Fingerprint quality control method of compound radix zanthoxyli composition

Abstract

The invention discloses a fingerprint quality control method of a compound radix zanthoxyli composition. The method comprises the following steps: carrying out gradient elution and determination by using a high performance liquid chromatograph; the high performance liquid chromatography conditions are as follows: a chromatographic column is a C18 chromatographic column; the detection wavelength is 250 nm; a mobile phase A is methanol, a mobile phase B is a 0.1% triethylamine solution containing 0.1% of formic acid by weight, and gradient elution is adopted. The HPLC fingerprint spectrum of the compound radix zanthoxyli composition is established through chromatographic condition optimization and a test sample preparation method. The method is simple and accurate to operate and good in repeatability, and the quality consistency of the compound radix zanthoxyli composition can be well controlled.

Description

Technical Field

[0001] The invention relates to the field of quality control of traditional Chinese medicines, and in particular to a fingerprint quality control method for a compound Zanthoxylum bungeanum composition. Background Art

[0002] Compound Liangmianzhen Lozenges are the exclusive products of Guangxi Zhuang Autonomous Region Huahong Pharmaceutical Group Co., Ltd. Its prescription consists of Xiqing fruit, butterfly fruit, Sophora flavescens, peppermint oil, menthol, Liangmianzhen, and licorice. Among them, Xiqing fruit has the effects of clearing heat, promoting body fluid, and detoxification; it is mainly used to treat yin deficiency diphtheria, pharyngitis, bacillary dysentery, etc. Butterfly fruit has the effect of clearing heat and detoxification, and is mainly used to treat hoarseness, pharyngitis, and tonsillitis. Sophora flavescens clears heat and detoxifies, and relieves swelling and relieves sore throat. It is used for fire toxicity accumulation, milk moth throat paralysis, sore throat, sore gums, and sores in the mouth and tongue. It can promote blood circulation and remove blood stasis, promote qi and relieve pain, dispel wind and dredge collaterals, and detoxify and reduce swelling. Liangmianzhen has the effects of promoting blood circulation and removing blood stasis, promoting qi and relieving pain, dispelling wind and dredge collaterals, detoxifying and reducing swelling, and is used for injuries caused by falls, stomachache, toothache, etc. Menthol has the effects of dispelling wind, clearing heat, and detoxification. It is mainly used to treat headache, red eyes, exogenous wind-heat, sore throat and teeth, itchy skin, etc. Peppermint oil is an aromatic, flavoring and carminative medicine, which can be used to produce a cooling sensation on the skin or mucous membrane to relieve discomfort and pain; these medicinal materials have the effect of clearing heat and detoxifying. Although there have been some reports on the quality control of this preparation, there is still a lack of qualitative analysis of the fingerprint of the chemical composition of this preparation. Summary of the invention

[0003] The object of the present invention is to provide a fingerprint quality control method for a compound Zanthoxylum bungeanum composition. The present invention establishes an HPLC fingerprint of the compound Zanthoxylum bungeanum composition by optimizing chromatographic conditions and preparing a sample. The method is simple to operate, accurate, and has good repeatability, and can better control the quality consistency of the compound Zanthoxylum bungeanum composition.

[0004] The technical contents of the present invention are as follows:

[0005] The fingerprint quality control method of the compound Zanthoxylum bungeanum composition;

[0006] including high performance liquid chromatography for gradient elution and determination;

[0007] The HPLC conditions are:

[0008] Chromatographic column: C18 column;

[0009] Detection wavelength: 250nm;

[0010] Mobile phase: Mobile phase A is methanol, and mobile phase B is a 0.1% triethylamine solution containing 0.1% formic acid by weight. The percentages here are weight percentages. Gradient elution is used. The elution procedure is as follows:

[0011]

[0012]

[0013] The specifications of the C18 chromatographic column are: 250×4.6 mm, 5 μm; the temperature of the chromatographic column is 25-35° C.; and the flow rate of the mobile phase is 0.7-1.2 mL / min.

[0014] The C18 chromatographic column is phenomenex C 18 Chromatographic column.

