A method for detecting traditional Chinese medicine compound fingerprint based on continuous extraction strategy

Abstract

The present application belongs to the technical field of traditional Chinese medicine compound component analysis and detection, and particularly relates to a traditional Chinese medicine compound fingerprint detection method based on a continuous extraction strategy. The method is realized through the following steps: firstly, the test sample of Jingfang granules is continuously extracted using extraction solvents with different polarities to prepare test solutions of different polarity parts; an internal standard solution is prepared; the test sample solution is detected for fingerprint by using ultra-high performance liquid chromatography-mass spectrometry, quantitatively characterized, and each component is subjected to attribution analysis. The present application compares different combined solvent extraction methods through qualitative and quantitative analysis, determines the best continuous extraction strategy, optimizes the extraction method, establishes the UPLC-MS fingerprint detection method of Jingfang granules based on the continuous extraction strategy, analyzes 112 compounds in total, and provides technical support for the precise quality evaluation of Jingfang granules.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine compound component analysis and detection, and particularly relates to a traditional Chinese medicine compound fingerprint detection method based on a continuous extraction strategy. BACKGROUND

[0002] Jingfang granules are traditional Chinese medicine preparations prepared by extraction, processing and concentration using modern pharmaceutical technology. Modern pharmacological studies have shown that Jingfang granules have the effects of relieving fever, relieving pain and anti-inflammation. Jingfang granules are mainly used for treating wind-cold type of common cold, fever with aversion to cold, nasal obstruction and rhinorrhea, cough with white sputum, and sore throat with sore throat.

[0003] Jingfang granules are prepared from 11 kinds of traditional Chinese medicines, including Schizonepeta, Saposhnikovia, Bupleurum, Chuanxiong, Notopterygium, Angelica pubescens, Peucedanum, Poria cocos, Platycodon, Citrus aurantium and Glycyrrhiza, in a certain proportion. The taste of Jingfang granules is mainly pungent and bitter. Pungent taste has the effects of dispersing, promoting blood circulation and resolving dampness, while bitter taste can clear heat and resolve toxins, and dry dampness and descend qi. Schizonepeta in the prescription has the effects of relieving the exterior and dispelling wind, and can also resolve rash and eliminate sore. Saposhnikovia has the effects of dispelling wind and relieving the exterior, and can also overcome dampness and relieve pain. The combination of the two can dispel wind and relieve the exterior, and can also be used as the monarch drug. Bupleurum has the effects of relieving the exterior and resolving heat. Chuanxiong has the effects of promoting blood circulation and relieving pain. Notopterygium has the effects of relieving the exterior and dispelling wind and dampness. Angelica pubescens has the effects of dispelling wind and dampness and relieving pain. The combination of Notopterygium and Angelica pubescens can dispel wind and dampness, and relieve headache and body pain. The three can be used as the ministerial drug. Peucedanum has the effects of dispelling wind and heat, and can also descend qi and resolve phlegm. Poria cocos has the effects of promoting water and resolving dampness. Platycodon has the effects of promoting lung and resolving phlegm, and can also carry drugs upward. Citrus aurantium has the effects of resolving phlegm and promoting qi. Glycyrrhiza can be used as the assistant drug, and has the effects of regulating the other drugs and tonifying the spleen and stomach. The combination of the above-mentioned drugs can relieve the exterior and dispel wind, and can also relieve the interior and dispel dampness, and can be used to treat wind-cold type of common cold, and can also be used to treat headache and body pain, nasal obstruction and rhinorrhea, and sore throat with sore throat. Modern pharmacological studies have shown that Jingfang granules have the effects of relieving fever, relieving pain and anti-inflammation, and can be used to treat wind-cold type of common cold, headache and body pain, and nasal obstruction and rhinorrhea.

[0004] Jingfang granules contain a large number of drugs, and the chemical components contained therein are complex and diverse. Formulating a complete analysis strategy and analyzing the components in detail can provide technical support for the research on the material basis of efficacy of Jingfang granules and the improvement of quality standards.

[0005] Jiang Li et al. established a solid phase extraction-ultra performance liquid chromatography (SPE-UPLC) method for simultaneously determining the contents of epimedoside and 5-O-methylvisamminol glycoside in Jingfang granules. Liu Wen et al. established an HPLC method for simultaneously determining the contents of epimedoside, asafetida acid, naringin, menthone, osthole and isoimperatorin in Jingfang granules.

