A method for quality control of fingerprint spectrum of compound Zanthoxylum bungeanum composition

Abstract

This invention discloses a fingerprint quality control method for a compound *Zanthoxylum nitidum* composition, comprising gradient elution and determination using high-performance liquid chromatography (HPLC). The HPLC conditions are: column: C18 column; detection wavelength: 250 nm; mobile phase: mobile phase A is methanol, and mobile phase B is a 0.1% triethylamine solution containing 0.1% formic acid by weight, using gradient elution. This invention establishes the HPLC fingerprint of the compound *Zanthoxylum nitidum* composition through optimized chromatographic conditions and sample preparation methodology. This method is simple to operate, accurate, and has good repeatability, and can effectively control the quality consistency of the compound *Zanthoxylum nitidum* composition.

Description

Technical Field

[0001] This invention relates to the field of quality control of traditional Chinese medicine, specifically to a fingerprint spectrum quality control method for a compound Zanthoxylum bungeanum composition. Background Technology

[0002] Compound Zanthoxylum nitidum lozenges are an exclusive product of Guangxi Zhuang Autonomous Region Huahong Pharmaceutical Group Co., Ltd. Its formula consists of Terminalia chebula, Butterfly fruit, Sophora tonkinensis root, peppermint oil, menthol, Zanthoxylum nitidum, and licorice. Terminalia chebula has the effects of clearing heat and promoting body fluid production, and detoxifying; it is mainly used to treat diphtheria due to yin deficiency, pharyngitis, and bacillary dysentery. Butterfly fruit has the effects of clearing heat and detoxifying, and is mainly used to treat hoarseness, pharyngitis, and tonsillitis. Sophora tonkinensis root clears heat and detoxifies, reduces swelling and soothes the throat. It is used for fire toxin accumulation, tonsillitis, sore throat, swollen gums, and oral ulcers. It also invigorates blood circulation, removes blood stasis, promotes qi circulation and relieves pain, dispels wind and unblocks meridians, and detoxifies and reduces swelling. Zanthoxylum nitidum has the effects of invigorating blood circulation, removing blood stasis, promoting qi circulation and relieving pain, dispelling wind and unblocking meridians, and detoxifying and reducing swelling; it is used for injuries from falls, stomach pain, and toothache. Menthol has the effects of dispelling wind, clearing heat, and detoxifying. It is mainly used to treat headaches, red eyes, external wind-heat, sore throat, toothache, and itchy skin. Peppermint oil is an aromatic, flavoring, and carminative agent that can be used on the skin or mucous membranes to produce a cooling sensation to relieve discomfort and pain; these medicinal materials all have the effect of clearing heat and detoxifying. Although there are some reports on the quality control of this preparation, qualitative analysis of its chemical composition by fingerprinting is still lacking. Summary of the Invention

[0003] The purpose of this invention is to provide a fingerprint chromatographic quality control method for a compound Zanthoxylum bungeanum composition. This invention establishes an HPLC fingerprint chromatogram for the compound Zanthoxylum bungeanum composition through chromatographic condition optimization and sample preparation methodology. This method is simple to operate, accurate, and has good repeatability, enabling effective control of the quality consistency of the compound Zanthoxylum bungeanum composition.

[0004] The technical content of this invention is as follows:

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

[0006] This includes gradient elution and determination using high-performance liquid chromatography;

[0007] The high-performance liquid chromatography conditions are as follows:

[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. Gradient elution is used, and the elution program is as follows:

[0011]

[0012]

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

[0014] The C18 column mentioned is a phenomenex C18 column. 18 Chromatographic column.

[0015] The method for preparing the test solution includes the following steps:

[0016] Accurately weigh 0.15-0.3g of compound Zanthoxylum nitidum tablets viscous extract, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the extract, sonicate for 40min, cool, accurately weigh the extract again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0017] Alternatively, take compound Zanthoxylum bungeanum tablets, grind them into a fine powder, accurately weigh 1-3g of the powdered sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the sample, sonicate for 40min, cool, accurately weigh the sample again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0018] The preparation method of the compound Zanthoxylum nitidum lozenge paste includes the following steps:

[0019] The above seven ingredients are: Sophora tonkinensis and Zanthoxylum nitidum are crushed into coarse powder, and an appropriate amount of 75% ethanol is added. After soaking in the solvent for 12 hours, the flow rate is controlled and the solution is percolated until no alkaloid reaction occurs. The ethanol is recovered from the solution and concentrated under reduced pressure to obtain a clear extract. Terminalia chebula is decocted twice with water, 1.5 hours each time. The decoctions are combined, filtered, and the filtrate is concentrated to obtain a clear extract. After standing for 12 hours, the supernatant is collected and concentrated to obtain a clear extract. Butterfly fruit and licorice are decocted twice with water, 1.5 hours each time. The decoctions are combined, filtered, and the filtrate is concentrated to obtain a clear extract. The above three clear extracts are combined to obtain the final product.

