CN113759062B

CN113759062B - Method for rapidly screening different producing areas of rhizoma acori graminei

Info

Publication number: CN113759062B
Application number: CN202110735303.8A
Authority: CN
Inventors: 张志强; 贾扬花; 史国华; 付静
Original assignee: Beijing Tcmages Pharmaceutical Co Ltd
Current assignee: Beijing Tcmages Pharmaceutical Co Ltd
Priority date: 2021-06-30
Filing date: 2021-06-30
Publication date: 2023-03-21
Anticipated expiration: 2041-06-30
Also published as: CN113759062A

Abstract

The invention provides a method for rapidly screening different producing areas of rhizoma acori graminei, which comprises the following steps: acquiring a characteristic map of the rhizoma acori graminei by adopting a high performance liquid chromatography; the characteristic map at least comprises characteristic peaks corresponding to beta-asarone and alpha-asarone; calculating a relative peak area A1 of a characteristic peak corresponding to the alpha-asarone and a reference peak S by taking a chromatographic peak of the beta-asarone as the reference peak S; when A1 is more than 0.1, the origin of the grass-leaved sweetflag is Hubei; when A1 is less than 0.01, the producing area of rhizoma Acori Graminei is Anhui; when A1 is 0.01-0.1, the producing area of rhizoma Acori Graminei is Hunan or Sichuan; the characteristic map also comprises a third characteristic peak, the relative peak area of the third characteristic peak is A2, in the grassleaf sweelflag rhizome of which the A1 is 0.01-0.1, the producing area of the grassleaf sweelflag rhizome of which the A2 is less than 0.2 is Hunan, and the producing area of the grassleaf sweelflag rhizome of which the A2 is more than or equal to 0.2 is Sichuan. The method can quickly and accurately screen the producing area of the rhizoma acori graminei and is very simple and convenient to operate.

Description

Method for rapidly screening different producing areas of rhizoma acori graminei

Technical Field

The invention relates to the field of traditional Chinese medicine identification, in particular to a method for rapidly screening different producing areas of rhizoma acori graminei.

Background

Rhizoma Acori Graminei has effects of inducing resuscitation, eliminating phlegm, refreshing mind, improving intelligence, eliminating dampness and promoting appetite. Mainly contains volatile oil, alkaloid, organic acid and carbohydrate, and is mainly produced in Hubei, hunan, sichuan and Anhui provinces. The active ingredients of the traditional Chinese medicinal materials are different due to differences of factors such as regions, climates, varieties and the like, and even if the same traditional Chinese medicinal material is used, the active ingredients are different. Therefore, screening the same medicinal materials in different producing areas has important practical significance for improving the quality of the traditional Chinese medicine.

At present, the differentiation and identification research of the grassleaf sweelflag rhizome producing area is less, and the method mainly comprises a gas phase method and a liquid phase method. Wherein, the gas phase mainly distinguishes producing areas according to the content information of the volatile oil components; for example: zou Junbo, a method for evaluating the quality of rhizoma Acori Tatarinowii by one-test-multiple-evaluation method, which adopts gas chromatography-mass spectrometry to detect 61 volatile components in rhizoma Acori Tatarinowii, and identifies the producing area of rhizoma Acori Tatarinowii by a formula consisting of the total amount of volatile oil and the content of 5 volatile components; although the method can realize the distinguishing of the producing areas of the grassleaf sweelflag rhizome, the operation and the treatment are complex, and the time and the labor are consumed. The liquid phase is mainly used for the quality control research of fingerprint or content measurement indexes of the deoiling water decoction and the like; for example: chen Xiaolou, HPLC fingerprinting study of water decoction of rhizoma Acori Tatarinowii in oil and water decoction, which only uses water soluble substances as the subject and does not distinguish the origin of rhizoma Acori Tatarinowii.

Therefore, when the method in the prior art is used for origin analysis, the problem of complex processing method exists, and the method may miss the substance information of the traditional Chinese medicine, so that the screened different substances cannot effectively represent the group difference of different origins, and the identification result has errors.

