Radix aucklandiae qi-regulating pill mixed powder fingerprint spectrum and multi-component content determination method

Abstract

The invention provides a method for determining the content of multiple components in mixed powder of costustoot qi-regulating pills. The invention also provides a detection method of the fingerprint spectrum of the mixed powder of the elecampane qi-regulating pill. The invention further provides a quality detection method of the radix aucklandiae qi-regulating pill mixed powder. The invention further provides a screening method of the fingerprint spectrum of multiple medicinal materials in the radix aucklandiae qi-regulating pill mixed powder. According to the costustoot qi-regulating pill mixed powder fingerprint spectrum and the multi-component content determination method provided by the invention, the HPLC fingerprint spectrum of the costustoot qi-regulating pill mixed powder is established, the contents of the nine components are determined, and a scientific basis is provided for controlling the quality of the medicine.

Description

Technical Field

[0001] The invention belongs to the technical field of Chinese medicine component detection, and relates to a fingerprint spectrum of a mixed powder of Muxiang Shunqi Pills and a method for determining the contents of multiple components, and specifically relates to a method for determining the contents of 9 chemical components in the mixed powder of Muxiang Shunqi Pills: hesperidin, naringin, neohesperidin, naringrutin, costunolide, dehydrocostusolide, magnolol, honokiol and atractylodesin, and a fingerprint spectrum determined based on the 9 components. Background Art

[0002] Muxiang Shunqi Pills originated from Muxiang Shunqi Powder in the Medical General Purpose of Ye Wenling in the Ming Dynasty. The prescription was included in the Ming Dynasty's Zhengzhi Zhunsheng Leifang and Jingyue Quanshu, and the Qing Dynasty's Shenshi Zunshengshu. Muxiang Shunqi Pills are a compound preparation made of 11 medicinal materials, including Muxiang, vinegar-cured cyperus, Magnolia officinalis, licorice, Atractylodes macrocephala (fried), Areca nut, Amomum villosum, dried tangerine peel, green peel (fried), Citrus aurantium (fried), and ginger. It has the effects of promoting qi and removing dampness, strengthening the spleen and harmonizing the stomach. In the prescription, Muxiang and Cyperus rotundus soothe the liver and regulate qi, harmonize the stomach and relieve pain, and are the main drugs. Magnolia officinalis and green peel promote qi and dry dampness, disperse knots and eliminate accumulation; Citrus aurantium and Areca nut promote qi and relieve stagnation; dried tangerine peel and Amomum villosum regulate qi and remove dampness and harmonize the middle; Atractylodes macrocephala dries dampness and strengthens the spleen, and are the minister drugs. Ginger descends qi and harmonizes the stomach, and is an adjuvant. Licorice replenishes qi and nourishes the middle, and harmonizes all the drugs, and is the guiding drug. It is clinically used to treat diseases such as chest tightness, nausea and vomiting, belching, poor appetite, and abdominal distension and pain caused by dampness and turbidity blocking the middle part of the body and disharmony between the stomach and spleen.

[0003] The mixed powder of Muxiang Shunqi Pills is obtained by crushing 10 medicinal materials (except ginger) including Muxiang, Cyperus rotundus, Magnolia officinalis, Licorice, Atractylodes macrocephala (fried), Areca nut, Amomum villosum, Tangerine peel, Citrus aurantium (fried), and Citrus aurantium (fried) into fine powder, sieving, and mixing. Therefore, the mixed powder of Muxiang Shunqi Pills is not the same Chinese medicine composition as Muxiang Shunqi Pills. The two have different medicinal material compositions, different corresponding ingredients, and different fingerprints.

[0004] At present, only Muxiang Shunqi Pills are included in the 2020 edition of the "Chinese Pharmacopoeia", but only the determination of the content of magnolol and magnolol in Muxiang Shunqi Pills is recorded. The existing literature also discloses the quality method research of costus lactones, glycyrrhizic acid, hesperidin and other ingredients in Muxiang Shunqi Pills. The existing literature does not disclose the determination method for the various components in the mixed powder of Muxiang Shunqi Pills and their fingerprint spectra. Therefore, it is necessary to further study and discuss the various components in the mixed powder of Muxiang Shunqi Pills and their fingerprint spectra. Summary of the invention

[0005] In view of the above-mentioned disadvantages of the prior art, the purpose of the present invention is to provide a fingerprint spectrum and a multi-component content determination method for the mixed powder of Aucklandiae and Saussureae Pill. By adopting optimized conditions, a content determination method for 9 chemical components in the mixed powder of Aucklandiae and Saussureae Pill, namely hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol and atractylodin, and a fingerprint spectrum determined based on these 9 index components are established, aiming to comprehensively analyze and evaluate the quality of the mixed powder of Aucklandiae and Saussureae Pill and provide a scientific basis for controlling the quality of this medicine.

[0006] To achieve the above purpose and other related purposes, the first aspect of the present invention provides a multi-component content determination method for the mixed powder of Aucklandiae and Saussureae Pill, including the following steps:

[0007] 1) Preparation of the test solution: Add the mixed powder sample of Aucklandiae and Saussureae Pill to methanol for dissolution, perform ultrasonic extraction, cool it, filter, and take the subsequent filtrate to obtain the test solution.

[0008] 2) Preparation of the reference solution: Add one or more reference substances of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol, atractylodin to methanol and make up the volume to obtain the reference solution.

[0009] 3) Determination: Use high performance liquid chromatography (HPLC) to separately determine the test solution in step 1) and the reference solution in step 2), and calculate the content of one or more components of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol, atractylodin in the test solution by the external standard method.

[0010] Preferably, in step 1), the mixed powder of Aucklandiae and Saussureae Pill is a powder sample.

[0011] Preferably, in step 1), the mixed powder sample of Aucklandiae and Saussureae Pill should be accurately weighed before adding methanol.

[0012] Preferably, in step 1), the weight should be weighed after adding methanol.

[0013] Preferably, in step 1), the ratio of the weight (g) of the mixed powder sample of Aucklandiae and Saussureae Pill added to the volume (mL) of the methanol added is 0.25:20 - 30, specifically such as 0.25:20 - 23, 0.25:23 - 27, 0.25:27 - 30, for example 0.25:20, 0.25:22, 0.25:25, 0.25:28, 0.25:30, and preferably 0.25:25.

[0014] Preferably, in step 1), the ultrasonic extraction time is 50 - 70 min, specifically such as 50 - 55 min, 55 - 65 min, 65 - 70 min, for example 50 min, 53 min, 55 min, 57 min, 60 min, 62 min, 65 min, 68 min, 70 min, and preferably 60 min.

[0015] Preferably, in step 1), the power of the ultrasonic extraction is 240 - 260 W, preferably 250 W; the frequency of the ultrasonic extraction is 35 - 45 kHz, preferably 40 kHz.

[0016] Preferably, in step 1), the cooling is to stand still and cool to room temperature. The room temperature is 20 - 30 °C.

[0017] Preferably, in step 1), the filtration is membrane filtration.

[0018] More preferably, the membrane is a 0.45 μm membrane.

[0019] The subsequent filtrate is the filtrate collected after pouring out the initial filtrate.

[0020] Preferably, in step 2), the reference substance solution can be directly prepared into a mixed reference substance solution, or can be prepared by first adding a first solvent to form a reference substance stock solution and then adding a second solvent for step - by - step dilution.

[0021] Preferably, in step 2), the CAS number of hesperidin is 520 - 26 - 3, the CAS number of naringin is 10236 - 47 - 2, the CAS number of neohesperidin is 13241 - 33 - 3, the CAS number of naringin rutinoside is 14259 - 46 - 2, the CAS number of costunolide is 553 - 21 - 9, the CAS number of dehydrocostunolide is 477 - 43 - 0, the CAS number of magnolol is 528 - 43 - 8, the CAS number of honokiol is 35354 - 74 - 6, and the CAS number of atractylodin is 55290 - 63 - 6.

[0022] Preferably, in step 21), the content range of hesperidin in the reference solution is 19.0415 - 1218.6547 μg / mL, preferably 152.3318 μg / mL; the content range of naringin is 6.3765 - 408.0938 μg / mL, preferably 51.0117 μg / mL; the content range of neohesperidin is 5.5021 - 352.1346 μg / mL, preferably 44.0168 μg / mL; the content range of naringin rutinoside is 3.3786 - 216.2306 μg / mL, preferably 27.0288 μg / mL; the content range of costunolide is 1.7585 - 112.5444 μg / mL, preferably 14.0681 μg / mL; the content range of dehydrocostus lactone is 2.5037 - 160.2338 μg / mL, preferably 20.0292 μg / mL; the content range of magnolol is 0.8790 - 56.2550 μg / mL, preferably 7.0319 μg / mL; the content range of honokiol is 1.3146 - 84.1341 μg / mL, preferably 10.5168 μg / mL; the content range of atractylodin is 1.0218 - 32.6976 μg / mL, preferably 4.0872 μg / mL.