[0020] The preparation method of the compound Zanthoxylum bungeanum lozenges includes the following steps:

[0021] The above seven ingredients are: * **Sophora tonkinensis** and *Zanthoxylum nitidum**, pulverized into coarse powder, mixed with an appropriate amount of 75% ethanol, partially soaked in the solvent for 12 hours, then percolated at a controlled flow rate until no alkaloid reaction occurs in the percolate. The ethanol is recovered from the percolate, and the mixture is concentrated under reduced pressure to obtain a clear extract. * **Terminalia chebula** is decocted twice with water, 1.5 hours each time, the decoctions are combined, filtered, and the filtrate is concentrated to obtain a clear extract. After standing for 12 hours, the supernatant is collected and concentrated to obtain a clear extract. * **Butterfly fruit** and *Glycyrrhiza uralensis** are decocted twice with water, 1.5 hours each time, the decoctions are combined, filtered, and the filtrate is concentrated to obtain a clear extract. The above three clear extracts are combined, and sucrose, mannitol, sodium carboxymethyl cellulose, stevioside, and sodium copper chlorophyllin are added, mixed well, dried, pulverized into a fine powder, then povidone solution is added, mixed well, granulated, peppermint oil, menthol, and magnesium stearate are added, mixed well, and compressed into tablets to produce 1000 tablets; or coated with a film.

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

[0023] Take gallic acid reference standard, dissolve it in 50% methanol to prepare a solution of 30-70 μg / mL.

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

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

[0026]

[0027]

[0028] The method for quality control of the fingerprint spectrum of the compound Zanthoxylum bungeanum composition includes the following:

[0029] (1) Preparation of the test solution:

[0030] Accurately weigh 0.15-0.3g of compound Zanthoxylum nitidum tablets viscous extract, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the extract, sonicate for 40min, cool, accurately weigh the extract again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0031] Alternatively, take compound Zanthoxylum bungeanum tablets, grind them into a fine powder, accurately weigh 1-3g of the finely ground sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the sample, sonicate for 40min, cool, accurately weigh the sample again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0032] (2) Preparation of reference solution:

[0033] Take gallic acid reference standard, dissolve it in 50% methanol to prepare a solution of 30-70 μg / mL;

[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, and mobile phase B is a 0.1% triethylamine solution containing 0.1% formic acid by weight. Gradient elution is used, and the elution program is as follows:

[0038]

[0039]

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

[0041] (5) Using the methods described in (1)-(3) as the fingerprint spectrum test means for the compound Zanthoxylum bungeanum composition to be tested, prepare the fingerprint spectrum of the sample to be tested;

[0042] (6) The fingerprint spectrum of the compound Zanthoxylum bungeanum to be tested is compared with the standard fingerprint spectrum of step (4), and the similarity is greater than 0.9.

[0043] The compound Zanthoxylum bungeanum composition is made from the following medicinal materials: Terminalia chebula, Butterfly fruit, Sophora tonkinensis root, peppermint oil, menthol, Zanthoxylum bungeanum, and licorice.

[0044] The beneficial effects of this invention are:

[0045] This invention realizes a fingerprint spectrum quality control mode and quality consistency control method for compound Zanthoxylum bungeanum compositions, which can accurately control and evaluate the quality of compound Zanthoxylum bungeanum compositions, and make up for the shortcomings of current quality control methods. The method of this invention is simple to operate, accurate, and has good repeatability, and has good application prospects. Attached Figure Description

[0046] Figure 1 The HPLC superimposed chromatograms (S1~S16) and the control chromatogram (R) of 16 batches of compound Zanthoxylum bungeanum tablets are shown.

[0047] Figure 2 The images show the HPLC chromatograms of gallic acid reference standard and compound Zanthoxylum bungeanum tablets. Detailed Implementation

[0048] The present invention will be further illustrated below by way of examples. It should be understood that the examples of the present invention are for illustrative purposes and not for limiting the present invention.

