Thin-layer chromatography identification method of traditional Chinese medicine granules for treating nasosinusitis

Abstract

The invention relates to a thin-layer chromatography identification method of traditional Chinese medicine granules for treating nasosinusitis, and belongs to the technical field of traditional Chinese medicine analysis. Comprising the following steps: step 1, preparing a test solution; 2, preparing a reference medicinal material solution; 3, preparing a negative control sample; 4, preparing a single medicine sample; and 5, performing thin-layer chromatography. According to the method, complex reagents, instruments or equipment are not needed, the six traditional Chinese medicinal materials including houttuynia cordata, scutellaria baicalensis, ligusticum wallichii, spina gleditsiae, liquorice and magnolia flower in the traditional Chinese medicine granules for treating nasosinusitis can be rapidly identified, the identification result is vivid and visual and easy to identify, and a convenient and practical way is provided.

Description

TECHNICAL FIELD

[0001] The present application relates to a thin layer chromatography identification method of a traditional Chinese medicine granule for treating sinusitis, and belongs to the technical field of traditional Chinese medicine analysis. BACKGROUND

[0002] Sinusitis is a common nasal disease caused by viruses, bacteria or fungi. According to the duration, sinusitis can be divided into acute sinusitis and chronic sinusitis. Common symptoms include nasal congestion, purulent discharge, headache, and reduced or even lost sense of smell. Sinusitis is not contagious or hereditary, and the general population is at risk of developing the disease, but children and the elderly have a higher incidence rate due to lower systemic resistance.

[0003] Sinusitis is known as "nasal dampness" in traditional Chinese medicine. Traditional Chinese medicine has its own characteristics and advantages in treating sinusitis, and some Chinese patent medicines with certain effects have emerged. The present inventors have prepared a traditional Chinese medicine granule with significant therapeutic effect on sinusitis after long-term experiments. The traditional Chinese medicine granule is prepared from nine traditional Chinese medicines, including Houttuynia cordata, Herba Patriniae, Gypsum Fibrosum, Scutellaria baicalensis, Magnolia biondii, Gleditsia sinensis, Chuanxiong Rhizome, Angelica dahurica and Glycyrrhiza uralensis.

[0004] Thin layer chromatography analysis is an important experimental technique for rapid separation and qualitative analysis of small amounts of substances. The equipment and operation process are relatively simple, and the sample usage and time are less. However, the existing thin layer analysis method is mainly for single herb analysis, and there is no report on thin layer identification analysis method for the traditional Chinese medicine granule for treating sinusitis.

[0005] In summary, it is urgent to develop a targeted thin layer chromatography identification method to overcome the above-mentioned defects. SUMMARY

[0006] The purpose of the present application is to provide a thin layer chromatography identification method of a traditional Chinese medicine granule for treating sinusitis.

[0007] The technical solution of the present application to solve the above technical problems is as follows: a thin layer chromatography identification method of a traditional Chinese medicine granule for treating sinusitis, which identifies Houttuynia cordata, Scutellaria baicalensis, Chuanxiong Rhizome, Gleditsia sinensis, Glycyrrhiza uralensis and Magnolia biondii in the traditional Chinese medicine granule for treating sinusitis, comprising the following steps:

[0008] Step 1: preparing a test solution;

[0009] Step 2: preparing a control medicine solution;

[0010] Step 3: preparing a negative control sample;

[0011] Step 4: preparing a single herb sample;

[0012] Step 5: thin layer chromatography: the thin layer chromatography test of Houttuynia cordata, Scutellaria baicalensis, Chuanxiong Rhizoma, Gleditsia sinensis and licorice uses silica gel G thin layer plate, the thin layer chromatography test of Magnolia biondii Pamp uses silica gel H thin layer plate, Houttuynia cordata, Magnolia biondii Pamp, Gleditsia sinensis, licorice are sprayed with color developing agent, heated to develop color spots, and observed under 365nm ultraviolet light;

[0013] In the test sample chromatogram, if the same color fluorescent spots are shown at the corresponding positions of the control medicinal material chromatogram, single medicinal material and control sample, and no same color fluorescent spots are shown at the corresponding positions of the negative control, then it is indicated that the test sample contains Magnolia biondii Pamp, Scutellaria baicalensis, Houttuynia cordata, Gleditsia sinensis, licorice and Chuanxiong Rhizoma.

[0014] In step 3 of the present application, the negative control sample is used to confirm that the selected control sample is specific to a certain medicinal material in the nine medicinal materials, and the thin layer chromatography result needs to ensure that the negative control does not contain the same fluorescent color spots at the same position of the control sample.

[0015] In step 5 of the present application, Chuanxiong Rhizoma and Scutellaria baicalensis can develop color without spraying color developing agent under the observation of 365nm ultraviolet light, and do not need to be sprayed with color developing agent.

[0016] The beneficial effects of the thin layer chromatography identification method of the traditional Chinese medicine granules for treating sinusitis of the present application are as follows:

[0017] 1. The present application can identify Houttuynia cordata, Scutellaria baicalensis, Magnolia biondii Pamp, Chuanxiong Rhizoma, Gleditsia sinensis and licorice, and the main spots of the sample chromatogram and the control medicinal material chromatogram are clear in color under normal temperature conditions, the separation degree is good, there is no tailing phenomenon, and the background is not interfered. The sample chromatogram shows the same color spots at the corresponding positions of the control medicinal material, single medicinal material and control sample, and the same positions of the negative control do not show the same color fluorescent spots, which indicates that the thin layer identification method has good durability under normal temperature conditions.

[0018] 2. The present application does not need complex reagents, instruments or equipment, can quickly identify the six kinds of medicinal materials in the traditional Chinese medicine granules for treating sinusitis, the identification result is visual and easy to identify, and a convenient and practical way is provided.

[0019] On the basis of the above technical scheme, the present application can also be improved as follows.

[0020] Further, the traditional Chinese medicine granules for treating sinusitis are prepared by the following method:

[0021] The following weight parts of raw materials are weighed: Houttuynia cordata 250 parts, Herba Patriniae 250 parts, Gypsum Fibrosum 250 parts, Scutellaria baicalensis 166.7 parts, Magnolia biondii Pamp 166.7 parts, Gleditsia sinensis 166.7 parts, Chuanxiong Rhizoma 100 parts, Radix Angelicae Dahuricae 100 parts and licorice 50 parts.

[0022] Distillation extraction: steam distillation extraction of Houttuynia cordata, Herba Patriniae and Magnolia biondii, collecting the residue, the water solution after distillation and the volatile oil;

[0023] Decoction concentration: mixing the residue with Scutellaria baicalensis, Gypsum fibrosum, Gleditsia sinensis, Rhizoma Chuanxiong, Radix Angelicae dahuricae and Glycyrrhiza uralensis to obtain decoction materials, adding water to the decoction materials and decocting to obtain decoction liquid, combining the decoction liquid with the water solution after distillation, evaporating and concentrating to obtain concentrated liquid;

[0024] Purification: adding anhydrous ethanol to the concentrated liquid, standing, recovering the ethanol to obtain extract;

[0025] Granulation: adding excipients to the extract, mixing uniformly, granulating and drying, spraying the volatile oil to obtain the traditional Chinese medicine granules for treating sinusitis.