[0006] Liang Hongbao et al. used UPLC-Q-Exactive MS technology to detect the non-volatile chemical components in Jingfang granules in positive and negative ion modes, and through mass spectrum information combined with reference substances, related literature and database retrieval, a total of 85 compounds were identified.

[0007] The above research methods have some shortcomings: (1) High performance liquid chromatography is only for the detection of coumarin, organic acid and flavonoid components in the components of Fangfeng, Jujube, Duhuo and Chuanhu in Jingfang granules. It cannot comprehensively evaluate the quality of traditional Chinese medicine preparation and prove the consistency of product quality. (2) Since Jingfang granules contain 11 kinds of traditional Chinese medicines, and the reported chemical components are 85 kinds, simple use of one kind of solvent extraction will cause partial loss or incomplete extraction of some components, so that the later component detection cannot truly reflect the types and content of the components contained in the sample. Therefore, it is of great significance to establish a new quality control method for Jingfang granules. SUMMARY

[0008] In view of the problems existing in the prior art, the present application provides a traditional Chinese medicine compound fingerprint detection method based on a continuous extraction strategy. The method prepares sample solution of Jingfang granules according to different polarities, uses gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry and other component analysis technologies, obtains Jingfang granule fingerprint detection method, reflects the types and quantities of chemical components in the compounds, and evaluates the quality of the traditional Chinese medicine compound as a whole. It can truly realize the effective characterization, comprehensive evaluation and overall control of the internal quality of traditional Chinese medicine, establish a new quality control method, and solve the problem that simple use of one kind of extraction method cannot accurately control the quality of traditional Chinese medicine and evaluate the stability of quality.

[0009] The technical scheme adopted by the present application to achieve the above-mentioned purpose is:

[0010] The present application provides a traditional Chinese medicine compound fingerprint detection method based on a continuous extraction strategy, comprising the following steps:

[0011] (1) Preparation of sample solution: the sample Jingfang granules are continuously extracted by different polar solvents to prepare different polar part sample solutions;

[0012] (2) Preparation of internal standard solution: 1.00mg of puerarin is selected as the internal standard, 1ml of 70% methanol solution is added, and ultrasonic is performed for 2min. The internal standard solution is added to the sample to prepare a sample solution containing the internal standard with a concentration of 10ug / ml.

[0013] (3) Sample detection: the sample solution is detected by ultra-high performance liquid chromatography-mass spectrometry to perform fingerprint detection, quantitative characterization and attribution analysis of each component.

[0014] Further, in step (1), the different polarity extraction solvent is ethyl acetate-95% ethanol-30% ethanol.

[0015] The specific process for preparing the test sample solution is as follows:

[0016] Take Jingfang Granules 0.50043 g, accurately weigh, add ethyl acetate 20 mL, weigh, heat reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, add 10 ml of 70% methanol to dissolve;

[0017] Take the filtrate, add 95% ethanol 20 mL, weigh, heat reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, add 70% methanol to dissolve;

[0018] Take the filtrate, add 30% ethanol 20 mL, weigh, heat reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, add 70% methanol to dissolve;

[0019] Combine the above 70% methanol solution, evaporate to dryness, add 70% methanol to make up to 10 mL in a volumetric flask, mix well, then transfer to a centrifuge tube, centrifuge for 5 min (speed 10000 r / min), take an appropriate amount of supernatant, pass through a 0.22 μm microporous filter membrane into a sample bottle, store at 4°C for standby.

[0020] The specific process of step (1) can also be:

[0021] Take Jingfang Granules 0.50043 g, accurately weigh, add ethyl acetate 20 mL, weigh, heat reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, add 10 ml of 70% methanol to dissolve;

[0022] Take the filtrate, add 95% ethanol 20 mL, weigh, heat reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, add 70% methanol to dissolve;

[0023] Take the filtrate, add 70% ethanol 20 mL, weigh, heat reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, add 70% ethanol to make up to 10 mL in a volumetric flask;

[0024] Take the filtrate, add 30% ethanol 20 mL, weigh, heat reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, add 30% ethanol to make up to 10 mL in a volumetric flask;

[0025] Take the filtrate, add water 20 mL, weigh, heat reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, add water to make up to 10 mL in a volumetric flask;

[0026] After mixing the extracted samples, transfer them to centrifuge tubes and centrifuge for 5 minutes (10,000 r / min). Take an appropriate amount of the supernatant, filter it through a 0.22 μm microporous membrane into a sample vial, and store it at 4°C for later use.