Disclosure of Invention

Therefore, the technical problem to be solved by the invention is to overcome the problems that the identification method in the prior art is complex in operation and cannot accurately identify different producing areas; thereby providing a method for rapidly screening different producing areas of the rhizoma acori graminei.

A method for rapidly screening different producing areas of rhizoma acori graminei comprises the following steps:

acquiring a characteristic map of the rhizoma acori graminei by adopting a high performance liquid chromatography; the characteristic map at least comprises characteristic peaks corresponding to beta-asarone and alpha-asarone;

calculating a relative peak area A1 of a characteristic peak corresponding to the alpha-asarone and a reference peak S by taking a chromatographic peak of the beta-asarone as the reference peak S; when A1 is greater than 0.1, the producing area of rhizoma acori graminei is Hubei; when A1 is less than 0.01, the producing area of rhizoma Acori Graminei is Anhui; when A1 is 0.01-0.1, the origin of rhizoma Acori Graminei is Hunan or Sichuan.

The relative retention time of the characteristic peak corresponding to the alpha-asarone is 1.069 +/-10%; the characteristic spectrum at least also comprises a third characteristic peak, and the relative retention time of the third characteristic peak is 1.030 +/-10%;

the relative peak area of the third characteristic peak is A2, in the grassleaf sweelflag rhizome of which the A1 is 0.01-0.1, the producing area of the grassleaf sweelflag rhizome of which the A2 is less than 0.2 is Hunan, and the producing area of the grassleaf sweelflag rhizome of which the A2 is more than or equal to 0.2 is Sichuan;

or/and the separation degree of the characteristic peak is more than or equal to 1.2.

The chromatographic conditions of the high performance liquid chromatography are as follows:

the detection wavelength is 272-278nm; methanol is taken as a mobile phase A, water or phosphoric acid aqueous solution is taken as a mobile phase B, and elution is carried out according to any one of the following gradient 1-4, wherein:

gradient 1:

a1 is 10 to 25;

gradient 2:

t1 is 20-23, T2 is 28-31, T3 is 30-33;

gradient 3:

t4 is 31-32, A2 is 25-26;

gradient 4:

t5 is 32-37 and A3 is 55-56.

The column temperature of a chromatographic column in the high performance liquid chromatography is 30-40 ℃; the chromatographic column is

Chromatography column or

A chromatography column having a specification of 2.1 x 100mm,1.6 μm; the flow rate is 0.18-0.22ml/min, and the theoretical plate number is not lower than 8000 according to the peak of beta-asarone.

The chromatographic column in the high performance liquid chromatography is

Chromatographic column, detection wavelength of 350nm for 0-2min, and detection wavelength of 275nm for the rest of time; the column temperature of the chromatographic column is 40 ℃; the specification of the chromatographic column is 2.1*100mm,1.6 μm; the flow rate is 0.20ml/min, and the number of theoretical plates is not less than 8000 according to the peak of beta-asarone.

The sample in the sample solution for high performance liquid chromatography is standard decoction of rhizoma Acori Graminei, rhizoma Acori Graminei decoction pieces, or extract prepared from rhizoma Acori Graminei or decoction pieces by different water extraction methods; the sample can also be rhizoma Acori Graminei granule.

The preparation process of the test solution comprises the following steps: adding solvent into standard decoction of rhizoma Acori Graminei, processing, and filtering to obtain filtrate, which is the sample solution.

The preparation process of the standard rhizoma acori graminei decoction comprises the following steps: soaking rhizoma Acori Calami decoction pieces for 30 min, decocting twice, adding water 9-12 times of the decoction pieces for the first time, decocting for 20 min, adding water 6-9 times of the decoction pieces for the second time, decocting for 15 min, filtering the medicinal liquid, and mixing the filtrates. The amount of water added for the second decoction is generally 3 times less than that added for the first decoction.

In the preparation process of the test solution, the treatment is uniform mixing treatment or ultrasonic treatment; during ultrasonic treatment, the treatment time is 10min, the power of ultrasonic treatment is 250W, and the frequency is 40kHz; the concentration of the test solution is 0.4g/ml, and the solvent is methanol, ethanol or water.

The solvent is a methanol water solution with the volume concentration of 60%.