[0026] The further beneficial effects of the above are: the traditional Chinese medicine granules for treating sinusitis have the effects of dispelling wind and clearing heat, resolving toxin and pus, removing dampness and dredging the nasal cavity. They are used for treating rhinitis, acute and chronic sinusitis caused by wind-heat attacking from outside and internal accumulation of phlegm-damp and toxin, with symptoms of nasal obstruction, purulent nasal discharge, headache, reduced sense of smell, yellowish tongue fur and floating and uneven pulse.

[0027] The traditional Chinese medicine granules for treating sinusitis overcome the problems of large dosage, inaccuracy of dosage, inconvenience of taking and non-commercialization of traditional Chinese medicine decoction, effectively retain the volatile components of the medicine, ensure the stability, have the effects of accurate dosage, stable quality, convenience of taking, good eating effect, safety of eating and suitability for commercial mass production, and can significantly relieve the symptoms of patients with acute and chronic sinusitis.

[0028] Further, the traditional Chinese medicine granules for treating sinusitis are prepared by the following method:

[0029] Weigh the following raw materials: 250g of Houttuynia cordata, 250g of Herba Patriniae, 250g of Gypsum fibrosum, 166.7g of Scutellaria baicalensis, 166.7g of Magnolia biondii, 166.7g of Gleditsia sinensis, 100g of Rhizoma Chuanxiong, 100g of Radix Angelicae dahuricae and 50g of Glycyrrhiza uralensis;

[0030] Distillation extraction: steam distillation extraction of Houttuynia cordata, Herba Patriniae and Magnolia biondii, adding water to immerse the medicine surface by 4-6cm, soaking for 30min, then distilling for 5h, collecting the residue, the water solution after distillation and the volatile oil;

[0031] Decoction concentration: mixing the residue with Scutellaria baicalensis, Gypsum fibrosum, Gleditsia sinensis, Rhizoma Chuanxiong, Radix Angelicae dahuricae and Glycyrrhiza uralensis, adding water and decocting three times, adding 8 times of water for the first time and decocting for 80min, adding 6 times of water for the second time and decocting for 60min, adding 6 times of water for the third time and decocting for 60min, filtering and combining the three times of decoction liquid and the water solution after distillation; evaporating and concentrating to 3L to obtain concentrated liquid;

[0032] Purification: the concentrated solution is placed at room temperature, anhydrous ethanol is added to reach 70% alcohol content, stirred, stand for 24h, take the supernatant, recover ethanol, concentrate to 500mL, obtain the extract;

[0033] Granulation: the extract is dried to obtain extract powder, then excipients are added to the extract powder, mixed uniformly and processed by granulation and drying, sprayed with volatile oil, to obtain the traditional Chinese medicine granules for treating sinusitis.

[0034] Further, the excipients are any one or more of starch, dextrin and microcrystalline cellulose, and the binder used is pure water.

[0035] Further, the specific method of thin layer chromatography identification of the Houttuynia cordata is as follows:

[0036] Step 1.1.1 Preparation of test solution: take 3g of the traditional Chinese medicine granules for treating sinusitis, dissolve in 5mL water, take 2mL, dissolve in 20mL water, extract with ethyl acetate, combine the ethyl acetate liquid, evaporate to dryness, dissolve the residue in 1mL methanol as the test solution;

[0037] Step 2.1.1 Preparation of control medicinal material solution: take 5g of Houttuynia cordata control medicinal material, prepare the Houttuynia cordata control medicinal material solution according to the method of step 1.1.1 for preparing the test solution;

[0038] Step 3.1.1 Preparation of negative control sample: take eight kinds of medicinal materials of Herba Patriniae, Gypsum Fibrosum, Radix Scutellariae, Flos Magnoliae, Rhizoma Chuanxiong, Radix Angelicae Dahuricae, Radix Sophorae and Radix Glycyrrhizae, prepare the extract according to the preparation method of the traditional Chinese medicine granules for treating sinusitis, take 2mL of the extract, prepare the Houttuynia cordata negative control sample according to the method of step 1.1.1 for preparing the test solution;

[0039] Step 4.1.1 Preparation of Houttuynia cordata single medicinal material sample:

[0040] Decoction and concentration: take 250g of Houttuynia cordata, soak in water, distill, collect the residue, the water solution after distillation and the volatile oil;

[0041] Add the residue to water and decoct, filter and combine the decoction liquid, and combine with the water solution after distillation, heat and concentrate to 3L to obtain the concentrated solution;

[0042] Purification: the concentrated solution is placed at room temperature, anhydrous ethanol is added to reach 70% alcohol content, stirred, stand for 24h, take the supernatant, recover ethanol, concentrate to 500mL, obtain the extract;

[0043] Mix the extract with the volatile oil, take 3mL, prepare the Houttuynia cordata single medicinal material solution according to the method of step 1.1.1 for preparing the test solution;

[0044] Step 5.1.1 Thin layer chromatography: Take the barker glycoside control, add methanol to make a solution containing 1 mg per 1 mL, as the barker control solution;

[0045] Take 5 μL of the barker control solution, the test solution of step 1.1.1, the Houttuynia cordata control medicinal material solution of step 2.1.1, the Houttuynia cordata negative control solution of step 3.1.1 and the Houttuynia cordata single-ingredient medicinal material solution of step 4.1.1 respectively, and point them on the same silica gel G thin layer plate. Use a mixture of ethyl acetate, toluene and formic acid in a volume ratio of 15:3:1 as the developing agent, pre-saturate for 30 min, develop, take out, air dry, spray with 10% sulfuric acid ethanol, place in a 105°C oven until the spots are clear, and observe under a 365 nm ultraviolet light lamp;

[0046] If the test chromatogram contains fluorescent spots of the same color at the positions corresponding to the control medicinal material, the control, and the single-ingredient medicinal material chromatograms, and the same fluorescent color spots are not contained at the same positions of the negative control, then it contains Houttuynia cordata.

[0047] The above further beneficial effects are: using the above method, it can quickly and accurately identify whether the traditional Chinese medicine granules for treating sinusitis contain Houttuynia cordata.

