] Figure 4 This is a superimposed chromatogram of the Mianma Guanzhong medicinal material in Example 1 of the present invention;

[0039] Figure 5 It is the superimposed chromatogram of the Mianma Guanzhong decoction pieces in Example 1 of the present invention;

[0040] Figure 6 It is a superimposed chromatogram of the standard decoction of Mianma Guanzhong in Example 1 of the present invention;

[0041] Figure 7 It is a chromatogram comparing Rhizoma Cyrthospermi Mianmai, Rhizoma Cyrthospermi Bge and Rhizoma Cyrthospermi in Example 2 of the present invention. DETAILED DESCRIPTION

[0042] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below with reference to the accompanying drawings and specific implementation methods.

[0043] In order to comprehensively reflect the quality information of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions, and to comprehensively and effectively control the quality of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions, the present invention provides a method for constructing a characteristic spectrum of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions, which is described in detail below:

[0044] Example 1: Establishment of a characteristic spectrum of Rhizoma Cynomorii and determination of its medicinal materials, decoction pieces, standard decoctions and formula granule samples

[0045] 1. Instruments, reagents and test drugs

[0046] Instruments: Waters ultra-high performance liquid chromatograph (H-Class, Waters Corporation); Shimadzu ultra-high performance liquid chromatograph (Lx-40cx, Shimadzu Corporation); Waters CORTECS C18 chromatographic column (3.0 mm × 150 mm, 2.7 μm); one-ten-thousandth balance (ME204E, Mettler-Toledo); one-hundred-thousandth balance (ABT220-5DM, Guangzhou Dexiang Technology Co., Ltd.); one-millionth balance (XP26, Mettler-Toledo); electronic balance (JJ600, Changshu Shuangjie Testing Instrument Factory); CNC ultrasonic cleaner (KQ500DE, Kunshan Ultrasonic Instrument Co., Ltd.); constant temperature water bath (HWS28, Shanghai Yiheng Technology Co., Ltd.); ultrapure water system (Villi-Q Direct, Merck & Co., Ltd.).

[0047] Reagents: Methanol (Tianjin Fuyu Fine Chemical Co., Ltd.) was of analytical grade, phosphoric acid (Tianjin Komiou Chemical Reagent Co., Ltd.) and methanol (Merck Co., Ltd.) were of chromatographic grade; water was ultrapure water (self-made in the laboratory).

[0048] Test drugs: protocatechuic acid (batch number: 110809-201906, content: 97.7%, China Food and Drug Inspection Institute); protocatechuic aldehyde (batch number: 110810-201909, content: 99.6%, China Food and Drug Inspection Institute); neochlorogenic acid (batch number: DSTDX001503, content: 99.6%, Chengdu Lottemei Pharmaceutical Technology Co., Ltd.); chlorogenic acid (batch number: 110753-202119, content: 96.3%, China Food and Drug Inspection Institute); Coptis chinensis reference medicinal materials (batch number: 121034-202104, China Food and Drug Inspection Institute); Coptis chinensis formula granules (batch numbers: KL01, KL02, KL03; source: Guangdong Yifang Pharmaceutical Co., Ltd.). The source information of Coptis chinensis medicinal materials and standard decoctions is shown in Table 1.

[0049] Table 1 Sample source information

[0050]

[0051] 2. Methods and Results

[0052] 2.1 Chromatographic conditions

[0053] A Waters CORTECS C18 chromatographic column (3.0 mm×150 mm, 2.7 μm) was used; methanol was used as mobile phase A, 0.1% phosphoric acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in Table 2; the flow rate was 0.40 ml per minute; the column temperature was 25°C; the detection wavelength was 300 nm; and the injection volume was 1 μl.

[0054] Table 2 Gradient elution table

[0055]

[0056] 2.2 Preparation of reference solution

[0057] Take about 2 g of the control medicinal material of Rhizoma Cynomorii, place it in a stoppered conical flask, add 30 ml of water, heat and reflux for 30 minutes, filter, evaporate to dryness, add 20 ml of 70% methanol to the residue, ultrasonically treat for 30 minutes, filter, and take the filtrate as the control medicinal material reference solution.

