Determination of the content of seven components in Jinteng Qingbi Granules by a single-method multi-evaluation method and its application

Abstract

This invention belongs to the field of traditional Chinese medicine preparation analysis, specifically disclosing a one-test-multiple-evaluation method for determining the content of seven components in Jinteng Qingbi granules and its application. Using sinomenine as an internal standard, a one-test-multiple-evaluation method is used to establish relative correction factors between sinomenine and gallic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C in Jinteng Qingbi granules. The contents of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C in Jinteng Qingbi granules are calculated using these correction factors. This invention's method is highly practical, simple to operate, cost-effective, and provides accurate and reliable test results, overcoming the shortcomings of existing methods for quality testing of Jinteng Qingbi granules and providing a new approach for quality control and intrinsic quality evaluation of Jinteng Qingbi granules.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of traditional Chinese medicine preparation, and particularly relates to a quality detection method of Jinteng Qingbi granules, especially a method for determining the content of seven components in Jinteng Qingbi granules by one measurement multiple evaluation and application thereof. BACKGROUND

[0002] Jinteng Qingbi granules are prepared from 11 traditional Chinese medicines, i.e. honeysuckle, kadsura, oldenlandia, figwort, rehmannia, monotropistra, pyrola, licorice, angelica, white peony root and centipede, and are a sole Chinese patent medicine of Lunan Houpu Pharmaceutical Co., Ltd. (Guo Yao Zhu Z20123065). The main effects of Jinteng Qingbi granules are clearing heat and resolving toxin, promoting blood circulation to remove stasis, and relieving arthralgia. Jinteng Qingbi granules are used for treating active stage of rheumatoid arthritis, and belong to the syndrome of toxic heat accumulation and damp-heat blocking collaterals. The symptoms include joint swelling, pain, morning stiffness, fever, red skin, high fever, profuse sweating, dry mouth, yellow urine, red tongue, yellow and greasy fur, and rapid pulse.

[0003] Jinteng Qingbi granules take honeysuckle and kadsura as monarch drugs, oldenlandia, monotropistra, pyrola, angelica and white peony root as ministerial drugs, figwort, rehmannia and licorice as auxiliary drugs, and centipede as a ministerial drug. The combination of the drugs has the effects of clearing heat and resolving toxin, promoting blood circulation to remove stasis, and relieving arthralgia. Honeysuckle is rich in volatile oil, flavonoids and organic acids. The experimental results show that the water decoction, oral liquid and injection of honeysuckle have the effects of relieving fever, anti-inflammation, regulating immune function and the like. Kadsura contains effective components such as sinomenine and magnolia base, and has the effects of anti-inflammation, analgesia, anti-rheumatism and immunosuppression, and is commonly used in treating rheumatoid arthritis. Angelica, white peony root, oldenlandia, pyrola, monotropistra, figwort, rehmannia, licorice and centipede have the effects of tonifying blood, relieving pain, strengthening muscles and bones, and clearing heat and resolving toxin. Pharmacological studies show that centipede can improve microcirculation, and has the effects of analgesia and anti-inflammation. The combination of the drugs has the effects of clearing heat and resolving toxin, promoting blood circulation to remove stasis, relieving arthralgia and relieving pain.

[0004] At present, there is no report on the content determination of Jinteng Qingbi granules, and the preparation lacks effective quality detection and evaluation methods. Jinteng Qingbi granules are traditional Chinese medicine compound preparations, and the chemical components are complex. Therefore, comprehensive quality control is needed to ensure the safety and effectiveness of clinical medication. The traditional external standard method for multi-component quantification has problems such as numerous control samples and high cost. The one measurement multiple evaluation (QAMS) method uses one internal reference in the sample to establish the relative correction factor of other components, and then calculates the content of other components. The method can overcome the problem of lack of control samples in the determination of multi-component content of traditional Chinese medicine preparations, and save detection time and cost. Therefore, the application of one measurement multiple evaluation method in Jinteng Qingbi granules can achieve the purpose of effectively and accurately quantifying more effective components in Jinteng Qingbi granules with less use of control samples, and has a wide application prospect. SUMMARY

[0005] This invention provides a method for determining the content of seven active ingredients in Jinteng Qingbi granules using a multi-analysis approach, and its application. The technical solution of this invention is as follows:

[0006] A method for determining the content of seven components in Jinteng Qingbi Granules using a single-test, multiple-evaluation approach was developed. Qingteng alkaloid was used as an internal standard. A relative correction factor was established between Qingteng alkaloid and gallic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C in Jinteng Qingbi Granules. The contents of gallic acid, neochlorogenic acid, Qingteng alkaloid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C in Jinteng Qingbi Granules were calculated using this correction factor.