[0049] Example 1

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

[0051] (1) Preparation of the test solution:

[0052] Accurately weigh 0.5g of compound Zanthoxylum nitidum tablets thick paste, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the contents, sonicate for 40min, cool, accurately weigh the contents again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0053] Alternatively, take compound Zanthoxylum bungeanum tablets, grind them into a fine powder, accurately weigh 1g of the finely ground sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the sample, sonicate for 40min, cool, accurately weigh the sample again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0054] (2) Preparation of reference solution:

[0055] Take gallic acid reference standard, dissolve it in 50% methanol to prepare a 30 μg / mL solution;

[0056] (3) Chromatographic conditions:

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

[0058] Detection wavelength: 250nm;

[0059] Mobile phase: Mobile phase A is methanol, and mobile phase B is a 0.1% triethylamine solution containing 0.1% formic acid by weight. Gradient elution is used, and the elution program 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 spectrum: Take 20 μL each of the reference solution and 16 batches of compound Zanthoxylum bungeanum as test solution, inject them into the high performance liquid chromatograph, measure and record the chromatogram, and determine gallic acid in the chromatogram of the test sample as the reference peak; detect at 250 nm to obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, and thus obtain the standard fingerprint spectrum formed by the 20 common characteristic peaks;

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

[0063]

[0064]

[0065] (5) Using the methods described in (1)-(3) as the fingerprint spectrum test means for the compound Zanthoxylum bungeanum composition to be tested, prepare the fingerprint spectrum of the sample to be tested;

[0066] (6) The fingerprint spectrum of the compound Zanthoxylum bungeanum to be tested is compared with the standard fingerprint spectrum of step (4), and the similarity is greater than 0.97.

[0067] Example 2

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

[0069] (1) Preparation of the test solution:

[0070] Accurately weigh 0.3g of compound Zanthoxylum nitidum tablets thick paste, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the contents, sonicate for 40min, cool, accurately weigh the contents again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0071] Alternatively, take compound Zanthoxylum bungeanum tablets, grind them into a fine powder, accurately weigh 3g of the finely ground sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the sample, sonicate for 40min, cool, accurately weigh the sample again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0072] (2) Preparation of reference solution:

[0073] Take gallic acid reference standard, dissolve it in 50% methanol to prepare a 70 μg / mL solution;

[0074] (3) Chromatographic conditions:

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

[0076] Detection wavelength: 250nm;

[0077] Mobile phase: Mobile phase A is methanol, and mobile phase B is a 0.1% triethylamine solution containing 0.1% formic acid by weight. Gradient elution is used, and the elution program 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 spectrum: Take 20 μL each of the reference solution and 16 batches of compound Zanthoxylum bungeanum as test solution, inject them into the high performance liquid chromatograph, measure and record the chromatogram, and determine gallic acid in the chromatogram of the test sample as the reference peak; detect at 250 nm to obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, and thus obtain the standard fingerprint spectrum formed by the 20 common characteristic peaks;

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

[0081]

[0082]

[0083] (5) Using the methods described in (1)-(3) as the fingerprint spectrum test means for the compound Zanthoxylum bungeanum composition to be tested, prepare the fingerprint spectrum of the sample to be tested;

[0084] (6) The fingerprint spectrum of the compound Zanthoxylum bungeanum to be tested is compared with the standard fingerprint spectrum of step (4), and the similarity is greater than 0.97.

[0085] Example 3

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

[0087] (1) Preparation of the test solution:

[0088] Accurately weigh 0.2g of compound Zanthoxylum nitidum tablets viscous extract, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the contents, sonicate for 40min, cool, accurately weigh the contents again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0089] Alternatively, take compound Zanthoxylum bungeanum tablets, grind them into a fine powder, accurately weigh 2g of the finely ground sample, place it in a container, accurately add 10mL of 50% methanol, accurately weigh the sample, sonicate for 40min, cool, accurately weigh the sample again, replenish the lost weight with 50% methanol, shake well, centrifuge, take the supernatant, and filter it through a 0.45μm microporous membrane to obtain the final product.