[0027] Furthermore, in step (3), the detection conditions for liquid chromatography-mass spectrometry are as follows: chromatographic conditions: Therma thermoelectric column (100 mm × 2.1 mm, 2.7 μm), mobile phase is 0.1% formic acid aqueous solution (A)-methanol (B), flow rate is 0.3 mL / min, column temperature is 30℃, injection volume is 2 µL, and gradient elution is performed.

[0028] The gradient elution procedure described above is as follows:

[0029]

[0030] Furthermore, the mass spectrometry conditions are as follows: electrospray ionization (ESI) source, positive and negative ion monitoring modes; positive ion spray voltage: 3500 V, negative ion spray voltage: 3000 V, sheath gas is nitrogen, pressure is 35 arb; auxiliary gas is nitrogen, pressure is 10 arb; capillary temperature is 350℃, and scanning mass range is m / z 100~1000.

[0031] The beneficial effects of this invention are as follows:

[0032] (1) Jingfang granules are composed of multiple Chinese herbal medicines and belong to a complex compound preparation. It is difficult to conduct a comprehensive analysis of the components. In this invention, the test solution is prepared by continuous extraction with extraction solvents of different polarities (ethyl acetate, 95% ethanol, 70% ethanol, 30% ethanol and water). The test solution is prepared by using different polar fractions to characterize the component characteristics of Jingfang granules.

[0033] (2) This invention determined the optimal continuous extraction strategy by comparing different combination solvent extraction methods through qualitative and quantitative analysis, optimized the extraction method, and established a UPLC-MS fingerprint detection method for Jingfang granules based on the continuous extraction strategy. A total of 112 compounds were analyzed, providing technical support for the accurate quality evaluation of Jingfang granules. Attached Figure Description

[0034] Figure 1 Total ion chromatogram (positive ion mode) of UPLC-Q-Exactive-Orbitrap-MS for ethyl acetate extraction of *Jingfang* granules.

[0035] Figure 2 Total ion chromatogram (negative ion mode) of UPLC-Q-Exactive-Orbitrap-MS for ethyl acetate extraction of Jingfang granules.

[0036] Figure 3 Total ion chromatogram (positive ion mode) of UPLC-Q-Exactive-Orbitrap-MS for 95% ethanol extraction of Jingfang granules;

[0037] Figure 4 Total ion chromatogram (negative ion mode) of UPLC-Q-Exactive-Orbitrap-MS for 95% ethanol extraction of Jingfang granules;

[0038] Figure 5 Total ion chromatogram (positive ion mode) of UPLC-Q-Exactive-Orbitrap-MS for 70% ethanol extraction of Jingfang granules;

[0039] Figure 6 Total ion chromatogram (negative ion mode) of UPLC-Q-Exactive-Orbitrap-MS for 70% ethanol extraction of Jingfang granules;

[0040] Figure 7 Total ion chromatogram (positive ion mode) of UPLC-Q-Exactive-Orbitrap-MS for 30% ethanol extraction of Jingfang granules;

[0041] Figure 8 Total ion chromatogram (negative ion mode) of UPLC-Q-Exactive-Orbitrap-MS for 30% ethanol extraction of Jingfang granules;

[0042] Figure 9 Total ion chromatogram (positive ion mode) for water extraction of *Jingfang* granules by UPLC-Q-Exactive-Orbitrap-MS.

[0043] Figure 10 Total ion chromatogram (negative ion mode) for water extraction of *Jingfang* granules by UPLC-Q-Exactive-Orbitrap-MS.

[0044] Figure 11 Total ion chromatogram (positive ion mode) of ethyl acetate-95% ethanol-30% ethanol extract of Jingfang granules was obtained by UPLC-Q-Exactive-Orbitrap-MS.

