A method for constructing a characteristic spectrum of a traditional Chinese medicine compound preparation for treating rhinitis

Abstract

The present invention provides a method for constructing a characteristic spectrum of a Chinese medicine compound preparation for Tongqiao rhinitis, and relates to the technical field of a characteristic spectrum analysis method for Chinese patent medicine. The method comprises the following steps: weighing a sample in proportion, grinding, extracting once with a solvent, heating and refluxing, and concentrating the extract to obtain residue 1; dissolving residue 1, extracting twice with a solvent, washing, and concentrating to obtain residue 2; taking residue 2, dissolving with a solvent, and shaking well to obtain; weighing a reference substance in the same container, and preparing a solution with a solvent; performing high-performance liquid chromatography analysis on the test solution and the reference solution to obtain a high-performance liquid chromatography characteristic spectrum, determining a common peak according to the relative retention time with the reference substance, determining a characteristic peak by comparing with the reference substance, and establishing a high-performance liquid chromatography characteristic spectrum. The technical scheme of the present invention adds a qualitative function for several components of siler, magnolia, and mint; it is stable and reliable, has good reproducibility, and high precision, which makes up for the shortcomings of the existing quality control methods.

Description

Technical Field

[0001] The present invention relates to the technical field of characteristic spectrum analysis methods for Chinese patent medicines, and in particular to a method for constructing a characteristic spectrum of a Chinese medicine compound preparation for treating rhinitis. Background Art

[0002] The traditional Chinese medicine compound preparation for Tongqiao rhinitis contains 7 traditional Chinese medicines: Fangfeng, Huangqi, Xinyi, Baizhi, Mentha, Xanthium and Baizhu. Its indications are mainly to dispel wind, reduce inflammation, and clear the nasal passages. It is used for sinusitis, nasal congestion, runny nose, etc. It can also be used for the treatment of frontal headache, rhinitis, sinusitis and allergic rhinitis. The ingredients of traditional Chinese medicine compound preparations are complex and the quality is uneven. How to detect the content of active ingredients and then improve the preparation process to improve the efficacy is the focus of research.

[0003] High performance liquid chromatography (HPLC) is used to detect Chinese medicine components, with good separation effect, high sensitivity and fast analysis speed. For example, Chinese patent CN 116539763 A specifically relates to a detection method for simultaneously determining multiple index components in Chinese medicine lily. According to the progress of pharmacological and pharmacodynamic research on Chinese medicine lily, the index components that can be used as quality markers of Chinese medicine lily include ferulic acid, p-coumaric acid, gallic acid, dioscin, and lilyoside B, etc. By exploring the treatment method of the test sample, the same test sample treatment method and the same liquid chromatography method are achieved at dual wavelengths of 210nm and 300nm to achieve the beneficial effect of simultaneously detecting five index components of ferulic acid, p-coumaric acid, gallic acid, dioscin, and lilyoside B. Chinese patent CN107976498B discloses a method for detecting the functional effective ingredients of Sanguisorba officinalis and its application, comprising the steps of preparing standard solutions of rutin, schisandrin A and schisandrin B respectively, preparing test solution, HPLC detection and the like. The HPLC detection using gradient elution can simultaneously determine the three functional effective ingredients in Sanguisorba officinalis, which can more comprehensively reflect the efficacy of Sanguisorba officinalis.

[0004] The quality control methods of the traditional Chinese medicine compound preparations for Tongqiao rhinitis (including granules, tablets, and capsules) mainly include identification and content determination. The identification items mainly use thin layer chromatography to identify the medicinal materials of Angelica dahurica, Astragalus membranaceus, Xanthium sibiricum and Atractylodes macrocephala in the preparations, and the content determination items use high performance liquid chromatography (HPLC) to determine the content of imperatorin in Angelica dahurica and astragaloside I in Astragalus membranaceus. From the prescription composition, it can be found that the traditional Chinese medicine compound preparations for Tongqiao rhinitis are made of 7 traditional Chinese medicines such as Angelica dahurica and Astragalus membranaceus, but its quality standard only identifies 4 raw medicinal materials and HPLC detects the content of two indicator components in Angelica dahurica and Astragalus membranaceus. It is not comprehensive and accurate to control the quality of the traditional Chinese medicine compound preparations for Tongqiao rhinitis.