[0015] The preparation method of the test solution comprises the following steps:

[0016] Accurately weigh 0.15-0.3g of the compound Zanthoxylum lozenge paste, place it in a container, accurately add 10mL of 50% methanol, accurately weigh it, ultrasonically treat it for 40 minutes, let it cool, accurately weigh it again, make up the lost weight with 50% methanol, shake it well, centrifuge it, take the supernatant, and filter it through a 0.45μm microporous filter membrane to obtain the product;

[0017] Or grind the compound Zanthoxylum bungeanum tablets, accurately weigh 1-3g of the ground sample, put it in a container, accurately add 10mL of 50% methanol, accurately weigh the weight, ultrasonically treat for 40 minutes, cool, accurately weigh again, make up the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter through a 0.45μm microporous filter membrane to obtain the product.

[0018] The preparation method of the compound Zanthoxylum bungeanum thick paste comprises the following steps:

[0019] Of the above seven herbs, Sophora flavescens and Zanthoxylum bungeanum are crushed into coarse powder, and an appropriate amount of 75% ethanol is added. After partial solvent immersion for 12 hours, the flow rate is controlled to percolate until the percolated liquid has no alkaloid reaction, and the ethanol is recovered from the percolated liquid, and the liquid is concentrated under reduced pressure to become a clear paste; Xiqing fruit is decocted with water twice, each time for 1.5 hours, the decoctions are combined, filtered, and the filtrate is concentrated into a clear paste, which is allowed to stand for 12 hours, and the supernatant is concentrated into a clear paste; Butterfly fruit and liquorice are decocted with water twice, each time for 1.5 hours, the decoctions are combined, filtered, and the filtrate is concentrated into a clear paste; the above three clear pastes are combined to obtain the paste.

[0020] The preparation method of the compound Zanthoxylum bungeanum tablets comprises the following steps:

[0021] The above seven ingredients, Sophora flavescens and Zanthoxylum bungeanum, are crushed into coarse powder, and an appropriate amount of 75% ethanol is added. After soaking with a partial solvent for 12 hours, the flow rate is controlled to percolate until the percolated liquid has no alkaloid reaction, and the percolated liquid is recovered with ethanol, and the percolated liquid is concentrated under reduced pressure to form a clear paste; Xiqing fruit is decocted with water twice, each time for 1.5 hours, the decoction is combined, filtered, and the filtrate is concentrated into a clear paste, and the supernatant is concentrated into a clear paste after standing for 12 hours; butterfly fruit and licorice are decocted with water twice, each time for 1.5 hours, the decoction is combined, filtered, and the filtrate is concentrated into a clear paste. Combine the above three clear pastes, add sucrose, mannitol, sodium carboxymethyl cellulose, stevioside and sodium copper chlorophyll, mix well, dry, crush into fine powder, add povidone solution, mix well, granulate, add peppermint oil, menthol and magnesium stearate, mix well, press into tablets, and make 100 tablets to obtain; or film-coat to obtain.

[0022] The method for preparing the reference solution comprises the following steps:

[0023] Take gallic acid reference substance, add 50% methanol to dissolve it, and make a 30-70μg / mL solution.

[0024] Establishment of standard fingerprint: Take 20 μL of each of the reference solution and 16 batches of compound Zanthoxylum bungeanum as the test solution, inject into high performance liquid chromatograph, measure, record the chromatogram, and determine the gallic acid in the test sample chromatogram as the reference peak; detect at 250 nm, obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, so as to obtain a standard fingerprint formed by 20 common characteristic peaks.

[0025] The characteristic peaks of the standard fingerprint are as follows:

[0026]

[0027]

[0028] The fingerprint quality control method of the compound Zanthoxylum bungeanum composition comprises the following contents:

[0029] (1) Preparation of test solution:

[0030] Accurately weigh 0.15-0.3g of the compound Zanthoxylum lozenge paste, place it in a container, accurately add 10mL of 50% methanol, accurately weigh it, ultrasonically treat it for 40 minutes, let it cool, accurately weigh it again, make up the lost weight with 50% methanol, shake it well, centrifuge it, take the supernatant, and filter it through a 0.45μm microporous filter membrane to obtain the product;

