Fingerprint spectrum and content determination method of a changmai quenching wind preparation

Abstract

The present invention provides a fingerprint spectrum and content determination method of a calamus herb preparation for extinguishing wind, belonging to the technical field of traditional Chinese medicine. The construction method comprises the following steps: using high performance liquid chromatography to detect a test solution containing the calamus herb preparation for extinguishing wind, and generating a fingerprint spectrum from the detection results; using mobile phase A of acetonitrile and mobile phase B of 0.1% phosphoric acid aqueous solution; and using mobile phases A and B for gradient elution. The fingerprint spectrum can simultaneously detect multiple chemical components in the calamus herb preparation for extinguishing wind, with high separation, and can systematically, stably and fully reflect the quality of the product, thereby effectively controlling the quality of the calamus herb preparation for extinguishing wind. At the same time, the construction method is simple and efficient, can shorten the detection time, reduce the use of solvents, and thus greatly reduce the detection cost. It has high specificity and selectivity for the analysis of unknown and known components, and is more conducive to actual production applications.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a fingerprint spectrum and content determination method of a Changma Xifeng preparation. Background Art

[0002] Acorus ramie and wind-extinguishing preparations have the effects of calming the liver and extinguishing wind, tranquilizing the mind and resolving phlegm. They are clinically used to treat mild to moderate pediatric multiple tics, which belong to the syndrome of internal liver wind and phlegm in Traditional Chinese Medicine. Symptoms include involuntary twitching of the head, neck, five senses or limbs, abnormal sounds in the throat, irritability, frequent dreams and easy startle, red tongue with white greasy coating, and slippery and stringy pulse.

[0003] The existing quality standards for Changma Xifeng preparations include thin layer chromatography identification and paeoniflorin content determination methods for white peony root, Gastrodia elata and Polygala tenuifolia. The thin layer chromatography identification method cannot accurately control the quantitative quality of the product, and the content determination of paeoniflorin alone cannot comprehensively control and evaluate product quality.

[0004] The applicant has researched and established a fingerprint detection method for quality control of the medicinal flavors of the whole prescription using high performance liquid chromatography, covering the medicinal flavors of the whole prescription, in order to more comprehensively control product quality.

[0005] The established chromatographic method can quickly and accurately separate the seven active ingredients in the product, including niacin, gallic acid, gastrodin, sibirica sugar A6, paeoniflorin, paeoniflorin, and 3,6-dienasinoylsucrose, with good separation effect. At the same time, it can accurately quantify the active ingredients gastrodin, paeoniflorin and 3,6'-dienasinoylsucrose contained in Gastrodia elata, white peony root and Polygala tenuifolia. The linear relationship of each component is good and the recovery rate is good, which can meet the technical requirements of "one test and multiple evaluations".

[0006] In view of this, the present invention provides a fingerprint spectrum and content determination method of a Changmai Xifeng preparation. Summary of the Invention

[0007] The purpose of the present invention is to provide a fingerprint spectrum and content determination method of a Changmai quenching wind preparation.

[0008] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are adopted:

[0009] In a first aspect, the present invention provides a method for constructing a fingerprint of a Canma serrata preparation, the method comprising the following steps:

[0010] The test solution containing the calamus herba anti-wind preparation is detected by high performance liquid chromatography, and the test results are used to generate a fingerprint spectrum; the high performance liquid chromatography detection adopts mobile phase A of acetonitrile and mobile phase B of 0.1% phosphoric acid aqueous solution; the mobile phase A and the mobile phase B are used for gradient elution, and the conditions are as follows:

[0011]

[0012] The percentages in the table are the percentages of each mobile phase in the total volume of the mobile phase.

[0013] Furthermore, in the high performance liquid chromatography detection, the chromatographic column is an octadecylsilane bonded silica gel chromatographic column, WelCh Ultimate XB-C18; the specification of the octadecylsilane bonded silica gel chromatographic column is preferably 4.6 mm×250 mm, 5 μm;

[0014] Furthermore, in the high performance liquid chromatography test, the column temperature of the chromatographic column is 20 to 25°C;

[0015] Furthermore, the detection wavelength of the HPLC is 220 to 320 nm;

[0016] Furthermore, the injection volume of the HPLC is 5 to 20 μL, preferably 10 μL.

[0017] Furthermore, the flow rate of the mobile phase is 0.9-1.1 ml / min, preferably 1.0 ml / min.

