Method for constructing characteristic maps of Mianma Guanzhong medicinal materials, Chinese medicine formula granules, decoction pieces or standard decoctions and their applications

Abstract

The present invention discloses a method for constructing a characteristic spectrum of a Corydalis guanzhong medicinal material, formula granules, decoction pieces or standard decoction, comprising the following steps: preparing a reference solution, the reference solution comprising a control 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; taking a Corydalis guanzhong medicinal material, formula granules, decoction pieces or standard decoction to prepare a test solution; taking a preset amount of the reference solution and the test solution, injecting them into a liquid chromatograph, the liquid chromatograph using octadecylsilane bonded silica gel as a filler, methanol as mobile phase A, and phosphoric acid solution as mobile phase B for gradient elution to establish a characteristic spectrum. The present invention can provide a data basis for the quality control of Corydalis guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions, and ensure the stability and controllability of product quality.

Description

Technical Field

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

[0002] Mianma Guanzhong (Mianma Guanzhong) is the dried rhizome and petiole residues of the Dryopteris crassirhizoma Nakai fern, a member of the Dryopteris family. It is listed in the 2020 edition of the Chinese Pharmacopoeia. It has the effects of clearing heat, detoxifying, and expelling parasites. It is used to treat abdominal pain caused by parasites and sores.

[0003] In the existing technology, the research on Rhizoma Cyrthospermi has focused on the preparation method of water extracts 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 Cyrthospermi, such as mynaic acid ABB, mynacrosol ABBA and mynacrosol ABBP, which mainly play an anti-influenza virus role. At present, the 2020 edition of the Chinese Pharmacopoeia includes the Rhizoma Cyrthospermi medicinal material standard, using thin-layer identification and extracts as quality control methods, and there are no relevant regulations on characteristic spectra. "Study on HPLC Fingerprint of Rhizoma Cyrthospermi Formula Granules" uses high-performance liquid chromatography to establish the characteristic spectrum of Rhizoma Cyrthospermi formula granules and identify 12 common peaks, but there are problems such as fuzzy baselines and poor chromatographic peak separation. From an application perspective, there are many counterfeit Rhizoma Cyrthospermi, such as Rhizoma Cyrthospermi, Rhizoma Cyrthospermi, Rhizoma Cyrthospermi, Rhizoma Cyrthospermi, Rhizoma Cyrthospermi, Rhizoma Cyrthospermi, Rhizoma Cyrthospermi, Rhizoma Cyrthospermi, Rhizoma Cyrthospermi, etc. Currently, the literature reports only differentiate the medicinal materials based on their properties, lacking an objective and effective method for identification. Therefore, 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 the medicinal materials, formula granules, medicinal slices or standard decoctions of Mianma Guanzhong. The method has good reproducibility, accuracy and reliability, and can provide a data basis for the mass production quality control of the medicinal materials, formula granules, medicinal slices or standard decoctions of Mianma Guanzhong, thereby ensuring the stability and controllability of the product quality of the medicinal materials, formula granules, medicinal slices or standard decoctions of Mianma Guanzhong.

[0005] In order to solve the above technical problems, the present invention provides a method for constructing a characteristic spectrum of a medicinal material, formula granules, decoction pieces or standard decoction of Mianma Guanzhong, comprising the following steps: preparing a reference solution, wherein the reference solution includes a control medicinal material reference solution and a reference substance reference solution, wherein 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;

[0006] Prepare the test solution from the medicinal material, formula granules, slices or standard decoction of Mian Ma Guan Zhong;

[0007] Take a preset amount of reference solution and test solution and inject them into a liquid chromatograph. The liquid chromatograph uses octadecylsilane bonded silica gel as a filler, methanol as mobile phase A, and phosphoric acid solution as 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] 0-2 min, mobile phase A is 3%, mobile phase B is 97%;

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

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

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

[0013] From 60 to 70 minutes, the concentration of mobile phase A increased from 13% to 70%, and that of mobile phase B increased from 87% to 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 to 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, place 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 is used 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 Mian Ma Guan Zhong, 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 2 corresponds to protocatechuic acid, peak 4 corresponds to protocatechuic aldehyde, peak 5 corresponds to neochlorogenic acid, and peak 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, with the protocatechuic aldehyde peak as the reference peak S1, the relative retention times of peaks 1 to 3, 5 to 7 and the reference peak S1 are as follows: 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 Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions;

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

[0032] (3) Distinguish between the medicinal materials, formula granules, decoction pieces or standard decoctions of Mianma Guanzhong and counterfeit Guanzhong.