[0031] Or grind the compound Zanthoxylum bungeanum tablets, accurately weigh 1-3g of the grinded sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the weight, ultrasonically treat for 40 minutes, cool, accurately weigh again, make up the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter through a 0.45μm microporous filter membrane to obtain;

[0032] (2) Preparation of reference solution:

[0033] Take gallic acid reference substance, add 50% methanol to dissolve, and make a 30-70μg / mL solution;

[0034] (3) Chromatographic conditions:

[0035] Chromatographic column: C18 column; 250×4.6mm, 5μm; Column temperature: 25-35℃;

[0036] Detection wavelength: 250nm;

[0037] Mobile phase: Mobile phase A is methanol, mobile phase B is 0.1% triethylamine solution containing 0.1% formic acid by weight percentage, gradient elution is adopted, and the elution procedure is as follows:

[0038]

[0039]

[0040] (4) Establishment of standard fingerprint: Take 20 μL of reference solution and 16 batches of compound Zanthoxylum bungeanum solution as test solution, inject into high performance liquid chromatograph, measure, record chromatogram, determine gallic acid in the test solution chromatogram as reference peak; detect at 250 nm, obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, and thus obtain the standard fingerprint formed by the 20 common characteristic peaks;

[0041] (5) using the methods described in (1) to (3) as a means of testing the fingerprint of the compound Zanthoxylum bungeanum composition to be tested, to prepare a fingerprint of the sample to be tested;

[0042] (6) Comparing the fingerprint of the tested compound Zanthoxylum bungeanum composition with the standard fingerprint of step (4), the similarity is greater than 0.9.

[0043] The compound Zanthoxylum bungeanum composition is prepared from the following medicinal materials: citronella, butterfly fruit, sophora flavescens, peppermint oil, menthol, Zanthoxylum bungeanum and liquorice.

[0044] Beneficial effects of the present invention:

[0045] The present invention realizes the fingerprint quality control mode and quality consistency control method of the compound Zanthoxylum bungeanum composition, can accurately control and evaluate the quality of the compound Zanthoxylum bungeanum composition, and makes up for the shortcomings of the current quality control method. The method of the present invention is simple to operate, accurate, and has good repeatability, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 HPLC superimposed spectra (S1-S16) and reference spectra (R) of 16 batches of compound Liangmianzhen lozenge samples;

[0047] Figure 2 HPLC charts of gallic acid reference substance and compound Zanthoxylum bungeanum tablets. DETAILED DESCRIPTION

[0048] The present invention is further described below by way of examples. It should be understood that the examples of the present invention are intended to illustrate the present invention rather than to limit the present invention.

[0049] Example 1

[0050] The fingerprint quality control method of the compound Zanthoxylum bungeanum composition includes the following contents:

[0051] (1) Preparation of test solution:

[0052] Accurately weigh 0.5 g of the compound Zanthoxylum bungeanum thick paste, place it in a container, accurately add 10 mL of 50% methanol, accurately weigh it, ultrasonicate it for 40 min, let it cool, accurately weigh it again, make up the lost weight with 50% methanol, shake it well, centrifuge it, take the supernatant, and filter it through a 0.45 μm microporous filter membrane to obtain the product;

[0053] Or grind the compound Zanthoxylum bungeanum tablets, accurately weigh 1g of the grinded sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the weight, ultrasonically treat for 40 minutes, cool, accurately weigh again, make up the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter through a 0.45μm microporous filter membrane to obtain;

[0054] (2) Preparation of reference solution:

[0055] Take the gallic acid reference substance, add 50% methanol to dissolve it, and make a 30μg / mL solution;

[0056] (3) Chromatographic conditions:

[0057] Chromatographic column: C18 column; 250×4.6mm, 5μm; column temperature is 25℃;

[0058] Detection wavelength: 250nm;

[0059] Mobile phase: Mobile phase A is methanol, mobile phase B is 0.1% triethylamine solution containing 0.1% formic acid by weight percentage, gradient elution is adopted, and the elution procedure is as follows:

[0060] Time (min) Methanol(%) 0.1% formic acid-0.1% triethylamine (%) 0-15 5 95 15-80 5-40 95-60 80-90 40-55 60-45 90-100 55-60 45-40