[0018] Furthermore, it also includes preparing a reference solution and subjecting the reference solution to high performance liquid chromatography detection to determine the attribution of each peak in the test solution according to the retention time of the reference solution; the reference solution includes niacin, gallic acid, gastrodin, sibirica sugar A6, paeoniflorin, paeoniflorin and further 3,6'-diesinapoylsucrose.

[0019] Furthermore, the construction method includes the following steps:

[0020] (1) Preparation of test solution: Dissolve the Acorus equisetifolia wind-extinguishing preparation in 50% methanol;

[0021] (2) HPLC detection: using a WelCh Ultimate XB-C18 column; the mobile phase was acetonitrile-0.1% phosphoric acid aqueous solution; the column temperature was 25°C; the flow rate was 1.0 mL / min; and the detection wavelength was 254 nm;

[0022] The gradient elution program is as follows:

[0023]

[0024] The percentages in the table are the percentages of each mobile phase in the total volume of the mobile phase.

[0025] Furthermore, step (1) further comprises ultrasonication for 30 minutes (power 100W, frequency 40kHz), followed by cooling, shaking, and filtering.

[0026] Furthermore, the step (1) also includes that the specification of the Wel Ch Ultimate XB-C18 chromatographic column is 4.6 mm×250 mm, 5 μm.

[0027] In a second aspect, the present invention provides a fingerprint of a Canma quenching wind preparation, which includes 7 common characteristic peaks, with common characteristic peak No. 12 as the reference peak, and the relative retention times of the 7 common characteristic peaks are:

[0028] The common characteristic peak of No. 1 is niacin, with a relative retention time of 0.122 ± 10%;

[0029] The common characteristic peak of No. 4 is gallic acid, with a relative retention time of 0.330 ± 10%;

[0030] The common characteristic peak of No. 5 was gastrodin, with a relative retention time of 0.357 ± 10%;

[0031] The common characteristic peak of No. 9 was Siberian Polygala sugar A6, with a relative retention time of 0.744 ± 10%;

[0032] The common characteristic peak of No. 10 was paeoniflorin, with a relative retention time of 0.898 ± 10%;

[0033] The common characteristic peak of No. 12 is paeoniflorin, with a relative retention time of 1.000;

[0034] The common characteristic peak of No. 14 is 3,6-diesinapoylsucrose, with a relative retention time of 1.784±10%.

[0035] Furthermore, the fingerprint of the Acorus ramie wind-extinguishing preparation also includes 9 common characteristic peaks, with the common characteristic peak No. 12 as the reference peak. The relative retention times of the 9 common characteristic peaks are:

[0036] The relative retention time of the common characteristic peak No. 2 was 0.186 ± 10%;

[0037] The relative retention time of the common characteristic peak No. 3 was 0.195 ± 10%;

[0038] The relative retention time of the common characteristic peak No. 6 was 0.530 ± 10%;

[0039] The relative retention time of the common characteristic peak No. 7 is 0.610±10%

[0040] The relative retention time of the common characteristic peak No. 8 was 0.672 ± 10%;

[0041] The relative retention time of the common characteristic peak No. 11 was 0.948 ± 10%;

[0042] The relative retention time of the common characteristic peak No. 13 was 1.040 ± 10%;

[0043] The relative retention time of the common characteristic peak No. 15 was 1.952 ± 10%;

[0044] The relative retention time of the common characteristic peak No. 16 was 2.068±10%.

[0045] In a third aspect, the present invention provides a fingerprint spectrum of a Cane Herba Expelling Wind preparation as a standard fingerprint spectrum for use in the detection and quality control of the Cane Herba Expelling Wind preparation.

[0046] Compared with the prior art, the beneficial effects of the present invention include at least:

[0047] The present invention provides the establishment and application of a fingerprint detection method for a Canma-Xiangfeng preparation. This fingerprint can simultaneously detect multiple chemical components in the Canma-Xiangfeng preparation with high resolution, systematically and stably reflecting product quality and effectively controlling the quality of the Canma-Xiangfeng preparation. Furthermore, the construction method is simple and efficient, shortening detection time and reducing solvent usage, thereby significantly lowering detection costs. The method also demonstrates high specificity and selectivity for the analysis of both unknown and known components, making it more suitable for practical production applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 :Niacin Reference Substance Spectrum

