A quality detection method for the traditional Chinese medicine compound Qingkui Yuyang Decoction

Abstract

The present invention discloses a quality detection method for a traditional Chinese medicine compound Qingkui Yuyang Decoction. The UPLC-Q-Exactive Orbitrap-MS / MS analysis method is used to qualitatively analyze the chemical components of the six medicinal materials in the Qingkui Yuyang Decoction, namely, coptis chinensis, frankincense, sanguisorba officinalis, indigo naturalis, catechu, and dragon's blood. Then, the high performance liquid chromatography method is used to determine the contents of epicatechin, catechin, rutin, coptisine, and berberine in the Qingkui Yuyang Decoction. The UPLC-Q-Exactive Orbitrap-MS / MS analysis method has the advantages of high resolution, good mass accuracy, and strong qualitative ability, and can perform extensive and accurate detection of the chemical components in complex traditional Chinese medicine compounds. The high performance liquid chromatography can provide detection results with high accuracy, high sensitivity, and good repeatability, is simple and easy to operate, and has passed methodological verification, with good linearity. The precision, stability, repeatability, and recovery rate all meet the requirements. The combination of qualitative and quantitative analysis lays the foundation for the establishment of the quality standard for Qingkui Yuyang Decoction.

Description

Technical Field

[0001] The invention belongs to the technical field of drug detection, and particularly relates to a quality detection method for a traditional Chinese medicine compound Qingkui Yuyang prescription. Background Art

[0002] Qingkui Yuyang Decoction is a clinically proven formula composed of coptis root, frankincense, sanguisorba officinalis, indigo naturalis, catechu, and draconis. Coptis root clears heat and dampness, purges fire and detoxifies; frankincense invigorates blood circulation, promotes qi circulation, and relieves pain; sanguisorba officinalis cools blood and stops bleeding, detoxifies, and heals ulcers; indigo naturalis clears heat and detoxifies, cools blood and eliminates freckles; catechu invigorates blood circulation and stops pain, stops bleeding, and promotes tissue regeneration; and draconis officinalis primarily invigorates blood circulation and relieves pain, resolves blood stasis, stops bleeding, and heals ulcers and promotes tissue regeneration. Theoretically, these herbs combined can clear heat and dampness, cool blood and detoxify, and invigorate blood circulation and promote tissue regeneration, thereby promoting ulcer healing.

[0003] As a traditional Chinese medicine, Qingkui Yuyang Recipe requires appropriate preparation techniques to produce a suitable dosage form for clinical use. Therefore, this recipe is prepared using traditional techniques. The main active ingredients of Qingkui Yuyang Recipe are derived from six medicinal herbs: coptis chinensis, frankincense, sanguisorba officinalis, indigo naturalis, catechu, and dragon's blood. To investigate the quality of Qingkui Yuyang Recipe, an efficient detection method was required for both qualitative and quantitative analysis. Summary of the Invention

[0004] The purpose of the present invention is to design a quality detection method for a traditional Chinese medicine compound Qingkui Yuyang prescription, which can perform extensive and accurate detection of chemical components in a complex traditional Chinese medicine compound.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0006] A quality detection method for a traditional Chinese medicine compound Qingkui Yuyang prescription is characterized in that the traditional Chinese medicine compound Qingkui Yuyang prescription is composed of six medicinal ingredients: epicatechin, catechin, rutin, coptisine, and berberine. The specific detection method is as follows:

[0007] S1: Qualitative analysis of six medicinal herbs, namely, coptis root, frankincense, sanguisorba officinalis, indigo naturalis, catechu, and dragon's blood, was performed using UPLC-Q-Exactive Orbitrap-MS / MS.

[0008] Qualitative analysis includes the following processes:

[0009] Step 1: Chromatographic and mass spectrometric conditions;

[0010] Step 2: Preparation of mixed reference solution:

[0011] Take appropriate amounts of berberine, citric acid, quercetin, catechin, isoquercetin, gallic acid, epicatechin, rutin, chlorogenic acid, coptisine, indigo and indirubin reference substances, place them in a brown volumetric flask, dissolve and dilute with methanol, and make up to 25 mL to prepare a 200 μg / mL reference substance stock solution. Take appropriate amounts of each reference substance stock solution and dilute it to a mixed reference substance solution of about 5 μg / mL.