[0045] Figure 12 Total ion chromatogram (negative ion mode) of ethyl acetate-95% ethanol-30% ethanol extract of Jingfang granules was obtained by UPLC-Q-Exactive-Orbitrap-MS.

[0046] Figure 13 Spectrum of puerarin internal standard;

[0047] Figure 14 Spectrum of clarithromycin internal standard assay;

[0048] Figure 15 The chromatogram of tinidazole internal standard was investigated.

[0049] Figure 16 Chloramphenicol internal standard assay chromatogram;

[0050] Figure 17 Spectrum of puerarin internal standard and blank sample. Detailed Implementation

[0051] The technical solution of the present invention will be further explained and described below through specific embodiments.

[0052] Example 1

[0053] 1. Materials and Methods

[0054] 1.1 Instruments and Materials

[0055] 1.1.1 Instruments

[0056] Table 1 List of Main Instruments

[0057]

[0058] 1.1.2 Materials

[0059] Jingfang granules (Shandong New Era Pharmaceutical Co., Ltd.), Thermo thermoelectric column (100 mm × 2.1 mm, 2.7 μm), purified water (Guangzhou Watsons Food & Beverage Co., Ltd.), methanol and formic acid were of mass spectrometry grade (Thermo Fisher Scientific, USA), and other reagents were of analytical grade.

[0060] 1.2 Methods

[0061] 1.2.1 Detection conditions for liquid chromatography-mass spectrometry (LC-MS)

[0062] Chromatographic conditions: Therma thermoelectric column (100 mm × 2.1 mm, 2.7 μm), mobile phase 0.1% formic acid aqueous solution (A)-methanol (B), flow rate 0.3 mL / min, column temperature 30℃, injection volume 2 µL, gradient elution according to Table 2.

[0063] Table 2 Gradient elution conditions

[0064]

[0065] Electrospray ionization (ESI) source, positive and negative ion monitoring modes; positive ion spray voltage: 3500 V, negative ion spray voltage: 3000 V, sheath gas is nitrogen, pressure is 35 arb; auxiliary gas is nitrogen, pressure is 10 arb; capillary temperature is 350℃, scanning quality range is m / z 100~1000.

[0066] 1.2.2 Preparation of the test solution

[0067] 1.2.2.1 Preparation of liquid chromatography-mass spectrometry (LC-MS) test solution

[0068] Accurately weigh 0.50039g of Jingfang granules, add 20 mL of ethyl acetate, weigh, heat under reflux for 40 min, cool, make up the weight loss, shake well, filter, evaporate the filtrate to dryness, and dilute to 10 mL in a volumetric flask with methanol.

[0069] Take the filter residue, add 20 mL of 95% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add 95% ethanol to make up to 10 mL in a volumetric flask.

[0070] Take the filter residue, add 20 mL of 70% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add 70% ethanol to make up to 10 mL in a volumetric flask.

[0071] Take the filter residue, add 20 mL of 30% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add 30% ethanol to make up to 10 mL in a volumetric flask.

[0072] Take the filter residue, add 20 mL of water, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add water to make up to 10 mL in a volumetric flask.

[0073] After mixing the extracted samples, transfer them to centrifuge tubes and centrifuge for 5 minutes (10,000 r / min). Take an appropriate amount of the supernatant, filter it through a 0.22 μm microporous membrane into a sample vial, and store it at 4°C for later use.

[0074] 1.2.3 Preparation of internal standard solution

[0075] Clarithromycin, tinidazole, chloramphenicol, and puerarin were selected as internal standards for investigation. The extraction ion chromatograms of the four internal standards in the sample solution are shown below. Figures 13-17As shown, clarithromycin was only identified in positive ion mode, possibly due to its easy degradation in acid bath solutions. Chloramphenicol could not be separated from the active ingredient peak in positive ion mode, and because it contains highly electronegative Cl-, it readily dehydrogenates to generate negative ions. Tinidazole showed no detected fragment ions in negative ion mode. Puerarin exhibited high peak intensity in both positive and negative ion modes and could be well separated from the active ingredient in the sample; therefore, puerarin was ultimately chosen as the internal standard solution: 1.00 mg of puerarin was weighed, added to 1 ml of 70% methanol solution, sonicated for 2 min, and the internal standard solution was added to the sample to prepare a 10 μg / ml sample solution containing the internal standard.