[0005] Based on the above problems, in view of the shortcomings of the existing quality control methods of the traditional Chinese medicine compound preparations for Tongqiao rhinitis, the researchers of the present invention are committed to providing a method for establishing a characteristic spectrum and an HPLC characteristic spectrum obtained by the method. It is characterized in that the traditional Chinese medicine compound preparation for Tongqiao rhinitis is prepared into a test solution, and the characteristic spectrum of the traditional Chinese medicine compound preparation for Tongqiao rhinitis is obtained through HPLC separation and detection, thereby providing a reliable basis for the intrinsic quality and authenticity identification of the traditional Chinese medicine compound preparation for Tongqiao rhinitis. Among the 7 medicinal flavors in the prescription, 5 medicinal flavors in the characteristic spectrum have been identified through standard products, negative samples, and medicinal material attribution, including fangfeng, astragalus, magnolia, angelica dahurica and mint, among which fangfeng, magnolia and mint are all medicinal flavors for which there are no qualitative and quantitative methods in the existing standards. According to industry requirements, the characteristic peaks of traditional Chinese medicine preparations should generally cover more than 2 / 3 of the characteristic peaks of medicinal flavors. The technical characteristic peaks of the present invention have covered 5 / 7 of the medicinal flavors, which is greater than 2 / 3 and meets the requirements. Summary of the invention

[0006] In view of the above problems, the present invention provides a method for constructing a characteristic spectrum of a Chinese herbal compound preparation for Tongqiao rhinitis. The Chinese herbal compound preparation for Tongqiao rhinitis is prepared into a test solution, and the characteristic spectrum of the Chinese herbal compound preparation for Tongqiao rhinitis is obtained through HPLC separation and detection, thereby providing a reliable basis for the intrinsic quality and authenticity identification of the Chinese herbal compound preparation for Tongqiao rhinitis.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0008] In one aspect, the present invention provides a method for constructing a characteristic spectrum of a traditional Chinese medicine compound preparation for treating rhinitis, comprising the following steps:

[0009] S1: Prepare the test solution: weigh the sample according to the proportion, grind, extract once with solvent, heat under reflux, and concentrate the extract to obtain residue 1; dissolve residue 1, extract twice with solvent, wash, and concentrate to obtain residue 2; dissolve residue 2 with solvent, shake well, and obtain;

[0010] S2: Prepare reference solution: weigh the reference substance in the same container and add solvent to prepare a solution with a concentration of 45-55 μg / mL;

[0011] S3: Performing HPLC analysis on the test solution and the reference solution to obtain a HPLC characteristic spectrum, determining common peaks according to the relative retention time with the reference solution, determining characteristic peaks by comparing with the reference solution, and establishing a HPLC characteristic spectrum;

[0012] The chromatographic conditions are:

[0013] The chromatographic column is a Phenomenex Luna C18 column (4.6 mm*250 mm, 5 μm);

[0014] The mobile phases were: methanol as mobile phase A and 0.1% formic acid water as mobile phase B;

[0015] Column temperature: 20-30°C;

[0016] Injection volume: 3-7 μL;

[0017] Flow rate: 0.5-1.5mL / min;

[0018] Detection wavelength: 270-280nm;

[0019] The elution gradient is as follows:

[0020] Time / min Mobile phase A / % Mobile phase B / % 0-10 20-25 75-80 10-(19-23) (25-29)→35 (75-71)→65 23-40 35 65 40-(50-85) (35-50)→80 (65-50)→20 .

[0021] Preferably, the solvent for the primary extraction described in S1 may be methanol.

[0022] Preferably, the mass volume ratio of the sample described in S1 to the solvent for the primary extraction is 1.0-1.5 g / 10 mL; further preferably, the mass volume ratio of the sample described in S1 to the solvent for the primary extraction is 1.25 g / 10 mL.