[0023] Preferably, in step 3), the chromatographic column in the high performance liquid chromatography is C 18 chromatographic column. More preferably, the chromatographic column in the high performance liquid chromatography is Waters Xbridge C 18 chromatographic column (4.6 mm × 250 mm, 5 μm).

[0024] Preferably, in step 3), the detector in the high performance liquid chromatography is a photodiode array detector (DAD).

[0025] Preferably, in step 3), in the high performance liquid chromatography, the column temperature is 20 - 30 °C, specifically such as 20 - 23 °C, 23 - 27 °C, 27 - 30 °C, preferably 25 °C.

[0026] Preferably, in step 3), the injection volume in the high performance liquid chromatography is 5 - 15 μL. More preferably, the injection volume in the high performance liquid chromatography is, for example, 5 μL, 6 μL, 7 μL, 8 μL, 9 μL, 10 μL, 11 μL, 12 μL, 13 μL, 14 μL, 15 μL, preferably 10 μL.

[0027] Preferably, in step 3), the flow rate in the high performance liquid chromatography is 0.5 - 2 mL / min, specifically such as 0.5 - 0.8 mL / min, 0.8 - 1.2 mL / min, 1.2 - 2 mL / min, preferably 1.0 mL / min.

[0028] Preferably, in step 3), the detection wavelength in the high performance liquid chromatography is selected from one or a combination of two of 205 - 215 nm and 265 - 275 nm, preferably 205 - 215 nm and 265 - 275 nm, more preferably 208 - 212 nm and 268 - 272 nm, and further preferably 210 nm and 270 nm.

[0029] More preferably, the detection wavelength changes with the time of the gradient elution program, and the specific change is as follows:

[0030] From 0 to 24.5 min, the detection wavelength is 205 - 215 nm, preferably 208 - 212 nm, and more preferably 210 nm;

[0031] From 24.5 to 29 min, the detection wavelength is 265 - 275 nm, preferably 268 - 272 nm, and more preferably 270 nm;

[0032] From 29 to 65 min, the detection wavelength is 205 - 215 nm, preferably 208 - 212 nm, and more preferably 210 nm.

[0033] Preferably, in step 3), in the high performance liquid chromatography, the mobile phase is acetonitrile - 0.09 - 0.11% phosphoric acid aqueous solution; in the acetonitrile - 0.09 - 0.11% phosphoric acid aqueous solution, phase A is acetonitrile and phase B is 0.09 - 0.11% phosphoric acid aqueous solution.

[0034] More preferably, in the high performance liquid chromatography, the mobile phase is acetonitrile - 0.1% phosphoric acid aqueous solution; in the acetonitrile - 0.1% phosphoric acid aqueous solution, phase A is acetonitrile and phase B is 0.1% phosphoric acid aqueous solution.

[0035] The 0.09 - 0.11% phosphoric acid aqueous solution is a phosphoric acid aqueous solution with a volume percentage of 0.09 - 0.11%. The 0.1% phosphoric acid aqueous solution is a phosphoric acid aqueous solution with a volume percentage of 0.1%.

[0036] Preferably, in step 3), in the high performance liquid chromatography, gradient elution is adopted and the analysis time is 65 min.

[0037] More preferably, the specific program of the gradient elution is as follows:

[0038] From 0 to 5 min, the volume ratio of phase A to phase B is 8 - 12:88 - 92 - 8 - 12:88 - 92;

[0039] From 5 to 20 min, the volume ratio of phase A to phase B is 8 - 12:88 - 92 - 18 - 22:78 - 82;

[0040] 20-40min, the volume ratio of phase A:phase B is 18-22:78-82-58-62:38-42;

[0041] 40-60min, the volume ratio of phase A:phase B is 58-62:38-42-58-62:38-42;

[0042] At 60-65min, the volume ratio of phase A:phase B is 58-62:38-42-88-92:8-12.

[0043] Further preferably, the specific procedure of the gradient elution is:

[0044] 0-5min, volume ratio of phase A:phase B was 10:90-10:90;

[0045] 5-20min, volume ratio of phase A:phase B is 10:90-20:80;

[0046] 20-40min, volume ratio of phase A:phase B is 20:80-60:40;

[0047] 40-60min, volume ratio of phase A:phase B is 60:40-60:40;

[0048] 60-65min, the volume ratio of phase A:phase B is 60:40-90:10.

[0049] Preferably, in step 3), the external standard method refers to: respectively taking a series of different volumes of the reference solution of step 2), respectively preparing a series of solutions of different concentrations, and using a high performance liquid chromatograph for sample analysis to obtain the linear relationship between the concentration and peak area of ​​the 9 components in the reference solution, and using the chromatographic peak area of ​​each component to correspond to its corresponding concentration, drawing the corresponding standard working curve, and calculating the regression equation of each standard working curve. Then, the test solution is detected by a high performance liquid chromatograph, and the chromatographic peak areas of the 9 components in the test solution are respectively substituted into the regression equation of each standard working curve, and the content of the corresponding component can be calculated.

[0050] More preferably, in the standard working curve, the peak area of ​​each component is used as the ordinate, and the concentration of each component in the reference solution is used as the abscissa.

[0051] The second aspect of the present invention provides a method for detecting the fingerprint of Muxiang Shunqi Pills mixed powder, comprising the following steps:

[0052] A) Preparation of test solution: the same as step 1) of the method for determining the content of multiple components in Muxiang Shunqi Pills mixed powder;

[0053] B) Preparation of reference solution: The same as step 2) in the method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussureae Formula

[0054] C) Determination: Using high performance liquid chromatography (HPLC) under the same chromatographic conditions as in step 3) of the method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussureae Formula, respectively determine the test solution in step A) and the reference solution in step B), obtain the fingerprint of the test solution and the fingerprint of the reference solution, compare the fingerprint of the test solution with the fingerprint of the reference solution, and assign and locate the target components in the fingerprint of the test solution, so as to obtain the fingerprint of the mixed powder of Aucklandiae and Saussureae Formula.

[0055] Preferably, in step C), when comparing the fingerprint of the test solution with the fingerprint of the reference solution, according to the known characteristic peaks in the fingerprint of the reference solution, through the relative retention time, identify the corresponding characteristic peaks in the fingerprint of the test solution, so as to assign and locate the target components in the fingerprint of the test solution.

[0056] The third aspect of the present invention provides a method for quality inspection of the mixed powder of Aucklandiae and Saussureae Formula, including obtaining the fingerprint of the mixed powder of Aucklandiae and Saussureae Formula by using the detection method of the fingerprint of the mixed powder of Aucklandiae and Saussureae Formula described above, and comparing the similarity between the obtained fingerprint of the mixed powder of Aucklandiae and Saussureae Formula and the control fingerprint of the mixed powder of Aucklandiae and Saussureae Formula obtained under the same fingerprint detection conditions.

[0057] Preferably, when comparing the similarity between the measured fingerprint of the mixed powder of Aucklandiae and Saussureae Formula and the control fingerprint of the mixed powder of Aucklandiae and Saussureae Formula in the present invention, the software "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicines" 2012 edition issued by the Pharmacopoeia Commission of the People's Republic of China is used for comparison. More preferably, the similarity between the measured fingerprint of the mixed powder of Aucklandiae and Saussureae Formula and the control fingerprint of the mixed powder of Aucklandiae and Saussureae Formula in the present invention is ≥0.99.

[0058] More preferably, when matching the common peaks of the fingerprint of the mixed powder of Aucklandiae and Saussureae Formula and the control fingerprint of the mixed powder of Aucklandiae and Saussureae Formula, the average method is adopted, the time window width is 0.20, and multi-point calibration matching is used to generate the fingerprint and the control fingerprint.

[0059] Preferably, the control fingerprint of the mixed powder of Aucklandiae and Saussureae Pill is obtained under the same conditions as those of the detection method of the fingerprint of the mixed powder of Aucklandiae and Saussureae Pill. The control fingerprint of the mixed powder of Aucklandiae and Saussureae Pill includes 14 common fingerprint peaks. Taking the 4th peak as the reference peak (S peak, relative retention time is 1.0000), the relative retention times of the other 13 peaks are in turn the 1st peak (0.4791±0.0072), the 2nd peak (0.9667±0.0025), the 3rd peak (0.9879±0.0008), the 5th peak (1.0190±0.0006), the 6th peak (1.1268±0.0021), the 7th peak (1.3982±0.0120), the 8th peak (1.4860±0.0136), the 9th peak (1.6094±0.0143), the 10th peak (1.6716±0.0139), the 11th peak (1.7219±0.0235), the 12th peak (1.7444±0.0257), the 13th peak (1.7671±0.0218), and the 14th peak (2.2611±0.0187).

[0060] The specific data of the control fingerprint of the Guipi Pills can be seen in Figure 3 .