[0049] Figure 2 :Gallic acid reference substance spectrum

[0050] Figure 3 :Gastrodin reference substance atlas

[0051] Figure 4 :Siberian Polygala A6 reference material spectrum

[0052] Figure 5 :Paeoniflorin Reference Material Spectrum

[0053] Figure 6 :Paeoniflorin reference substance atlas

[0054] Figure 7 :3,6'-Dierucyl Sucrose Reference Standard Spectrum

[0055] Figure 8 :Mixed reference sample spectrum

[0056] Figure 9 : Sample atlas

[0057] Figure 10 :Radix Paeoniae Alba Extract

[0058] Figure 11:Gastrodia elata extract profile

[0059] Figure 12 :Acorus calamus extract spectrum

[0060] Figure 13 :Profile of Polygala tenuifolia Extract

[0061] Figure 14 : Profile of Mother of Pearl Extract

[0062] Figure 15 :Mixed reference sample spectrum—220nm

[0063] Figure 16 : Sample spectrum—220nm

[0064] Figure 17 :Mixed reference sample spectrum—320nm

[0065] Figure 18 : Sample spectrum - 320nm

[0066] Figure 19 : Comparative fingerprint of Changma Xifeng Tablet

[0067] Figure 20 : Fingerprints of 20 batches of test samples. DETAILED DESCRIPTION

[0068] The embodiments of the present invention will be described in detail below with reference to the examples, but those skilled in the art will appreciate that the following examples are intended only to illustrate the present invention and should not be construed as limiting the scope of the invention. Where specific conditions are not specified in the examples, the methods were performed according to conventional conditions or the conditions recommended by the manufacturer. Where the manufacturers of the reagents or instruments are not specified, they are all conventional products that can be purchased commercially.

[0069] The features and performance of the present invention are further described in detail below with reference to the embodiments:

[0070] The method for constructing the fingerprint spectrum of the Acorus ramie wind-extinguishing preparation comprises the following steps:

[0071] ⑴ Mobile phase type: acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B

[0072] (2) Chromatographic column: Wel Ch Ultramate XB-C18, 4.6mm×250mm, 5μm

[0073] ⑶ Flow rate: 0.8ml / min

[0074] ⑷ Column temperature: 25℃~35℃

[0075] ⑸Detection wavelength: 254nm for full fingerprint spectrum; 220nm for content determination of Gastrodia elata and White Peony Root; 320nm for content determination of Polygala tenuifolia.

[0076] ⑹ Gradient elution ratio is:

[0077]

[0078] ⑺ Preparation method of reference solution: Take an appropriate amount of paeoniflorin reference substance, accurately weigh it, add 50% methanol to make a solution containing 1 mg per 1 ml, shake well, and obtain.

[0079] ⑻ Preparation of mixed reference solution: Take appropriate amount of nicotinic acid, gallic acid, gastrodin, Siberian polygala sugar A6, paeoniflorin, paeoniflorin, and 3,6'-dieserinoylsucrose reference substances, weigh them accurately, add 50% methanol to dissolve them, and prepare solutions containing 0.1 mg of gastrodin, paeoniflorin, paeoniflorin, 1,2,3,4,6-O-galloylglucose, 3,6'-dieserinoylsucrose, and gallic acid per 1 ml.

[0080] ⑼ Remove the film coating of this product, grind it into powder, take about 1.0 g, place it in a stoppered conical flask, add 25 ml of 50% methanol, and ultrasonically treat it for 30 minutes (power 100 W, frequency 40 kHz). Let it cool, shake it well, filter it, and take the filtrate to obtain the product.

[0081] ⑽ Determination method: Accurately pipette 10 μl of reference solution, mixed reference solution and test solution into liquid chromatograph, measure and record the chromatogram for 90 minutes.

[0082] The fingerprint of the test sample should present 16 chromatographic peaks corresponding to the fingerprint of the control, among which peak No. 12 corresponds to the retention time of the reference peak; the similarity between the fingerprint of the test sample and the fingerprint of the control is calculated according to the similarity evaluation system of the chromatographic fingerprint of traditional Chinese medicine, and should not be less than 0.90.

[0083] The contents of gastrodin, paeoniflorin and 3,6'-diesinapoylsucrose in the test samples were calculated by the external standard method.

[0084] 1. Establishment of fingerprint construction method for Changmai quenching wind preparation

[0085] (1) Preparation of reference solution for peak identification: Take appropriate amount of nicotinic acid, gallic acid, gastrodin, sibirica sugar A6, paeoniflorin, paeoniflorin, and 3,6'-dieserinoylsucrose reference substances, weigh them accurately, add 50% methanol to dissolve them, and prepare solutions containing 0.1 mg of gastrodin, paeoniflorin, paeoniflorin, 1,2,3,4,6-O-galloylglucose, 3,6'-dieserinoylsucrose, and gallic acid per ml.