As another way to obtain the test solution, the preparation process of the test solution is as follows: extracting rhizoma Acori Calami decoction pieces powder with 12-15 times of 60% -80% methanol for 40-60 min, and collecting filtrate.

The technical scheme of the invention has the following advantages:

1. the invention provides a method for rapidly screening different producing areas of acorus gramineus, which comprises the steps of taking acorus gramineus medicinal materials, decoction pieces, standard decoction, water extracts and the like obtained from different producing areas as samples to be detected, preparing the samples into a sample solution, carrying out gradient elution by adopting a high performance liquid chromatography to obtain a characteristic map at least comprising characteristic peaks corresponding to beta-asarone and alpha-asarone, calculating a relative peak area A1 of the characteristic peak corresponding to the alpha-asarone by taking the beta-asarone characteristic peak as a reference peak S, and effectively identifying and distinguishing the producing areas of the samples to be detected according to the quantity value of the A1; specifically, the method comprises the following steps: when A1 is greater than 0.1, the producing area of rhizoma acori graminei is Hubei; when A1 is less than 0.01, the producing area of rhizoma Acori Graminei is Anhui; when A1 is 0.01-0.1, the producing area of rhizoma Acori Graminei is Hunan or Sichuan;

according to the invention, a plurality of batches of medicinal materials in known production places are adopted for identification, and the comparison between the identification result and the actual production places shows that the identification result is completely consistent with the actual production places, so that the identification result of the invention has high accuracy, and the identification process is very simple and rapid.

2. The invention further adds a third characteristic peak, and the third characteristic peak can more accurately judge the grassleaf sweelflag rhizome of which the producing area is Hunan or Sichuan; specifically, when the relative retention time of the characteristic peak corresponding to the alpha-asarone is 1.069 +/-10%; the relative retention time of the third characteristic peak is 1.030 +/-10%; further judging the producing area of the grass-leaved sweetflag by the relative peak area of the third characteristic peak, wherein the producing area of the grass-leaved sweetflag with A2 less than 0.2 is Hunan, and the producing area of the grass-leaved sweetflag with A2 more than or equal to 0.2 is Sichuan.

3. The UPLC method adopted by the invention can quickly distinguish the production places of the acorus gramineus and has the advantages of quickness, accuracy, stability and the like; the invention does not need quantitative characterization and calculation of reference substances; the data does not need to be imported into software for secondary analysis, and time and labor are saved.

4. In the UPLC method, the detection and analysis of the standard decoction are aimed at, and the aimed substance information is comprehensive, so that the method can be popularized and applied to the screening of the raw material producing area of the rhizoma acori graminei formula granules and the raw materials of related preparations thereof, and has wider application range.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a control map of 18 batches of Acorus gramineus soland of different origins in example 1 of the present invention.

FIG. 2 is a fingerprint under chromatographic conditions of point 1 in example 2 of the present invention.

FIG. 3 is a fingerprint under chromatographic conditions at point 2 in example 2 of the present invention.

FIG. 4 is a fingerprint under chromatographic conditions at point 3 in example 2 of the present invention.

FIG. 5 is a fingerprint under chromatographic conditions at point 4 in example 2 of the present invention.

FIG. 6 is a fingerprint under chromatographic conditions at point 5 in example 2 of the present invention.

FIG. 7 is a fingerprint under chromatographic conditions at point 6 in example 2 of the present invention.

FIG. 8 is a fingerprint under chromatographic conditions of point 7 in example 2 of the present invention.

FIG. 9 is a fingerprint under chromatographic conditions at point 8 in example 2 of the present invention.

FIG. 10 is a feature map of Acorus gramineus soland of batch No. 190320-628100-01 of the present invention.

Detailed Description

The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.

The instrument comprises the following steps: waters ACQUITY

H-Class ultra-high performance liquid chromatograph, PDA Detector; TUV Detector ultraviolet Detector, empower 3 chromatographic workstation, ME104E electronic balance (Mettler Torlo), JY2002 electronic balance (Mettler Torlo) DZKW-4 electronic constant temperature water bath (Beijing Zhongxing Wei instruments Co., ltd.); KQ-300DB model digital control ultrasonic cleaner (Kunshan ultrasonic instruments Co., ltd.).