[0061] (4) Establishment of standard fingerprint: Take 20 μL of reference solution and 16 batches of compound Zanthoxylum bungeanum solution as test solution, inject into high performance liquid chromatograph, measure, record chromatogram, determine gallic acid in the test solution chromatogram as reference peak; detect at 250 nm, obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, and thus obtain the standard fingerprint formed by the 20 common characteristic peaks;

[0062] The characteristic peaks of the standard fingerprint are as follows:

[0063]

[0064]

[0065] (5) using the methods described in (1) to (3) as a means of testing the fingerprint of the compound Zanthoxylum bungeanum composition to be tested, to prepare a fingerprint of the sample to be tested;

[0066] (6) Comparing the fingerprint of the tested compound Zanthoxylum bungeanum composition with the standard fingerprint of step (4), the similarity is greater than 0.97.

[0067] Example 2

[0068] The fingerprint quality control method of the compound Zanthoxylum bungeanum composition includes the following contents:

[0069] (1) Preparation of test solution:

[0070] Accurately weigh 0.3 g of the compound Zanthoxylum bungeanum paste, place it in a container, accurately add 10 mL of 50% methanol, accurately weigh it, ultrasonicate it for 40 min, let it cool, accurately weigh it again, make up the lost weight with 50% methanol, shake it well, centrifuge it, take the supernatant, and filter it through a 0.45 μm microporous membrane to obtain the product;

[0071] Or grind the compound Zanthoxylum bungeanum tablets, accurately weigh 3g of the grinded sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the weight, ultrasonically treat for 40 minutes, cool, accurately weigh again, make up the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter through a 0.45μm microporous membrane to obtain;

[0072] (2) Preparation of reference solution:

[0073] Take the gallic acid reference substance, add 50% methanol to dissolve it, and make a 70μg / mL solution;

[0074] (3) Chromatographic conditions:

[0075] Chromatographic column: C18 column; 250×4.6mm, 5μm; column temperature is 35℃;

[0076] Detection wavelength: 250nm;

[0077] Mobile phase: Mobile phase A is methanol, mobile phase B is 0.1% triethylamine solution containing 0.1% formic acid by weight percentage, gradient elution is adopted, and the elution procedure is as follows:

[0078] Time (min) Methanol(%) 0.1% formic acid-0.1% triethylamine (%) 0-15 5 95 15-80 5-40 95-60 80-90 40-55 60-45 90-100 55-60 45-40

[0079] (4) Establishment of standard fingerprint: Take 20 μL of reference solution and 16 batches of compound Zanthoxylum bungeanum solution as test solution, inject into high performance liquid chromatograph, measure, record chromatogram, determine gallic acid in the test solution chromatogram as reference peak; detect at 250 nm, obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, and thus obtain the standard fingerprint formed by the 20 common characteristic peaks;

[0080] The characteristic peaks of the standard fingerprint are as follows:

[0081]

[0082]

[0083] (5) using the methods described in (1) to (3) as a means of testing the fingerprint of the compound Zanthoxylum bungeanum composition to be tested, to prepare a fingerprint of the sample to be tested;

[0084] (6) Comparing the fingerprint of the tested compound Zanthoxylum bungeanum composition with the standard fingerprint of step (4), the similarity is greater than 0.97.

[0085] Example 3

[0086] The fingerprint quality control method of the compound Zanthoxylum bungeanum composition includes the following contents:

[0087] (1) Preparation of test solution:

[0088] Accurately weigh 0.2 g of the compound Zanthoxylum bungeanum thick paste, place it in a container, accurately add 10 mL of 50% methanol, accurately weigh it, ultrasonicate it for 40 min, let it cool, accurately weigh it again, make up the lost weight with 50% methanol, shake it well, centrifuge it, take the supernatant, and filter it through a 0.45 μm microporous filter membrane to obtain the product;

[0089] Or grind the compound Zanthoxylum bungeanum tablets, accurately weigh 2g of the grinded sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the weight, ultrasonically treat for 40 minutes, cool, accurately weigh again, make up the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter through a 0.45μm microporous membrane to obtain;