[0048] Further, the specific method of thin layer chromatography identification of the Scutellaria baicalensis is:

[0049] Step 1.1.2 Preparation of test solution: Take 3 g of the traditional Chinese medicine granules for treating sinusitis, dissolve in 5 mL of water, take 2 mL, add 20 mL of methanol, ultrasonic treatment, filter, evaporate the filtrate to dryness, dissolve the residue in 2 mL of methanol as the test solution;

[0050] Step 2.1.2 Preparation of control medicinal material solution: Take 1 g of Scutellaria baicalensis control medicinal material, and prepare a Scutellaria baicalensis control medicinal material solution according to the method of step 1.1.2 for preparing the test solution;

[0051] Step 3.1.2 Preparation of negative control sample: Take Houttuynia cordata, Herba Patriniae, Gypsum Fibrosum, Magnolia biondii, Chuanxiong Rhizoma, Radix Angelicae Dahuricae, Radix Sophorae and Radix Glycyrrhizae eight kinds of medicinal materials, and prepare an extract according to the preparation method of the traditional Chinese medicine granules for treating sinusitis. Take 2 mL of the extract, and prepare a Scutellaria baicalensis negative control solution according to the method of step 1.1.2 for preparing the test solution;

[0052] Step 4.1.2 Preparation of Scutellaria baicalensis single-ingredient sample:

[0053] Concentration by decoction: Take 166.7 g of Scutellaria baicalensis, add water and decoct, filter and combine the decoction liquid, heat and concentrate to 3 L to obtain a concentrated liquid;

[0054] Purification: the concentrated solution was placed at room temperature, anhydrous ethanol was added to reach 70% alcohol content, stirred, placed, the supernatant was taken, the ethanol was recovered, concentrated to 500 mL, and the extract was obtained;

[0055] Take 3 mL of the extract to prepare the test solution according to the method of step 1.1.2 to prepare the single herb solution of Scutellaria baicalensis Georgi;

[0056] Step 5.1.2 Thin layer chromatography: take the baicalin reference substance, add methanol to prepare a solution containing 1 mg per 1 mL as the baicalin reference solution;

[0057] Take 3 μL of the baicalin reference solution, the test solution of step 1.1.2, the Scutellaria baicalensis Georgi reference drug solution of step 2.1.2, the Huangceng negative control solution of step 3.1.2 and the single herb solution of Scutellaria baicalensis Georgi of step 4.1.2 respectively, and point them on the same silica gel G thin layer plate. Use a mixture of ethyl acetate, acetone, formic acid and water in a volume ratio of 5:3:1:1 as the developing agent, pre-saturate for 30 min, develop, take out, dry, place in a 105°C oven until the spots are clear, and place under a 365 nm ultraviolet light to observe;

[0058] In the test chromatogram, if the same color fluorescent spots appear at the corresponding positions of the reference drug, the reference substance and the single herb, while the same color fluorescent spots do not appear at the same positions of the negative control, then it contains Scutellaria baicalensis Georgi.

[0059] The further beneficial effect of using the above method is that it can quickly and accurately identify whether the traditional Chinese medicine granules for treating sinusitis contain Scutellaria baicalensis Georgi.

[0060] Further, the specific method of thin layer chromatography identification of said Ligusticum chuanxiong Hort. is:

[0061] Step 1.1.3 Preparation of test solution: take 3 g of the traditional Chinese medicine granules for treating sinusitis, dissolve in 5 mL of water, take 1 mL, add 10 mL of ethyl acetate, shake, stand, take the ethyl acetate layer solution, remove the ethyl acetate, dissolve the product in 1 mL of ethyl acetate as the test solution;

[0062] Step 2.1.3 Preparation of reference drug solution: take 0.5 g of Ligusticum chuanxiong Hort. reference drug, prepare Ligusticum chuanxiong Hort. reference drug solution according to the method of step 1.1.3 for preparing test solution;

[0063] Step 3.1.3 Preparation of negative control sample: take Houttuynia cordata Thunb., Herba Patriniae, Gypsum Fibrosum, Scutellaria baicalensis Georgi, Magnolia biondii Pamp., Radix Sophorae Flavescentis, Radix Angelicae Dahuricae and Radix Glycyrrhizae eight kinds of medicinal materials, prepare the extract according to the preparation method of the traditional Chinese medicine granules for treating sinusitis, take 10 mL of the extract, and prepare Ligusticum chuanxiong Hort. negative control solution according to the preparation method of step 1.1.3 for preparing test solution;

[0064] Step 4.1.3 Preparation of Ligusticum chuanxiong Hort. single herb sample:

[0065] Decoction concentration: take 100g of Huangqi, add water and decoct, filter and combine the decoction liquid, heat and concentrate to 3L to obtain a concentrated liquid;

[0066] Purification: place the concentrated liquid at room temperature, add anhydrous ethanol to reach an alcohol content of 70%, stir well, stand, take the supernatant, recover the ethanol, concentrate to 500mL to obtain an extract;

[0067] Take 3mL of the extract and prepare a Chuanxiong single-ingredient solution according to the method of preparing the test solution in step 1.1.3.

[0068] Step 5.1.3 thin layer chromatography:

[0069] Take the ferulic acid reference substance, add methanol to prepare a solution containing 1mg per 1mL as a ferulic acid reference solution;

[0070] Take 6μL of the ferulic acid standard solution, 10μL of the test solution of step 1.1.3, 10μL of the Chuanxiong reference medicinal material solution of step 2.1.3, 10μL of the Chuanxiong negative control substance solution of step 3.1.3 and 10μL of the Chuanxiong single-ingredient solution of step 4.1.3, respectively, and spot them on the same silica gel G thin layer plate. Use a mixture of toluene: ethyl acetate: formic acid at a volume ratio of 5:3:0.5 as the developing agent, presaturate for 30min, develop, take out, air dry, spray with 10% sulfuric acid ethanol, place in a 105℃ oven until the spots are clear, and place under a 365nm ultraviolet light to observe;

[0071] In the test chromatogram, if the same color fluorescent spots appear at the same positions as the reference medicinal material, the reference substance and the single-ingredient solution, but not in the negative control, then it contains Chuanxiong.

[0072] The further beneficial effect of the above is that the above method can quickly and accurately identify whether the traditional Chinese medicine granules for treating sinusitis contain Chuanxiong.

[0073] Further, the specific method of thin layer chromatography identification of the Gleditsiae Spina is:

[0074] Step 1.1.4 preparation of test solution: take 3g of the traditional Chinese medicine granules for treating sinusitis, dissolve in 5mL of water, take 2mL, add 10mL of methanol, ultrasonic treat for 30min, filter, evaporate the filtrate to dryness, dissolve the residue in 20mL of water, extract with ethyl acetate, combine the ethyl acetate liquid, evaporate to dryness, dissolve the residue in 3mL of methanol as the test solution;

[0075] Step 2.1.4 Preparation of the control medicinal material solution: Take 2 g of the control Gleditsia sinensis L. medicinal material, add 50 mL of water, and boil for 45 min. Filter, evaporate the filtrate to dryness, add 20 mL of water to the residue to dissolve it, and extract it with ethyl acetate for 2 times, 10 mL each time. Evaporate the ethyl acetate liquid to dryness, add 2 mL of methanol to the residue to dissolve it, and use it as the Gleditsia sinensis L. control medicinal material solution;

[0076] Step 3.1.4 Preparation of the negative control sample: Take the eight medicinal materials of Houttuynia cordata Thunb., Patrinia scabiosaa Fisch., Gypsum fibrosum Curn, Scutellaria baicalensis Georgi, Magnolia biondii Pamp, Ligusticum chuanxiong Hort, Angelica dahurica Benth, and Glycyrrhiza uralensis Fisch., and prepare the extract according to the preparation method of the Chinese medicine granules for treating sinusitis; take 2 mL of each extract, and prepare the Gleditsia sinensis L. negative control solution according to the preparation method of the test solution in step 1.1.4.

[0077] Step 4.1.4 Preparation of the Gleditsia sinensis L. single-ingredient medicine sample:

[0078] Concentration by decoction: take 166.7 g of Gleditsia sinensis L., add water, and decoct it. Filter and combine the decoction liquid, and heat it to concentrate it to 3 L to obtain the concentrated liquid.