[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. The characteristic spectrum can fully display the chemical composition characteristics of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions, has rich characteristic peak information, a stable baseline, and good peak separation, and can comprehensively reflect the quality information of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions, thereby achieving the purpose of comprehensively and effectively controlling the quality of Mianma Guanzhong medicinal materials, formula granules, decoction pieces or standard decoctions. It can also achieve 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 This is a chromatogram of the durability of the characteristic spectrum of the Mianma Guanzhong formula granules in Example 1 of the present invention - investigated at different column temperatures;

[0036] Figure 2 This 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 rates;

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

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

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

[0040] Figure 6 This 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 Bianchii and Rhizoma Cyrthospermi in Example 2 of the present invention. DETAILED DESCRIPTION

[0042] In order to make the objectives, 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 the medicinal materials, formula granules, decoction pieces or standard decoctions of Mianma Guanzhong, and to achieve comprehensive and effective quality control of the medicinal materials, formula granules, decoction pieces or standard decoctions of Mianma Guanzhong, the present invention provides a method for constructing a characteristic spectrum of the medicinal materials, formula granules, decoction pieces or standard decoctions of Mianma Guanzhong, which is described in detail below:

[0044] Example 1: Establishment of a characteristic spectrum of Rhizoma Cyrthospermi 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); 1 / 10,000 balance (ME204E, Mettler-Toledo); 1 / 100,000 balance (ABT220-5DM, Guangzhou Dexiang Technology Co., Ltd.); 1 / 100,000 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).

[0047] Reagents: Methanol (Tianjin Fuyu Fine Chemical Co., Ltd.) was of analytical grade, phosphoric acid (Tianjin Kemiou Chemical Reagent Co., Ltd.) and methanol (Merck Co., Ltd.) were of chromatographic grade; water was ultrapure water (prepared 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 Lotte Beauty Pharmaceutical Technology Co., Ltd.); chlorogenic acid (batch number: 110753-202119, content: 96.3%, China Food and Drug Inspection Institute); Mianma Guanzhong reference medicinal material (batch number: 121034-202104, China Food and Drug Inspection Institute); Mianma Guanzhong formula granules (batch numbers: KL01, KL02, KL03; source: Guangdong Yifang Pharmaceutical Co., Ltd.). The source information of Mianma Guanzhong 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 column (3.0 mm × 150 mm, 2.7 μm) was used; methanol was used as mobile phase A, and 0.1% phosphoric acid solution was used as mobile phase B, and gradient elution was performed according to the requirements 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 Cyrthospermi, 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 them in a 50 ml volumetric flask, and 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 the reference substance solution.

[0059] 2.3 Preparation of test solution

[0060] (1) Medicinal materials and decoction pieces: Take 2 g of the powder (pass through 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. Let it cool, shake well, filter, and take the filtrate.

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

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

[0063] 2.4 Assay

[0064] Accurately pipette 1 μl of reference solution and test solution respectively, inject into 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 determined under "2.3". Except that the column temperature is 23℃, 25℃ and 27℃ respectively, the other chromatographic conditions are the same as those specified under "2.1". Sample injection and analysis are performed. The protocatechuic aldehyde chromatographic peak is used as the reference peak S1. The relative retention time and relative peak area of ​​peaks 1 to 3, 5 to 7 and S1 are calculated. The chlorogenic acid chromatographic peak is used as the reference peak S2. The relative retention time and relative peak area of ​​peaks 9 to 12 and S2 are calculated. 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 showed that the RSD values ​​of relative retention time and relative peak area were both less than 3.0%, indicating that the method had 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 under "2.3". Except that the flow rates were 0.38ml / min, 0.40ml / min and 0.42ml / min respectively, other chromatographic conditions were the same as those specified under "2.1". Sample injection and analysis were performed. The protocatechuic aldehyde chromatographic peak was used as the reference peak S1. The relative retention time and relative peak area of ​​peaks 1 to 3, peaks 5 to 7 and S1 were calculated. The chlorogenic acid chromatographic peak was used as the reference peak S2. The relative retention time and relative peak area of ​​peaks 9 to 12 and S2 were calculated. The relative retention time and relative peak area of ​​each characteristic peak and S peak were calculated, and the RSD value was calculated. The results are shown as follows: Figure 2 The results showed that the RSD values ​​of relative retention time and relative peak area were both less than 5.0%, indicating that the method had good durability for different flow rates.