The grass-leaved sweetflag medicinal material is specifically shown in table 1:

TABLE 1

Reagent: methanol (merck) is chromatographically pure, methanol and ethanol are analytically pure, and water is distilled water of Drech.

Example 1

A method for rapidly screening different producing areas of rhizoma acori graminei comprises the following specific processes:

1. preparation of standard decoction:

processing the rhizoma Acori Graminei of different origins in Table 1 according to processing standard, making into decoction pieces, and preparing into standard agent of rhizoma Acori Graminei. Soaking rhizoma Acori Graminei decoction pieces for 30 min, decocting twice, adding water 9 times of the decoction pieces into the first decoction, decocting for 20 min, adding water 6 times of the decoction pieces into the second decoction, decocting for 15 min, filtering the decoction, and mixing the filtrates.

2. Obtaining the characteristic map of the standard decoction

2.1 preparing test solution

Taking 10ml of standard decoction in a 25ml volumetric flask, adding methanol until the scales are uniformly shaken, and taking the subsequent filtrate as a test solution.

2.2 chromatography methods

The following chromatographic conditions were used for detection:

octadecylsilane chemically bonded silica is used as a filler; by using

A column (2.1 x 100mm,1.6 μm) with methanol as mobile phase a and water as mobile phase B, which was subjected to gradient elution according to the elution procedure specified in table 2 below; the detection wavelength is 350nm in 0-2min and 275nm in 2-35 min; the column temperature is 40 ℃, the flow rate is 0.2ml/min, and the number of theoretical plates is not lower than 8000 according to the peak of beta-asarone.

TABLE 2

2ul of the sample solution is injected into a liquid chromatograph, and the results shown in figure 1 and table 3 are obtained after the measurement. Wherein, fig. 1 is a characteristic map corresponding to 18 batches of rhizoma acori graminei medicinal materials in table 1; table 3 shows the peak areas of the characteristic peaks corresponding to the characteristic peaks in 18 batches of Acorus tatarinowii Schott.

TABLE 3

Wherein peak 4 is β -asarone, peak 5 is the third characteristic peak, and peak 6 is α -asarone.

3. Calculating and judging the producing area according to the detection result

3.1, calculating the relative peak areas of the standard rhizoma acori graminei decoction peaks of different producing areas according to the results in the table 3, wherein the calculation results are shown in the table 4.

TABLE 4

Here, the S peak is the peak 4, the relative peak area of the peak 5 is represented by A2, and the relative peak area of the peak 6 is represented by A1.

3.2, judging the producing areas of the grassleaf sweelflag rhizome according to the results in the table 4, and the judgment rule is shown in the table 5.

TABLE 5

The results of table 4 are combined with the judgment rules of table 5 above to show that: 190418-432100-15, 190418-432100-16

190418-432100-17, 190418-432100-18 are in Hubei province; the production areas of 190325-411226-07, 190325-422117-08, 190325-414500-09 and 190325-413200-10 are provinces of Hunan province, 190320-628100-01, 190320-636150-02, 190325-638400-11, 190325-620500-12, 190325-636600-13 and 190325-646500-14 are provinces of Sichuan province, 190708-236400-19, 190708-236600-20, 190708-234300-21 and 190708-236701-22 are provinces of Anhui province.

As can be seen from the comparison of the above judgment results with the actual production areas reported in Table 1, the method of the present invention is suitable for the rapid differentiation of the production areas of Acorus tatarinowii Schott, and the operation results are very accurate.

Example 2

The difference between this example and example 1 is that only one batch of acorus gramineus soland is used for detection under different chromatographic conditions, which is as follows:

taking the rhizoma acori graminei with the number of 190320-628100-01, and respectively detecting according to the following chromatographic conditions:

1. by using

Chromatographic column replacement

The chromatographic column, other conditions were the same as in example 1, and the obtained fingerprint is shown in fig. 2, wherein the resolution R and relative peak area of β -asarone, α -asarone and third characteristic peak are shown in table 9.

2. The sample solution was prepared by using water instead of methanol under the same conditions as in example 1, and the obtained fingerprint was shown in fig. 3, in which the degrees of separation R and the relative peak areas of β -asarone, α -asarone and the third characteristic peak are shown in table 9.