[0079] Purification: place the concentrated liquid at room temperature, add anhydrous ethanol to make the alcohol content reach 70%, stir it, stand it, take the supernatant, recover the ethanol, and concentrate it to 500 mL to obtain the extract. Take 3 mL of the extract, and prepare the Ligusticum chuanxiong Hort single-ingredient medicine solution according to the method for preparing the test solution in step 1.1.4.

[0080] Step 5.1.4 Thin layer chromatography: take the taxifolin control, add methanol to prepare a solution containing 1 mg per 1 mL, and use it as the taxifolin control solution.

[0081] Take 5 μL of the taxifolin control solution, the test solution in step 1.1.4, the Gleditsia sinensis L. control medicinal material solution in step 2.1.4, the Gleditsia sinensis L. negative control solution in step 3.1.4, and the Gleditsia sinensis L. single-ingredient medicine solution in step 4.1.4, respectively, and spot them on the same silica gel G thin layer plate. Use a mixture of petroleum ether, ethyl acetate, and formic acid with a volume ratio of 10:10:0.5 as the developing agent, presaturate it for 30 min, develop it, take it out, dry it, spray it with 10% sulfuric acid ethanol, heat it at 105°C until the spots are clear, and observe it under a 365 nm ultraviolet light. In the test sample chromatogram, if fluorescent spots appear at positions corresponding to the control, the control medicinal material, and the single-ingredient medicine chromatogram, but no fluorescent spots appear at the same positions of the negative control, then it contains Gleditsia sinensis L.

[0082] The further beneficial effects of the above method are that it can quickly and accurately identify whether the Chinese medicine granules for treating sinusitis contain Gleditsia sinensis L.

[0083] Further, the specific method for the thin layer chromatography identification of the Glycyrrhiza uralensis Fisch. is as follows:

[0084] Step 1.1.5 Preparation of test solution: Take 3g of the Chinese medicine granules for treating sinusitis, add 60mL of water to completely dissolve, stand, take supernatant, extract once with 40mL of ether, discard the ether liquid, add n-butanol to the lower layer, combine the n-butanol liquid, wash with water, discard the water liquid, evaporate the n-butanol liquid to dryness, add 5mL of methanol to completely dissolve the residue, and use as the test solution;

[0085] Step 2.1.5 Preparation of control medicinal material solution: Take 3g of licorice control medicinal material, add 60mL of water to completely dissolve, stand, take supernatant, extract once with 40mL of ether, discard the ether liquid, add n-butanol to the lower layer, combine the n-butanol liquid, wash with water, discard the water liquid, evaporate the n-butanol liquid to dryness, add 5mL of methanol to completely dissolve the residue, and use as the licorice control medicinal material solution;

[0086] Step 3.1.5 Preparation of negative control sample: Take eight kinds of medicinal materials of Houttuynia cordata, Patrinia, Gypsum, Scutellaria, Magnolia, Sophora, Chuanxiong and Angelica, and prepare the extract according to the preparation method of the Chinese medicine granules for treating sinusitis, take 3mL of the extract, and prepare the licorice negative control solution according to the preparation method of the test solution of step 1.1.5;

[0087] Step 4.1.5 Preparation of licorice single herb sample:

[0088] Concentration by decoction: Take 50g of licorice, add water and decoct, filter and combine the decoction liquid, heat and concentrate to 3L to obtain the concentrated liquid;

[0089] Purification: Place the concentrated liquid at room temperature, add anhydrous ethanol to reach an alcohol content of 70%, stir well, stand, take supernatant, recover ethanol, and concentrate to 500mL to obtain the extract;

[0090] Take 3mL of the extract, and prepare the licorice single herb solution according to the preparation method of the test solution of step 1.1.5;

[0091] Step 5.1.5 Thin layer chromatography: Take the glycyrrhizin control and ammonium glycyrrhizinate control, add methanol to prepare a solution containing 2mg per 1mL, and use as the glycyrrhizin control solution and ammonium glycyrrhizinate control solution;

[0092] Take 5 μL each of the glycyrrhizin reference solution, glycyrrhizic acid ammonium reference solution, the test solution from step 1.1.5, the licorice reference herb solution from step 2.1.5, the licorice negative control solution from step 3.1.5, and the licorice single herb solution from step 4.1.5, and spot them separately onto the same silica gel G thin-layer plate. Use a mixture of ethyl acetate:formic acid:glacial acetic acid:water (v / v) as the developing solvent, pre-saturate for 30 min, develop, remove, air dry, spray with 10% sulfuric acid ethanol, heat at 105℃ until the spots are clear, and examine under a 365 nm UV lamp. If the test sample chromatogram shows fluorescent spots of the same color at the corresponding positions as the glycyrrhizin reference solution, glycyrrhizic acid ammonium reference solution, licorice reference herb solution, and licorice single herb chromatogram, while the negative control does not show fluorescent spots of the same color at the same positions, then licorice is present.

[0093] The further beneficial effect of using the above method is that it can quickly and accurately identify whether the Chinese medicine granules for treating sinusitis contain licorice.

[0094] Furthermore, the specific method for thin-layer chromatography identification of Magnolia biondii is as follows:

[0095] Step 1.1.6 Preparation of test solution: Take 3g of the Chinese medicine granules for treating sinusitis, dissolve them in 5mL of water, take 3mL of the solution, add 20mL of chloroform, sonicate, cool, extract, evaporate to dryness and concentrate, add 5mL of methanol to dissolve the residue, and use it as the test solution.

[0096] Step 2.1.6 Preparation of reference medicinal material solution: Take 2g of Magnolia biondii reference medicinal material and prepare the Magnolia biondii reference medicinal material solution according to the method for preparing the test sample solution in step 1.1.6;

[0097] Step 3.1.6 Preparation of negative control samples: Take eight medicinal materials, namely Houttuynia cordata, Patrinia scabiosaefolia, gypsum, Scutellaria baicalensis, Gleditsia sinensis thorns, Ligusticum chuanxiong, Angelica dahurica and Glycyrrhiza uralensis, and prepare them into an extract according to the preparation method of the Chinese medicine granules for treating sinusitis described above.

[0098] Take 3 mL of the extract and prepare the Magnolia biondii negative control sample according to the method for preparing the test solution in step 1.1.6;

[0099] Step 4.1.6 Preparation of Magnolia biondii single herb sample:

[0100] Decoction and concentration: Take 166.7g of Magnolia biondii, soak it in water, distill it, and collect the dregs, the distilled aqueous solution and the volatile oil;

[0101] Add the dregs of the medicine to water and decoct them. Filter and combine the decoctions, and combine them with the distilled aqueous solution. Heat and concentrate to 3L to obtain a concentrated solution.