[0007] The determination was performed using high-performance liquid chromatography (HPLC), and the specific method is as follows:

[0008] Preparation of Reference Solution A: Accurately weigh gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C reference standards, add methanol to completely dissolve them, prepare a mixed reference solution, filter through a 0.22 μm microporous membrane, and collect the filtrate for later use.

[0009] B. Preparation of the test solution: Accurately weigh Jinteng Qingbi granules, accurately add ethanol solution, sonicate to dissolve, filter through a 0.22μm microporous membrane, and take the filtrate to obtain the test solution.

[0010] C. Chromatographic conditions: Column: Octadecylsilane-bonded silica gel as the packing material; Mobile phase: Methanol-phosphoric acid aqueous solution;

[0011] Elute using the following gradient elution procedure:

[0012]

[0013] The chromatographic column is selected from one of the following: Agilent ZORBAX SB-C18, Agilent Eclipse XDB-C18, and ThermoSecientific BDS HYPERSIL C18. The column has dimensions of 4.6 mm × 250 mm and a diameter of 5 μm.

[0014] The concentration of the phosphoric acid aqueous solution in the mobile phase is 0.1%-1.0%; preferably 0.3%-0.5%.

[0015] The column temperature is 25-35℃, preferably 30℃.

[0016] Preferably, the detection wavelength for gallic acid and sinomenine is 260-270 nm, and the detection wavelength for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C is 320-340 nm.

[0017] Preferably, the detection wavelength for gallic acid and sinomenine is 262 nm, and the detection wavelength for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C is 330 nm.

[0018] The mobile phase flow rate is 0.7-1.1 ml / min; preferably 1.0 mL / min.

[0019] The injection volume is 5-20 μL; preferably 10 μL.

[0020] The solvent used to prepare the test solution was a 70% ethanol solution. The ultrasonic treatment power for preparing the test solution was 250W, the frequency was 50kHz, and the ultrasonic treatment time was 30min.

[0021] The above-mentioned method of determining the content of components in Jinteng Qingbi Granules 7 using a single test and multiple evaluation method is applied to the quality control and evaluation of Jinteng Qingbi Granules.

[0022] This invention relates to the determination of multiple indexes in Jinteng Qingbi granules, and establishes a QAMS method that uses one of its components as an internal reference to achieve simultaneous determination of multiple components, achieving the following beneficial technical effects:

[0023] First, sinomenine, which is inexpensive, readily available, and stable, was selected as an internal reference, and a relative correction factor was established between sinomenine and the six main active ingredients in Jinteng Qingbi Granules.

[0024] Secondly, the accuracy of the QAMS method was verified by quantitative analysis of the components of different batches of Jinteng Qingbi granules. The reproducibility of the method was also examined under different brands of instruments, different chromatographic columns, and different column temperatures and flow rates. The results showed that the RAD values ​​between the QAMS and external standard methods were both less than 2%. This method can calculate the content of gallic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C reference standards using relative correction factors. This method effectively reduces costs and is accurate and reliable, providing a basis for improving the quality control standards of Jinteng Qingbi granules and comprehensively evaluating their quality.