3. The elution procedure was different from example 1 and the detection wavelength was 275nm, the column temperature was 30 ℃ and the specific elution procedure is shown in table 6 below:

TABLE 6

The fingerprint obtained under the above chromatographic conditions is shown in fig. 4, wherein the resolution R and the relative peak area of the β -asarone, α -asarone and the third characteristic peak are shown in table 9.

4. The elution procedure was varied and used with 0.1% phosphoric acid solution as mobile phase B, column temperature 40 ℃, and the rest of the chromatographic conditions were the same as in example 1, and the specific elution procedure is shown in table 7 below:

TABLE 7

The fingerprint obtained under the above chromatographic conditions is shown in fig. 5, wherein the resolution R and the relative peak area of the β -asarone, α -asarone and third characteristic peak are shown in table 9.

5. The chromatographic conditions were the same as in point 4 above except that water was replaced with a 0.1% phosphoric acid solution, and the fingerprint obtained under the chromatographic conditions is shown in FIG. 6, in which the degrees of separation R and the relative peak areas of the beta-asarone, alpha-asarone and third characteristic peak are shown in Table 9.

6. The percentage of the initial gradient of water of mobile phase B in the elution procedure was modified to 85% and the detection wavelength was 275nm, and the rest was the same as in example 1, and the fingerprint obtained under the chromatographic conditions is shown in FIG. 7, in which the degrees of separation R and the relative peak areas of the beta-asarone, alpha-asarone and third characteristic peak are shown in Table 9.

7. The percentage of the initial gradient of water of mobile phase B in the elution procedure was modified to 75%, and the rest was the same as in example 1, and the fingerprint obtained under the chromatography conditions is shown in FIG. 8, in which the resolution R and the relative peak area of the beta-asarone, alpha-asarone and third characteristic peak are shown in Table 9.

8. The elution procedure differs from example 1 and uses a 0.1% phosphoric acid solution as mobile phase B and a detection wavelength of 275nm, the specific elution procedure is shown in table 8 below:

TABLE 8

The fingerprint obtained under the above chromatographic conditions is shown in fig. 9, wherein the resolution R and the relative peak area of the β -asarone, α -asarone and the third characteristic peak are shown in table 9.

TABLE 9

According to the detection results, the accuracy of identification can be effectively ensured only by ensuring that the separation degree R of the characteristic peak is more than or equal to 1.2, and the effect is obvious.

By comparing the fingerprint spectrum with the fingerprint spectrum of the embodiment 1, the integral chromatogram peak base line of the fingerprint spectrum of the embodiment 1 is better, the peak height is proper, and the integral chromatogram is more attractive, so that the optimal chromatographic condition adopted by the embodiment 1 can be determined.

Example 3

This example demonstrates the methodology of the chromatographic conditions used in example 1, including:

1. repeatability: a Waters UPLC H-Class TUV detector is adopted, 10g of calamus standard decoction solution (batch number: 190320-628100-01) is taken, 6 parts are taken, the characteristic spectrum is obtained by measuring according to the specified chromatographic conditions, the relative peak area and the relative retention time are calculated by taking a No. 4 peak (beta-asarone) as a reference peak, and RSD is calculated. The results are shown in tables 10 and 11

TABLE 10 retention time and relative retention time table for repetitive examination of Acorus tatarinowii Schott

TABLE 11 repeatability test peak area and relative peak area table for grassleaf sweelflag rhizome

And (3) knotting: the results of the repeatability experiments show that the relative retention time RSD of 6 identification peaks of six repeatability experiment samples is in the range of 0.01-0.1%, and the RSD of the relative peak area is in the range of 0.23-2.52%, which indicates that the characteristic spectrum has good repeatability.