[0076] 2 Results

[0077] 2.1 Establishment of total ion chromatogram

[0078] The gas chromatography-mass spectrometry (GC-MS) and total ion chromatogram of the Jingfang particles are shown below. Figures 1-10 Total ion chromatograms of liquid chromatography-mass spectrometry (LC-MS) under positive and negative ion modes for the Jingfang particles are shown in [reference needed]. Figures 11-12 Preliminary observations revealed that the instrument is relatively stable and produces a clear total ion chromatogram, making it suitable for data analysis with highly reliable results.

[0079] Example 2

[0080] Operating steps:

[0081] 1. Materials and Methods

[0082] 1.1 Instruments and Materials

[0083] Same as Example 1

[0084] 1.2 Methods

[0085] 1.2.1 Detection conditions for liquid chromatography-mass spectrometry

[0086] Same as Example 1

[0087] 1.2.2 Preparation of the test solution

[0088] 1.2.2.1 Preparation of liquid chromatography-mass spectrometry (LC-MS) test solution

[0089] Take 0.50043g of Jingfang granules, accurately weigh them, add 20mL of ethyl acetate, weigh them, heat under reflux for 40min, cool them, make up the weight loss, shake well, filter them, evaporate the filtrate to dryness, and add 10ml of 70% methanol to dissolve them.

[0090] Take the filter residue, add 20 mL of 95% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add 10 mL of 70% methanol to dissolve it.

[0091] Take the filter residue, add 20 mL of 30% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add 10 mL of 70% methanol to dissolve it.

[0092] Combine the above 70% methanol solutions, evaporate to dryness, add 70% methanol to a 10mL volumetric flask, mix well, transfer to a centrifuge tube, centrifuge for 5min (10000 r / min), take an appropriate amount of supernatant and filter it through a 0.22μm microporous membrane into a sample vial, and store at 4℃ for later use.

[0093] 1.2.3 The preparation of the internal standard solution is the same as in Example 1;

[0094] 1.2.4 Identification of chemical composition of Jingfang granules

[0095] Same as Example 1

[0096] 2 Results

[0097] 2.1 Establishment of total ion chromatogram

[0098] Total ion chromatograms of liquid chromatography-mass spectrometry (LC-MS) under positive and negative ion modes for Jingfang particles are shown below. Figures 11-12 Preliminary observations revealed that the instrument is relatively stable and produces a clear total ion chromatogram, making it suitable for data analysis with highly reliable results.

[0099] Example 1

[0100] Because the schizonepeta granules contain a variety of components, Example 1 selected extraction solvents of different polarities (ethyl acetate, 95% ethanol, 70% ethanol, 30% ethanol, and water) for investigation. Example 1 used liquid chromatography-mass spectrometry (LC-MS) to preliminarily analyze and identify the chemical components of different extraction fractions of the schizonepeta granules. Based on the precise relative molecular mass and mass spectrometry fragment structure information, a total of 27 compounds were analyzed from the ethyl acetate fraction of the schizonepeta granules, 107 compounds from the 95% ethanol fraction, 15 compounds from the 70% ethanol fraction, 18 compounds from the 30% ethanol fraction, and 8 compounds from the water fraction. After removing duplicate components from each fraction, a total of 110 compounds were identified in the schizonepeta granules, which is 25 more than previously reported. The detection results of different fractions are shown in Table 3.

[0101] Example 2

[0102] Case 2: Although the extraction method provided in Example 1 could comprehensively characterize the chemical components of Jingfang granules, the stepwise extraction operation was cumbersome. Through research, Example 2 ultimately selected continuous extraction using ethyl acetate-95% ethanol-30% ethanol, ultimately identifying 112 compounds in Jingfang granules, two more components (floxacin / isofloxacin) than in Example 1. The reason for this may be that Scheme 2 used 70% methanol as the final dissolution and volume adjustment solvent, and flavonoid glycosides are more soluble in 70% methanol than 95% ethanol, thus increasing their solubility in the test solution. Among them, 61 non-volatile components were discovered and detected in Jingfang granules for the first time. The composition of each component by this method is summarized in Table 3.