[0086] (2) Preparation of mixed reference solution: Accurately pipette appropriate amount of the above 6 reference stock solutions, add 50% methanol to make a mixed solution containing 100 μg of each reference solution per ml.

[0087] ⑶ Preparation of test solution: Remove the film coating of the product, grind it into powder, take about 1.0 g, place it in a stoppered conical flask, add 25 ml of 50% methanol, and ultrasonically treat it for 30 minutes (power 100 W, frequency 40 kHz). Let it cool, shake it well, filter it, and take the filtrate to obtain the product.

[0088] (4) Chromatographic conditions and system suitability test: Thermo Scientific U3000 high-performance liquid chromatograph, Well Ch Ultramate XB-C18 column; acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, flow rate of 0.8 ml / min; column temperature of 30°C; detection wavelengths of 254 nm, 220 nm, and 320 nm. The number of theoretical plates calculated based on the paeoniflorin peak should be no less than 5000.

[0089] The high performance liquid chromatography method adopts gradient elution, and the elution procedure is:

[0090] 0-6 min, mobile phase A 2%, mobile phase B 98%;

[0091] 6-15 min, mobile phase A 2%-5%, mobile phase B 98%-95%;

[0092] 15-20 min, mobile phase A 5%-10%, mobile phase B 95%-90%;

[0093] 20-25 min, mobile phase A 10%-12%, mobile phase B 90%-88%;

[0094] 25-65 min, mobile phase A 12%-17%, mobile phase B 88%-83%;

[0095] 65-95 min, mobile phase A 17-28%, mobile phase B 83%-72%;

[0096] 95-97 min, mobile phase A 28%-90%, mobile phase B 72%-10%;

[0097] 97-103 min, mobile phase A 90%, mobile phase B 10%;

[0098] 103-105 min, mobile phase A 90%-2%, mobile phase B 10%-98%;

[0099] 105-120 min, mobile phase A 2%, mobile phase B 98%;

[0100] (5) The detection wavelengths of the high performance liquid chromatography method are three wavelengths, 254 nm, 220 nm and 320 nm.

[0101] The fingerprint was detected at a wavelength of 254 nm, the content of paeoniflorin and gastrodin was detected at a wavelength of 220 nm, and the content of 3,6'-diesinapoylsucrose was detected at a wavelength of 320 nm.

[0102] ⑹ Accurately aspirate 10 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, measure, and record the chromatogram for 60 minutes.

[0103] ⑺See attached chromatogram Figure 1-9 .

[0104] ⑻Relative retention time of each common peak (min)

[0105]

[0106]

[0107] ⑼ Similarity between the fingerprints of each batch of test sample and the control

[0108] batch number Similarity batch number Similarity 221001 0.988 230505 0.998 221017 0.992 230509 1.000 221101 0.993 230901 0.996 221201 0.996 230904 0.996 221208 0.999 230910 0.996 230401 0.999 231101 0.994 230402 0.999 231102 0.995 230405 0.999 231103 0.994 230501 0.999 240201 0.998 230503 0.999 240304 0.995

[0109] ⑽ Figure 9 The chromatogram of the test product is Figures 1 to 8 The chromatographic peaks in the chromatogram of the test sample are peak 1: niacin, peak 4: gallic acid, peak 5: gastrodin, peak 9: sibirica sugar A6, peak 10: paeoniflorin, peak 12: paeoniflorin (S), peak 14: 3,6-diesinapoylsucrose, peaks 2-3, 6-8, 11, 13, 15-16: unknown peaks, and the peaks of the chromatogram have good separation effect without tailing or crossing, which realizes the simultaneous detection of the fingerprint spectrum and the effective contents of the three medicinal flavors of white peony root, gastrodia and polygala, and can be used for a comprehensive quality evaluation of the changma wind-extinguishing preparation.

[0110] 2. Chromatographic peak attribution of fingerprint of Acorus ramie preparation

[0111] ⑴ Experimental method: Take the single-herb extract of white peony root, the single-herb extract of Gastrodia elata, the single-herb extract of Polygala tenuifolia, the single-herb extract of Acorus calamus, and the single-herb extract of mother-of-pearl respectively, and treat them according to the test solution preparation method of Example 1 to obtain the corresponding single-herb test solution.