[0090] (2) Preparation of reference solution:

[0091] Take the gallic acid reference substance, add 50% methanol to dissolve it, and make a 50μg / mL solution;

[0092] (3) Chromatographic conditions:

[0093] Chromatographic column: C18 column; 250×4.6mm, 5μm; Column temperature: 30℃;

[0094] Detection wavelength: 250nm;

[0095] Mobile phase: Mobile phase A is methanol, mobile phase B is 0.1% triethylamine solution containing 0.1% formic acid by weight percentage, gradient elution is adopted, and the elution procedure is as follows:

[0096] Time (min) Methanol(%) 0.1% formic acid-0.1% triethylamine (%) 0-15 5 95 15-80 5-40 95-60 80-90 40-55 60-45 90-100 55-60 45-40

[0097] (4) Establishment of standard fingerprint: Take 20 μL of reference solution and 16 batches of compound Zanthoxylum bungeanum solution as test solution, inject into high performance liquid chromatograph, measure, record chromatogram, determine gallic acid in the test solution chromatogram as reference peak; detect at 250 nm, obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, and thus obtain the standard fingerprint formed by the 20 common characteristic peaks;

[0098] The characteristic peaks of the standard fingerprint are as follows:

[0099]

[0100]

[0101] (5) using the methods described in (1) to (3) as a means of testing the fingerprint of the compound Zanthoxylum bungeanum composition to be tested, to prepare a fingerprint of the sample to be tested;

[0102] (6) Comparing the fingerprint of the tested compound Zanthoxylum bungeanum composition with the standard fingerprint of step (4), the similarity is greater than 0.97.

[0103] Example 4 Study on the Fingerprint of Compound Zanthoxylum bungeanum

[0104] 1. Materials

[0105] 1.1 Instrument

[0106] High performance liquid chromatograph (Agilent1100, DAD detector), 1 / 10,000 balance (Sartorius Scientific Instruments (Beijing) Co., Ltd.), 1 / 100,000 balance (Sartorius Scientific Instruments (Beijing) Co., Ltd.), ultrasonic cleaner (Yumeng Shenzhen Fangao Microelectronics Co., Ltd.), HC-3018R high-speed refrigerated centrifuge (Anhui Zhongke Zhongjia Scientific Instruments Co., Ltd.)

[0107] 1.2 Drug testing

[0108] Methanol (chromatographic grade, Merck), formic acid (analytical grade, Tianjin Fuyu Fine Chemical Co., Ltd.), triethylamine (Tianjin Damao Chemical Reagent Factory), methanol (analytical grade, Tianjin Jindong Tianzheng Fine Chemical Reagent Factory), Watsons distilled water (Guangzhou Watsons Food and Beverage Co., Ltd.), gallic acid reference substance (batch number: WP23071413, manufacturer: Sichuan Weikeqi Biotechnology Co., Ltd.), 16 batches of compound Liangmianzhen lozenges, batch numbers are: 20220505, 20220506, 20220507, 20220508, 20220509, 20220501, 20220503, 20220507, 20220508, 20220509, 20220501, 20220503, 20220505, 20220506, 20220507, 20220508, 20220509 ... 11001, 20210801, 20220201, 20220401, 20211202, 20220302, 20220502, 20220501, 20211101, 20211201, 20220301, 20220402, 20220503, 20220504, 20230201 (provided by Guangxi Zhuang Autonomous Region Huahong Pharmaceutical Group Co., Ltd.), numbers S1 to S16, see Table 1.

[0109] Table 1 16 batches of compound prescription Liangmianzhen lozenges serial numbers and batch information

[0110]

[0111] 2. Methods and Results

[0112] 2.1 Chromatographic conditions

[0113] A phenomenex C18 column (250×4.6 mm, 5 μm) was used, the mobile phase was methanol (A) 0.1% formic acid-0.1% triethylamine aqueous solution (B), gradient elution, column temperature: 30°C, flow rate: 1 mL / min, injection volume: 20 μL, detection wavelength: 250 nm.

