Fingerprint spectrum and content determination method of acorus calamus dizziness relieving preparation

Abstract

The invention provides a fingerprint spectrum and content determination method of a calamus dizziness relieving preparation, and belongs to the technical field of traditional Chinese medicines. The construction method comprises the following steps: detecting a test solution containing the calamus wind-quenching preparation by utilizing high performance liquid chromatography, and generating a fingerprint spectrum according to a detection result; an adopted mobile phase A is acetonitrile, and a mobile phase B is a 0.1% phosphoric acid aqueous solution; and carrying out gradient elution by using the mobile phases A and B. The fingerprint spectrum can be used for simultaneously detecting various chemical components in the calamus parviflora preparation, is high in separation degree, and can systematically, stably and fully reflect the quality of a product, so that the quality of the calamus parviflora preparation can be effectively controlled. Meanwhile, the construction method is simple and efficient, the detection time can be shortened, the use of a solvent is reduced, the detection cost is further greatly reduced, and the method has high specificity and selectivity on analysis of unknown and known components and is more beneficial to actual production and application.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicines, and in particular to a fingerprint spectrum and a content determination method of an Acorus ramie extinguishing wind preparation. Background Art

[0002] Acorus emaxifeng preparation has the effects of calming the liver and extinguishing wind, tranquilizing the nerves and resolving phlegm. It is clinically used to treat mild to moderate pediatric multiple tics, which belongs 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 Acorus emaxifeng 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 perform accurate quantitative quality control on 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 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 an Acorus ramie extinguishing wind preparation. Summary of the invention

[0007] The purpose of the present invention is to provide a fingerprint spectrum and content determination method of an Acorus ramie extinguishing wind preparation.

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

[0009] In a first aspect, the present invention provides a method for constructing a fingerprint of an Acorus ramie extinguishing wind preparation, the method comprising the following steps:

[0010] The test solution containing the Acorus ramie wind-extinguishing preparation is detected by high performance liquid chromatography, and the detection result is used to generate a fingerprint spectrum; the mobile phase A used in the high performance liquid chromatography detection is acetonitrile, and the mobile phase B is a 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 HPLC detection, the column temperature of the chromatographic column is 20 to 25°C;

[0015] Further, the detection wavelength of the HPLC is 220-320 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 the high performance liquid chromatography test to determine the attribution of each peak in the test solution according to the retention time of the reference solution; the reference solution includes nicotinic acid, gallic acid, gastrodin, sibirica polygala A6, paeoniflorin, paeoniflorin and further 3,6'-dicerinoylsucrose.

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

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

[0021] (2) HPLC detection: using a Well Ch 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, the step (1) further comprises ultrasonication for 30 minutes (power 100 W, frequency 40 kHz), followed by cooling, shaking and filtering.

[0026] Furthermore, the step (1) also includes that the specification of the Well 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 an Acorus ramie extinguishing wind preparation, the fingerprint comprising 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-dicerinoylsucrose, and the relative retention time is 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, and 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 of 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 of No. 8 was 0.672 ± 10%;

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

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

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

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

[0045] In a third aspect, the present invention provides a fingerprint spectrum of an Acorus emaciated wind-extinguishing preparation as an application of a standard fingerprint spectrum in the detection and quality control of the Acorus emaciated wind-extinguishing 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 an Acorus ramie extinguishing wind preparation, wherein the fingerprint can simultaneously detect multiple chemical components in the Acorus ramie extinguishing wind preparation, has a high degree of separation, can systematically and stably fully reflect the quality of the product, and can effectively control the quality of the Acorus ramie extinguishing wind preparation. 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, and has a high degree of specificity and selectivity for the analysis of unknown and known components, which is more conducive to actual 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 spectrum

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

[0052] Figure 5 :Paeoniflorin Reference Material Spectrum

[0053] Figure 6 :Paeoniflorin reference substance spectrum

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

[0055] Figure 8 :Mixed reference sample spectrum

[0056] Fig. 9 :Test sample atlas

[0057] Fig.10 :Peony root extract spectrum

[0058] Fig.11:Gastrodia elata extract profile

[0059] Fig.12 :Acorus calamus extract profile

[0060] Fig.13 :Profile of Polygala root extract

[0061] Fig.14 :Nacre extract profile

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

[0063] Fig.16 : Sample spectrum—220nm

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

[0065] Fig.18 : Sample spectrum—320nm

[0066] Fig.19 :Comparative fingerprint of Changma Xifeng Tablets

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

[0068] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. If specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0069] The features and properties of the present invention are further described in detail below in conjunction with the embodiments:

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

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

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

[0073] ⑶Flow rate: 0.8ml / min

[0074] ⑷ Column temperature: 25℃~35℃

[0075] ⑸ Detection 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:

[0077]

[0078] ⑺ Preparation method of reference solution: Take an appropriate amount of paeoniflorin reference substance, weigh it accurately, 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, sibirica polygala A6, paeoniflorin, peonidin, and 3,6'-dicorinoyl sucrose reference substances, weigh them accurately, add 50% methanol to dissolve them, and prepare solutions containing 0.1 mg of gastrodin, paeoniflorin, 1,2,3,4,6-O-galloyl glucose, 3,6'-dicorinoyl sucrose, and gallic acid per 1 ml.

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

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

[0082] The fingerprint of the test sample should present 16 chromatographic peaks corresponding to the reference fingerprint, 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 reference fingerprint calculated by the similarity evaluation system of Chinese medicine chromatographic fingerprint should not be less than 0.90.

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

[0084] 1. Establishment of fingerprint construction method of Acorus ramie extinguishing wind preparation

[0085] ⑴ Preparation of reference solution for peak identification: Take appropriate amount of nicotinic acid, gallic acid, gastrodin, sibirica polygala A6, paeoniflorin, paeoniflorin, and 3,6'-dicerinoylsucrose reference substances, weigh them accurately, add 50% methanol to dissolve them, and prepare solutions containing 0.1 mg of gastrodin, paeoniflorin, 1,2,3,4,6-O-galloylglucose, 3,6'-dicerinoylsucrose, and gallic acid per 1 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 substance per ml.

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

[0088] (4) Chromatographic conditions and system suitability test: Thermo U3000 high performance liquid chromatograph, chromatographic column Well Chloride XB-C18; acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, flow rate of 0.8 ml per minute; column temperature of 30°C; detection wavelengths of 254 nm, 220 nm and 320 nm. The theoretical plate number 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] 103-105min, 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 wavelength of the high performance liquid chromatography method is three wavelengths, 254nm, 220nm and 320nm.

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

[0102] ⑹ Accurately pipette 10 μl of the reference solution and the test solution respectively, inject 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 of fingerprints of each batch of test sample and 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 Fig. 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, peaks 6-8, peak 11, peak 13, peaks 15-16: unknown peaks, and the peaks of the chromatogram have good separation effect, without tailing or crossover, etc., which realizes the simultaneous detection of the fingerprint spectrum and the effective ingredients of three medicinal flavors of white peony, gastrodia and polygala, and can carry out a comprehensive quality evaluation of the Acorus emaciated wind-extinguishing preparation.

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

[0111] (1) 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 each single herb test solution 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 (peonidin), peak 11 (unknown peak), peak 12 (peonidin), 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 cover the four medicinal ingredients in the whole prescription. This method can achieve 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 the wind-extinguishing preparation of Acorus imastatus

[0118] ⑴Test method: Take about 26.2 mg of 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, as the gastrodin reference substance stock solution. Accurately pipette the gastrodin reference substance stock solution and add 50% methanol solution to dilute into solutions containing 20 μg, 60 μg, 100 μg, 200 μg, and 300 μg per 1 ml as the gastrodin linear reference substance solution.

[0119] Take about 26 mg of paeoniflorin reference substance (batch number: 110736-202246, content: 96.7%), accurately weigh, place in a 50 ml volumetric flask, add 50% methanol solution and dilute to the scale, shake well, as the paeoniflorin reference substance stock solution. Accurately pipette the paeoniflorin reference substance stock solution and add 50% methanol solution to dilute into solutions containing 20μg, 40μg, 100μg, 200μg, and 300μg per 1 ml as the paeoniflorin linear reference substance solution.

[0120] Take about 26 mg of 3,6'-bisinapoyl sucrose reference substance (batch number: 111848-202307, content: 96.7%), accurately weigh, place in a 50 ml volumetric flask, add 50% methanol solution and dilute to the mark, shake well, as the paeoniflorin reference substance stock solution. Accurately pipette the reference substance stock solution and add 50% methanol solution to dilute to 25 μg, 50 μg, 100 μg, 200 μg, 300 μg per 1 ml solution, as the 3,6'-bisinapoyl sucrose linear reference substance solution.

[0121] (2) The above-mentioned 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 calamus herba anti-wind preparation

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

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

[0132] ⑶ Preparation of reference solution: Accurately weigh appropriate amounts of gastrodin, paeoniflorin and 3,6'-dicerinoylsucrose reference substances, add 50% methanol to prepare a mixed solution containing 100 μg of gastrodin, 200 μg of paeoniflorin and 100 μg of 3,6'-dicerinoylsucrose per 1 ml.