[0025] Finally, the method of the present invention is highly practical, simple to operate, cost-effective, and the test results are accurate and reliable, making up for the deficiencies of the quality control method of Jinteng Qingbi Granules, thus providing a new option for the quality control and intrinsic quality evaluation of Jinteng Qingbi Granules. Attached Figure Description

[0026] Figure 1 This is the chromatogram of the mixed reference standard from Example 1 at 262 nm;

[0027] Figure 2 This is the chromatogram of the mixed reference standard from Example 1 at 330 nm;

[0028] Figure 3 This is the chromatogram of the test sample from Example 1 at 262 nm;

[0029] Figure 4 This is the chromatogram of the test sample from Example 1 at 330 nm;

[0030] Figure 5 This is the chromatogram of the test sample from Example 2 at 262 nm;

[0031] Figure 6 This is the chromatogram of the test sample from Example 2 at 330 nm;

[0032] Figure 7 This is the chromatogram of the test sample in Example 3 at 262 nm;

[0033] Figure 8 This is the chromatogram of the test sample in Example 3 at 330 nm;

[0034] Figure 9 This is the chromatogram of the test sample from Comparative Example 1 at 262 nm;

[0035] Figure 10 This is the chromatogram of the test sample in Comparative Example 1 at 330 nm;

[0036] Figure 11 This is the chromatogram of the test sample in Comparative Example 2 at 262 nm;

[0037] Figure 12 This is the chromatogram of the test sample in Comparative Example 2 at 330 nm.

[0038] In the above attached diagram, 1 is gallic acid; 2 is neochlorogenic acid; 3 is sinomenine; 4 is chlorogenic acid; 5 is cryptochlorogenic acid; 6 is isochlorogenic acid B; and 7 isochlorogenic acid C. Detailed Implementation

[0039] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following examples will be used for illustrative purposes. Where specific experimental steps or conditions are not specified in the examples, they can be performed according to the conventional experimental steps or conditions described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available.

[0040] SHIMADZU Essentia SIL-16 liquid chromatograph (Shimadzu Corporation, Japan), SK5210LHC ultrasonic cleaner (Shanghai Kedao Ultrasonic Instrument Co., Ltd.), SECURA125-1CN electronic balance (Sartorius Scientific Instruments Co., Ltd.).

[0041] Reference standards gallic acid (110831-201204) and chlorogenic acid (110753-201314) were purchased from the National Institutes for Food and Drug Control; reference standards neochlorogenic acid (DSTDX001501), cryptochlorogenic acid (DSTDY003501), isochlorogenic acid B (DSTDY003701), and isochlorogenic acid C (DSTDY003801) were purchased from Chengdu Desite Biotechnology Co., Ltd.; and reference standard sinomenine (22080212) was purchased from Beijing Better Renkang Biomedical Technology Co., Ltd.

[0042] Jinteng Qingbi Granules were provided by Lunan Houpu Pharmaceutical Co., Ltd., and sample information is shown in Table 1. Methanol was of chromatographic grade and purchased from Merck AG, Germany; anhydrous ethanol was purchased from Sinopharm Chemical Reagent Co., Ltd.; phosphoric acid was purchased from Laiyang Kangde Chemical Co., Ltd.; and purified water was purchased from Hangzhou Wahaha Group Co., Ltd.

[0043] Table 1. Sample Information of Jinteng Qingbi Granules

[0044]

[0045]

[0046] Example 1: Determination of the content of 7 components in Jinteng Qingbi Granules

[0047] 1.1 Preparation of mixed reference solution

[0048] Accurately weigh gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C reference standards sequentially and place them in a volumetric flask. Add methanol to completely dissolve them, and prepare a mixed reference solution containing 0.450 mg, 0.400 mg, 0.808 mg, 0.404 mg, 0.400 mg, 0.406 mg, and 0.400 mg of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C per mL, respectively. Filter the solution through a 0.22 μm microporous membrane and collect the filtrate for later use.

[0049] 1.2 Preparation of the test solution

[0050] Accurately weigh approximately 0.5g of Jinteng Qingbi granules and place them in a stoppered conical flask. Accurately add 10mL of 70% ethanol, seal tightly, and sonicate (250W, 50kHz) for 30min to dissolve. Filter the solution through a 0.22μm microporous membrane and collect the filtrate to obtain the test solution.

[0051] 1.3 Chromatographic conditions

[0052] Chromatographic column: Agilent ZORBAX SB-C18 (4.6×250mm, 5μm); mobile phase: methanol-0.5% phosphoric acid aqueous solution; flow rate: 1.0mL / min; detection wavelength: 262nm, 330nm; column temperature: 30℃; injection volume: 10μL;

[0053] Elution was performed according to the following gradient table:

[0054]

[0055] 1.4 Chromatogram

[0056] Accurately pipette the mixed reference solution and the test solution into the high-performance liquid chromatography system, measure and record the chromatograms; the chromatograms of the mixed reference solution and the test solution at 262 nm and 330 nm are shown in [Figure number missing]. Figures 1-4 .