[0114] The elution procedure is as follows:

[0115]

[0116]

[0117] 2.2 Solution preparation

[0118] 2.2.1 Preparation of reference solution

[0119] Accurately weigh 0.01256 g of gallic acid reference substance, place it in a 250 mL volumetric flask, add 50% methanol to dissolve and dilute to the mark to prepare a 50 μg / mL reference substance solution.

[0120] 2.2.2 Preparation of test solution

[0121] Take an appropriate amount of sample, grind it into powder, take about 1.44g, weigh it accurately, put it in a stoppered conical flask, accurately add 10mL of 50% methanol, weigh it, treat it with ultrasound for 40min, let it cool, weigh it again, make up the lost weight with 50% methanol, shake it well, centrifuge it (8000r, 5min), take the supernatant, and filter it through a 0.45μm microporous filter membrane to obtain the product.

[0122] 2.3 Methodological investigation

[0123] 2.3.1 Precision test

[0124] Prepare one portion of the test solution according to the method under "2.2.2". Under the chromatographic conditions of "2.1", inject the sample six times continuously. Take gallic acid as the reference peak (S). Calculate the relative retention time and relative peak area of ​​20 common peaks. The relative retention time RSD is <0.473%, and the relative peak area RSD is <3.983%, indicating that the instrument has good precision.

[0125] 2.3.2 Repeatability test

[0126] Take 6 portions of sample with batch number 20220502, prepare the test solution according to the method under "2.2.2", and inject each sample once under the chromatographic conditions of "2.1". Take gallic acid as the reference peak (S), calculate the relative retention time and relative peak area of ​​20 common peaks, the relative retention time RSD <1.203%, the relative peak area RSD <4.979%, indicating that the method has good repeatability.

[0127] 2.3.3 Stability test

[0128] Prepare one portion of the test solution according to the method under "2.2.2", store it at room temperature under the chromatographic conditions of "2.1", and measure it at 0, 2, 4, 6, 8, 12, and 24 h, respectively. Take gallic acid as the reference peak (S), calculate the relative retention time and relative peak area of ​​20 common peaks, and the relative retention time RSD <1.232%, and the relative peak area RSD <4.891%, indicating that the test solution has good stability within 24 h.

[0129] 2.4 HPLC fingerprint establishment

[0130] Take 16 batches of compound Liangmianzhen lozenge sample solutions, inject and measure according to the chromatographic conditions under "2.1", record the chromatogram, import the data into the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)", use the chromatogram of sample S12 as the reference spectrum, use the average method to generate a common pattern, match the common peaks of the characteristic spectra of 16 batches of compound Liangmianzhen lozenges, and calibrate a total of 20 common peaks, see Figure 1 By comparing with the reference substance ( Figure 2 ) and identified the gallic acid component. The similarity calculation was performed on 16 batches of samples, and the similarity was above 0.97.

[0131] The above is an embodiment of the present invention. The above embodiments and the specific parameters in the embodiments are only for the purpose of clearly describing the invention verification process, and are not intended to limit the patent protection scope of the present invention. The patent protection scope of the present invention is still subject to its claims. Any equivalent structural changes made by using the contents of the description and drawings of the present invention should be included in the protection scope of the present invention.

Claims

1. A fingerprint quality control method for a compound Zanthoxylum bungeanum composition, characterized in that: including high performance liquid chromatography for gradient elution and determination; The HPLC conditions are: Chromatographic column: C18 column; Detection wavelength: 250nm; Mobile phase: Mobile phase A is methanol, mobile phase B is 0.1% triethylamine solution containing 0.1% formic acid by weight percentage, gradient elution is adopted, and the elution procedure is as follows: 。 2. The fingerprint quality control method of the compound Zanthoxylum bungeanum composition according to claim 1, characterized in that: The specifications of the C18 chromatographic column are: 250×4.6 mm, 5 μm; the temperature of the chromatographic column is 25-35° C.; and the flow rate of the mobile phase is 0.7-1.2 mL / min.

3. The fingerprint quality control method of the compound Zanthoxylum bungeanum composition according to claim 2, characterized in that: The C18 chromatographic column is phenomenex C 18 Chromatographic column.