[0012] Step 3: Preparation of test solution:

[0013] Weigh the six medicinal materials of Qingkui Yuyang Recipe according to the prescription, add water, extract, combine the filtrate, and concentrate under reduced pressure on a rotary evaporator; accurately pipette the concentrate, place it in a volumetric flask, dilute it with distilled water and make up to the mark, shake well, and the Qingkui Yuyang Recipe test solution is obtained;

[0014] After determining the chromatography-mass spectrometry conditions, appropriate amounts of the mixed reference solution and test solution were filtered through a 0.22 μm microporous filter membrane and introduced into a UPLC-Q-Exactive Orbitrap-MS / MS chromatography-mass spectrometer for detection. Based on the chromatographic peaks and mass spectrometric data of the reference solution, the compounds in the Qingkui Yuyang recipe were accurately identified. Other compounds were inferred by comparing the molecular ions and secondary fragment ions with the natural compound database.

[0015] S2: Determination of the contents of epicatechin, catechin, rutin, coptisine, and berberine in Qingkuiyuyang Decoction by HPLC;

[0016] The content determination method is as follows:

[0017] Step 1: Chromatographic conditions

[0018] The chromatographic column is Waters C18, 4.6×250 mm, 5 μm, mobile phase 0.05 mol·L -1 Potassium dihydrogen phosphate solution A: acetonitrile B, wavelength 280nm, flow rate 0.6mL / min, column temperature 30℃, injection volume 10μL, gradient elution.

[0019] Step 2: Preparation of test solution

[0020] Weigh the six medicinal ingredients of Qingkui Yuyang Decoction according to the prescription, add water, extract, combine the filtrate, and concentrate under reduced pressure on a rotary evaporator; accurately pipette the concentrate, place it in a volumetric flask, dilute it with distilled water and make up to the mark, shake well, and the Qingkui Yuyang Decoction test solution is obtained;

[0021] Step 3: Preparation of reference stock solution

[0022] Take appropriate amounts of catechin, epicatechin, rutin, coptisine, and berberine reference substances, place them in brown volumetric flasks, dissolve and dilute with methanol, and dilute to 5 mL to prepare 500 μg / mL stock solutions of each reference substance;

[0023] Step 4: Preparation of mixed reference solution

[0024] Take an appropriate amount of each reference substance stock solution and dilute it to a mixed reference substance solution containing about 36.72 μg / mL of catechin, about 9.15 μg / mL of epicatechin, about 10 μg / mL of rutin, about 1.2 μg / mL of coptisine, and about 6.4 μg / mL of berberine.

[0025] Furthermore, the Qingkui Yuyang prescription in S1 is composed of six medicinal ingredients: coptis chinensis 4g, frankincense 6g, sanguisorba officinalis 6g, indigo naturalis 3.2g, catechu 3.2g, and blood clotting agent 4g.

[0026] Furthermore, the HPLC method for measuring the content of the chromatographic column is Waters C18, 4.6×250 mm, 5 μm, mobile phase 0.05 mol·L -1 Potassium dihydrogen phosphate solution A: acetonitrile B, wavelength at 280 nm, flow rate at 0.6 mL / min, column temperature at 30°C, injection volume of 10 μL, gradient elution; from 0 to 10 min, mobile phase B was 10-20%; from 10 to 25 min, mobile phase B was 20%-30%; from 25 to 40 min, mobile phase B was 30%-35%; from 45 to 50 min, mobile phase B was 35%-40%; from 50 to 60 min, mobile phase B was 40%-10%.