[0090] (2) Preparation of reference solution:

[0091] Take gallic acid reference standard, dissolve it in 50% methanol to prepare a 50 μg / 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, and mobile phase B is a 0.1% triethylamine solution containing 0.1% formic acid by weight. Gradient elution is used, and the elution program 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 spectrum: Take 20 μL each of the reference solution and 16 batches of compound Zanthoxylum bungeanum as test solution, inject them into the high performance liquid chromatograph, measure and record the chromatogram, and determine gallic acid in the chromatogram of the test sample as the reference peak; detect at 250 nm to obtain 20 common characteristic peaks, calculate the relative retention time and relative peak area of ​​each common peak, and thus obtain the standard fingerprint spectrum formed by the 20 common characteristic peaks;

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

[0099]

[0100]

[0101] (5) Using the methods described in (1)-(3) as the fingerprint spectrum test means for the compound Zanthoxylum bungeanum composition to be tested, prepare the fingerprint spectrum of the sample to be tested;

[0102] (6) The fingerprint spectrum of the compound Zanthoxylum bungeanum to be tested is compared with the standard fingerprint spectrum of step (4), and the similarity is greater than 0.97.

[0103] Example 4: Fingerprint analysis of compound Zanthoxylum bungeanum tablets

[0104] 1. Materials

[0105] 1.1 Instruments

[0106] High-performance liquid chromatograph (Agilent 1100, DAD detector), 0.01% balance (Sartorius Scientific Instruments (Beijing) Co., Ltd.), 0.01% 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 Test Drugs

[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), Watson's distilled water (Guangzhou Watson's Food & Beverage Co., Ltd.), gallic acid reference standard (batch number: WP23071413, manufacturer: Sichuan Weikeqi Biotechnology Co., Ltd.), 16 batches of compound Zanthoxylum bungeanum tablets, batch numbers: 20220505, 202... 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.), numbered S1 to S16, see Table 1.

[0109] Table 1. Batch Number and Information of Compound Zanthoxylum bungeanum Tablets (16 batches)

[0110]

[0111] 2. Methods and Results

[0112] 2.1 Chromatographic conditions

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

[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 standard, place it in a 250 mL volumetric flask, dissolve it in 50% methanol and dilute to the mark to prepare a 50 μg / mL reference solution.

[0120] 2.2.2 Preparation of the test solution

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

[0122] 2.3 Methodological Examination

[0123] 2.3.1 Precision Test

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

[0125] 2.3.2 Repeatability Test

[0126] Six samples with batch number 20220502 were taken and test solutions were prepared according to the method in section “2.2.2”. Under the chromatographic conditions in section “2.1”, each sample was injected once. Gallic acid was used as the reference peak (S). The relative retention time and relative peak area of ​​20 common peaks were calculated. The relative retention time RSD was <1.203% and the relative peak area RSD was <4.979%, indicating that the method has good repeatability.

[0127] 2.3.3 Stability Test

[0128] Prepare one test solution according to the method in section "2.2.2", store it at room temperature under the chromatographic conditions in section "2.1", and measure it at 0, 2, 4, 6, 8, 12 and 24 h respectively. Using 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 <1.232% and the relative peak area RSD is <4.891%, indicating that the test solution has good stability within 24 h.

[0129] 2.4 Establishment of HPLC fingerprint chromatogram

[0130] Sixteen batches of Compound Zanthoxylum bungeanum tablets were sampled and analyzed under the chromatographic conditions described in section "2.1". Chromatograms were recorded, and the data were imported into the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)". Using the chromatogram of sample S12 as a reference, a common pattern was generated using the mean method. Common peaks in the characteristic chromatograms of the 16 batches of Compound Zanthoxylum bungeanum tablets were matched, and a total of 20 common peaks were identified. (See attached table). Figure 1 By comparing with the control ( Figure 2 The gallic acid component was identified through comparison. Similarity calculations were performed on 16 batches of samples, and the similarity scores were all above 0.97.

[0131] The above are embodiments of the present invention. The above embodiments and specific parameters are only for clearly illustrating 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 shall still be determined by its claims. Similarly, any equivalent structural changes made based on the description and drawings of the present invention shall also be included within the protection scope of the present invention.