[0133] (4) Accurately pipette 10 μl of the reference solution and the test solution respectively, inject into the liquid chromatograph, measure according to the chromatographic conditions of Example 1, calculate the test sample content by the external standard method, calculate the recovery rate, and 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'-dicerinoylsucrose

[0142]

[0143] ⑸According to the above recovery rate data, the recovery rates of gastrodin, paeoniflorin and 3,6'-dicerinoylsucrose 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] The links not described in detail in the present invention are all common knowledge that can be selected by ordinary technicians in the field. Although the present invention has been described in detail above with general descriptions and specific implementation schemes, it is obvious to ordinary technicians in the field that some modifications or improvements can be made on the basis of the present invention. Therefore, these modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. A method for constructing a fingerprint of an Acorus ramie extinguishing wind preparation, characterized in that: The construction method comprises the following steps: The test solution containing the Acorus ramie wind-extinguishing preparation is detected by high performance liquid chromatography, and the detection result is used to generate a fingerprint spectrum; the mobile phase A used in the high performance liquid chromatography detection is acetonitrile, and the mobile phase B is a 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.

2. The construction method according to claim 1, characterized in that: 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; And / or, in the high performance liquid chromatography detection, the column temperature of the chromatographic column is 20 to 25°C; And / or, the detection wavelength of the high performance liquid chromatography is 220 to 320 nm; And / or, the injection volume of the HPLC is 5 to 20 μL, preferably 10 μL.

3. The construction method according to claim 1 or 2, characterized in that: The flow rate of the mobile phase is 0.9-1.1 ml / min, preferably 1.0 ml / min.

4. The construction method according to any one of claims 1 to 3, characterized in that: It also includes preparing a reference solution, and subjecting the reference solution to the 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 nicotinic acid, gallic acid, gastrodin, sibirica polygala A6, paeoniflorin, paeoniflorin and / or 3,6'-dicerinoylsucrose.

5. The construction method according to any one of claims 1 to 4, characterized in that: The construction method comprises the following steps: (1) Preparation of test solution: Dissolve the Acorus ramieae 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.

6. The construction method according to claim 5, characterized in that: The step (1) further comprises ultrasonicating for 30 minutes (power 100 W, frequency 40 kHz), cooling, shaking and filtering.

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

8. A fingerprint spectrum of a calamus herb preparation for extinguishing wind, characterized in that: The fingerprint spectrum includes 7 common characteristic peaks, with common characteristic peak No. 12 as the reference peak. The relative retention times of the 7 common characteristic peaks are: The common characteristic peak of No. 1 is niacin, with a relative retention time of 0.122 ± 10%; The common characteristic peak of No. 4 is gallic acid, with a relative retention time of 0.330 ± 10%; The common characteristic peak of No. 5 was gastrodin, with a relative retention time of 0.357 ± 10%; The common characteristic peak of No. 9 was Siberian Polygala sugar A6, with a relative retention time of 0.744 ± 10%; The common characteristic peak of No. 10 was paeoniflorin, with a relative retention time of 0.898 ± 10%; The common characteristic peak of No. 12 is paeoniflorin, with a relative retention time of 1.000; The common characteristic peak of No. 14 is 3,6-dicerinoylsucrose, and the relative retention time is 1.784±10%.

9. The fingerprint spectrum according to claim 8, characterized in that: The fingerprint of the Acorus ramie extinguishing wind preparation also includes 9 common characteristic peaks, with the common characteristic peak No. 12 as the reference peak, and the relative retention times of the 9 common characteristic peaks are: The relative retention time of the common characteristic peak No. 2 was 0.186 ± 10%; The relative retention time of the common characteristic peak No. 3 was 0.195 ± 10%; The relative retention time of the common characteristic peak of No. 6 was 0.530 ± 10%; The relative retention time of the common characteristic peak No. 7 is 0.610±10% The relative retention time of the common characteristic peak of No. 8 was 0.672 ± 10%; The relative retention time of the common characteristic peak of No. 11 was 0.948 ± 10%; The relative retention time of the common characteristic peak of No. 13 was 1.040 ± 10%; The relative retention time of the common characteristic peak of No. 15 was 1.952 ± 10%; The relative retention time of the common characteristic peak No. 16 is 2.068±10%.

10. Application of the fingerprint of the Acorus emagidecao preparation according to claim 8 or 9 as a standard fingerprint in the detection and quality control of the Acorus emagidecao preparation.

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