[0027] Furthermore, the chromatography-mass spectrometry method is used, and the chromatographic column is ACQUITY UPLC BEH C 18 , 2.1mm×100mm, 1.7μm, with mobile phase B containing 0.1% formic acid-mobile phase A containing 0.1% formic acid as the mobile phase, flow rate of 0.3mL / min, column temperature of 30℃, injection volume of 10μL, and gradient elution; UPLC-Q-Exactive-Orbitrap MS / MS high-resolution mass spectrometry was used for mass spectrometry data acquisition, and the detection mode was Full MS-dd-MS 2 , positive and negative ion modes were scanned simultaneously, the scanning range m / z was 100-1500, the capillary temperature was 320℃; the evaporation temperature was 350℃; the spray voltage was 3500 / 3200V (+ / -); the auxiliary gas flow rate was 10Arb; the sheath gas flow rate was 35Arb; the collision energy was 20, 40, 60, 80%; and the full scan resolution was 120000.

[0028] Furthermore, the gradient elution is specifically as follows: from 0 to 5 minutes, the mobile phase B is 5%; from 5 to 20 minutes, the mobile phase B is increased from 5% to 95%; from 20 to 30 minutes, the mobile phase B is 95%; from 30 to 35 minutes, the mobile phase B is reduced from 95% to 5%; from 35 to 40 minutes, the mobile phase B is 5%.

[0029] Furthermore, the average contents of epicatechin, catechin, rutin, coptisine and berberine in the Qingkui Yuyang recipe were 11.36 mg / g, 7.36 mg / g, 5.06 mg / g, 6.62 mg / g and 3.17 mg / g, respectively.

[0030] The following beneficial effects can be achieved through the above technical solution:

[0031] The present invention first uses the UPLC-Q-Exactive Orbitrap-MS / MS analysis method to qualitatively analyze the chemical components of the six medicinal materials in the Qingkui Yuyang prescription, namely, coptis chinensis, frankincense, sanguisorba officinalis, indigo naturalis, catechu, and dragon's blood; then uses high-performance liquid chromatography to determine the contents of epicatechin, catechin, rutin, coptisine, and berberine in the Qingkui Yuyang prescription. The UPLC-Q-Exactive Orbitrap-MS / MS analysis method has the advantages of high resolution, good mass accuracy, and strong qualitative ability, and can perform extensive and accurate detection of chemical components in complex traditional Chinese medicine compounds; high-performance liquid chromatography can provide detection results with high accuracy, high sensitivity, and good repeatability, is simple and easy to operate, and has passed methodological validation, with good linearity, and precision, stability, repeatability, and recovery rate all meet regulations. The combination of qualitative and quantitative analysis lays the foundation for the establishment of the quality standard of the Qingkui Yuyang prescription. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The total ion current chromatograms of Qingkui Yuyang Formula in positive (A) and negative (B) ion modes are shown;

[0033] Figure 2 It is a comparative chromatogram of blank solution, reference solution and test solution;

[0034] Figure 3 This is the standard curve of epicatechin, catechin, rutin, coptisine and berberine. DETAILED DESCRIPTION

[0035] The following is attached Figure 1-3 The present invention will be further described in conjunction with the embodiments:

[0036] Example 1:

[0037] A quality detection method for Qingkui Yuyang prescription comprises the following steps: firstly performing qualitative analysis on six medicinal materials in the Qingkui Yuyang prescription, namely, coptis chinensis, frankincense, sanguisorba officinalis, indigo naturalis, catechu, and dragon's blood, using a UPLC-Q-Exactive Orbitrap-MS / MS analysis method; and then determining the contents of epicatechin, catechin, rutin, coptisine, and berberine in the Qingkui Yuyang prescription by high performance liquid chromatography.