[0023] Preferably, the temperature of the heating reflux in S1 is 60-70°C, and the time is 3.5-4.5h; further preferably, the temperature of the heating reflux in S1 is 65°C, and the time is 4h.

[0024] Preferably, the specific steps of dissolving the residue 1 described in S1 are: adding water to the residue 1, and heating it to 60°C-70°C to completely dissolve it; further preferably, the specific steps of dissolving the residue 1 described in S1 are: adding water to the residue 1, and heating it to 65°C to completely dissolve it.

[0025] Preferably, the solvent for the secondary extraction described in S1 can be a water-saturated n-butanol solution.

[0026] Preferably, the mass volume ratio of the sample described in S1 to the water-saturated n-butanol solution is 0.8-1.2 g / 10 mL; further preferably, the mass volume ratio of the sample described in S1 to the water-saturated n-butanol solution is 1 g / 10 mL.

[0027] Preferably, the specific steps of the secondary extraction described in S1 can be: adding a water-saturated n-butanol solution, shaking and extracting for 3-5 times, and combining the n-butanol solutions; further preferably, the specific steps of the secondary extraction described in S1 can be: adding a water-saturated n-butanol solution, shaking and extracting for 4 times, and combining the n-butanol solutions.

[0028] Preferably, the solvent used for washing in S1 can be an ammonia test solution.

[0029] Preferably, the preparation method of the ammonia test solution is: take 400 mL of concentrated ammonia solution, add water to make 1000 mL, and obtain the solution.

[0030] Preferably, the mass volume ratio of the sample described in S1 to the ammonia test solution is 0.8-1.2 g / 10 mL; further preferably, the mass volume ratio of the sample described in S1 to the ammonia test solution is 1 g / 10 mL.

[0031] Preferably, the specific operation of washing described in S1 is: fully washing with ammonia test solution for 1-3 times to remove the ammonia liquid; further preferably, the specific operation of washing described in S1 is: fully washing with ammonia test solution for 2 times to remove the ammonia liquid.

[0032] Preferably, the solvent for dissolving the residue 2 in S1 can be methanol, with a volume of 5 mL.

[0033] Preferably, the reference substance in S2 is selected from at least one of cimicifuga glycoside, calycosin glucoside, 5-O-methylvisaminol glycoside, and montana glycoside; further preferably, the reference substance in S2 is cimicifuga glycoside, calycosin glucoside, 5-O-methylvisaminol glycoside, and montana glycoside.

[0034] Preferably, the concentration described in S2 may be 50 μg / mL.

[0035] Preferably, the reference substance described in S3 is 5-O-methylvisaminol glycoside.

[0036] Preferably, the number of theoretical plates in the high performance liquid chromatography analysis process described in S3 is calculated based on the 5-O-methylvisaminol glycoside peak, and the number of theoretical plates is ≥3000.

[0037] Preferably, the characteristic peaks described in S3 include 7, and the medicinal taste attribution and relative retention of each common peak are:

[0038] Peak tR / min Compound Attribution 1 19.921 Cimicifuga glycosides Windproof 2 21.899 Isoflavone Glucoside Astragalus 3 30.693 / Magnolia 4(S) 38.526 5-O-Methylvisaminol Windproof 5 47.169 / Angelica dahurica 6 50.454 / Angelica dahurica 7 56.976 Ligusticum glabra Mint .

[0039] Preferably, the chromatographic conditions are:

[0040] The chromatographic column is a Phenomenex Luna C18 column (4.6 mm*250 mm, 5 μm);

[0041] The mobile phases were: methanol as mobile phase A and 0.1% formic acid water as mobile phase B;

[0042] Column temperature: 20-30°C;

[0043] Injection volume: 3-7 μL;

[0044] Flow rate: 0.5-1.5mL / min;

[0045] Detection wavelength: 275nm;

[0046] The elution gradient is as follows:

[0047] Time / min Mobile phase A / % Mobile phase B / % 0-10 25 75 10-16 25→29 75→71 16-23 29→35 71→65 23-40 35 65 40-50 35→50 65→50 50-85 50→80 50→20 .