[0061] More preferably, the control fingerprint of the mixed powder of Aucklandiae and Saussureae Pill is compared with the fingerprint of the reference substance solution. As shown in Figure 3 , 5 , the 2nd peak is determined as the fingerprint peak of naringin; the 3rd peak is determined as the fingerprint peak of naringin; the 4th peak is determined as the fingerprint peak of hesperidin; the 5th peak is determined as the fingerprint peak of neohesperidin; the 9th peak is determined as the fingerprint peak of magnolol; the 10th peak is determined as the fingerprint peak of costunolide; the 11th peak is determined as the fingerprint peak of dehydrocostus lactone; the 13th peak is determined as the fingerprint peak of honokiol; the 14th peak is determined as the fingerprint peak of atractylodin.

[0062] The fourth aspect of the present invention provides a method for screening the fingerprints of multiple herbs in the mixed powder of Aucklandiae and Saussureae Pill, including the following steps:

[0063] a) Preparation of the sample solution of single herb: Any one or more of the 10 herb samples of Aucklandia lappa, Cyperus rotundus L. var. xanthorrhiza (Bunge) Hand.-Mazz., Magnolia officinalis Rehd. et Wils., Glycyrrhiza uralensis Fisch., Atractylodes lancea (Thunb.) DC., Areca catechu L., Amomum villosum Lour., Citrus reticulata Blanco, Citrus reticulata Blanco cv. Tankan Hayata, and Poncirus trifoliata (L.) Raf. in the mixed powder of Aucklandiae and Saussureae Pill are added with methanol for dissolution, and prepared according to step A) of the detection method of the fingerprint of the mixed powder of Aucklandiae and Saussureae Pill to obtain at least one sample solution of single herb respectively;

[0064] b) Preparation of negative sample solutions: For the 10 medicinal material samples of Aucklandiae Radix, Cyperi Rhizoma Rotundatum, Magnoliae Officinalis Cortex, Glycyrrhizae Radix, Atractylodis Rhizoma Praeparatum, Arecae Semen, Amomi Fructus Villosi, Citri Reticulatae Pericarpium, Citri Reticulatae Viride Pericarpium, and Aurantii Fructus Immaturus Praeparatum, respectively lacking Aucklandiae Radix, Cyperi Rhizoma Rotundatum, Magnoliae Officinalis Cortex, Glycyrrhizae Radix, Atractylodis Rhizoma Praeparatum, Arecae Semen, Amomi Fructus Villosi, Citri Reticulatae Pericarpium, Citri Reticulatae Viride Pericarpium, and Aurantii Fructus Immaturus Praeparatum, they were prepared according to step A) of the detection method for the fingerprint of the mixed powder of Aucklandiae Radix and Saussurea Root Pills to obtain negative sample solutions lacking Aucklandiae Radix, Cyperi Rhizoma Rotundatum, Magnoliae Officinalis Cortex, Glycyrrhizae Radix, Atractylodis Rhizoma Praeparatum, Arecae Semen, Amomi Fructus Villosi, Citri Reticulatae Pericarpium, Citri Reticulatae Viride Pericarpium, and Aurantii Fructus Immaturus Praeparatum, respectively.

[0065] c) Determination: The fingerprint spectra of the single medicinal material sample solutions in step a) and the negative sample solutions in step b) were respectively determined by high performance liquid chromatography (HPLC) under the same chromatographic conditions as in step C) of the detection method for the fingerprint of the mixed powder of Aucklandiae Radix and Saussurea Root Pills.

[0066] d) Obtaining the reference fingerprint spectrum: The test solution prepared according to step A) of the detection method for the fingerprint of the mixed powder of Aucklandiae Radix and Saussurea Root Pills was used, and the reference fingerprint spectrum of the mixed powder of Aucklandiae Radix and Saussurea Root Pills was obtained by the same step C) as the detection method for the fingerprint of the mixed powder of Aucklandiae Radix and Saussurea Root Pills.

[0067] e) Quality inspection: The fingerprint spectra of the single medicinal material sample solutions and the negative sample solutions were compared with the reference fingerprint spectrum of the mixed powder of Aucklandiae Radix and Saussurea Root Pills. Through the relative retention time, the corresponding characteristic peaks of the single medicinal material sample solutions in the reference fingerprint spectrum of the mixed powder of Aucklandiae Radix and Saussurea Root Pills were identified, so as to attribute and locate the characteristic peaks in the fingerprint spectra of the single medicinal material sample solutions.

[0068] Preferably, in step a), the ratio of the weight (mg) of any one or more of the 10 medicinal material samples of Aucklandiae Radix, Cyperi Rhizoma Rotundatum, Magnoliae Officinalis Cortex, Glycyrrhizae Radix, Atractylodis Rhizoma Praeparatum, Arecae Semen, Amomi Fructus Villosi, Citri Reticulatae Pericarpium, Citri Reticulatae Viride Pericarpium, and Aurantii Fructus Immaturus Praeparatum added to the volume (mL) of methanol added is 25 - 27:20 - 30, specifically such as 25 - 27:20 - 23, 25 - 27:23 - 27, 25 - 27:27 - 30, for example 25 - 27:20, 25 - 27:22, 25 - 27:25, 25 - 27:28, 25 - 27:30, and preferably 25 - 27:25.

[0069] Preferably, in step a), the Aucklandiae Radix is the dried root of Aucklandia lappa Decne. of the Compositae family. The Cyperi Rhizoma Praeparatum cum Vinegar is the Cyperi Rhizoma processed with vinegar, and the Cyperi Rhizoma is the dried rhizome of Cyperus rotundus L. of the Cyperaceae family. The Magnoliae Officinalis Cortex is the dried trunk bark, root bark and branch bark of Magnolia officinalis Rehd. et Wils. or Magnolia officinalis Rehd. et Wils. var. biloba Rehd. et Wils. of the Magnoliaceae family. The Glycyrrhizae Radix et Rhizoma is the dried root and rhizome of Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L. of the Leguminosae family. The Atractylodis Rhizoma Praeparatum is the Atractylodis Rhizoma processed by frying, and the Atractylodis Rhizoma is the dried rhizome of Atractylodes lancea (Thunb.) DC. or Atractylodes chinensis (DC.) Koidz. of the Compositae family. The Arecae Semen is the dried ripe seed of Areca catechu L. of the Palmae family. The Amomi Fructus is the dried ripe fruit of Amomum villosum Lour., Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen or Amomum longiligulare T. L. Wu of the Zingiberaceae family. The Citri Reticulatae Pericarpium is the dried ripe pericarp of Citrus reticulata Blanco and its cultivated varieties of the Rutaceae family. The Citri Reticulatae Viride Pericarpium Praeparatum is the Citri Reticulatae Viride Pericarpium processed by frying, and the Citri Reticulatae Viride Pericarpium is the pericarp of the dried young fruit or immature fruit of Citrus reticulata Blanco and its cultivated varieties of the Rutaceae family. The Aurantii Fructus Immaturus Praeparatum is the Aurantii Fructus Immaturus processed by frying, and the Aurantii Fructus Immaturus is the dried immature fruit of Citrus aurantium L. and its cultivated varieties of the Rutaceae family.

[0070] Preferably, in step e), the present invention locates the attribution of characteristic peaks by comparing the fingerprint of the single-herb medicine sample solution with the control fingerprint of the mixed powder of Aucklandiae Flos. The analysis and processing are carried out using the software "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicines" (2012 Edition) released by the Pharmacopoeia Commission of the People's Republic of China, and the attribution of the characteristic peaks of each single-herb medicine in the mixed powder of Aucklandiae Flos is confirmed by the relative retention time of each characteristic peak on the control fingerprint of the mixed powder of Aucklandiae Flos. The specific results are shown in Figure 2 、 6 and Table 1.

[0071] Table 1 Attribution of Characteristic Peaks of Each Single-Herb Medicine in the Mixed Powder of Aucklandiae Flos

[0072]

[0073] As can be seen from Table 1, the fingerprint method of the present invention can characterize the characteristic peaks of 8 herbs (Aucklandiae Radix, Magnoliae Officinalis Cortex, Glycyrrhizae Radix et Rhizoma, Atractylodis Rhizoma Praeparatum, Arecae Semen, Citri Reticulatae Pericarpium, Citri Reticulatae Viride Pericarpium Praeparatum, Aurantii Fructus Immaturus Praeparatum) in the mixed powder of Aucklandiae Flos, but fails to characterize the index components of Cyperi Rhizoma Praeparatum cum Vinegar and Amomi Fructus.

[0074] All the water used in the present invention is purified water.

[0075] As described above, the present invention provides a fingerprint and multi-component content determination method for the mixed powder of Aucklandiae Flos, establishes an HPLC fingerprint of the mixed powder of Aucklandiae Flos, and determines 14 common characteristic peaks; and conducts HPLC-DAD content determination on 9 components such as hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostunolide, magnolol, honokiol and atractylodin. The method is fast and simple, aiming to comprehensively analyze and systematically and rapidly evaluate the drug quality of the mixed powder of Aucklandiae Flos, and provide a scientific basis for controlling the quality of this drug.