[0038] UPLC-Q-Exactive Orbitrap-MS / MS was used to qualitatively analyze the six medicinal ingredients in the Qingkui Yuyang formula (Coptis rhizome, Frankincense, Sanguisorba officinalis, Indigo naturalis, Catechu, and Sanguisorba officinalis). Compared with traditional HPLC and thin-layer chromatography qualitative methods, this method offers a wider mass detection range, higher sensitivity, and higher peak capacity, enabling large-scale qualitative analysis of complex traditional Chinese medicine formulas. The method includes the following:

[0039] (1) Chromatographic and mass spectrometry conditions

[0040] The chromatographic column is ACQUITY UPLC BEH C 18 The column was eluted with a column diameter of 2.1 mm × 100 mm, 1.7 μm. The mobile phase was acetonitrile containing 0.1% formic acid (mobile phase B) and water containing 0.1% formic acid (mobile phase A). The flow rate was 0.3 mL / min, the column temperature was 30°C, and the injection volume was 10 μL. Gradient elution was performed using a UPLC-Q-Exactive-Orbitrap MS / MS high-resolution mass spectrometer. The detection mode was Full MS-dd-MS2, with simultaneous positive and negative ion scanning over the m / z range of 100 to 1500. The capillary temperature was 320°C, the evaporation temperature was 350°C, the spray voltage was 3500 / 3200 V (+ / -), the auxiliary gas flow rate was 10 Arb, the sheath gas flow rate was 35 Arb, the collision energy was 20, 40, 60, and 80%, and the full scan resolution was 120,000.

[0041] Table 1 Gradient elution table

[0042]

[0043] (2) Preparation of mixed reference solution:

[0044] Take appropriate amounts of berberine, citric acid, quercetin, catechin, isoquercetin, gallic acid, epicatechin, rutin, chlorogenic acid, coptisine, indigo and indirubin reference substances, place them in a brown volumetric flask, dissolve and dilute with methanol, and make up to 25 mL to prepare a 200 μg / mL reference substance stock solution. Take appropriate amounts of each reference substance stock solution and dilute it to a mixed reference substance solution of about 5 μg / mL.

[0045] (3) Preparation of test solution

[0046] Weigh 27.6 g of the six medicinal ingredients of the Qingkui Yuyang Recipe according to the prescription, add 10 times the amount of water, and extract for 60 min. Extract three times. Combine the filtrates and concentrate under reduced pressure (60°C) on a rotary evaporator to 250 mL. Accurately transfer 1 mL of the concentrate to a 50 mL volumetric flask, dilute with distilled water, and bring to volume. Shake well to obtain the Qingkui Yuyang Recipe test solution (2.2 mg / mL).

[0047] In this study, UPLC-Q-Exactive Orbitrap-MS / MS technology was used to collect mass spectrometric information of the aqueous extract of Qingkui Yuyang Recipe in positive and negative ion modes. The total ion current chromatograms are shown in Figure 1 . Based on the self-built "Qingkui Yuyang Fang Compound" database and Xcalibur qualitative software, combined with the molecular ions and secondary fragment ions of the compounds, a total of 71 compounds in the traditional Chinese medicine compound Qingkui Yuyang Fang were compared, including 22 flavonoids, 17 alkaloids, 17 organic acid compounds, 4 triterpenoids, 2 terpenoids, 2 amino acid compounds and 7 other organic compounds. See Table 2. Among them, based on the chromatographic peaks and mass spectrometry data of the reference substances, 12 compounds were accurately identified, including: 5 flavonoids (quercetin, catechin, isoquercetin, epicatechin, rutin), 4 alkaloids (berberine, coptisine, indigo, indigotin) and 3 organic acid compounds (citric acid, gallic acid, chlorogenic acid). By comparing with the molecular ions and secondary fragment ions of the natural compound database, another 59 compounds were inferred.