[0102] Purification: the concentrated solution was placed at room temperature, anhydrous ethanol was added to reach 70% alcohol content, stirred, stood for 24h, the supernatant was taken, ethanol was recovered, concentrated to 500mL, and a extract was obtained;

[0103] The extract was mixed with volatile oil, 3mL was taken, and a magnolia officinalis single herb solution was prepared according to the method of step 1.1.1;

[0104] Step 5.1.6 thin layer chromatography: magnol b took the control solution, added methanol to prepare a solution containing 1mg per 1mL, as a magnol b control solution;

[0105] 5μL of the magnolia officinalis control solution, the test solution of step 1.1.6, the magnolia officinalis control drug solution of step 2.1.6, the magnolia officinalis negative control solution of step 3.1.6 and the magnolia officinalis single herb solution of step 4.1.6 were taken respectively and spotted on the same silica gel H thin layer plate, a chloroform: diethyl ether mixture with a volume ratio of 12:1 was used as the developing agent, pre-saturation was performed for 30min, development was carried out, and then the plate was taken out, dried, sprayed with 10% sulfuric acid ethanol, heated at 105℃ until the spots were clear, and observed under white light; in the test chromatogram, if the same colored fluorescent spots appeared at the corresponding positions of the magnol b control and the magnolia officinalis control drug, the magnolia officinalis single herb, and no same colored fluorescent spots appeared at the same position of the negative control, then it contained magnolia officinalis.

[0106] The further beneficial effects of the above are: the above method can quickly and accurately identify whether the traditional Chinese medicine granules for treating sinusitis contain magnolia officinalis. BRIEF DESCRIPTION OF DRAWINGS

[0107] Figure 1 The first figure is a thin layer chromatogram of chuanqiong in the embodiment of the present application. Among them, 1 is ferulic acid control, 2 is negative control, 3 is chuanqiong control drug, and 4 is chuanqiong single herb.

[0108] Figure 2 The second figure is a thin layer chromatogram of chuanqiong in the embodiment of the present application. Among them, 1 is ferulic acid control, and 2-6 are traditional Chinese medicine granules for treating sinusitis.

[0109] Figure 3 The thin layer chromatogram of magnolia officinalis in the embodiment of the present application. Among them, 1 is magnol b control, 2 is magnolia officinalis control drug, 3 is magnolia officinalis single herb, 4 is negative control, and 5 is traditional Chinese medicine granules for treating sinusitis.

[0110] Figure 4 The thin layer chromatogram of magnolia officinalis in the embodiment of the present application. Among them, 1 is magnol b control, 2 is magnolia officinalis control drug, 3 is magnolia officinalis single herb, 4 is negative control, and 5 is traditional Chinese medicine granules for treating sinusitis.

[0111] Figure 5 As an embodiment of the present application, the thin layer chromatogram of Huangqi is shown in the figure. Among them, 1 is Huangqi single herb, 2 is Huangqi reference medicinal material, 3 is Huangqi glycoside reference substance, 4 is negative control, and 5-9 are all traditional Chinese medicine granules for treating rhinosinusitis.

[0112] Figure 6 As an embodiment of the present application, the thin layer chromatogram of Huangqi is shown in the figure. Among them, 1 is Huangqi single herb, 2 is Huangqi reference medicinal material, 3 is Huangqi glycoside reference substance, 4 is negative control, and 5-9 are all traditional Chinese medicine granules for treating rhinosinusitis.

[0113] Figure 7 As an embodiment of the present application, the thin layer chromatogram of Huangqi is shown in the figure. Among them, 1 is Huangqi single herb, 2 is Huangqi reference medicinal material, 3 is Huangqi glycoside reference substance, 4 is negative control, and 5-9 are all traditional Chinese medicine granules for treating rhinosinusitis. DETAILED DESCRIPTION

[0114] The principles and characteristics of the present application are described below in conjunction with embodiments, and the examples are used only to explain the present application and are not intended to limit the scope of the present application.

[0115] Silica gel high-performance G plate (batch number: 200119460614709), manufactured by Qingdao Haoyang Chemical Co., Ltd.; silica gel high-performance H plate (batch number: 220524061212702), manufactured by Qingdao Haoyang Chemical Co., Ltd.; reference medicinal material sources: Houttuynia cordata (batch number: DZYC-Y-007), Scutellaria baicalensis (batch number: DZYC-H-021), Chuanxiong rhizome (batch number: DZYC-C-004), Gleditsia sinensis (batch number: DZYC-Z-001), and Glycyrrhiza uralensis (batch number: DZYC-G-005) were all purchased from Chengdu Ruifensidende Dan Biological Technology Co., Ltd. Reference substances: quercetin (batch number: RDD-HO1111805008), baicalin (batch number: RDD-HO1602105028), ferulic acid (batch number: RDD-A00211812016), taxifolin (batch number: RDD-E00101905022), monammonium glycyrrhizinate (batch number: RDD-G00302309006), glycyrrhizin (batch number: RDD-G00911812016), and magnolin (batch number: RFS-M03111808031) were all purchased from Chengdu Ruifensidende Dan Biological Technology Co., Ltd.

[0116] Example 1: Preparation of traditional Chinese medicine granules for treating rhinosinusitis

[0117] The following weights of raw materials were weighed: Houttuynia cordata 250g, Herba Patriniae 250g, Gypsum fibrosum 250g, Scutellaria baicalensis 166.7g, Magnolia biondii 166.7g, Gleditsia sinensis 166.7g, Chuanxiong rhizome 100g, Radix Angelicae Dahuricae 100g, and Glycyrrhiza uralensis 50g.

[0118] Distillation extraction: Houttuynia cordata, Herba Patriniae and Magnolia biondii, immerse the medicine surface in water by 4cm-6cm, soak for 30min, then distill extract for 5h, collect the residue, the water solution after distillation and the volatile oil.

[0119] Decoction concentration: mix the residue with Scutellaria baicalensis, Gypsum fibrosum, Gleditsia sinensis, Chuanxiong Rhizoma, Angelica dahurica, Glycyrrhiza uralensis, add water and decoct for three times, the first time add 8 times water and decoct for 80min, the second time add 6 times water and decoct for 60min, the third time add 6 times water and decoct for 60min, filter and combine the three decoction liquid, then combine with the water solution after distillation in step 1.2, evaporate and concentrate to 3L, obtain the concentrated liquid.

[0120] Purification: put the concentrated liquid to room temperature, add anhydrous ethanol to make the alcohol content reach 70%, stir well, stand for 24h, take the supernatant, recover the ethanol, concentrate to 500mL, obtain the extract.

[0121] Granule preparation: add excipients to the extract, the excipients are any one or more than two of starch, dextrin and microcrystalline cellulose, use pure water as the binder, mix well and granulate and dry, spray the volatile oil, then the traditional Chinese medicine granules for treating sinusitis are obtained.

[0122] Example 2: TLC identification of Houttuynia cordata

[0123] The specific method is:

[0124] Step 1.1.1 Preparation of test solution: take 3g of the traditional Chinese medicine granules for treating sinusitis in example 1, dissolve in 5mL water, take 2mL, dissolve in 20mL water, extract with ethyl acetate for 2 times, 15mL each time, combine the ethyl acetate liquid, evaporate to dryness, dissolve the residue in 1mL methanol as the test solution.

[0125] Step 2.1.1 Preparation of control drug solution: take 5g of Houttuynia cordata control drug, prepare the Houttuynia cordata control drug solution according to the method of step 1.1.1.