[0076] Among them, the present invention provides a method for detecting the fingerprint of the mixed powder of Aucklandiae and Saussureae Pill, which has 14 common peaks in total. The total area of the common peaks reaches more than 90% of the total area, and the characteristic peaks of 8 out of the 10 medicinal materials, except for Aucklandiae Radix Auklandiae Praeparata cum Vinegar and Amomi Fructus, are well characterized in the fingerprint and their attribution is confirmed. This method can separately compare the similarities of the fingerprints of 20 batches of mixed powder samples of Aucklandiae and Saussureae Pill. Since there is no increase or decrease in the number of peaks, the fingerprint similarities are relatively high (all ≥0.99). This method can truly and comprehensively evaluate the drug quality of the mixed powder of Aucklandiae and Saussureae Pill, and has good reproducibility and stability. The results of the methodological verification are all good, which can meet the needs of the quality control research of the mixed powder of Aucklandiae and Saussureae Pill.

[0077] In addition, the present invention provides a method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussureae Pill, which can detect the contents of 9 components (hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol and Atractylodis Rhizoma) in 20 batches of mixed powder samples of Aucklandiae and Saussureae Pill. This method uses the wavelength transformation method to simultaneously determine 9 active components in Aucklandiae and Saussureae Pill in a single chromatogram, reducing the test cost and time; using an acetonitrile-phosphoric acid water system as the mobile phase for gradient elution, the chromatographic peaks of each component have good shapes, the resolution is greater than 2, and the peak signal intensity is higher than that of other systems. The method has high sensitivity, good reproducibility and strong specificity, and is simple to operate. The results of the methodological investigation are good.

[0078] In the above method, since the mixed powder of Aucklandiae and Saussureae Pill is composed of 10 Chinese medicinal materials, namely Aucklandiae Radix, Aucklandiae Radix Auklandiae Praeparata cum Vinegar, Magnoliae Officinalis Cortex, Glycyrrhizae Radix et Rhizoma, Atractylodis Rhizoma Praeparata, Arecae Semen, Amomi Fructus, Citri Reticulatae Pericarpium, Citri Reticulatae Viride Pericarpium and Aurantii Fructus Immaturus Pericarpium, except for Zingiberis Rhizoma Recens, and contains complex chemical components, the representative components of the monarch drug Aucklandiae Radix, costunolide and dehydrocostus lactone; the representative components of the ministerial drug Magnoliae Officinalis Cortex, magnolol and honokiol; the representative components of the ministerial drugs Citri Reticulatae Pericarpium and Aurantii Fructus Immaturus Pericarpium, hesperidin, naringin, neohesperidin and naringin rutinoside; and the representative component of the ministerial drug Atractylodis Rhizoma, atractylodin are selected as the index components of the mixed powder of Aucklandiae and Saussureae Pill. Among them, the main pharmacological components of the monarch drug Aucklandiae Radix are costunolide and dehydrocostus lactone, and these components have effects on promoting gastrointestinal motility and protecting the gastric mucosa. The main pharmacological components of the ministerial drugs Magnoliae Officinalis Cortex, Citri Reticulatae Viride Pericarpium, Aurantii Fructus Immaturus Pericarpium, Arecae Semen, Citri Reticulatae Pericarpium, Amomi Fructus and Atractylodis Rhizoma Praeparata, magnolol, honokiol, hesperidin, naringin, neohesperidin, naringin rutinoside and atractylodin have a wide range of pharmacological effects on anti-ulcer, promoting digestion and anti-inflammatory. These 9 components have a strong correlation with the indications of the mixed powder of Aucklandiae and Saussureae Pill, such as chest oppression, nausea and vomiting, belching and anorexia, and abdominal distension and pain caused by retention of dampness and turbidity and disharmony between the stomach and spleen, and can reflect the internal quality of the mixed powder of Aucklandiae and Saussureae Pill, and can be used as important indicators for quantitative analysis of the activity analysis of the mixed powder of Aucklandiae and Saussureae Pill. Description of the Drawings

[0079] Figure 1Shown is the comparison liquid chromatogram of the reference substances and samples of 9 chemical components in the mixed powder of Aucklandiae and Saussureae Pill of the present invention. Among them, A: the liquid chromatogram of the sample; B: the liquid chromatogram of the reference substance; 1 is naringin rutinoside; 2 is naringin; 3 is hesperidin; 4 is neohesperidin; 5 is magnolol; 6 is costunolide; 7 is dehydrocostus lactone; 8 is honokiol; 9 is atractylodin.

[0080] Figure 2 Shown is the liquid chromatogram of the sample and the sample lacking components in the mixed powder of Aucklandiae and Saussureae Pill of the present invention. Among them, 2 is naringin rutinoside; 3 is naringin; 4 is hesperidin; 5 is neohesperidin; 9 is magnolol; 10 is costunolide; 11 is dehydrocostus lactone; 13 is honokiol; 14 is atractylodin.

[0081] Figure 3 Shown is the HPLC reference fingerprint chromatogram of the mixed powder of Aucklandiae and Saussureae Pill of the present invention, which has 14 characteristic peaks. Among them, 2 is naringin rutinoside; 3 is naringin; 4 is hesperidin; 5 is neohesperidin; 9 is magnolol; 10 is costunolide; 11 is dehydrocostus lactone; 13 is honokiol; 14 is atractylodin.

[0082] Figure 4 Shown is the HPLC superposed fingerprint chromatogram of 20 batches of the mixed powder of Aucklandiae and Saussureae Pill of the present invention. Among them, S1 - S20 are the HPLC fingerprint chromatograms of 20 batches of the mixed powder samples of Aucklandiae and Saussureae Pill, and S21 is the HPLC reference fingerprint chromatogram of the mixed powder of Aucklandiae and Saussureae Pill.

[0083] Figure 5 Shown is the comparison diagram of the fingerprint chromatogram of the mixed reference substances and the liquid chromatogram of the index components in the mixed powder of Aucklandiae and Saussureae Pill of the present invention. Among them, A: the fingerprint chromatogram of the mixed reference substances; B: the liquid chromatogram of the index components. Among them, 2 is naringin rutinoside; 3 is naringin; 4 is hesperidin; 5 is neohesperidin; 9 is magnolol; 10 is costunolide; 11 is dehydrocostus lactone; 13 is honokiol; 14 is atractylodin.

[0084] Figure 6 Shown is the attribution diagram of the characteristic peaks of the fingerprint chromatograms of each single herb in the mixed powder of Aucklandiae and Saussureae Pill of the present invention. Among them, 2 is naringin rutinoside; 3 is naringin; 4 is hesperidin; 5 is neohesperidin; 9 is magnolol; 10 is costunolide; 11 is dehydrocostus lactone; 13 is honokiol; 14 is atractylodin. Detailed Embodiments

[0085] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the protection scope of the present invention.

[0086] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0087] The reagents and instruments used in the following examples are as follows:

[0088] 1. Reagents

[0089] Reference substances: hesperidin (batch number: 110721-202220, mass fraction 97.2%, China Food and Drug Inspection Institute), naringin (batch number: 110722-202116, mass fraction 93.5%, China Food and Drug Inspection Institute), neohesperidin (batch number: 111857-202305, mass fraction 99.6%, China Food and Drug Inspection Institute), naringin (batch number: 23081421, mass fraction 98.0%), costunolide (batch number: 111524-202312, mass fraction 99. 6%, China Food and Drug Inspection Institutes), dehydrocostus lactone (batch number: 74299-48-2, mass score 99.3%, Shanghai Shidande Standard Technical Services Co., Ltd.), magnolol (batch number: 528-43-8, mass score 99.2%, Shanghai Shidande Standard Technical Services Co., Ltd.), honokiol (batch number: 110730-201905, mass score 99.8%, China Food and Drug Inspection Institutes), atractylodesin (batch number: 111924-202207, mass score 99.7%, China Food and Drug Inspection Institutes).

[0090] Samples: 20 batches of Muxiang Shunqi Pills mixed powder samples, all produced and provided by Shanghai Hutchison Pharmaceuticals Co., Ltd. The specific batch number information is shown in Table 2 below.

[0091] Medicinal materials: costus root, vinegar-cured cyperus rotundus, magnolia bark, licorice, stir-fried atractylodes, areca catechu, amomum villosum, tangerine peel, stir-fried green tangerine peel, and stir-fried aurantium are all commonly used medicinal materials and provided by Shanghai Hutchison Pharmaceuticals Co., Ltd.

[0092] Reagents: methanol (analytical grade AR, Sinopharm Chemical Reagent Co., Ltd.), acetonitrile, methanol, phosphoric acid (chromatographic grade, TEDIA, USA), ultrapure water was prepared by Milli-Q ultrapure water treatment system.

[0093] Table 2

[0094] Serial number Batch number Serial number Batch number 1 230701-1 11 230703-3 2 230701-2 12 230703-4 3 230701-3 13 230704-1 4 230701-4 14 230704-2 5 230702-1 15 230704-3 6 230702-2 16 230704-4 7 230702-3 17 230205-1 8 230702-4 18 230205-2 9 230703-1 19 230205-3 10 230703-2 20 230205-4

[0095] 2. Instruments

[0096] Agilent 1260 high performance liquid chromatograph (OpenLAB CDS2.1 chromatography workstation, G1312B binary pump, G1322A automatic degasser, G7116A column oven, G7115ADAD detector, G7129A autosampler, Agilent, USA); SB-5200DTD ultrasonic cleaner (Ningbo Xinzhi Biotechnology Co., Ltd.); AL204 and XS205 analytical electronic balances (METTLER TOLEDO Instrument Shanghai Co., Ltd.); Milli-Q Advantage A10 ultrapure water system (Merck Millipore, Germany).