[0057] 2. Methodological Examination

[0058] 2.1 Examination of Linear Relationships

[0059] Accurately pipette the mixed reference solution from section "1.1", and sequentially dilute it to different mass concentrations. Inject the solution according to the chromatographic conditions described in section "1.3". Plot a standard curve with the reference concentration (X) as the abscissa and the peak area (Y) as the ordinate, and establish a linear regression equation, as shown in Table 2. The results show that gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C exhibit good linearity within their respective ranges.

[0060] Table 2. Linear equations for the 7 components of Jinteng Qingbi Granules

[0061]

[0062] 2.2 Precision test

[0063] The mixed reference solution from section "1.1" was injected six times consecutively under the chromatographic conditions described in section "1.3," and the peak areas of the corresponding chromatographic components were determined. The RSD values ​​were calculated. The RSDs for the peak areas of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C were 0.54%, 0.53%, 0.52%, 0.53%, 0.53%, 0.51%, and 0.50%, respectively, indicating that the instrument has good precision.

[0064] 2.3 Repeatability Test

[0065] Six samples (sample number S1) of Jinteng Qingbi granules from the same batch, each approximately 0.5g, were randomly selected. Test solutions were prepared according to the method described in section "1.2," and the samples were injected according to the chromatographic conditions described in section "1.3." The peak areas of the seven components were determined, and the RSD values ​​were calculated. The RSDs for the peak areas of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C were 0.98%, 1.84%, 1.45%, 1.78%, 1.72%, 1.98%, and 1.73%, respectively, indicating good repeatability of the method.

[0066] 2.4 Stability Test

[0067] Accurately weigh 0.5g of Jinteng Qingbi Granules (sample number S1), and prepare the test solution according to the method in section "1.2". Under the chromatographic conditions in section "1.3", inject the sample sequentially at 0h, 2h, 4h, 8h, 12h, and 24h after the test solution preparation is completed. Measure the peak areas of the corresponding chromatographic components and calculate the RSD values. The RSDs of the peak areas of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C were 0.76%, 0.06%, 0.21%, 0.07%, 0.26%, 0.73%, and 1.05%, respectively, indicating that the test sample has good stability within 24h.

[0068] 2.5 Recovery rate study

[0069] Accurately weigh 0.25g of Jinteng Qingbi Granules (sample number S1) with known content, and accurately add equal amounts of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C reference standards. Prepare the test solution according to the method in section "1.2". Determine the peak area of ​​the corresponding chromatographic peaks of the above 7 components according to the chromatographic conditions in section "1.3", and calculate their RSD values ​​and recovery rates. The RSDs of the peak areas of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C were 1.65%, 0.50%, 2.50%, 1.28%, 1.52%, 1.65%, and 1.65%, respectively, with average recoveries of 97.95%, 103.59%, 103.15%, 96.93%, 95.81%, 96.51%, and 95.01%.

[0070] 2.6 Determination of Relative Correction Factor

[0071] Using the multi-point correction method, the mixed reference solution prepared by the method in section "1.1" was precisely pipetted and injected for determination according to the chromatographic conditions in section "1.3". Using sinomenine as an internal standard, the relative correction factors (f) for the other six components were calculated. k / s ).

[0072] f k / s =(C k A s ) / (C s A k )

[0073] In the formula, C k For the content of other ingredients, A k For the peak areas of other components, C s For the content of internal standard, A s The value represents the peak area of ​​the internal standard. The results are shown in Table 3.

[0074] Table 3. Relative Correction Factors for Each Component of Jinteng Qingbi Granules

[0075]

[0076] 2.6.1 Durability test of relative correction factor

[0077] 2.6.1.1 Instruments and chromatographic columns

[0078] This experiment investigated the effects of Shimadzu SIL-16 and Huapu 6000 chromatographs, as well as Agilent ZORBAX SB-C18 columns (4.6×250mm, 5μm), Agilent Eclipse XDB-C18 columns (4.6×250mm, 5μm), and ThermoSecientific BDS HYPERSIL C18 columns (4.6×250mm, 5μm) on the relative correction factor. The results (Table 4) show that the RSDs of the relative correction factors for different instruments and columns all met the requirements (all less than 5%), indicating no significant impact.