[0126] Step 3.1.1 Preparation of negative control sample: take eight kinds of medicinal materials of Herba Patriniae, Gypsum fibrosum, Scutellaria baicalensis, Magnolia biondii, Chuanxiong Rhizoma, Angelica dahurica, Gleditsia sinensis and Glycyrrhiza uralensis, prepare the extract according to the preparation method of the traditional Chinese medicine granules for treating sinusitis, take 2mL of the extract, prepare the Houttuynia cordata negative control sample according to the method of step 1.1.1.

[0127] Step 4.1.1 Preparation of Houttuynia cordata single herb sample:

[0128] Distillation extraction: take 250g of Houttuynia cordata, cut into pieces, immerse the medicine surface in water by 4cm-6cm, soak for 30min, then distill for 5h, collect the residue, the water solution after distillation and the volatile oil.

[0129] The dregs were added to water and decocted three times, the first time with 8 times the amount of water for 80 min, the second time with 6 times the amount of water for 60 min, and the third time with 6 times the amount of water for 60 min. The three decoction liquids were filtered and combined, and the distilled aqueous solution was combined. The mixture was heated and concentrated to 3 L to obtain a concentrated liquid.

[0130] Purification: The concentrated liquid was brought to room temperature, anhydrous ethanol was added to reach an alcohol content of 70%, stirred, and left to stand for 24 h. The supernatant was taken, the ethanol was recovered, and the concentrate was concentrated to 500 mL to obtain an extract.

[0131] The extract was mixed with volatile oil, and 3 mL was taken to prepare a Houttuynia cordata Thunb. single herb solution according to the method of Step 1.1.1.

[0132] Step 5.1.1 Thin-layer chromatography: Take the Houttuynia cordata Thunb. reference substance, add methanol to prepare a solution containing 1 mg per 1 mL as the Houttuynia cordata Thunb. reference substance solution.

[0133] According to the test of Chinese Pharmacopoeia 2020 edition 0502, 5 μL of the Houttuynia cordata Thunb. reference substance solution, the test solution of Step 1.1.1, the Houttuynia cordata Thunb. control medicinal material solution of Step 2.1.1, the Houttuynia cordata Thunb. negative control substance solution of Step 3.1.1, and the Houttuynia cordata Thunb. single herb solution of Step 4.1.1 were taken respectively and spotted on the same silica gel G thin-layer plate. A mixture of ethyl acetate, toluene and formic acid with a volume ratio of 15:3:1 was used as the developing agent, pre-saturated for 30 min, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol, heated at 105°C until the spots were clear, and observed under a 365 nm ultraviolet light.

[0134] In the test sample chromatogram, if the same color fluorescent spots are shown at the corresponding positions of the control medicinal material, the reference substance, and the single herb medicinal material chromatogram, and no spots of the same fluorescent color are present at the same positions of the negative control, then Houttuynia cordata Thunb. is contained.

[0135] Example 3: Thin-layer chromatography identification of Scutellaria baicalensis Georgi

[0136] The specific method is as follows:

[0137] Step 1.1.2 Preparation of test solution: Take 3 g of the traditional Chinese medicine granules for treating rhinitis and dissolve it in 5 mL of water. Then, take 2 mL of the solution and add 20 mL of methanol. Ultrasonically treat for 30 min, filter, evaporate the filtrate to dryness, and dissolve the residue in 2 mL of methanol as the test solution.

[0138] Step 2.1.2 Preparation of control medicinal material solution: Take 1 g of Scutellaria baicalensis Georgi control medicinal material and prepare a Scutellaria baicalensis Georgi control medicinal material solution according to the method of Step 1.1.2.

[0139] Step 3.1.2 Preparation of negative control sample: Take Houttuynia cordata, Patrinia, Gypsum, Magnolia, Chuanxiong, Bai Zhi, Radix Sophorae and Radix Glycyrrhizae, and prepare the extract according to the preparation method of the Chinese medicine granules for treating rhinosinusitis. Take 2 mL of the extract and prepare the Scutellaria baicalensis negative control solution according to the method of step 1.1.2.

[0140] Step 4.1.2 Preparation of Scutellaria baicalensis single herb sample:

[0141] Concentration by decoction: Take 166.7 g of Scutellaria baicalensis and cut it into pieces. First, add 8 times the amount of water and decoct for 80 min. Second, add 6 times the amount of water and decoct for 60 min. Third, add 6 times the amount of water and decoct for 60 min. Filter and combine the three decoction liquids. Heat and concentrate to 3 L to obtain the concentrated liquid.

[0142] Purification: Place the concentrated liquid at room temperature. Add anhydrous ethanol to a content of 70%. Stir well. Stand for 24 h. Take the supernatant. Recover the ethanol. Concentrate to 500 mL to obtain the extract.

[0143] Take 3 mL of the extract and prepare the Scutellaria baicalensis single herb solution according to the method of step 1.1.2.

[0144] Step 5.1.2 Thin layer chromatography: Take the Scutellaria baicalensis glycoside reference substance and add methanol to prepare a solution containing 1 mg per 1 mL as the Scutellaria baicalensis glycoside reference solution.

[0145] According to the test of Chinese Pharmacopoeia 2020 edition 0502, take 3 μL of the Scutellaria baicalensis glycoside reference solution, the test solution of step 1.1.2, the Scutellaria baicalensis control herb solution of step 2.1.2, the Scutellaria baicalensis negative control solution of step 3.1.2 and the Scutellaria baicalensis single herb solution of step 4.1.2, respectively, and point them on the same silica gel G thin layer plate. Use a mixture of ethyl acetate, acetone, formic acid and water with a volume ratio of 5:3:1:1 as the developing agent, presaturate for 30 min, develop, take out, air dry, place at 105°C until the spots are clear, and place under a 365 nm ultraviolet light to observe.

[0146] In the test sample chromatogram, if the same colored fluorescent spots appear at the positions corresponding to the control herb, the reference substance and the single herb, while the same colored fluorescent spots do not appear at the same positions in the negative control, then Scutellaria baicalensis is contained.

[0147] Example 4: Thin layer chromatography identification of Chuanxiong

[0148] The specific method is:

[0149] Step 1.1.3 Preparation of test solution: Take 3g of the Chinese medicine granules for treating rhinosinusitis, dissolve in 5mL of water, take 1mL, add 10mL of ethyl acetate, shake, stand, take the ethyl acetate layer solution, remove the ethyl acetate, dissolve the obtained product in 1mL of ethyl acetate as the test solution;

[0150] Step 2.1.3 The method for preparing the control medicinal material solution is: take 0.5g of the control medicinal material of Chuanxiong Rhizoma, and prepare the control medicinal material solution of Chuanxiong Rhizoma according to the method for preparing the test solution in step 1.1.3;

[0151] Step 3.1.3 Preparation of negative control sample: Take Houttuynia cordata, Herba Patriniae, Gypsum Fibrosum, Scutellaria baicalensis, Magnolia biondii, Radix Sophorae, Radix Angelicae Dahuricae and Radix Glycyrrhizae eight kinds of medicinal materials, and prepare the extract according to the preparation method of the Chinese medicine granules for treating rhinosinusitis. Take 10mL of the extract, and prepare the Chuanxiong Rhizoma negative control solution according to the preparation method of the test solution in step 1.1.3;

[0152] Step 4.1.3 Preparation of Chuanxiong Rhizoma single herb sample:

[0153] Decoction concentration: Take 100g of Scutellaria baicalensis, cut it into pieces, add 8 times the amount of water to decoct for 80min for the first time, add 6 times the amount of water to decoct for 60min for the second time, and add 6 times the amount of water to decoct for 60min for the third time. Filter and combine the three decoction liquids, heat and concentrate to 3L to obtain the concentrated liquid.