[0097] Example 1

[0098] 1. Sample pretreatment

[0099] Preparation of test solution: Take 0.25 g of the powder of the mixed powder sample of Muxiang Shunqi Pills from batch 230701-1, accurately weigh it, put it in a 50 mL conical flask with a stopper, accurately add 25 mL of methanol, stopper it, weigh it, perform ultrasonic extraction (power 250 W, frequency 40 kHz) for 60 minutes, let it stand and cool to room temperature, filter it through a 0.45 μm microporous filter membrane, and take the filtrate to obtain the test solution 1#.

[0100] Preparation of reference solution: Accurately weigh the reference substances of hesperidin, naringin, neohesperidin, naringin, costunolide, dehydrocostunolide, magnolol, honokiol and atractylodesin respectively, place them in the same 100mL volumetric flask, dissolve them in methanol and dilute to the mark, shake well to prepare the reference solution.

[0101] In the reference solution, the content of hesperidin ranged from 19.0415 to 1218.6547 μg / mL; the content of naringin ranged from 6.3765 to 408.0938 μg / mL; the content of neohesperidin ranged from 5.5021 to 352.1346 μg / mL; the content of naringin ranged from 3.3786 to 216.2306 μg / mL; the content of costunolide ranged from The content range of dehydrocostus lactone was 1.7585~112.5444μg / mL; the content range of magnolol was 2.5037~160.2338μg / mL; the content range of magnolol was 0.8790~56.2550μg / mL; the content range of honokiol was 1.3146~84.1341μg / mL; the content range of atractylodesin was 1.0218~32.6976μg / mL.

[0102] 2. Chromatographic conditions

[0103] The chromatographic conditions of the HPLC method were as follows: the chromatographic column was Waters Xbridge C18 Chromatographic column (4.6 mm × 250 mm, 5 μm); the detector is a photodiode array detector (DAD); the column temperature is 25 °C; the injection volume is 10 μL; the flow rate is 1.0 mL / min.

[0104] Analysis was carried out in a multi-wavelength manner, and the detection wavelengths were 210 nm and 270 nm. Specifically: from 0 - 24.5 min, the detection wavelength was 210 nm; from 24.5 - 29 min, the detection wavelength was 270 nm; from 29 - 65 min, the detection wavelength was 210 nm.

[0105] The mobile phase was acetonitrile - 0.1% phosphoric acid aqueous solution. Among them, phase A was acetonitrile and phase B was 0.1% phosphoric acid aqueous solution; the analysis time was 65 min; gradient elution was used.

[0106] The specific program of gradient elution was as follows:

[0107] From 0 - 5 min, the volume ratio of phase A to phase B was 10:90 - 10:90;

[0108] From 5 - 20 min, the volume ratio of phase A to phase B was 10:90 - 20:80;

[0109] From 20 - 40 min, the volume ratio of phase A to phase B was 20:80 - 60:40;

[0110] From 40 - 60 min, the volume ratio of phase A to phase B was 60:40 - 60:40;

[0111] From 60 - 65 min, the volume ratio of phase A to phase B was 60:40 - 90:10.

[0112] 3. Determination

[0113] The external standard method was adopted. A series of reference solution with different volumes were respectively taken and prepared into a series of solutions with different concentrations. They were analyzed by injection using a high performance liquid chromatograph to plot the standard working curve. Then the obtained test solution was analyzed by injection using a high performance liquid chromatograph, and the analysis results were substituted into the standard working curve to obtain the contents of 9 components in the test solution.

[0114] Specifically, a series of reference substance solutions with different volumes were respectively taken and prepared into a series of solutions with different concentrations. High performance liquid chromatography was used for injection analysis to obtain the linear relationship between the concentrations of 9 components and the peak areas in the reference substance solutions. Taking the chromatographic peak area of each component corresponding to its respective concentration, the corresponding standard working curve was plotted, and the regression equation of each standard working curve was calculated. Then, the test solution was detected by high performance liquid chromatography. The chromatographic peak areas of the 9 components in the obtained test solution were respectively substituted into the regression equations of the respective standard working curves, and the contents of the corresponding components could be calculated. The specific chromatogram is shown in Figure 1 。

[0115] Example 2

[0116] 1. Sample pretreatment

[0117] Preparation of test solution: Weigh accurately 0.25 g of the powder of the mixed powder sample of the Aucklandiae Flos and Lignum Aquilariae Resinatum Pills of batch 230701-2, place it in a 50 mL stoppered conical flask, accurately add 23 mL of methanol, stopper it, weigh, ultrasonically extract (power 255 W, frequency 42 kHz) for 58 minutes, let it stand and cool to room temperature, filter through a 0.45 μm microporous membrane, and take the subsequent filtrate to obtain test solution 2#.

[0118] Preparation of reference substance solution: Weigh accurately the reference substances of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostunolide, magnolol, honokiol, and atractylodin, place them in the same 100 mL volumetric flask, dissolve with methanol and dilute to the scale, shake well to prepare the reference substance solution. The concentration range of the reference substance solution is the same as that in step 1 of Example 1.

[0119] 2. Chromatographic conditions

[0120] The chromatographic conditions for high performance liquid chromatography are as follows: The chromatographic column is a Waters Xbridge C 18 chromatographic column (4.6 mm × 250 mm, 5 μm); the detector is a photodiode array detector (DAD); the column temperature is 23 °C; the injection volume is 12 μL; the flow rate is 0.8 mL / min.

[0121] Analysis is carried out in a multi-wavelength manner, and the detection wavelengths are 208 nm and 272 nm. Specifically: from 0 to 24.5 min, the detection wavelength is 208 nm; from 24.5 to 29 min, the detection wavelength is 272 nm; from 29 to 65 min, the detection wavelength is 208 nm.

[0122] The mobile phase is acetonitrile - 0.09% phosphoric acid aqueous solution. Among them, phase A is acetonitrile and phase B is 0.09% phosphoric acid aqueous solution; the analysis time is 65 min; gradient elution.

[0123] The specific procedure for gradient elution is as follows:

[0124] From 0 to 5 min, the volume ratio of phase A to phase B is 9:91 - 9:91;

[0125] From 5 to 20 min, the volume ratio of phase A to phase B is 9:91 - 19:81;

[0126] From 20 to 40 min, the volume ratio of phase A to phase B is 19:81 - 59:41;

[0127] From 40 to 60 min, the volume ratio of phase A to phase B is 59:41 - 59:41;

[0128] From 60 to 65 min, the volume ratio of phase A to phase B is 59:41 - 91:9.

[0129] 3. Determination

[0130] The specific determination process is the same as that in step 3 of Example 1.

[0131] Example 3

[0132] 1. Sample pretreatment

[0133] Preparation of the test solution: Weigh accurately 0.25 g of the powder of the mixed powder sample of the Aucklandia and Saussurea Pill (batch No. 230701 - 3), place it in a 50 - mL stoppered conical flask, accurately add 27 mL of methanol, stopper it, weigh it, extract ultrasonically (power 245 W, frequency 38 kHz) for 62 minutes, let it stand and cool to room temperature, filter through a 0.45 - μm microporous membrane, and take the subsequent filtrate to obtain the test solution 3#.

[0134] Preparation of the reference solution: Weigh accurately the reference substances of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostunolide, magnolol, honokiol, and atractylodin respectively, place them in the same 100 - mL volumetric flask, dissolve with methanol and dilute to the scale, shake well to prepare the reference solution. The concentration range of the reference solution is the same as that in step 1 of Example 1.

[0135] 2. Chromatographic conditions

[0136] The chromatographic conditions for high - performance liquid chromatography are as follows: The chromatographic column is Waters Xbridge C 18 chromatographic column (4.6 mm × 250 mm, 5 μm); the detector is a photodiode array detector (DAD); the column temperature is 27 °C; the injection volume is 8 μL; the flow rate is 1.2 mL / min.

[0137] Analysis is carried out by using a multi-wavelength method, and the detection wavelengths are 212 nm and 268 nm. The specific situation is as follows: from 0 - 24.5 min, the detection wavelength is 212 nm; from 24.5 - 29 min, the detection wavelength is 268 nm; from 29 - 65 min, the detection wavelength is 212 nm.

[0138] The mobile phase is acetonitrile - 0.11% phosphoric acid aqueous solution, where phase A is acetonitrile and phase B is 0.11% phosphoric acid aqueous solution; the analysis time is 65 min; gradient elution is used.