[0048] Further preferably, the chromatographic conditions are:

[0049] The chromatographic column is a Phenomenex Luna C18 column (4.6 mm*250 mm, 5 μm);

[0050] The mobile phases were: methanol as mobile phase A and 0.1% formic acid water as mobile phase B;

[0051] Column temperature: 25°C;

[0052] Injection volume: 5 μL;

[0053] Flow rate: 1 mL / min;

[0054] Detection wavelength: 275nm;

[0055] The elution gradient is as follows:

[0056] Time / min Mobile phase A / % Mobile phase B / % 0-10 25 75 10-16 25→29 75→71 16-23 29→35 71→65 23-40 35 65 40-50 35→50 65→50 50-85 50→80 50→20 .

[0057] Compared with the prior art, the present invention has the following beneficial effects:

[0058] 1. Compared with the records in the Chinese Pharmacopoeia, the technical solution of the present invention has added the qualitative function of several ingredients such as siler, magnolia and mint;

[0059] 2. Compared with traditional thin-layer chromatography identification, the integrity and fuzziness of the characteristic spectrum of traditional Chinese medicine just meet the requirements of traditional Chinese medicine quality control. Compared with the quality control method of a single component or indicator component, it is more scientific and comprehensive. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 It is a high performance liquid chromatography characteristic spectrum with 7 characteristic peaks;

[0061] Figure 2 Overlapping chromatograms for identification of characteristic spectra; S1 is the control drug of Angelica dahurica, S2 is the mixed control solution of calycosin glucoside, 5-O-methylvisaminol glucoside, and monanthin, S3 is the mixed control solution of cimicifuga glycoside and magnolin, among which magnolin has interference in the test solution and is therefore not listed as a characteristic component, and S4 is the test solution of batch 220929;

[0062] Figure 3The HPLC overlapping chromatograms of 7 batches of samples, the sample batch numbers are shown in Table 3;

[0063] Figure 4 Overlapping chromatograms of HPLC under different wavelength conditions; the wavelengths from top to bottom are 275nm, 335nm, 315nm, 294nm, and 249nm;

[0064] Figure 5 The HPLC overlapping chromatograms are obtained under the conditions of three mobile phase systems: methanol-1% phosphoric acid water, acetonitrile-0.1% phosphoric acid water, and methanol-acetonitrile-0.1% phosphoric acid water; S1 is methanol-0.1% phosphoric acid water (30:70), S2 is methanol-acetonitrile-0.1% phosphoric acid water (25:5:70), S3 is methanol-acetonitrile-0.1% phosphoric acid water (15:5:80), S4 is acetonitrile-0.1% phosphoric acid water (5:95), S5 is acetonitrile-0.1% phosphoric acid water (20:80), and the elution time is 110 min; S6 is acetonitrile-0.1% phosphoric acid water (20:80), and the elution time is 120 min;

[0065] Figure 6 The HPLC overlapping chromatograms under different composition ratios of the mobile phase system of methanol, acetonitrile and 0.1% phosphoric acid·2% isopropanol water; S1 is methanol-0.1% phosphoric acid·2% isopropanol (25:75), with a detection wavelength of 290 nm, S2 is methanol-0.1% phosphoric acid·2% isopropanol (25:75), with a detection wavelength of 294 nm, S3 is methanol-0.1% phosphoric acid·2% isopropanol (20:80), with a detection wavelength of 294 nm, and S4 is acetonitrile-0.1% phosphoric acid·2% isopropanol (25:75), with a detection wavelength of 294 nm;

[0066] Figure 7 The HPLC overlapping chromatograms under the conditions of different ratios of mobile phase system of methanol-acetonitrile-0.1% formic acid water; S1 is methanol-acetonitrile-0.1% formic acid water (20:5:75), S2 is methanol-acetonitrile-0.1% formic acid water (15:10:75), and the detection wavelength is 275nm;

[0067] Figure 8 This is the HPLC characteristic spectrum of pretreatment method 1;

[0068] Fig. 9 It is the HPLC characteristic spectrum of pretreatment method 2;