4. The fingerprint quality control method of the compound Zanthoxylum bungeanum composition according to claim 1, wherein the preparation method of the test solution comprises the following steps: Accurately weigh 0.15-0.3g of the compound Zanthoxylum lozenge paste, place it in a container, accurately add 10mL of 50% methanol, accurately weigh it, ultrasonically treat it for 40 minutes, let it cool, accurately weigh it again, make up the lost weight with 50% methanol, shake it well, centrifuge it, take the supernatant, and filter it through a 0.45μm microporous filter membrane to obtain the product; Or grind the compound Zanthoxylum bungeanum tablets, accurately weigh 1-3g of the ground sample, put it in a container, accurately add 10mL of 50% methanol, accurately weigh the weight, ultrasonically treat for 40 minutes, cool, accurately weigh again, make up the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter through a 0.45μm microporous filter membrane to obtain the product.

5. The fingerprint quality control method of the compound Zanthoxylum bungeanum composition according to claim 4, characterized in that: The method for preparing the reference solution comprises the following steps: Take gallic acid reference substance, add 50% methanol to dissolve it, and make a 30-70μg / mL solution.

6. The fingerprint quality control method of the compound Zanthoxylum bungeanum composition according to claim 5, characterized in that: Establishment of standard fingerprint: Take 20 μL of each of the reference solution and 16 batches of compound Zanthoxylum bungeanum as the test solution, inject into high performance liquid chromatograph, measure, record the chromatogram, and determine the gallic acid in the test sample chromatogram as the reference peak; detect at 250 nm, obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, so as to obtain a standard fingerprint formed by 20 common characteristic peaks.

7. The fingerprint quality control method of the compound Zanthoxylum bungeanum composition according to claim 6, characterized in that: The characteristic peaks of the standard fingerprint are as follows:

8. The fingerprint quality control method of the compound Zanthoxylum bungeanum composition according to claim 1, characterized in that: Includes the following: (1) Preparation of test solution: Accurately weigh 0.15-0.3g of the compound Zanthoxylum lozenge paste, place it in a container, accurately add 10mL of 50% methanol, accurately weigh it, ultrasonically treat it for 40 minutes, let it cool, accurately weigh it again, make up the lost weight with 50% methanol, shake it well, centrifuge it, take the supernatant, and filter it through a 0.45μm microporous filter membrane to obtain the product; Or grind the compound Zanthoxylum bungeanum tablets, accurately weigh 1-3g of the grinded sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the weight, ultrasonically treat for 40 minutes, cool, accurately weigh again, make up the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter through a 0.45μm microporous filter membrane to obtain; (2) Preparation of reference solution: Take the gallic acid reference substance, add 50% methanol to dissolve it, and make a 30-70μg / mL solution; (3) Chromatographic conditions: Chromatographic column: C18 column; 250×4.6mm, 5μm; Column temperature: 25-35℃; Detection wavelength: 250nm; Mobile phase: Mobile phase A is methanol, mobile phase B is 0.1% triethylamine solution containing 0.1% formic acid by weight percentage, gradient elution is adopted, and the elution procedure is as follows: (4) Establishment of standard fingerprint: Take 20 μL of reference solution and 16 batches of compound Zanthoxylum bungeanum solution as test solution, inject into high performance liquid chromatograph, measure, record chromatogram, determine gallic acid in the test solution chromatogram as reference peak; detect at 250 nm, obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, and thus obtain the standard fingerprint formed by the 20 common characteristic peaks; (5) using the methods described in (1) to (3) as a means of testing the fingerprint of the compound Zanthoxylum bungeanum composition to be tested, to prepare a fingerprint of the sample to be tested; (6) Comparing the fingerprint of the tested compound Zanthoxylum bungeanum composition with the standard fingerprint of step (4), the similarity is greater than 0.

9.

9. The fingerprint quality control method of the compound Zanthoxylum bungeanum composition according to any one of claims 1 to 8, characterized in that: The compound Zanthoxylum bungeanum composition is prepared from the following medicinal materials: citronella, butterfly fruit, sophora flavescens, peppermint oil, menthol, Zanthoxylum bungeanum and liquorice.

Images