[0048] Table 2 Chemical composition information of Qingkui Yuyang prescription

[0049]

[0050]

[0051]

[0052] Example 2 A method for determining the content of Qingkui Yuyang Recipe, comprising the following:

[0053] High-performance liquid chromatography (HPLC) was used to determine the contents of epicatechin, catechin, rutin, coptisine, and berberine in the Qingkui Yuyang formula. Compared to traditional ultraviolet spectrophotometry, this method offers high accuracy, high sensitivity, and excellent repeatability. It is simple to perform and can be used to determine the contents of multiple components in complex traditional Chinese medicine formulas. Related studies on content determination are as follows:

[0054] (1) Chromatographic conditions

[0055] The chromatographic column is Waters C18 (4.6 × 250 mm, 5 μm), mobile phase: 0.05 mol L -1 Potassium dihydrogen phosphate solution (A): acetonitrile (B), wavelength at 280 nm, flow rate at 0.6 mL / min, column temperature at 30°C, injection volume of 10 μL, gradient elution, time program as shown in Table 3.

[0056] Table 3 Gradient elution table

[0057]

[0058] (2) Preparation of test solution

[0059] According to the prescription, 27.6 g of the six medicinal materials of Qingkui Yuyang Decoction were weighed, 10 times the amount of water was added, and the mixture was extracted for 60 min. Extraction was repeated three times, and the filtrates were combined and concentrated to 250 mL under reduced pressure on a rotary evaporator; 1 mL of the concentrate was accurately transferred to a 50 mL volumetric flask, diluted with distilled water and adjusted to the scale, and shaken to obtain the Qingkui Yuyang Decoction test solution (2.2 mg / mL).

[0060] (3) Preparation of reference substance stock solution

[0061] Take appropriate amounts of epicatechin, catechin, rutin, coptisine and berberine reference substances, place them in brown volumetric flasks, dissolve and dilute them with methanol, and make up to 20 mL to prepare 500 μg / mL stock solutions of each reference substance.

[0062] (4) Preparation of mixed reference solution

[0063] Take an appropriate amount of each reference substance stock solution and dilute it to a mixed reference substance solution containing about 36.72 μg / mL of catechin, about 9.15 μg / mL of epicatechin, about 10 μg / mL of rutin, about 1.2 μg / mL of coptisine, and about 6.4 μg / mL of berberine.

[0064] Example 3 Investigation of the linear range

[0065] A series of mixed reference solutions of epicatechin, catechin, rutin, coptisine and berberine were prepared respectively, and the samples were injected and measured according to the liquid phase method of "Claim 3". The peak area was used as the ordinate (Y) and the solution concentration (μg / mL) was used as the abscissa (X) for linear plotting. The linear equations of each reference solution were obtained. The results are shown in Tables 4 and Figure 3 , indicating that the linear relationship of each reference solution is good.

[0066] Table 4 Linearity and range

[0067]

[0068] Example 4 Precision Experiment

[0069] The mixed reference solution was continuously injected 6 times according to the liquid phase method of "Claim 3", and the peak area RSDs of epicatechin, catechin, rutin, coptisine and berberine were calculated to be 0.5%, 0.3%, 1.2%, 2.8% and 1.9%, respectively, indicating that the instrument precision of this method is good.

[0070] Example 5 Stability Test

[0071] After the test solution was placed at room temperature for 0, 4, 8, 12, 24 and 48 hours, the sample was injected and measured according to the liquid phase method of "Claim 3", and the peak area RSDs of epicatechin, catechin, rutin, coptisine and berberine were calculated to be 2.6%, 1.4%, 9.9%, 6.4% and 3.7%, respectively, indicating that the test solution was stable within 48 hours.

[0072] Example 6 Repeatability

[0073] Six test solutions were prepared in parallel and sampled and measured according to the liquid phase method of "Claim 3". The RSDs of the contents of epicatechin, catechin, rutin, coptisine and berberine were calculated to be 4.1%, 4.8%, 8.5%, 5.3% and 3.4%, respectively, indicating that the method had good repeatability.

[0074] Example 7 Sample recovery test

[0075] Precisely pipette a test solution containing known epicatechin, catechin, rutin, coptisine, and berberine contents, and add a predetermined amount of a mixed reference solution. Six parallel aliquots of these solutions were prepared and assayed using the liquid phase method described in Claim 3. The results are shown in Table 5. The average recoveries for epicatechin, catechin, rutin, coptisine, and berberine were 91.97%, 97.94%, 112.09%, 104.74%, and 95.65%, respectively, demonstrating the good accuracy of this method.