[0079] Table 4. Effects of different instruments and chromatographic columns on relative correction factors.

[0080]

[0081]

[0082] 2.6.1.2 Column Temperature

[0083] This experiment investigated the effects of different column temperatures (25℃, 28℃, 30℃, 32℃, and 35℃) on the relative correction factor. The experimental results (Table 5) show that different column temperatures had no significant effect on the relative correction factor.

[0084] Table 5. Effect of different column temperatures on relative correction factors

[0085]

[0086] 2.6.1.3 Volumetric Flow Rate

[0087] This experiment investigated different volumetric flow rates of 0.7, 0.8, 0.9, 1.0, and 1.1 mL·min. -1 The effect on the relative correction factor. As shown in the experimental results (Table 6), different volumetric flow rates have no effect on the relative correction factor.

[0088] Table 6. Effect of different volumetric flow rates on relative correction factor

[0089]

[0090] 2.7 Chromatographic Peak Localization

[0091] This experiment employed the relative retention value method (relative retention value = retention time of analyte / retention time of internal standard) to locate chromatographic peaks under different chromatographic instruments and columns. The experimental results (Table 7) show that all components exhibited good stability under different chromatographic instruments and columns, with RSDs all less than 5%, making them suitable for chromatographic peak localization of analytes.

[0092] Table 7. Relative retention values ​​under different chromatograms and columns.

[0093]

[0094] 2.8 Sample content determination

[0095] Nineteen batches of Jinteng Qingbi Granules samples were collected, and test solutions were prepared according to the method described in section "1.2". The chromatographic conditions described in section "1.3" were used for determination. The content of each component was calculated using both the external standard method and the QAMS method. The content of the seven components showed little difference among the batches of samples. The results are shown in Table 8. The RAD values ​​of both the external standard method and QAMS were less than 2%, indicating that the results of the two methods are similar, and the QAMS method can be used for content determination. The contents of sinomenine, gallic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, and isochlorogenic acid C in 19 batches of samples were determined by QAMS to be 24.986–42.087, 7.014–13.908, 8.883–14.601, 21.676–30.777, 12.194–19.118, 6.524–10.757, and 7.480–12.473, respectively.

[0096] Table 8. Results of component content determination for 19 batches of Jinteng Qingbi Granules

[0097]

[0098]

[0099] Example 2: Determination of the content of 7 components in Jinteng Qingbi Granules

[0100] 1.1 Preparation of test solution

[0101] See Example 1.

[0102] 1.2 Chromatographic conditions

[0103] Chromatographic column: Agilent Eclipse XDB-C18 (4.6×250mm, 5μm); mobile phase: methanol-0.3% phosphoric acid aqueous solution; flow rate: 1.0mL / min; detection wavelength: 262nm, 330nm; column temperature: 30℃; injection volume: 10μL;

[0104] Elution was performed according to the following gradient table:

[0105]

[0106] 1.3 Chromatogram

[0107] Accurately pipette the test solution and inject it into the high-performance liquid chromatography system. Measure and record the chromatogram; the chromatograms of the test solution at 262 nm and 330 nm are shown below. Figures 5-6 .

[0108] Example 3: Determination of the content of 7 components in Jinteng Qingbi Granules

[0109] The content of seven components in Jinteng Qingbi Granules was determined using a multi-component analysis method, including the following steps:

[0110] 1.1 Preparation of test solution

[0111] See Example 1.

[0112] 1.2 Chromatographic conditions

[0113] Chromatographic column: Thermo Secientific BDS HYPERSIL C18 (4.6×250mm, 5μm); mobile phase: methanol-0.1% phosphoric acid aqueous solution; flow rate: 1.0mL / min; detection wavelength: 262nm, 330nm; column temperature: 30℃; injection volume: 10μL;

[0114] Elution was performed according to the following gradient table:

[0115]

[0116] 1.3 Chromatogram

[0117] Accurately pipette the test solution and inject it into the high-performance liquid chromatography system. Measure and record the chromatogram; the chromatograms of the test solution at 262 nm and 330 nm are shown below. Figures 7-8 .