[0058] Accurately weigh 2.301 mg of protocatechuic acid reference substance, 2.805 mg of protocatechuic aldehyde reference substance, 2.031 mg of neochlorogenic acid reference substance, and 2.338 mg of chlorogenic acid reference substance, place in a 50 ml volumetric flask, add 70% methanol to make a solution containing 44.9615 μg of protocatechuic acid, 55.8756 μg of protocatechuic aldehyde, 40.4575 μg of neochlorogenic acid, and 45.0299 μg of chlorogenic acid per 1 ml as a reference substance solution.

[0059] 2.3 Preparation of test solution

[0060] (1) Medicinal materials and decoction pieces: Take 2 g of the powder (passed through a No. 3 sieve) and place it in a stoppered conical flask. Add 20 ml of 70% methanol and ultrasonically treat (power 300 W, frequency 40 kHz) for 30 minutes. Cool, shake well, filter, and obtain the filtrate.

[0061] (2) Standard decoction: Take an appropriate amount of this product, grind it into powder, take 1 g, put it in a stoppered conical flask, add 20 ml of 70% methanol, ultrasonically treat (power 300 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate.

[0062] (3) Formula granules: Take an appropriate amount of the product, grind it into powder, take 1 g, place it in a stoppered conical flask, add 20 ml of 70% methanol, and treat it with ultrasound (power 300 W, frequency 40 kHz) for 30 minutes. Cool it, shake it well, filter it, and take the filtrate to obtain the product.

[0063] 2.4 Determination method

[0064] Accurately pipette 1 μl of the reference solution and the test solution respectively, inject into the liquid chromatograph, and measure to obtain the result.

[0065] 3. Durability inspection

[0066] (1) Investigation of different column temperatures

[0067] Take Mianma Guanzhong Formula Granules (KL01) and prepare the test solution according to the test solution preparation method specified in "2.3". Except for the column temperature of 23℃, 25℃ and 27℃, the other chromatographic conditions are the same as those specified in "2.1". Sample injection and analysis are performed. The protocatechuic aldehyde chromatographic peak is used as the reference peak S1 to calculate the relative retention time and relative peak area of ​​peaks 1 to 3, 5 to 7 and S1. The chlorogenic acid chromatographic peak is used as the reference peak S2 to calculate the relative retention time and relative peak area of ​​peaks 9 to 12 and S2. The relative retention time and relative peak area of ​​each characteristic peak and S peak are calculated, and the RSD value is calculated. The results are as follows: Figure 1 The results show that the RSD values ​​of relative retention time and relative peak area are both less than 3.0%, indicating that the method has good durability at different column temperatures.

[0068] (2) Investigation of different flow rates

[0069] Take Mianma Guanzhong Formula Granules (KL01) and prepare the test solution according to the test solution preparation method determined in "2.3". Except that the flow rates are 0.38ml / min, 0.40ml / min and 0.42ml / min respectively, other chromatographic conditions are the same as those specified in "2.1". Sample injection analysis, using the protocatechuic aldehyde chromatographic peak as the reference peak S1, calculate the relative retention time and relative peak area of ​​peaks 1 to 3, peaks 5 to 7 and S1 peak, using the chlorogenic acid chromatographic peak as the reference peak S2, calculate the relative retention time and relative peak area of ​​peaks 9 to 12 and S2 peak, calculate the relative retention time and relative peak area of ​​each characteristic peak and S peak, and calculate the RSD value. The results are as follows: Figure 2 The results show that the RSD values ​​of relative retention time and relative peak area are both less than 5.0%, indicating that the method has good durability for different flow rates.

[0070] 4. Sample determination

[0071] 4.1 Determination of formula granule samples

[0072] Take 3 batches of Mianma Guanzhong formula granules (batch numbers: KL01, KL02, KL03), prepare the test solution according to the test solution preparation method determined in "2.3", accurately pipette the above test solution, and inject and measure according to the chromatographic conditions in "2.1". The characteristic spectrum overlay of the 3 batches of samples is shown in Figure 3 The relative retention time and relative peak area of ​​each characteristic peak and S peak were calculated and shown in Tables 3 and 4.