[0139] The specific program of gradient elution is as follows:

[0140] From 0 - 5 min, the volume ratio of phase A to phase B is 11:89 - 11:89;

[0141] From 5 - 20 min, the volume ratio of phase A to phase B is 11:89 - 21:79;

[0142] From 20 - 40 min, the volume ratio of phase A to phase B is 21:79 - 61:39;

[0143] From 40 - 60 min, the volume ratio of phase A to phase B is 61:39 - 61:39;

[0144] From 60 - 65 min, the volume ratio of phase A to phase B is 61:39 - 89:11.

[0145] 3. Determination

[0146] The specific determination process is the same as step 3 in Example 1.

[0147] Example 4

[0148] Precisely weigh the reference substances of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol, and atractylodin respectively, place them in the same 100 mL volumetric flask, dissolve with methanol and dilute to the scale, shake well, and prepare a reference substance solution containing 216.231 μg of naringin rutinoside, 408.094 μg of naringin, 1218.655 μg of hesperidin, 352.135 μg of neohesperidin, 56.25 μg of magnolol, 112.544 μg of costunolide, 160.234 μg of dehydrocostus lactone, 84.13 μg of honokiol, and 32.698 μg of atractylodin per 1 mL.

[0149] Precisely pipette 0.156 mL, 0.312 mL, 0.625 mL, 1.25 mL, 2.5 mL, and 5 mL of the reference substance stock solution respectively, and make up the volume to 10 mL. Inject 10 μL according to the chromatographic conditions in Step 2 of Example 1 for determination. With the peak area (Y) as the ordinate and the concentration (X) as the abscissa, plot the standard curve and perform regression calculation. The linear relationships of each component are good. The specific results are shown in Table 3.

[0150] Table 3

[0151]

[0152] And successively dilute the above reference substance solution to samples with different concentrations, pipette 10 μL for determination. Take the reference substance concentration when the signal-to-noise ratio of the peak area is 3 times (S / N = 3) as the detection limit (LOD), and take the reference substance concentration when the signal-to-noise ratio of the peak area is 10 times (S / N = 10) as the quantitation limit (LOQ). The specific results are shown in Table 4 below.

[0153] Table 4

[0154]

[0155] Example 5

[0156] 1. Method specificity

[0157] Take the mixed powder sample of Aucklandia and Saussurea Pill of Batch 230701-1, and prepare the test solution according to Step 1 of Example 1. At the same time, take the reference substances of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostunolide, magnolol, honokiol, and atractylodin, and prepare the reference substance solution according to Step 1 of Example 1.

[0158] Take the samples of 10 medicinal materials including Aucklandia lappa, Cyperus rotundus L. var. xanthorrhiza, Magnolia officinalis Rehd. et Wils., Glycyrrhiza uralensis Fisch., Atractylodes lancea (Thunb.) DC., Areca catechu L., Amomum villosum Lour., Citrus reticulata Blanco, Citrus reticulata Blanco cv. Tangerina, and Citrus aurantium L. var. amara Engl. without Aucklandia lappa, Cyperus rotundus L. var. xanthorrhiza, Magnolia officinalis Rehd. et Wils., Glycyrrhiza uralensis Fisch., Atractylodes lancea (Thunb.) DC., Areca catechu L., Amomum villosum Lour., Citrus reticulata Blanco, Citrus reticulata Blanco cv. Tangerina, and Citrus aurantium L. var. amara Engl. respectively, and prepare the negative sample solutions without Aucklandia lappa, Cyperus rotundus L. var. xanthorrhiza, Magnolia officinalis Rehd. et Wils., Glycyrrhiza uralensis Fisch., Atractylodes lancea (Thunb.) DC., Areca catechu L., Amomum villosum Lour., Citrus reticulata Blanco, Citrus reticulata Blanco cv. Tangerina, and Citrus aurantium L. var. amara Engl. according to Step 1 of Example 1.

[0159] Determine the test solution, reference substance solution, and negative sample solution according to the chromatographic conditions in Step 2 of Example 1. The results are shown in Figure 1 、 2 . From Figure 1 、 2 , it can be seen that the method specificity is good.

[0160] 2. Precision

[0161] Take the mixed powder sample of Aucklandia and Saussurea Pill with batch number 230701-1, prepare 1 portion of the test solution according to step 1 in Example 1, perform the detection according to step 2 in the above Example 1, inject samples continuously for 6 times respectively for analysis, and record the peak areas of the chromatographic peaks of each component. The results show that the RSDs of the peak areas of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol and atractylodin are all less than 3.0%, indicating good instrument precision.

[0162] 2. Stability

[0163] Take the mixed powder sample of Aucklandia and Saussurea Pill with batch number 230701-1, prepare 1 portion of the test solution according to step 1 in Example 1, perform the detection according to step 2 in the above Example 1, inject samples at 0, 2, 4, 8, 12, 24 h, and 48 h respectively, record the chromatogram, and measure the peak area. The results show that the RSDs of the peak areas of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol and atractylodin are all less than 3.0%, indicating good stability of the test solution within 48 h.

[0164] 4. Repeatability

[0165] Take the mixed powder sample of Aucklandia and Saussurea Pill with batch number 230701-1, prepare 6 portions of the test solution according to step 1 in Example 1, accurately weigh 6 portions, perform the detection according to step 2 in the above Example 1, record the chromatogram, and record the peak areas of the chromatographic peaks of each component. The results show that the RSDs of the peak areas of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol and atractylodin are all less than 3.0%, indicating good repeatability of the method.

[0166] 3. Recovery

[0167] Take the mixed powder sample of Aucklandia and Saussurea Pill with batch number 230701-1, about 0.125 g for each portion, accurately weigh, add reference substances of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol and atractylodin at three different concentration levels respectively, take 3 portions for each mass concentration, prepare the test solution according to step 1 in Example 1, perform the detection according to step 2 in the above Example 1, record the chromatogram, calculate the recovery of each component according to the measured amount and the added amount, and the results are shown in Table 5. As can be seen from Table 5, it shows good accuracy of the method.

[0168] Table 5

[0169]

[0170]

[0171]

[0172] Example 6

[0173] Take 20 batches of Muxiang Shunqi Pills mixed powder samples, prepare the test solution according to step 1 in Example 1, and analyze them according to the chromatographic conditions of step 2 in Example 1, record the chromatograms, and calculate the contents of 9 index components in the Muxiang Shunqi Pills mixed powder by external standard method. The results are shown in Table 6.

[0174] Table 6 Contents of 9 ingredients in 20 batches of Muxiang Shunqi Pills mixed powder (mg g-1)

[0175]

[0176] Example 7

[0177] The various testing conditions in the method for determining the content of multiple components in Muxiang Shunqi Pills mixed powder were investigated, and the specific conditions are as follows:

[0178] 1. Comparison of extraction solvents

[0179] N, N-dimethylformamide (DMF) and methanol were used as solvents, respectively, and the test solution was prepared according to step 1 in Example 1, and the samples were analyzed according to the chromatographic conditions of step 2 in Example 1, and the contents of 9 index components in the mixed powder of Muxiang Shunqi Pills were calculated by external standard method. The results showed that there was no significant difference in the results of the two solvents with the number of peaks, interference between peaks, separation degree and quantitative peak area as indicators. Since DMF is more toxic than methanol, methanol is selected as the extraction solvent, which is not only less toxic than DMF, but also simpler and easier to obtain than DMF.

[0180] 2. Sample quantity

[0181] 0.1, 0.25, 0.5, 1.0g of the mixed powder sample of Muxiang Shunqi Pills were prepared according to step 1 in Example 1 to obtain the test solution, and the samples were respectively injected and analyzed according to the chromatographic conditions of step 2 in Example 1, and the contents of the 9 index components in the mixed powder of Muxiang Shunqi Pills were calculated by the external standard method. The results showed that at a sampling amount of 0.25g, the mixed powder sample of Muxiang Shunqi Pills could be completely extracted, the chromatographic peaks of each index component had a good peak shape, the number of peaks was not short, and the ratio of the index component content to the sampling amount was the highest, that is, at a sampling amount of 0.25g, the accuracy of the results could be guaranteed at a smaller sampling amount.

[0182] 3. Mobile phase

[0183] The mobile phases were selected as acetonitrile-0.1% phosphoric acid aqueous solution, acetonitrile-methanol-0.1% phosphoric acid aqueous solution, and methanol-0.1% phosphoric acid aqueous solution, respectively. The test solution was prepared according to step 1 in Example 1, and the samples were injected and analyzed according to the chromatographic conditions of step 2 in Example 1, and the contents of the 9 index components in the mixed powder of Muxiang Shunqi Pills were calculated by the external standard method. The results showed that the acetonitrile-0.1% phosphoric acid water system was used as the mobile phase for gradient elution, and the chromatographic peaks of each component had good peak shapes, the separation degrees were all greater than 2, and the peak signal intensity was higher than that of other systems. It can be seen that selecting acetonitrile-0.1% phosphoric acid aqueous solution as the mobile phase can achieve high sensitivity, good reproducibility, and strong specificity.

[0184] Example 8

[0185] 1. Sample pretreatment

[0186] Preparation of the test solution: The preparation process of the test solution is the same as that in step 1 of Example 1.

[0187] Preparation of reference solution: The preparation process of reference solution is the same as that in step 1 of Example 1.