[0103] Table 3 Summary of the analysis results of 112 chemical components of Jingfang Granules

[0104]

[0105]

[0106]

[0107]

[0108]

Claims

1. A method for detecting the fingerprint spectrum of traditional Chinese medicine compound prescriptions based on a continuous extraction strategy, characterized in that, Includes the following steps: (1) Preparation of test solution: The test sample Jingfang granules were continuously extracted using extraction solvents of different polarities to prepare test solutions of different polarities; (2) Preparation of internal standard solution: Puerarin was selected as the internal standard. Puerarin was weighed, added to methanol solution, and ultrasonically mixed. The internal standard solution was added to the test sample solution to prepare a sample solution containing the internal standard. (3) Sample detection: The fingerprint spectrum of the test solution was detected by ultra-high performance liquid chromatography coupled with mass spectrometry, quantitative characterization was performed, and the components were assigned to each component. The specific process of step (1) is as follows: Take 0.50043g of Jingfang granules, accurately weigh them, add 20 mL of ethyl acetate, weigh them, heat under reflux for 40 min, cool them, make up the weight loss, shake them well, filter them, evaporate the filtrate to dryness, add 10 mL of 70% methanol to dissolve them, and obtain a 70% methanol solution. Take the filter residue, add 20 mL of 95% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, add 70% methanol to dissolve it, and obtain a 70% methanol solution. Take the filter residue, add 20 mL of 30% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, add 70% methanol to dissolve it, and obtain a 70% methanol solution. Combine the 70% methanol solutions, evaporate to dryness, add 70% methanol to a 10mL volumetric flask, mix well, transfer to a centrifuge tube, centrifuge for 5 min at 10000 r / min, take the supernatant and filter it through a 0.22μm microporous membrane into a sample vial, store at 4℃ for later use; In step (3), the detection conditions for liquid chromatography-mass spectrometry (LC-MS) are as follows: Chromatographic conditions: Therma thermoelectric column, with dimensions of 100 mm × 2.1 mm and 2.7 μm; mobile phase A is 0.1% formic acid aqueous solution, mobile phase B is methanol, flow rate is 0.3 mL / min, column temperature is 30℃, injection volume is 2 µL, and gradient elution is performed; The gradient elution procedure is as follows: The mass spectrometry conditions are as follows: electrospray ionization source, positive and negative ion monitoring modes; positive ion spray voltage: 3500 V, negative ion spray voltage: 3000 V, sheath gas is nitrogen, pressure is 35 arb; auxiliary gas is nitrogen, pressure is 10 arb; capillary temperature is 350℃, and scanning mass range is m / z 100~1000.

2. The method for detecting fingerprint spectra of traditional Chinese medicine compound prescriptions according to claim 1, characterized in that, The specific process of step (1) is as follows: Take 0.50039g of Jingfang granules, accurately weigh them, add 20 mL of ethyl acetate, weigh them, heat under reflux for 40 min, cool them, make up the weight loss, shake well, filter them, evaporate the filtrate to dryness, and dilute to 10 mL in a volumetric flask with methanol. Take the filter residue, add 20 mL of 95% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add 95% ethanol to make up to 10 mL in a volumetric flask. Take the filter residue, add 20 mL of 70% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add 70% ethanol to make up to 10 mL in a volumetric flask. Take the filter residue, add 20 mL of 30% ethanol, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add 30% ethanol to make up to 10 mL in a volumetric flask. Take the filter residue, add 20 mL of water, weigh it, heat under reflux for 40 min, cool it, make up the weight loss, shake it well, filter it, evaporate the filtrate to dryness, and add water to make up to 10 mL in a volumetric flask. Extract the sample, mix well, transfer to a centrifuge tube, centrifuge for 5 min at 10000 r / min, take the supernatant and filter it through a 0.22 μm microporous membrane into a sample vial, and store at 4℃ for later use.

3. The method for detecting fingerprint spectra of traditional Chinese medicine compound prescriptions according to claim 1, characterized in that, In step (2), the concentration of the internal standard in the sample solution containing the internal standard is 10 ug / ml; the volume fraction of the methanol solution is 70%; and the ratio of puerarin to methanol solution is 1 mg: 1 mL.

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