[0154] Purification: Place the concentrated liquid at room temperature, add anhydrous ethanol to make the alcohol content reach 70%, stir evenly, stand for 24h, take the supernatant, recover the ethanol, and concentrate to 500mL to obtain the extract.

[0155] Take 3mL of the extract, and prepare the Chuanxiong Rhizoma single herb solution according to the preparation method of the test solution in step 1.1.3.

[0156] Step 5.3.1 Thin layer chromatography:

[0157] Take the ferulic acid reference substance, add methanol to prepare a solution containing 1mg per 1mL as the ferulic acid reference solution;

[0158] According to the test in Chinese Pharmacopoeia 2020 edition general test 0502, respectively take 6μL of the ferulic acid standard solution, 10μL of the test solution in step 1.1.3, 10μL of the Chuanxiong Rhizoma control medicinal material solution in step 2.1.3, 10μL of the Chuanxiong Rhizoma negative control solution in step 3.1.3 and 10μL of the Chuanxiong Rhizoma single herb solution in step 4.1.3, and point them on the same silica gel G thin layer plate. Use a mixture of toluene: ethyl acetate: formic acid with a volume ratio of 5:3:0.5 as the developing agent, presaturate for 30min, develop, take out, dry, place in a 105℃ oven until the spots are clear, and place under a 365nm ultraviolet light for inspection;

[0159] If the same color fluorescent spots appear in the chromatogram of the test sample at the corresponding positions of the chromatograms of the control medicinal material, the control substance and the single-ingredient medicinal material, and the same color fluorescent spots do not appear at the same positions of the negative control, then the test sample contains Chuanqiong.

[0160] Example 5: TLC identification of Gleditsia sinensis L.

[0161] The specific method is as follows:

[0162] Step 1.1.4 Preparation of the test sample solution: 3 g of the granules for treating rhinosinusitis was dissolved in 5 mL of water, and 2 mL of the solution was taken and added to 10 mL of methanol. The mixture was ultrasonically treated for 30 min, filtered, and the filtrate was evaporated to dryness. The residue was dissolved in 20 mL of water, and the solution was extracted with ethyl acetate for 2 times, 10 mL each time. The ethyl acetate extracts were combined, evaporated to dryness, and the residue was dissolved in 3 mL of methanol to obtain the test sample solution.

[0163] Step 2.1.4 Preparation of the control medicinal material solution: 2 g of the control medicinal material of Gleditsia sinensis L. was added to 50 mL of water, and boiled for 45 min. The solution was filtered, and the filtrate was evaporated to dryness. The residue was dissolved in 20 mL of water, and the solution was extracted with ethyl acetate for 2 times, 10 mL each time. The ethyl acetate extracts were combined, evaporated to dryness, and the residue was dissolved in 2 mL of methanol to obtain the control medicinal material solution of Gleditsia sinensis L.

[0164] Step 3.1.4 Preparation of the negative control sample: eight kinds of medicinal materials of Houttuynia cordata, Patrinia scabiosae fisch, Gypsum fibrosum, Scutellaria baicalensis, Magnolia biondii, Chuanqiong, Baizhi and Glycyrrhiza uralensis were prepared into extracts according to the preparation method of the granules for treating rhinosinusitis. 2 mL of each extract was taken and dissolved in 3 mL of methanol to obtain the negative control sample solution of Gleditsia sinensis L.

[0165] Step 4.1.4 Preparation of the single-ingredient medicinal sample of Gleditsia sinensis L.

[0166] Decoction and concentration: 166.7 g of Gleditsia sinensis L. was cut into pieces, and boiled with 8 times the amount of water for 80 min in the first time, 6 times the amount of water for 60 min in the second time, and 6 times the amount of water for 60 min in the third time. The three times of boiling solutions were combined, and concentrated to 3 L under heating to obtain the concentrated solution.

[0167] Purification: the concentrated solution was placed at room temperature, and anhydrous ethanol was added to reach an alcohol content of 70%. The mixture was stirred and stood for 24 h. The supernatant was taken, and the ethanol was recovered. The solution was concentrated to 500 mL to obtain the extract.

[0168] 3 mL of the extract was taken and dissolved in 3 mL of methanol to obtain the single-ingredient medicinal solution of Chuanqiong according to the method of Step 1.1.4.

[0169] Step 5.1.4 TLC: 1 mg of taxifolin was dissolved in 1 mL of methanol to obtain the taxifolin control substance solution.

[0170] According to the test of Chinese Pharmacopoeia 2020 edition 0502, respectively, aspirate Taxifolin reference solution, step 1.1.4 test solution, step 2.1.4 Gouqi Zhi control drug solution, step 3.1.4 Gouqi Zhi negative control solution and step 4.1.4 Gouqi Zhi single herb solution 5 μL, respectively, point to the same silica gel G thin layer plate, with volume ratio of 10:10:0.5 60-90 ℃ petroleum ether: ethyl acetate: formic acid mixture as developing agent, pre-saturation 30 min, development, take out, dry, spray with 10% sulfuric acid ethanol, placed in 105 ℃ heating to the spot obvious, placed under 365 nm ultraviolet light observation; if the test sample chromatogram, if the corresponding position of the control sample, control sample and single herb chromatogram shows fluorescent spots, while the negative control does not show fluorescent spots at the same position, then it contains Gouqi Zhi.

[0171] Example 6: TLC identification of licorice

[0172] The specific method is:

[0173] Step 1.1.5 preparation of test solution: take the treatment of rhinitis of traditional Chinese medicine granules 3g, add water 60ml completely dissolved, stand, take supernatant, add ether 40ml extraction 1 times, discard the ether liquid, the lower layer is added to n-butanol extraction 2 times, 30ml each time, combined n-butanol liquid, washed with water 3 times, discard the water, n-butanol liquid dry, the residue is added to methanol 5ml completely dissolved, as test solution.

[0174] Step 2.1.5 preparation of control drug solution: take licorice control drug 3g, add water 60ml completely dissolved, stand, take supernatant, add ether 40ml extraction 1 times, discard the ether liquid, the lower layer is added to n-butanol extraction 2 times, 30ml each time, combined n-butanol liquid, washed with water 3 times, discard the water, n-butanol liquid dry, the residue is added to methanol 5ml completely dissolved, as licorice control drug solution.

[0175] Step 3.1.5 preparation of negative control sample: take the fishy grass, Baijiangcao, Gypsum fibrosum, Huangqin, Xinyi, Gouqi Zhi, Chuanqiong and Baizhi eight kinds of medicinal materials, according to the preparation method of the treatment of rhinitis of traditional Chinese medicine granules to make extract, take extract 3ml, according to the method of step 1.1.5 preparation of test solution, preparation of licorice negative control solution.