[0069] Fig.10 It is the high performance liquid chromatography characteristic spectrum of pretreatment method three (technical solution of the present invention). DETAILED DESCRIPTION

[0070] In order to make the technical means, creative features, purpose and effect of the present invention easy to understand, the present invention is further explained below in conjunction with specific embodiments, but the following embodiments are only preferred embodiments of the present invention, not all. Based on the embodiments in the implementation mode, other embodiments obtained by those skilled in the art without making creative work all belong to the protection scope of the present invention. It is worth noting that the raw materials used in the present invention are all common commercial products, and their sources are not specifically limited. The technology and scientific terms used in the embodiments have the meanings commonly understood by those of ordinary skill in the art to which the present invention belongs.

[0071] 1. Technical solution

[0072] 1) Preparation method of test solution: Take about 4g of Tongqiao Biyan Granules, grind them into powder, weigh them accurately, put them in a Soxhlet extractor, add 50mL of methanol, heat and reflux for 4 hours, recover the solvent from the extract and concentrate it to dryness, add 10mL of water to the residue, dissolve it with slight heat, shake and extract it with water-saturated n-butanol for 4 times, 40mL each time, combine the n-butanol solutions, wash them thoroughly with ammonia test solution for 2 times, 40mL each time, discard the ammonia solution, recover the solvent from the n-butanol solution to dryness, dissolve the residue with methanol and transfer it to a 5mL volumetric flask, add methanol to the mark, shake well, and obtain.

[0073] (2) Preparation of mixed reference solution: Take appropriate amount of cimicifuga glycoside, calycosin glucoside, 5-O-methylvisaminol glycoside, and monanthus glycoside, accurately weigh them, place them in the same volumetric flask, and add methanol to make a solution containing 50 μg per 1 mL.

[0074] (3) performing high performance liquid chromatography analysis on the test solution and the reference solution to obtain a high performance liquid chromatography characteristic spectrum, determining common peaks based on the relative retention time with the reference solution, determining characteristic peaks using the reference solution, and establishing a high performance liquid chromatography characteristic spectrum;

[0075] Chromatographic conditions: octadecylsilane bonded silica gel filler (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm), methanol as mobile phase A, 0.1% formic acid water as mobile phase B; gradient elution as specified in the following table, column temperature 25°C, detection wavelength 275 nm, flow rate 1.0 mL / min, injection volume 5 μL, theoretical plate number calculated based on 5-O-methylvisaminol glycoside peak, not less than 3000.

[0076] Table 1. Elution gradient

[0077]

[0078]

[0079] The characteristic spectrum of the traditional Chinese medicine compound preparation for Tongqiao rhinitis is based on the chromatographic peak of 5-O-methylvisaminol glycoside. The characteristic spectrum of the traditional Chinese medicine compound preparation for Tongqiao rhinitis includes 7 common peaks. The medicinal taste attribution and relative retention of each common peak are as follows:

[0080] Table 2. Medicinal taste attribution and relative retention time of each common peak

[0081]

[0082] Three of the components were identified through reference substances, including calycosin from Astragalus, 5-O-methylvisamol from Saposhnikovia divaricata, and montanol from Mentha. By comparing with the medicinal materials, one chromatographic peak was identified as a component of Magnolia officinalis, and two chromatographic peaks were identified as components of Angelica dahurica. Figure 2 shown.

[0083] In the present invention, the Tongqiao rhinitis preparations are all commercially available drugs, and the sample batch numbers are:

[0084] Table 3. Sample batch numbers

[0085] sample batch number S1 220923 S2 220929 S3 221023 S4 220415 S5 221212 S6 221035 S7 221223

[0086] The HPLC characteristic spectra of 7 batches of samples are as follows: Figure 3 shown.

[0087] 2. Method optimization

[0088] 1) Investigation of different detection wavelengths

[0089] Keeping other conditions unchanged, only changing the detection wavelength to 249nm, 275nm, 294nm, 315nm, 335nm, the results are as follows Figure 4 As shown in the figure, the chromatographic peak distribution and separation are compared. In general, the characteristic peak at 275nm is more obvious and has better separation. In addition, at 275nm, the characteristic components in magnolia have maximum absorption. Combined with the 3D full scan section data, 275nm was finally determined as the detection wavelength.