2. Intermediate precision: another Waters UPLC H-Class and TUV detector with the same model is adopted, 10g of calamus standard decoction solution (batch number: 190320-628100-01) is taken, 6 parts are taken, the characteristic spectrum is obtained by measuring according to the specified chromatographic conditions, and the relative peak area and the relative retention time are calculated by taking a No. 4 peak (beta-asarone) as a reference peak. And calculates the RSD. The results are shown in tables 12 and 13

TABLE 12 Tatarius sweetflag intermediate precision investigation retention time and relative retention time table

TABLE 13 middle precision survey of peak area and relative peak area table for grassleaf sweelflag rhizome

And (3) knotting: the intermediate precision experimental result shows that the relative retention time RSD of 6 identification peaks of six experimental samples is within the range of 0.01-0.12%, and the RSD of the relative peak area is within the range of 0.15-2.89%, which indicates that the intermediate precision of the characteristic spectrum is better.

Example 4

This example performed a durability validation of the chromatographic conditions of example 1, including:

1. stability of

A10 g sample of calamus standard decoction (batch No. 190320-628100-01) is taken, a test sample solution is prepared according to a specified method, the test is carried out according to the method in the embodiment 1 at 0 hour, 2 hour, 4 hour, 8 hour, 10 hour, 12 hour and 24 hour respectively, a characteristic spectrum is obtained, and as shown in figure 10, the No. 4 peak (beta-asarone) is taken as a reference peak, and the relative peak area and the relative retention time are calculated. And calculates the RSD. The results are shown in tables 14 and 15

TABLE 14 stability relative Retention time Table

TABLE 15 table of stability versus peak area

And (3) knotting: the stability experiment result shows that after the solution stability is examined for 24 hours, the relative retention time RSD of each characteristic peak is in the range of 0.01-0.38%, and the RSD of the relative peak area of each characteristic peak is in the range of 0.31-1.99%. The standard rhizoma acori graminei decoction adopted in the experiment has good stability within 24 hours.

2. Investigation of different flow rates

The preparation method comprises collecting rhizoma Acori Calami standard decoction solution (batch No. 190320-628100-01), and preparing the test sample according to the specified preparation method of the test sample solution. The measurements were carried out as described in example 1 using different flow rates of 0.18ml/min, 0.20ml/min and 0.22ml/min, respectively, and the durability of the test method was examined for the different flow rates, and the results are shown in tables 16 and 17.

TABLE 16 retention time results for different flow rate profiles

TABLE 17 different flow Rate characteristic Profile Peak area results Table

And (3) knotting: under different flow rate conditions, the relative retention time RSD% range of the characteristic peak is 0.04% -1.47%, and the RSD value is less than 3%; the relative peak area RSD range is 0.99% -3.04% and the value is less than 4%. The method is proved to have better durability for the flow rate between 0.18 and 0.22 ml/min.

3. Investigation of different column temperatures

A test sample solution is prepared from rhizoma Acori Graminei standard decoction solution (lot number: 190320-628100-01) by the specified test sample solution preparation method, and the test is carried out at different column temperatures of 38 deg.C, 40 deg.C and 42 deg.C respectively according to the method of example 1. The durability of the experimental method for different column temperatures was examined and the results are shown in tables 18 and 19.

TABLE 18 result table of retention time of different column temperature characteristic maps

TABLE 19 peak area result table of different column temperature characteristic maps

And (3) knotting: under different column temperature conditions, the relative retention time RSD% range of the characteristic peak is 0.02% -1.11%, and the RSD value is less than 2%; the relative peak area is 0.25-2.36% of the RSD range of other characteristic peaks except the peak 1. The method is proved to have better durability to the flow rate between 38 ℃ and 42 ℃ except the characteristic peak 1.

It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.

Claims

1. A method for rapidly screening different producing areas of rhizoma Acori Graminei is characterized in that an ultra-high performance liquid chromatography is adopted to obtain a characteristic map of rhizoma Acori Graminei; the characteristic map at least comprises characteristic peaks corresponding to beta-asarone and alpha-asarone;

calculating a relative peak area A1 of a characteristic peak corresponding to the alpha-asarone and a reference peak S by taking a chromatographic peak of the beta-asarone as the reference peak S; when A1 is greater than 0.1, the producing area of rhizoma acori graminei is Hubei; when A1 is less than 0.01, the producing area of rhizoma Acori Graminei is Anhui; when A1 is 0.01-0.1, the producing area of rhizoma Acori Graminei is Hunan or Sichuan;