Claims

1.A quality control method of a compound Zuijiaozhen composition, characterized in that: comprising gradient elution and determination by a high performance liquid chromatograph; The high performance liquid chromatography conditions are as follows: a C18 chromatographic column; a detection wavelength of 250 nm; a mobile phase A of methanol and a mobile phase B of 0.1% triethylamine solution containing 0.1% formic acid by weight percentage, gradient elution, and an elution procedure as follows: precisely weighing 0.15-0.3 g of a compound Zuijiaozhen tablet thick paste, placing it in a container, precisely adding 10 mL of 50% methanol, precisely weighing, ultrasonic treatment for 40 min, cooling, precisely weighing again, supplementing the lost weight with 50% methanol, shaking, centrifugation, taking the supernatant, and filtering through a 0.45 μm microporous filter to obtain; ； or finely grinding the compound Zuijiaozhen tablet, precisely weighing 1-3 g of the finely ground sample, placing it in a container, precisely adding 10 mL of 50% methanol, precisely weighing, ultrasonic treatment for 40 min, cooling, precisely weighing again, supplementing the lost weight with 50% methanol, shaking, centrifugation, taking the supernatant, and filtering through a 0.45 μm microporous filter to obtain; a control solution preparation method comprising the following steps: adding 50% methanol to dissolve the gallic acid control sample to prepare a 30-70 μg / mL solution to obtain; The compound Zuijiaozhen composition is made of the following medicinal materials: Xiqingguo, Budiaoguo, Shandougen, menthol oil, menthol, Zuijiaozhen, and Gancao. 2.The quality control method of the compound Zuijiaozhen composition according to claim 1, characterized in that: the C18 chromatographic column has a specification of 250×4.6 mm, 5 μm, and a column temperature of 25-35 ℃; and the mobile phase flow rate is 0.7-1.2 mL / min. 3.The quality control method of the compound Zuijiaozhen composition according to claim 2, characterized in that: 4.The quality control method of the compound Zuijiaozhen composition according to claim 1, characterized in that: The C18 chromatographic column is a phenomenex C 18 Chromatographic column. standard fingerprint spectrum establishment: 20 μL of the control solution and 20 μL of 16 batches of compound Zuijiaozhen composition are combined as test sample solutions, injected into the high performance liquid chromatograph, determined, and the chromatogram is recorded to determine the gallic acid in the test sample chromatogram as the reference peak; detection at 250 nm gives 20 common characteristic peaks, and the relative retention time and relative peak area of each common peak are calculated to obtain the standard fingerprint spectrum formed by the 20 common characteristic peaks. 5.The quality control method of the compound Zuijiaozhen composition according to claim 4, characterized in that: the characteristic peaks of the standard fingerprint spectrum are as follows: including the following contents: 。 6.The method of claim 1, wherein the compound liangxuanyin composition fingerprint quality control method is characterized by, (1) test sample solution preparation: precisely weighing 0.15-0.3 g of a compound Zuijiaozhen tablet thick paste, placing it in a container, precisely adding 10 mL of 50% methanol, precisely weighing, ultrasonic treatment for 40 min, cooling, precisely weighing again, supplementing the lost weight with 50% methanol, shaking, centrifugation, taking the supernatant, and filtering through a 0.45 μm microporous filter to obtain; ​ or take two compound two-sided needle containing pieces of fine, precision 1-3 g of fine sample, placed in a container, precision 50% methanol 10 mL, precision weight, ultrasonic treatment for 40 min, cool, and then precision weight, with 50% methanol to make up for the weight loss, shake, centrifuge, take the supernatant, 0.45 μm microporous filter membrane, obtained; (2) Preparation of reference solution: Take the gallic acid reference substance, add 50% methanol to dissolve, make a solution of 30-70 μg / mL, obtained; (3) Chromatographic conditions: Chromatographic column: C18 chromatographic column; 250 x 4.6 mm, 5 μm; the column temperature is 25-35℃; Detection wavelength: 250 nm; Mobile phase: mobile phase A is methanol, mobile phase B is 0.1% triethylamine solution containing 0.1% formic acid by weight, using gradient elution, elution program as follows: (4) The establishment of standard fingerprint: take the reference solution and 16 batches of compound two-sided needle combination as test sample solution, 20 μL each, inject high performance liquid chromatograph, determine, record chromatogram, determine the gallic acid in the test sample chromatogram as the reference peak; detection at 250 nm, 20 common characteristic peaks are obtained, the relative retention time and relative peak area of each common peak are calculated, and the standard fingerprint formed by 20 common characteristic peaks is obtained; (5) The methods described in (1)-(3) are used as the testing means of the fingerprint of the compound two-sided needle composition to be tested, and the fingerprint of the sample to be tested is prepared; (6) Compare the fingerprint of the compound two-sided needle composition to be tested with the standard fingerprint of step (4), and the similarity is greater than 0.9.

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