[0070] 4. Sample Measurement

[0071] 4.1 Determination of Formula Granule Samples

[0072] Take 3 batches of Mianma Guanzhong granules (batch number: KL01, KL02, KL03), prepare the test solution according to the test solution preparation method determined in "2.3", accurately aspirate 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 Table 3 to Table 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 the medicinal material of Mianma Guanzhong (pass through No. 3 sieve) respectively, prepare the test solution according to the test solution preparation method determined in "2.3", accurately aspirate 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 Characteristic spectrum of the medicinal material of Mianma Guanzhong Sample determination (relative retention time)

[0080]

[0081]

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

[0083]

[0084] 4.3 Decoction Sample Determination

[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 aspirate 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 Mian Ma Guan Zhong respectively, prepare the test solution according to the test solution preparation method determined in "2.3", accurately aspirate 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 The relative retention time and relative peak area of ​​each characteristic peak and S peak were calculated and shown in Tables 9 and 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 Mianma Guanzhong medicinal material and its counterfeit products Ziqi Guanzhong and Guanzhong

[0099] Take the counterfeit Ziqi Guanzhong and Guanzhong in appropriate amounts, 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 Guanzhong is used as a reference. The results are shown in FIG. Figure 7 It can be seen from the figure that peaks 1, 3, 5, 9, 10, 11, and 12 of batch 1 of Ziqi Cyperus and batch 2 of Cyperus are all missing. Therefore, the characteristic spectrum of the present invention can be used to distinguish Mianma Cyperus from other counterfeit Ziqi Cyperus and Cyperus.

[0100] The above is a preferred embodiment of the invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the invention. These improvements and modifications are also considered to be within the scope of protection of the present invention.

Claims

1. A method for constructing a characteristic spectrum of a medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cyrthospermi, characterized in that: The following steps are involved: Prepare a reference solution, the reference solution including a control medicinal material reference solution and a reference substance reference solution, the reference substance reference solution including a protocatechuic acid reference substance, a protocatechuic aldehyde reference substance, a neochlorogenic acid reference substance, and a chlorogenic acid reference substance; Prepare the test solution from the medicinal material, formula granules, slices or standard decoction of Mian Ma Guan Zhong; The test solution was prepared by the following method: Grind the medicinal materials, formula granules, slices or standard decoction of Rhizoma Cynomorii into powder, weigh 1g-2g, place 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. Take a preset amount of reference solution and test solution and inject them into a liquid chromatograph. The liquid chromatograph uses a Waters CORTECS C18 column with a column length of 150 mm, an inner diameter of 3 mm, and a particle size of 2.7 μm. Methanol is used as mobile phase A and phosphoric acid solution is used as mobile phase B for gradient elution to establish a characteristic spectrum. The concentration of the phosphoric acid solution is 0.05% to 0.15%, and the ultraviolet detection wavelength of the liquid chromatograph is 300nm to 310nm; The gradient elution was performed according to the following procedure: 0-2 min, mobile phase A was 3%, mobile phase B was 97%; 2 min to 16 min, mobile phase A from 3% to 4%, mobile phase B from 97% to 96%; 16-27 min, mobile phase A from 4% to 10%, mobile phase B from 96% to 90%; 27 min to 60 min, mobile phase A from 10% to 13%, mobile phase B from 90% to 87%; From 60 to 70 minutes, mobile phase A increased from 13% to 70%, and mobile phase B increased from 87% to 30%; 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.

2. The method for constructing a characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cyrthospermi as claimed in claim 1, characterized in that: The preset volume of the reference solution and the test solution is 0.5 μL~1.5 μL; The column temperature of the chromatographic column is 23°C to 27°C; When performing the gradient elution, the flow rate of the mobile phase is 0.38-0.42 mL / min.

3. The method for constructing a characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cyrthospermi 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 is used as the reference substance solution.

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

5. The method for constructing a characteristic spectrum of the medicinal material, formula granules, decoction pieces or standard decoction of Rhizoma Cyrthospermi as claimed in claim 1, characterized in that: In the characteristic spectrum of the described Mianma Guanzhong medicinal material, formula granules, decoction pieces or standard decoction, with the protocatechuic aldehyde peak as the reference peak S1, the relative retention times of peaks 1 to 3, 5 to 7 and the reference peak S1 are as follows: 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.

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