[0073] Table 3 Characteristic spectrum of Mianma Guanzhong formula granules sample determination (relative retention time)

[0074]

[0075] Table 4 Characteristic spectrum of Mianma Guanzhong formula granules sample determination (relative peak area)

[0076]

[0077] 4.2 Determination of medicinal material samples

[0078] Take the powder of Mianma Guanzhong medicinal material (pass through No. 3 sieve) respectively, prepare the test solution according to the test solution preparation method determined in "2.3", accurately pipette the above test solution respectively, and inject and measure according to the chromatographic conditions in "2.1". The characteristic spectrum overlay of 15 batches of samples is shown in Figure 4 The relative retention time and relative peak area of ​​each characteristic peak and S peak were calculated and shown in Tables 5 and 6.

[0079] Table 5 Determination of characteristic spectrum samples of Mianma Guanzhong medicinal materials (relative retention time)

[0080]

[0081]

[0082] Table 6 Determination of characteristic spectrum samples of Mianma Guanzhong medicinal materials (relative peak area)

[0083]

[0084] 4.3 Determination of decoction pieces samples

[0085] Take the powder of Mianma Guanzhong slices (pass through No. 3 sieve) respectively, prepare the test solution according to the test solution preparation method determined in "2.3", accurately pipette the above test solution respectively, and inject and measure according to the chromatographic conditions in "2.1". The characteristic spectrum overlay of 15 batches of samples is shown in Figure 5 The relative retention time and relative peak area of ​​each characteristic peak and S peak were calculated and shown in Tables 7 and 8.

[0086] Table 7 Characteristic spectrum of Mianma Guanzhong decoction pieces sample determination (relative retention time)

[0087]

[0088] Table 8 Characteristic spectrum of Mianma Guanzhong decoction pieces sample determination (relative peak area)

[0089]

[0090]

[0091] 4.4 Determination of standard decoction samples

[0092] Take the standard decoction of Mianma Guanzhong respectively, prepare the test solution according to the test solution preparation method determined in "2.3", accurately pipette the above test solution respectively, and inject and measure according to the chromatographic conditions in "2.1". The characteristic spectrum overlay of 15 batches of samples is shown in Figure 6 , calculate the relative retention time and relative peak area of ​​each characteristic peak and S peak and see Table 9 to Table 10.

[0093] Table 9 Characteristic spectrum of standard decoction of Mianma Guanzhong Sample determination (relative retention time)

[0094]

[0095]

[0096] Table 10 Characteristic spectrum of standard decoction of Mianma Guanzhong Sample determination (relative peak area)

[0097]

[0098] Example 2: Application of the present invention in the identification of the medicinal material of Mianma cyrtomium and its counterfeit products, Ziqi cyrtomium and cyrtomium

[0099] Take the counterfeit Asteraceae and Cynomorium, and prepare the test solution according to the test solution preparation method under "2.3" in the above Example 1. According to the chromatographic conditions under "2.1" in Example 1, the characteristic spectrum of the control medicinal material of Mianma Cynomorium is used as a reference. The results are shown in Figure 7 It can be seen from the figure that peaks 1, 3, 5, 9, 10, 11, and 12 of batch 1 of Ziqi Cynomorium and batch 2 of Cynomorium are all missing. Therefore, the characteristic spectrum of the present invention can be used to distinguish Mianma Cynomorium from other counterfeit Ziqi Cynomorium and Cynomorium.

[0100] The above is a preferred embodiment of the invention. It should be pointed out that a person skilled in the art can make several improvements and modifications without departing from the principle of the invention. These improvements and modifications are also considered to be within the scope of protection of the invention.

Claims

1. A method for constructing a characteristic spectrum of a medicinal material, a formula granule, a decoction piece or a standard decoction of Rhizoma Cynomorii, characterized in that: The following steps are involved: Prepare a reference solution, the reference solution includes a reference medicinal material reference solution and a reference substance reference solution, the reference substance reference solution includes a protocatechuic acid reference substance, a protocatechuic aldehyde reference substance, a neochlorogenic acid reference substance, and a chlorogenic acid reference substance; Take the medicinal material of Mianma Guanzhong, formula granules, decoction pieces or standard decoction to prepare the test solution; Preset amounts of reference solution and test solution were taken and injected into a liquid chromatograph, which used octadecylsilane bonded silica gel as a filler, methanol as a mobile phase A, and phosphoric acid solution as a mobile phase B for gradient elution to establish a characteristic spectrum.