[0112] (2) According to the fingerprint analysis method of Example 1, inject the test solution of each single herb into the high performance liquid chromatograph for analysis to determine the peak attribution. See the attached chromatogram Figure 8-14 .

[0113] ⑶Research results

[0114] Peak attribution study results

[0115]

[0116] ⑷ By the attachment Figure 8-14 It can be seen that peak 4 (gallic acid), peak 9 (Siberian polygala sugar A6), peak 10 (peoniflorin), peak 11 (unknown peak), peak 12 (peoniflorin), and peak 13 (unknown peak) are characteristic peaks exclusive to white peony, peak 5 (gastrodin) is a characteristic peak exclusive to Gastrodia elata, peak 7 (unknown peak), peak 14 (3,6'-dieserinoylsucrose), peak 15 (unknown peak) and peak 16 (unknown peak) are characteristic peaks exclusive to Polygala tenuifolia, peak 1 (niacin) is a common peak of white peony, Gastrodia elata, Acorus tatarinowii, and Polygala tenuifolia, peak 2 (unknown peak) is a common peak of Gastrodia elata and Polygala tenuifolia, peak 3, peak 6 and peak 8 (unknown peak) are all common peaks of Gastrodia elata, Acorus tatarinowii, and Polygala tenuifolia, and peak 9 (Siberian polygala sugar A6) is a common peak of white peony and Polygala tenuifolia. The 16 common peaks covered the four medicinal ingredients in the whole prescription. This method can realize the quality control of the four medicinal ingredients in the prescription: white peony root, gastrodia elata, calamus tatarinowii and polygala tenuifolia.

[0117] 3. Linearity test of Acorus ramie wind-extinguishing preparation

[0118] ⑴Test method: Take approximately 26.2 mg of the gastrodin reference substance (batch number: 110807-202010, content: 95.5%), accurately weigh, place in a 50 ml volumetric flask, add 30% methanol solution and dilute to the scale, shake well, and use as the gastrodin reference substance stock solution. Accurately pipette the gastrodin reference substance stock solution and add 50% methanol solution to dilute to solutions containing 20 μg, 60 μg, 100 μg, 200 μg, and 300 μg per ml as the gastrodin linear reference solution.

[0119] Accurately weigh approximately 26 mg of paeoniflorin reference substance (batch number: 110736-202246, content: 96.7%) and place in a 50 mL volumetric flask. Add 50% methanol solution and dilute to the mark. Shake well to prepare this as the paeoniflorin reference substance stock solution. Accurately pipette the paeoniflorin reference substance stock solution and dilute it to 50% methanol solution to produce solutions containing 20 μg, 40 μg, 100 μg, 200 μg, and 300 μg per mL, respectively. These solutions serve as the paeoniflorin linearity reference solutions.

[0120] Accurately weigh approximately 26 mg of 3,6'-diesinapoylsucrose reference standard (batch number: 111848-202307, content: 96.7%) and place in a 50 ml volumetric flask. Add 50% methanol solution and dilute to the mark. Shake well to prepare this as the paeoniflorin reference standard stock solution. Accurately pipette the reference standard stock solution and dilute it to 50% methanol solution to prepare solutions containing 25 μg, 50 μg, 100 μg, 200 μg, and 300 μg per ml, respectively. These will serve as the 3,6'-diesinapoylsucrose linear reference standard solutions.

[0121] (2) The above series of concentration reference solution were tested according to the chromatographic conditions of Example 1, and a standard curve was drawn with the reference concentration as the abscissa and the peak area as the ordinate, and the linear regression equation and correlation coefficient were calculated.

[0122] ⑶ Linear correlation of gastrodin

[0123]

[0124] ⑷ Linear correlation of paeoniflorin

[0125]

[0126] ⑸3,6'-Dierucyl sucrose linear correlation

[0127]

[0128] ⑹The above data show that this method has a good linear correlation in detecting the contents of gastrodin, paeoniflorin and 3,6'-diesinapoylsucrose.

[0129] 4. Accuracy test of the changmai wind-extinguishing preparation

[0130] (1) Test method: This example uses reference substances for sample addition recovery test. Gastrodin, paeoniflorin and 3,6'-dieserinoylsucrose reference substances are added to the test sample with known gastrodin, paeoniflorin and 3,6'-dieserinoylsucrose content for determination, and the recovery rate of each reference substance is calculated.