[0188] 2. Chromatographic conditions

[0189] The chromatographic conditions of the HPLC are the same as those of the HPLC in step 2 of Example 1.

[0190] 3. Determination

[0191] The high performance liquid chromatography method with the chromatographic conditions in the above step 2 is used to measure the test solution and the reference solution in the above step 1 respectively, and the fingerprint of the test solution and the fingerprint of the reference solution are obtained. The fingerprint of the test solution is compared with the fingerprint of the reference solution, and the corresponding characteristic peaks in the fingerprint of the test solution are identified by relative retention time according to the known characteristic peaks in the fingerprint of the reference solution, so as to attribute and locate the index components in the fingerprint of the test solution and obtain the fingerprint of the mixed powder of Muxiang Shunqi Pills.

[0192] Example 9

[0193] The detection method of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula established in Example 8 above was used to detect 20 batches of the mixed powder of Aucklandiae and Saussureae Formula, and the fingerprint spectra of the test solution and the reference solution were obtained. The fingerprint spectrum data of the obtained test samples were imported into the software "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicines" 2012 version released by the Pharmacopoeia Commission of the People's Republic of China. Taking sample 230701-1 as the reference spectrum, by the average method, the time window width was set to 0.20, and multi-point calibration was used to match the fingerprint spectra to generate the fingerprint spectrum and the reference fingerprint spectrum. The fingerprint spectrum of the test sample was compared with the reference fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula obtained under the same fingerprint spectrum detection conditions, and the similarity of the fingerprint spectrum of each batch of the mixed powder of Aucklandiae and Saussureae Formula was calculated. Taking peak 4 (hesperidin) as the reference peak S, 14 common peaks were calibrated, as shown in Figure 3 . The results of comparing the similarities of the fingerprint spectra of 20 batches of test samples with the reference fingerprint spectrum showed that the similarities of the fingerprint spectra of 20 batches of the mixed powder of Aucklandiae and Saussureae Formula were all greater than 0.99, indicating that the overall quality of each batch of samples was relatively stable. The superimposed map of the fingerprint spectra of 20 batches of the mixed powder of Aucklandiae and Saussureae Formula is shown in Figure 4 . The specific results are shown in Table 7 below.

[0194] Table 7

[0195]

[0196]

[0197] Example 10

[0198] The detection method of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula established in Example 8 above was used to detect the mixed powder of Aucklandiae and Saussureae Formula. According to the obtained fingerprint spectra of the test solution and the reference solution, 14 common fingerprint peaks were identified, and 9 of them were assigned by comparison with reference substances. The 14 common characteristic peaks and their numbers of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula are as shown in Figure 3 、 5As shown, the proportion of 14 common peaks in the total peak area reached over 90%. Among them, the elution time of peak No. 4 was moderate, the resolution was good, and the peak area was large. Therefore, peak No. 4 was selected as the reference peak (S peak), and the relative retention times of other peaks were calculated. Thus, with peak No. 4 as the reference peak (S peak, relative retention time being 1.0000), the relative retention times of the other 13 peaks were successively as follows: peak No. 1 (0.4791 ± 0.0072), peak No. 2 (0.9667 ± 0.0025), peak No. 3 (0.9879 ± 0.0008), peak No. 5 (1.0190 ± 0.0006), peak No. 6 (1.1268 ± 0.0021), peak No. 7 (1.3982 ± 0.0120), peak No. 8 (1.4860 ± 0.0136), peak No. 9 (1.6094 ± 0.0143), peak No. 10 (1.6716 ± 0.0139), peak No. 11 (1.7219 ± 0.0235), peak No. 12 (1.7444 ± 0.0257), peak No. 13 (1.7671 ± 0.0218), peak No. 14 (2.2611 ± 0.0187).

[0199] By comparison with Figure 5 it was determined that peak No. 2 was the fingerprint peak of naringin rutinoside; peak No. 3 was the fingerprint peak of naringin; peak No. 4 was the fingerprint peak of hesperidin; peak No. 5 was the fingerprint peak of neohesperidin; peak No. 9 was the fingerprint peak of magnolol; peak No. 10 was the fingerprint peak of costunolide; peak No. 11 was the fingerprint peak of dehydrocostus lactone; peak No. 13 was the fingerprint peak of honokiol; peak No. 14 was the fingerprint peak of atractylodin..

[0200] Example 11

[0201] 1. Precision

[0202] Take the mixed powder sample of Aucklandia and Saussurea Pill of batch 230701-1, prepare 1 portion of the test solution according to the procedure in step 1 of Example 8, perform detection according to the procedure in step 2 of Example 8 above, inject samples continuously for 6 times, record the chromatogram, with peak No. 4 (hesperidin) as the reference peak, calculate that the RSD% of the relative retention times of 14 common peaks are all less than 2.0%, and the RSD of the relative peak areas are all less than 3.0%, indicating good precision of the instrument.

[0203] 2. Repeatability

[0204] Take the mixed powder sample of Aucklandia and Saussurea Pill of batch 230701-1, prepare 6 portions of the test solution in parallel according to the procedure in step 1 of Example 8, perform detection according to the procedure in step 2 of Example 8 above, record the chromatogram, with peak No. 4 (hesperidin) as the reference peak, calculate that the RSD% of the relative retention times of 14 common peaks are all less than 2.0%, and the RSD of the relative peak areas are all less than 3.0%, and the results show good repeatability of the method.

[0205] 3. Stability

[0206] Take the mixed powder sample of Muxiang Shunqi Pills from batch 230701-1, prepare 1 portion of the test solution according to step 1 in Example 8, and detect according to step 2 in the above Example 8, inject samples at 0, 2, 4, 8, 12, and 24 h, record the chromatogram, take peak No. 4 (hesperidin) as the reference peak, calculate the relative retention time RSD% of 14 common peaks, all of which are less than 2.0%, and the relative peak area RSD% are all less than 3.0%, indicating that the test solution has good stability within 24 h.

[0207] Example 12

[0208] The test solution was prepared by using step 1 in Example 8 above.

[0209] Take 26.32 mg each of 10 medicinal material powders, including costusroot, vinegar-soaked cyperus, magnolia bark, liquorice, stir-fried atractylodes, areca catechu, amomum villosum, tangerine peel, stir-fried green peel, and stir-fried aurantium, add 25 mL of methanol, and adopt step 1 in the above Example 8 to prepare 10 single medicinal material sample solutions.

[0210] Using step 2 in Example 7, the test solution and 10 single medicinal material sample solutions were measured respectively to obtain fingerprints of the test solution and 10 single medicinal material sample solutions. The attribution was confirmed by the retention time of the chromatographic peak of each single medicinal material sample solution and the chromatographic peak of the fingerprint of Muxiang Shunqi Pills mixed powder, as shown in detail. Figure 6 , Table 1.

[0211] Depend on Figure 6 As shown in Table 1, the characteristic peaks of 8 out of 10 medicinal materials are well represented in the fingerprint spectrum, and their attribution is confirmed.

[0212] In summary, the present invention provides a fingerprint spectrum of a mixed powder of Muxiang Shunqi Pills and a method for determining the content of multiple components, establishes an HPLC fingerprint spectrum of the mixed powder of Muxiang Shunqi Pills, and determines the content of 9 components, which provides a scientific basis for controlling the quality of the drug. Therefore, the present invention overcomes various shortcomings in the prior art and has a high industrial utilization value.

[0213] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussureae Pill, comprising the following steps: 1) Preparation of the test solution: Add the sample of the mixed powder of Aucklandiae and Saussureae Pill to methanol for dissolution, perform ultrasonic extraction, cool it, filter, and take the subsequent filtrate to obtain the test solution; 2) Preparation of the reference solution: Add one or more reference substances of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol, and atractylodin to methanol as the solvent and make up the volume to obtain the reference solution; 3) Determination: Use high performance liquid chromatography to separately determine the test solution in step 1) and the reference solution in step 2), and calculate the contents of one or more components of hesperidin, naringin, neohesperidin, naringin rutinoside, costunolide, dehydrocostus lactone, magnolol, honokiol, and atractylodin in the test solution by the external standard method.

2. The method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussurea Pill according to claim 1, wherein In step 1), any one or more of the following conditions are included: A1) The mixed powder of Aucklandiae and Saussureae Pill is a powder sample; A2) The ratio of the weight of the added mixed powder sample of Aucklandiae and Saussureae Pill to the volume of the added methanol is 0.25:20 - 30, g / mL; A3) The ultrasonic extraction time is 50 - 70 min; A4) The power of the ultrasonic extraction is 240 - 260 W; the frequency of the ultrasonic extraction is 35 - 45 kHz; A5) The filtration is membrane filtration; the membrane is a 0.45 μm membrane.