[0090] 2) Investigation of different mobile phase systems

[0091] The other conditions were kept unchanged, and only the mobile phase was changed. The comparison of three mobile phase systems, methanol-1% phosphoric acid water, acetonitrile-0.1% phosphoric acid water, and methanol-acetonitrile-0.1% phosphoric acid water, was investigated. Figure 5 ), comparison of different composition ratios of methanol, acetonitrile, 0.1% phosphoric acid, and 2% isopropanol water mobile phase systems ( Figure 6 ), Comparison of different composition ratios of methanol-acetonitrile-0.1% formic acid water mobile phase system ( Figure 7). The following conclusions can be drawn: adding formic acid inhibits the activity of silanol groups, and the tailing phenomenon of the chromatographic peak is significantly improved; using methanol as the organic phase, the peak shape is sharper, the separation time is shorter, and the separation effect is better than acetonitrile, so methanol-0.1% formic acid aqueous solution is finally selected as the mobile phase.

[0092] 3) Optimization of sample processing methods

[0093] Method 1: Take about 2.0 grams of sample, weigh accurately, add 25 mL of methanol, extract by ultrasonic for 40 minutes, filter, and use as the test solution. The detection wavelength is temporarily set at 275 nm for comparison. The results are as follows Figure 8 As shown,

[0094] There are many impurities, low chromatographic peak response, and poor separation, which can easily cause chromatographic column contamination and difficulty in cleaning the detector.

[0095] Method 2: The preparation method of the test solution is the same as that of the test solution in the test of astragaloside IV content under Tongqiao Rhinitis Granules in the 2020 edition of the Chinese Pharmacopoeia; specifically: take the product under the filling amount, grind it, take about 4g, accurately weigh it, put it in a Soxhlet extractor, add 50mL of methanol, heat and reflux for 4 hours, recover the solvent from the extract and concentrate it to dryness, add 10mL of water to the residue, heat it slightly to dissolve it, shake and extract it 4 times with water-saturated n-butanol, 40mL each time, combine the n-butanol solutions, and fill it with ammonia test solution. Wash twice, 40 mL each time, discard the ammonia solution, recover the solvent with n-butanol solution until dry, add 5 mL of water to the residue to dissolve it, cool it, pass it through a D101 macroporous adsorption resin column (inner diameter 1.5 cm, column height 12 cm), elute with 50 mL of water, discard the water, then elute with 30 mL of 40% ethanol, discard the eluate, and then elute with 80 mL of 70% ethanol, collect the eluate, evaporate it to dryness, dissolve it with methanol and transfer it to a 5 mL volumetric flask, add methanol to the scale, shake well, and obtain it.

[0096] The results are as follows Fig. 9 As shown in the figure, the separation of the chromatographic peaks at 19min-21min is poor, there are more interfering peaks, the pretreatment method is more complicated, and the convenience and repeatability are slightly poor.

[0097] Method 3: The technical solution of the present invention. Fig.10 As shown. Based on method 2, the macroporous resin column-elution procedure was deleted, and the test sample was prepared and the characteristic spectrum of water-soluble components was detected simultaneously. The results showed that compared with methods 1 and 2, the chromatographic peaks were more comprehensive, the chromatographic separation effect was better, the peak shape was better, and the response was higher ( Fig.10 ).

[0098] 3. Methodological investigation

[0099] 1) Precision experiment

[0100] 220929 batches of samples were taken, and the test solution was prepared according to the determined method. The samples were injected continuously for 6 times. Peak 4 was taken as the reference peak, and the relative retention times of common peaks 1-7 and the RSDs of the retained peaks were calculated to be less than 3%. At the same time, the similarities of each chromatographic fingerprint calculated by similarity software were all greater than 0.99, indicating that the instrument had good precision.