2. The method for constructing the characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii as claimed in claim 1, characterized in that: The gradient elution was carried out according to the following procedure: 0-2min, mobile phase A is 3%, mobile phase B is 97%; 2min~16min, mobile phase A from 3%→4%, mobile phase B from 97%→96%; 16min-27min, mobile phase A from 4% to 10%, mobile phase B from 96% to 90%; 27min~60min, mobile phase A from 10%→13%, mobile phase B from 90%→87%; 60min~70min, mobile phase A changes from 13%→70%, and mobile phase B changes from 87%→30%.

3. The method for constructing the characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii as claimed in claim 1, characterized in that: The concentration of the phosphoric acid solution is 0.05% to 0.15%.

4. The method for constructing the characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii as claimed in claim 1, characterized in that: The preset amount of the reference solution and the test solution is 0.5 μL to 1.5 μL; The liquid chromatograph uses octadecylsilane bonded silica gel as a filler, with a column length of 150 mm, an inner diameter of 3 mm, a particle size of 2.7 μm, and a column temperature of 23° C. to 27° C.; When performing gradient elution, the flow rate of the mobile phase is 0.38-0.42 mL / min; The ultraviolet detection wavelength of the liquid chromatograph is 300nm-310nm.

5. The method for constructing the characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii as claimed in claim 1, characterized in that: The test solution was prepared by the following method: Take the medicinal materials, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii, grind them into powder, weigh 1g-2g, put them in a stoppered conical flask, add 15mL-25mL of 65%-75% methanol, ultrasonically treat for 25min-35min, cool, shake well, filter, and take the filtrate to obtain the product.

6. The method for constructing the characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii as claimed in claim 1, characterized in that: The reference substance solution was prepared according to the following method: Take appropriate amount of protocatechuic acid reference substance, protocatechuic aldehyde reference substance, neochlorogenic acid reference substance, and chlorogenic acid reference substance, accurately weigh them, and add 70% methanol to make a mixed solution containing 50 μg of each per 1 ml, which serves as the reference substance solution.

7. The method for constructing the characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii as claimed in claim 1, characterized in that: The control medicinal material reference solution is prepared by the following method: Take about 1.5g to 2g of the control medicinal material of Rhizoma Cynomorii, place it in a stoppered conical flask, add 25mL to 35mL of water, heat and reflux for 25min to 35min, filter, evaporate to dryness, add 15mL to 25mL of 65% to 75% methanol to the residue, ultrasonically treat for 25min to 35min, filter, and take the filtrate as the control medicinal material reference solution.

8. The method for constructing the characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii as claimed in claim 1, characterized in that: The characteristic spectrum includes 12 common peaks, among which peak No. 2 corresponds to protocatechuic acid, peak No. 4 corresponds to protocatechuic aldehyde, peak No. 5 corresponds to neochlorogenic acid, and peak No. 8 corresponds to chlorogenic acid.

9. The method for constructing the characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cynomorii as claimed in claim 8, characterized in that: In the characteristic spectrum of the Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction, the protocatechuic aldehyde peak is used as the reference peak S1, and the relative retention times of peaks 1 to 3, 5 to 7 and the reference peak S1 are consistent with: peak 1 0.275-0.32, peak 2 0.585-0.597, peak 3 0.796-0.894, peak 5 1.253-1.292, peak 6 1.431-1.528, peak 7 1.516-1.609; Taking the chlorogenic acid peak as the reference peak S2, the relative retention times of peaks 9 to 12 and the reference peak S2 are consistent with: peak 9 1.022-1.040, peak 10 1.142-1.168, peak 11 1.177-1.223, peak 12 1.265-1.

328.

10. Use of the characteristic spectrum of the Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction obtained by the method for constructing the characteristic spectrum of the Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction according to any one of claims 1 to 9 in any one of (1) to (3): (1) Authenticity identification and ingredient testing of the medicinal materials, formula granules, decoction pieces or standard decoctions of Rhizoma Cynomorii; (2) Distinguishing between the medicinal materials, formula granules, decoction pieces or standard decoctions of Rhizoma Cynomorii and the counterfeit Rhizoma Cynomorii; (3) Differentiate between the medicinal materials, formula granules, decoction pieces or standard decoctions of Rhizoma Cynomorium and counterfeit Rhizoma Cynomorium.

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