the relative retention time of the characteristic peak corresponding to the alpha-asarone is 1.069 +/-10%; the characteristic map at least also comprises a third characteristic peak, and the relative retention time of the third characteristic peak is 1.030 +/-10%;

the chromatographic conditions of the ultra-high performance liquid chromatography are as follows:

the chromatographic column is CORTECS ^R UPLC ^R T3 chromatographic column or CORTECS ^R UPLC ^R A C18 chromatography column having a specification of 2.1 x 100mm,1.6 μm; methanol is taken as a mobile phase A, water or phosphoric acid aqueous solution is taken as a mobile phase B, and elution is carried out according to any one of the following gradient 1-4, wherein:

gradient 1:

time (minutes) A（%） B（%） 0～23 A1→49 (100-A1)→51 23～31 49→57 51→43 31～33 57→100 43→0

A1 is 10 to 25;

gradient 2:

time (minutes) A（%） B（%） 0～T1 10→49 90→51 T1～T2 49→57 51→43 T2～T3 57→100 43→0

T1 is 20, T2 is 28, T3 is 30;

gradient 3:

time (minutes) Mobile phase A (%) Mobile phase B (%) 0～10 0.5→2 99.5→98 10～12 2→30 98→70 12～20 30→45 70→55 20～35 45→60 55→40 35～37 60→100 40→0 37～39 100→0.5 0→99.5

；

Gradient 4:

time (minutes) Mobile phase A (%) Mobile phase B (%) 0～22 30→A3 70→(100-A3) 22～T5 A3→58 (100-A3)→42

T5 is 32, A3 is 55-56;

when the sample is rhizoma Acori Graminei standard decoction, the solvent of the sample solution is methanol, ethanol or water; when the sample is rhizoma acori graminei decoction piece powder, the solvent of the sample solution is 60-80% methanol.

2. The method for rapidly screening different producing areas of acorus gramineus according to claim 1, wherein the separation degree of characteristic peaks is more than or equal to 1.2.

3. The method as claimed in claim 1, wherein the detection wavelength of the ultra performance liquid chromatography is 272-278nm.

4. The method for rapidly screening different origins of acorus gramineus according to claim 3, wherein the column temperature of a chromatographic column in the ultra-high performance liquid chromatography is 28-42 ℃; the flow rate is 0.18-0.22ml/min, and the number of theoretical plates is not less than 8000 according to the peak of beta-asarone.

5. The method as claimed in claim 4, wherein the chromatographic column in the ultra-high performance liquid chromatography is CORTECS ^R UPLC ^R C18 chromatographic column, the detection wavelength of 0-2min is 350nm, and the detection wavelength of the rest time is 275nm; the column temperature of the chromatographic column is 38-42 ℃.

6. The method for rapidly screening different producing areas of acorus gramineus according to any one of claims 1 to 5, wherein the preparation process of the sample solution used in the ultra performance liquid chromatography is as follows: adding solvent into standard decoction of rhizoma Acori Graminei, processing, and filtering to obtain filtrate, which is the sample solution.

7. The method as claimed in claim 6, wherein the standard decoction of Acorus tatarinowii Schott is prepared by the following steps: soaking rhizoma Acori Calami decoction pieces for 30 min, decocting twice, adding water 9-12 times of the decoction pieces for the first time, decocting for 20 min, adding water 6-9 times of the decoction pieces for the second time, decocting for 15 min, filtering the medicinal liquid, and mixing the filtrates.

8. The method for rapidly screening different producing areas of acorus gramineus according to claim 6, wherein in the preparation process of the test solution, the treatment is uniform mixing treatment or ultrasonic treatment; during ultrasonic treatment, the treatment time is 10min, the power of ultrasonic treatment is 250W, and the frequency is 40kHz; the concentration of the test solution was 0.4g/ml.

9. The method for rapidly screening different producing areas of acorus gramineus according to any one of claims 1 to 5, wherein the preparation process of the sample solution used in the ultra performance liquid chromatography is as follows: extracting rhizoma Acori Calami decoction pieces powder with 12-15 times of 60% -80% methanol for 40-60 min, and collecting filtrate.