[0101] 2) Reproducibility experiment

[0102] Take 220929 batches of samples, accurately weigh 6 portions respectively, prepare the test solution according to the determined method, inject them separately, take peak 4 as the reference peak, calculate the relative retention time and relative peak area of ​​common peaks 1-7, the RSD is less than 3%, and the similarity of each chromatographic fingerprint calculated by similarity software is greater than 0.99, indicating that the method has good reproducibility.

[0103] 3) Stability test

[0104] 220929 batches of samples were taken, and the test solutions were prepared according to the determined method. The samples were injected at 0, 2, 4, 8, 12, 18, and 24 h, respectively. Peak 4 was taken as the reference peak, and the relative retention times of common peaks 1-7 and the RSDs of the retained peaks were calculated to be less than 3%. At the same time, the similarities of each chromatographic fingerprint calculated by similarity software were all greater than 0.99, indicating that the test solution had good stability within 24 hours.

[0105] In summary, the technical solution of the present invention achieves comprehensive evaluation and control of the quality of the Tongqiao rhinitis preparation by establishing a characteristic spectrum method for the Tongqiao rhinitis preparation and an HPLC characteristic spectrum prepared by the method, thereby ensuring the stability of product quality and the effectiveness and safety of clinical medication.

[0106] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A method for constructing a characteristic spectrum of a Chinese medicine compound preparation for treating rhinitis, characterized in that: The following steps are involved: S1: Prepare the test solution: weigh the sample according to the proportion, grind, extract once with solvent, heat under reflux, and concentrate the extract to obtain residue 1; dissolve residue 1, extract twice with solvent, wash, and concentrate to obtain residue 2; dissolve residue 2 with solvent, shake well, and obtain; S2: Prepare reference substance solution: weigh reference substances in the same container, add solvent to prepare a solution with a concentration of 45-55 μg / mL; the reference substances are cimicifuga glycoside, calycosin glucoside, 5-O-methylvisaminol glycoside and monanthus glycoside; S3: Performing HPLC analysis on the test solution and the reference solution to obtain a HPLC characteristic spectrum, determining common peaks according to the relative retention time with the reference solution, determining characteristic peaks by comparing with the reference solution, and establishing a HPLC characteristic spectrum; The characteristic peaks described in S3 include 7, and the medicinal taste attribution and relative retention of each common peak are: The chromatographic conditions are: The chromatographic column was a Phenomenex Luna C18 column with a length of 25 cm, an inner diameter of 4.6 mm, and a particle size of 5 μm; The mobile phases were: methanol as mobile phase A and 0.1% formic acid water as mobile phase B; Column temperature: 20-30°C; Injection volume: 3-7 μL; Flow rate: 0.5-1.5mL / min; Detection wavelength: 275nm; The elution gradient is as follows:

2. The construction method according to claim 1, characterized in that: The mass volume ratio of the sample described in S1 to the solvent for primary extraction is 1.0-1.5 g / 10 mL.

3. The construction method according to claim 1, characterized in that: The heating reflux temperature in S1 is 60-70° C. and the time is 3.5-4.5 h.

4. The construction method according to claim 1, characterized in that: The solvent for the secondary extraction described in S1 is a water-saturated n-butanol solution.

5. The construction method according to claim 4, characterized in that: The mass volume ratio of the sample described in S1 to the water-saturated n-butanol solution is 0.8-1.2 g / 10 mL.

6. The construction method according to claim 1, characterized in that: The solvent used for washing described in S1 is ammonia test solution.

7. The construction method according to claim 6, characterized in that: The mass volume ratio of the sample described in S1 to the ammonia test solution is 0.8-1.2 g / 10 mL.

8. The construction method according to claim 1, characterized in that: The concentration described in S2 is 50 μg / mL.

9. The construction method according to claim 1, characterized in that: The reference substance described in S3 is 5-O-methylvisaminol glycoside.

10. The construction method according to claim 1, characterized in that: The theoretical plate number in the HPLC analysis process described in S3 is calculated based on the 5-O-methylvisaminol glycoside peak, and the theoretical plate number is ≥3000.

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