[0131] ⑵ Preparation of test solution: First, prepare a stock solution containing gastrodin, paeoniflorin and 3,6'-diesinapoylsucrose reference substance, and add the stock solution of the reference substance to the test solution at a ratio of 1:0.5, 1:1 and 1:1.5 to prepare three test solution concentrations of low, medium and high.

[0132] ⑶ Preparation of reference solution: Accurately weigh appropriate amounts of gastrodin, paeoniflorin and 3,6'-dieserinoylsucrose reference substance, add 50% methanol to make a mixed solution containing 100 μg of gastrodin, 200 μg of paeoniflorin and 100 μg of 3,6'-dieserinoylsucrose per ml.

[0133] (4) Accurately aspirate 10 μl of each reference solution and test solution, inject them into the liquid chromatograph, and measure them according to the chromatographic conditions of Example 1. Calculate the test sample content by the external standard method and calculate the recovery rate. The results are shown in the table below.

[0134] Recovery rate calculation formula

[0135]

[0136] ① Recovery rate of gastrodin

[0137]

[0138]

[0139] ② Recovery rate of paeoniflorin

[0140]

[0141] ③Recovery rate of 3,6'-diesinapoylsucrose

[0142]

[0143] ⑸ According to the above recovery rate data, the recovery rates of gastrodin, paeoniflorin and 3,6'-diesinapoylsucrose detected by this method are all between 90% and 108%, which is in line with the requirements of the Pharmacopoeia "Guidelines for Validation of Analytical Methods".

[0144] Any aspects of the present invention not described in detail are common knowledge that can be selected at the discretion of a person skilled in the art. Although the present invention has been described in detail above using general descriptions and specific embodiments, it is obvious to a person skilled in the art that modifications or improvements can be made based on the present invention. Therefore, such modifications or improvements made without departing from the spirit of the present invention are within the scope of protection claimed in the present invention.

Claims

1. A method for constructing a fingerprint of a changmai wind-extinguishing preparation, characterized in that: The construction method comprises the following steps: The test solution containing the calamus herba anti-wind preparation is detected by high performance liquid chromatography, and the test results are used to generate a fingerprint spectrum; the high performance liquid chromatography detection adopts mobile phase A of acetonitrile and mobile phase B of 0.1% phosphoric acid aqueous solution; the mobile phase A and the mobile phase B are used for gradient elution, and the conditions are as follows: The percentages in the table are the percentages of each mobile phase in the total volume of the mobile phase; In the high performance liquid chromatography test, the chromatographic column is an octadecylsilane bonded silica gel chromatographic column, WelCh UltimateXB-C18; the specification of the octadecylsilane bonded silica gel chromatographic column is 4.6 mm×250 mm, 5 μm; In the high performance liquid chromatography detection, the column temperature of the chromatographic column is 20-25°C; The detection wavelength of the high performance liquid chromatography is 220 to 320 nm; The injection volume of the HPLC is 5 to 20 μL; The flow rate of the mobile phase is 0.9-1.1 ml / min; The construction method further includes preparing a reference solution and subjecting the reference solution to high performance liquid chromatography detection to determine the attribution of each peak in the test solution based on the retention time of the reference solution; the reference solution includes niacin, gallic acid, gastrodin, sibirica polygala sugar A6, paeoniflorin, paeoniflorin and 3,6'-diesinapoylsucrose.

2. The construction method according to claim 1, characterized in that The injection volume was 10 μL, and the flow rate of the mobile phase was 1.0 ml / min.

3. The construction method according to any one of claims 1 and 2, characterized in that: The construction method comprises the following steps: (1) Preparation of test solution: Dissolve the Acorus equisetifolia wind-extinguishing preparation in 50% methanol; (2) HPLC detection: WelCh Ultimate XB-C18 column was used; the mobile phase was acetonitrile-0.1% phosphoric acid aqueous solution, the column temperature was 25°C, the flow rate was 1.0 mL / min, and the detection wavelength was 254 nm; The gradient elution program is as follows: The percentages in the table are the percentages of each mobile phase in the total volume of the mobile phase.

4. The construction method according to claim 3, characterized in that The step (1) further comprises ultrasonication for 30 minutes at a power of 100 W and a frequency of 40 kHz, cooling, shaking, and filtering.

5. The construction method according to claim 3, characterized in that The step (1) also includes that the WelChUltimate XB-C18 chromatographic column has a specification of 4.6 mm×250 mm and 5 μm.

6. Application of the fingerprint construction method of the Changma Xifeng preparation according to any one of claims 1 to 5 in the detection and quality control of the Changma Xifeng preparation.

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