[0076] Table 5 Sample recovery test results

[0077]

[0078]

[0079] Example 8 Content Determination

[0080] Six parallel aliquots of the test solution were prepared and sampled and assayed according to the liquid phase method of Claim 3. The results are shown in Table 6. The average contents of epicatechin, catechin, rutin, coptisine, and berberine were calculated to be 11.36 mg / g, 7.36 mg / g, 5.06 mg / g, 6.62 mg / g, and 3.17 mg / g, respectively.

[0081] Table 6 Content determination results

[0082]

[0083] The above are all preferred embodiments of the present invention. For ordinary technicians in this technical field, without departing from the principle of the present invention, modifications to various equivalent forms of the present invention all fall within the scope of protection of the claims attached to this application. The above are all preferred embodiments of the present invention. For ordinary technicians in this technical field, without departing from the principle of the present invention, modifications to various equivalent forms of the present invention all fall within the scope of protection of the claims attached to this application. The above are all preferred embodiments of the present invention. For ordinary technicians in this technical field, without departing from the principle of the present invention, modifications to various equivalent forms of the present invention all fall within the scope of protection of the claims attached to this application.

Claims

1. A quality testing method for a traditional Chinese medicine compound Qingkui Yuyang Decoction, characterized in that: The traditional Chinese medicine compound Qingkui Yuyang prescription is composed of six medicinal ingredients: epicatechin, catechin, rutin, coptisine, and berberine. The specific detection method is as follows: S1: Qualitative analysis of six medicinal herbs, namely, coptis root, frankincense, sanguisorba officinalis, indigo naturalis, catechu, and dragon's blood, was performed using UPLC-Q-Exactive Orbitrap-MS / MS. Qualitative analysis includes the following processes: Step 1: Chromatographic and mass spectrometric conditions; Step 2: Preparation of mixed reference solution: Take appropriate amounts of berberine, citric acid, quercetin, catechin, isoquercetin, gallic acid, epicatechin, rutin, chlorogenic acid, coptisine, indigo, and indirubin reference substances, place them in a brown volumetric flask, dissolve and dilute with methanol, and dilute to 25 mL to prepare a 200 μg / mL reference substance stock solution. Take appropriate amounts of each reference substance stock solution and dilute to a mixed reference substance solution of approximately 5 μg / mL; Step 3: Preparation of test solution: Weigh the six medicinal materials of Qingkui Yuyang Recipe according to the prescription, add water, extract, combine the filtrate, and concentrate under reduced pressure on a rotary evaporator; accurately pipette the concentrate, place it in a volumetric flask, dilute it with distilled water and make up to the mark, shake well, and the Qingkui Yuyang Recipe test solution is obtained; After determining the chromatography-mass spectrometry conditions, appropriate amounts of the mixed reference solution and test solution were filtered through a 0.22 μm microporous filter membrane and introduced into a UPLC-Q-Exactive Orbitrap-MS / MS chromatography-mass spectrometer for detection. Based on the chromatographic peaks and mass spectrometric data of the reference solution, the compounds in the Qingkui Yuyang recipe were accurately identified. Other compounds were inferred by comparing the molecular ions and secondary fragment ions with the natural compound database. S2: Determination of the contents of epicatechin, catechin, rutin, coptisine, and berberine in Qingkuiyuyang Decoction by HPLC; The content determination method is as follows: Step 1: Chromatographic conditions The chromatographic column was Waters Symmetry® C18, 4.6 × 250 mm, 5 μm, and the mobile phase was 0.05 mol·L -1 Potassium dihydrogen phosphate solution A: acetonitrile B, wavelength 280 nm, flow rate 0.6 mL / min, column temperature 30 °C, injection volume 10 μL, gradient elution; Step 2: Preparation of test solution Weigh the six medicinal ingredients of Qingkui Yuyang Decoction according to the prescription, add water, extract, combine the filtrate, and concentrate under reduced pressure on a rotary evaporator; accurately transfer the concentrate to a volumetric flask, dilute with distilled water and adjust the volume to the mark, shake well, and the Qingkui Yuyang Decoction test solution is obtained; Step 3: Preparation of reference substance stock solution Take appropriate amounts of catechin, epicatechin, rutin, coptisine and berberine reference substances, place them in brown volumetric flasks, dissolve and dilute with methanol, and dilute to 5 mL to prepare 500 μg / mL stock solutions of each reference substance; Step 4: Preparation of mixed reference solution Take an appropriate amount of each reference substance stock solution and dilute it to a mixed reference substance solution containing about 36.72 μg / mL of catechin, about 9.15 μg / mL of epicatechin, about 10 μg / mL of rutin, about 1.2 μg / mL of coptisine, and about 6.4 μg / mL of berberine.