[0118] Comparative Example 1: Determination of the content of 7 components in Jinteng Qingbi Granules

[0119] 1.1 Preparation of test solution

[0120] See Example 1.

[0121] 1.2 Chromatographic conditions

[0122] Chromatographic column: Agilent ZORBAX SB-C18 (4.6×250mm, 5μm); mobile phase: methanol-0.1% formic acid aqueous solution; flow rate: 1.0mL / min; detection wavelength: 262nm, 330nm; column temperature: 30℃; injection volume: 10μL;

[0123] Elution was performed according to the following gradient table:

[0124]

[0125]

[0126] 1.3 Chromatogram

[0127] Accurately pipette the test solution and inject it into the high-performance liquid chromatography system. Measure and record the chromatogram; the chromatograms of the test solution at 262 nm and 330 nm are shown below. Figures 9-10 .

[0128] Comparative Example 2: Determination of the content of 7 components in Jinteng Qingbi Granules

[0129] 1.1 Preparation of test solution

[0130] See Example 1.

[0131] 1.2 Chromatographic conditions

[0132] Chromatographic column: Agilent ZORBAX SB-C18 (4.6×250mm, 5μm); mobile phase: acetonitrile-0.5% phosphoric acid aqueous solution; flow rate: 1.0mL / min; detection wavelength: 262nm, 330nm; column temperature: 30℃; injection volume: 10μL;

[0133] Elution was performed according to the following gradient table:

[0134]

[0135] 1.3 Chromatogram

[0136] Accurately pipette the test solution and inject it into the high-performance liquid chromatography system. Measure and record the chromatogram; the chromatograms of the test solution at 262 nm and 330 nm are shown below. Figures 11-12 .

Claims

1. A method for determining the contents of seven components in Jinduqingbi granules by QAMS, characterized in that, Using sinomenine as an internal standard, the relative correction factors between sinomenine and gallic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C in Jintongqingbi granules are established, and the contents of gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C in Jintongqingbi granules are calculated by correction factors; The high performance liquid chromatography method is used for determination, and the specific method is as follows: A. Preparation of reference substance solution: precisely weigh the gallic acid, neochlorogenic acid, sinomenine, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C reference substances, add methanol to completely dissolve them, prepare a mixed reference substance solution, pass through a microporous filter membrane, and take the filtrate for standby use; B. Preparation of test sample solution: precisely weigh Jintongqingbi granules, precisely add ethanol solution, ultrasonically dissolve them, pass through a microporous filter membrane, and take the filtrate to obtain the test sample solution; C. Chromatographic conditions: chromatographic column: octadecylsilane bonded silica gel as filler; mobile phase: methanol-phosphoric acid aqueous solution; According to the following gradient elution program: 。 2. The method of claim 1, wherein, The chromatographic column is selected from one of Agilent ZORBAX SB-C18, Agilent Eclipse XDB-C18 and Thermo Secientific BDS HYPERSIL C18.

3. The method of claim 2, wherein, The specification of the chromatographic column is 4.6mm×250mm, 5μm.

4. The method of claim 1, wherein, The concentration of the phosphoric acid aqueous solution in the mobile phase is 0.1%-1.0%.

5. The method of claim 4, wherein, The concentration of the phosphoric acid aqueous solution in the mobile phase is 0.3%-0.5%.

6. The method of claim 1, wherein, The chromatographic column temperature is 25-35℃.

7. The method of claim 1, wherein, The detection wavelength of gallic acid and sinomenine is 260-270nm, and the detection wavelength of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C is 320-340nm.

8. The method of claim 7, wherein, The detection wavelength of gallic acid and sinomenine is 262nm, and the detection wavelength of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C is 330nm.

9. The method of claim 1, wherein, The flow rate of the mobile phase is 0.7-1.1ml / min.

10. The method according to any one of claims 1-9 is used for quality detection and evaluation of Jintongqingbi granules.

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