3. The method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussurea Pill according to claim 1, characterized in that, In step 2), the content range of hesperidin in the reference solution is 19.0415 - 1218.6547 μg / mL; the content range of naringin is 6.3765 - 408.0938 μg / mL; the content range of neohesperidin is 5.5021 - 352.1346 μg / mL; the content range of naringin rutinoside is 3.3786 - 216.2306 μg / mL; the content range of costunolide is 1.7585 - 112.5444 μg / mL; the content range of dehydrocostus lactone is 2.5037 - 160.2338 μg / mL; the content range of magnolol is 0.8790 - 56.2550 μg / mL; the content range of honokiol is 1.3146 - 84.1341 μg / mL; the content range of atractylodin is 1.0218 - 32.6976 μg / mL.

4. The method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussurea Pill according to claim 1, characterized in that, In step 3), the high performance liquid chromatography includes any one or more of the following conditions: B1) The chromatographic column is C 18 Chromatographic column; B2) The detector is a photodiode array detector; B3) The column temperature is 20 - 30 °C; B4) The injection volume is 5 - 15 μL; B5) The flow rate is 0.5 - 2 mL / min; B6) The detection wavelength is selected from one or a combination of two of 205 - 215 nm and 265 - 275 nm; preferably, the detection wavelengths are 205 - 215 nm and 265 - 275 nm; B7) The mobile phase is acetonitrile - 0.09 - 0.11% phosphoric acid aqueous solution; in the acetonitrile - 0.09 - 0.11% phosphoric acid aqueous solution, phase A is acetonitrile, phase B is 0.09 - 0.11% phosphoric acid aqueous solution; In the high performance liquid chromatography, gradient elution is adopted and the analysis time is 65 min.

5. The method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussurea Pill according to claim 4, wherein It further includes any one or more of the following conditions: B11) In item B1), the chromatographic column is a Waters Xbridge C 18 chromatographic column, 4.6 mm × 250 mm, 5 μm; B61) In item B6), the detection wavelength changes with the time of the gradient elution program. The specific change is as follows: from 0 to 24.5 min, the detection wavelength is 205 - 215 nm; from 24.5 to 29 min, the detection wavelength is 265 - 275 nm; from 29 to 65 min, the detection wavelength is 205 - 215 nm. B81) In item B8), the specific program of the gradient elution is as follows: From 0 to 5 min, the volume ratio of phase A to phase B is 8 - 12:88 - 92 - 8 - 12:88 - 92; From 5 to 20 min, the volume ratio of phase A to phase B is 8 - 12:88 - 92 - 18 - 22:78 - 82; From 20 to 40 min, the volume ratio of phase A to phase B is 18 - 22:78 - 82 - 58 - 62:38 - 42; From 40 to 60 min, the volume ratio of phase A to phase B is 58 - 62:38 - 42 - 58 - 62:38 - 42; From 60 to 65 min, the volume ratio of phase A to phase B is 58 - 62:38 - 42 - 88 - 92:8 - 12.

6. A detection method for the fingerprint of the mixed powder of Aucklandiae and Saussureae Formula Pills, comprising the following steps: A) Preparation of the test solution: the same as step 1) of the method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussureae Formula Pills according to any one of claims 1 - 5; B) Preparation of the reference solution: the same as step 2) of the method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussureae Formula Pills according to any one of claims 1 - 5; C) Determination: Using high performance liquid chromatography under the same chromatographic conditions as in step 3) of the method for determining the contents of multiple components in the mixed powder of Aucklandiae and Saussureae Formula Pills according to any one of claims 1 - 5, respectively determine the test solution in step A) and the reference solution in step B), obtain the fingerprint of the test solution and the fingerprint of the reference solution, compare the fingerprint of the test solution with the fingerprint of the reference solution, and conduct attribution and positioning of the target components in the fingerprint of the test solution, so as to obtain the fingerprint of the mixed powder of Aucklandiae and Saussureae Formula Pills.

7. A quality detection method for the mixed powder of Aucklandiae and Saussureae Formula Pills, comprising obtaining the fingerprint of the mixed powder of Aucklandiae and Saussureae Formula Pills by using the detection method for the fingerprint of the mixed powder of Aucklandiae and Saussureae Formula Pills according to claim 6, and comparing the similarity between the obtained fingerprint of the mixed powder of Aucklandiae and Saussureae Formula Pills and the control fingerprint of the mixed powder of Aucklandiae and Saussureae Formula Pills obtained under the same fingerprint detection conditions.

8. The quality inspection method of the Aucklandiae and Saussureae Formula powder according to claim 8, characterized in that Under the same conditions as those in the detection method of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills described in claim 6, the control fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills is obtained. The control fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills includes 14 common fingerprint peaks. Taking the 4th peak as the reference peak S, the relative retention time is 1.0000; the relative retention times of the other 13 peaks are as follows: the 1st peak is 0.4791±0.0072, the 2nd peak is 0.9667±0.0025, the 3rd peak is 0.9879±0.0008, the 5th peak is 1.0190±0.0006, the 6th peak is 1.1268±0.0021, the 7th peak is 1.3982±0.0120, the 8th peak is 1.4860±0.0136, the 9th peak is 1.6094±0.0143, the 10th peak is 1.6716±0.0139, the 11th peak is 1.7219±0.0235, the 12th peak is 1.7444±0.0257, the 13th peak is 1.7671±0.0218, and the 14th peak is 2.2611±0.0187.

9. A method for screening the fingerprint spectra of multiple medicinal materials in the mixed powder of Aucklandiae and Saussureae Formula Pills, comprising the following steps: a) Preparation of the sample solution of single medicinal material: Any one or more of the 10 medicinal material samples of Aucklandia lappa, Cyperus rotundus L. var. suaveolens (Pursh) C. Y. Wu et Hsiao, Magnolia officinalis Rehd. et Wils., Glycyrrhiza glabra L., Atractylodes lancea (Thunb.) DC. var. lancea, Areca catechu L., Amomum villosum Lour., Citrus reticulata Blanco, Citrus reticulata Blanco cv. Chachiensis Hort., and Poncirus trifoliata (L.) Raf. in the mixed powder of Aucklandiae and Saussureae Formula Pills are added with methanol for dissolution, and prepared according to step A) of the detection method of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills described in claim 6 to obtain at least one sample solution of single medicinal material respectively; b) Preparation of the negative sample solution: The 10 medicinal material samples of Aucklandia lappa, Cyperus rotundus L. var. suaveolens (Pursh) C. Y. Wu et Hsiao, Magnolia officinalis Rehd. et Wils., Glycyrrhiza glabra L., Atractylodes lancea (Thunb.) DC. var. lancea, Areca catechu L., Amomum villosum Lour., Citrus reticulata Blanco, Citrus reticulata Blanco cv. Chachiensis Hort., and Poncirus trifoliata (L.) Raf. are respectively lacking Aucklandia lappa, Cyperus rotundus L. var. suaveolens (Pursh) C. Y. Wu et Hsiao, Magnolia officinalis Rehd. et Wils., Glycyrrhiza glabra L., Atractylodes lancea (Thunb.) DC. var. lancea, Areca catechu L., Amomum villosum Lour., Citrus reticulata Blanco, Citrus reticulata Blanco cv. Chachiensis Hort., and Poncirus trifoliata (L.) Raf., and are prepared according to step A) of the detection method of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills described in claim 6 to obtain the negative sample solutions lacking Aucklandia lappa, Cyperus rotundus L. var. suaveolens (Pursh) C. Y. Wu et Hsiao, Magnolia officinalis Rehd. et Wils., Glycyrrhiza glabra L., Atractylodes lancea (Thunb.) DC. var. lancea, Areca catechu L., Amomum villosum Lour., Citrus reticulata Blanco, Citrus reticulata Blanco cv. Chachiensis Hort., and Poncirus trifoliata (L.) Raf. respectively; c) Determination: The fingerprint spectra of the sample solution of single medicinal material in step a) and the negative sample solution in step b) are respectively determined by high performance liquid chromatography under the same chromatographic conditions as those in step C) of the detection method of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills described in claim 6; d) Obtaining the control fingerprint spectrum: The test solution prepared according to step A) of the detection method of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills described in claim 6 is used, and the control fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills is obtained by using the same step C) as that in the detection method of the fingerprint spectrum of the mixed powder of Aucklandiae and Saussureae Formula Pills described in claim 6; e) Quality inspection: Compare the fingerprints of the single medicinal sample solution and the negative sample solution with the control fingerprint of the Muxiang Shunqi Pills mixed powder. By the relative retention time, identify the corresponding characteristic peaks of the single medicinal sample solution in the control fingerprint of the Muxiang Shunqi Pills mixed powder, so as to attribute and locate the characteristic peaks in the fingerprint of the single medicinal sample solution.

10. The screening method for the fingerprint spectra of multiple medicinal materials in the mixed powder of Aucklandia and Saussurea Pill according to claim 9, characterized in that, In step a), the ratio of the weight of any one or more of the 10 medicinal material samples of costus root, vinegar-soaked cyperus, magnolia bark, liquorice, stir-fried atractylodes, areca nut, amomum villosum, tangerine peel, stir-fried green peel, and stir-fried aurantium in the mixed powder of the Muxiang Shunqi Pills to the volume of the added methanol is 25-27:20-30, mg / mL.

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