2. The quality detection method of a traditional Chinese medicine compound Qingkui Yuyang Decoction according to claim 1, characterized in that: The Qingkui Yuyang prescription in S1 is composed of six medicinal ingredients: 4g of coptis chinensis, 6g of frankincense, 6g of sanguisorba officinalis, 3.2g of indigo, 3.2g of catechu, and 4g of blood clotting agent.

3. The quality detection method of a traditional Chinese medicine compound Qingkui Yuyang Decoction according to claim 1, characterized in that: High-performance liquid chromatography (HPLC) was used for content determination. The column was Waters Symmetry® C18, 4.6 × 250 mm, 5 μm, and the mobile phase was 0.05 mol·L -1 Potassium dihydrogen phosphate solution A: acetonitrile B, wavelength at 280 nm, flow rate at 0.6 mL / min, column temperature at 30°C, injection volume of 10 μL, gradient elution; from 0 to 10 min, mobile phase B was 10-20%; from 10 to 25 min, mobile phase B was 20%-30%; from 25 to 40 min, mobile phase B was 30%-35%; from 45 to 50 min, mobile phase B was 35%-40%; from 50 to 60 min, mobile phase B was 40%-10%.

4. The quality detection method of a traditional Chinese medicine compound Qingkui Yuyang Decoction according to claim 1, characterized in that: Chromatography-mass spectrometry method, the chromatographic column is ACQUITY UPLC BEH C 18 , 2.1mm×100mm, 1.7μm, with mobile phase B containing 0.1% formic acid-mobile phase A containing 0.1% formic acid as the mobile phase, flow rate of 0.3mL / min, column temperature of 30℃, injection volume of 10μL, and gradient elution. UPLC-Q-Exactive-Orbitrap MS / MS high-resolution mass spectrometry was used for mass spectrometry data acquisition, and the detection mode was Full MS-dd-MS 2 , positive and negative ion modes were scanned simultaneously, the scanning range m / z was 100~1500, the capillary temperature was 320℃; the evaporation temperature was 350℃; the spray voltage was 3500 / 3200V (+ / -); the auxiliary gas flow rate was 10Arb; the sheath gas flow rate was 35Arb; the collision energy was 20, 40, 60, 80%; and the full scan resolution was 120000.

5. The quality detection method of a traditional Chinese medicine compound Qingkui Yuyang Decoction according to claim 4, characterized in that: The gradient elution is specifically as follows: from 0 to 5 minutes, the mobile phase B is 5%; from 5 to 20 minutes, the mobile phase B is increased from 5% to 95%; from 20 to 30 minutes, the mobile phase B is 95%; from 30 to 35 minutes, the mobile phase B is reduced from 95% to 5%; from 35 to 40 minutes, the mobile phase B is 5%.

6. The quality detection method of a traditional Chinese medicine compound Qingkui Yuyang Decoction according to claim 1, characterized in that: The average contents of epicatechin, catechin, rutin, coptisine and berberine in Qingkui Yuyang prescription were 11.36 mg / g, 7.36 mg / g, 5.06 mg / g, 6.62 mg / g and 3.17 mg / g, respectively.

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