[0176] Step 4.1.5 preparation of licorice single herb sample:

[0177] Decoction concentration: take 50g licorice cut, add 8 times the amount of water to decoction 80min, add 6 times the amount of water to decoction 60min, add 6 times the amount of water to decoction 60min, filter and combine three times of decoction liquid, heat concentration to 3L, obtain the concentrated solution.

[0178] Purification: the concentrated solution was placed at room temperature, anhydrous ethanol was added to reach 70% alcohol content, stirred, stood for 24h, the supernatant was taken, the ethanol was recovered, concentrated to 500mL, and the extract was obtained.

[0179] Take 3mL of the extract, and prepare the test sample solution according to the method of step 1.1.5 to prepare the Glycyrrhiza solution.

[0180] Step 5.1.5 Thin layer chromatography: take the glycyrrhizin reference substance and ammonium glycyrrhizinate reference substance, add methanol to prepare a solution containing 2mg per 1mL as the glycyrrhizin reference substance solution and ammonium glycyrrhizinate reference substance solution;

[0181] According to the test of Chinese Pharmacopoeia 2020 edition 0502, 5μL of glycyrrhizin reference substance solution, ammonium glycyrrhizinate reference substance solution, test sample solution of step 1.1.5, Glycyrrhiza officinalis reference drug solution of step 2.1.5, negative control sample solution of Glycyrrhiza officinalis of step 3.1.5 and Glycyrrhiza officinalis single herb solution of step 4.1.5 were taken respectively and spotted on the same silica gel G thin layer plate. The developing agent was a mixture of ethyl acetate, formic acid, glacial acetic acid and water with a volume ratio of 15:1:1:2, pre-saturated for 30min, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol, heated to 105℃ until the spots were clear, and observed under 365nm ultraviolet light; In the test sample chromatogram, if the same color fluorescent spots appear at the corresponding positions of glycyrrhizin reference substance, ammonium glycyrrhizinate reference substance, Glycyrrhiza officinalis reference drug and Glycyrrhiza officinalis single herb, and the same color fluorescent spots do not appear at the same position of the negative control, then it contains Glycyrrhiza officinalis.

[0182] Example 7: Thin layer chromatography identification of Magnolia

[0183] The specific method is:

[0184] The method for preparing the test sample solution of step 1.1.6 is: take 3g of the traditional Chinese medicine granules for treating rhinosinusitis, dissolve in 5mL of water, take 3mL, add 20mL of chloroform, ultrasonic treatment for 30min, cool down, extract and evaporate to dryness, add 5mL of methanol to dissolve as the test sample solution.

[0185] Step 2.1.6 Preparation of reference drug solution: take 2g of Magnolia officinalis reference drug, and prepare the Magnolia officinalis reference drug solution according to the method of step 1.1.6 for preparing the test sample solution.

[0186] Step 3.1.6 Preparation of negative control sample: take Houttuynia cordata, Herba Patriniae, Gypsum Fibrosum, Scrophularia ningpoensis, Radix Sophorae, Chuanxiong, Radix Angelicae Dahuricae and Radix Glycyrrhizae eight kinds of medicinal materials, and prepare the extract according to the preparation method of the traditional Chinese medicine granules for treating rhinosinusitis;

[0187] Take 3mL of the extract, and prepare the test sample solution according to the method of step 1.1.6 to prepare the Glycyrrhiza solution.

[0188] Step 4.1.6 Preparation of Magnolia officinalis single herb sample:

[0189] Decoction concentration: Take 166.7 g of Magnolia officinalis, cut into pieces, add water to immerse the medicinal surface by 4-6 cm, soak for 30 min, and then distill for 5 h. Collect the residue, the water solution after distillation, and the volatile oil.

[0190] Add the residue to water and decoct three times. The first time, add 8 times the amount of water and decoct for 80 min. The second time, add 6 times the amount of water and decoct for 60 min. The third time, add 6 times the amount of water and decoct for 60 min. Filter and combine the three decoction solutions, and combine with the water solution after distillation. Heat and concentrate to 3 L to obtain a concentrated solution.

[0191] Purification: Place the concentrated solution at room temperature, add anhydrous ethanol to reach an alcohol content of 70%, stir well, stand for 24 h, take the supernatant, recover the ethanol, and concentrate to 500 mL to obtain an extract.

[0192] Mix the extract with the volatile oil, take 3 mL, and prepare a Magnolia officinalis single herb solution according to the method of Step 1.1.6 for preparing a test solution.

[0193] Step 5.1.6 Thin layer chromatography: Take magnolin reference substance, add methanol to prepare a solution containing 1 mg per 1 mL as a magnolin reference solution.

[0194] According to the test of Chinese Pharmacopoeia 2020 edition general test 0502, take 5 μL of the Magnolia officinalis reference solution, the test solution of Step 1.1.6, the Magnolia officinalis reference medicinal material solution of Step 2.1.6, the Magnolia officinalis negative reference solution of Step 3.1.6, and the Magnolia officinalis single herb solution of Step 4.1.6, respectively, and point them on the same silica gel H thin layer plate. Use a mixture of chloroform and diethyl ether with a volume ratio of 12:1 as the developing agent, presaturate for 30 min, develop, take out, air dry, spray with 10% sulfuric acid ethanol, heat at 105°C until the spots are clear, and observe under white light; in the test solution chromatogram, if the same color fluorescent spots are shown at the corresponding positions of the magnolin reference solution and the Magnolia officinalis reference medicinal material and single herb chromatogram, and the same color fluorescent spots are not shown at the same positions of the negative control, then Magnolia officinalis is contained.

[0195] Figure 1 and Figure 2 are the thin layer chromatograms of Chuanqiong in the examples of the present application. As can be seen from the figures, the same color fluorescent spots are shown at the corresponding positions of the test solution chromatogram and the reference medicinal material, reference substance, and single herb material chromatogram, and the same color fluorescent spots are not shown at the same positions of the negative control, indicating that Chuanqiong is contained.

[0196] Figure 3The TLC chromatogram of the Magnolia biondii Pamp. in the embodiment of the present application is shown in the figure. As shown in the figure, the fluorescent spots of the same color appear on the corresponding positions of the test sample chromatogram, the magnolia bark extract, the Magnolia biondii Pamp. control sample and the single-ingredient medicine chromatogram, while the negative control sample does not show the fluorescent spots of the same color on the same positions, indicating that the Magnolia biondii Pamp. is contained.

[0197] Figure 4 The TLC chromatogram of the Houttuynia cordata Thunb. in the embodiment of the present application is shown in the figure. As shown in the figure, the fluorescent spots of the same color appear on the corresponding positions of the test sample chromatogram, the control sample, the single-ingredient medicine chromatogram, while the negative control sample does not show the fluorescent spots of the same color on the same positions, indicating that the Houttuynia cordata Thunb. is contained.

[0198] Figure 5 The TLC chromatogram of the Scutellaria baicalensis Georgi in the embodiment of the present application is shown in the figure. As shown in the figure, the fluorescent spots of the same color appear on the corresponding positions of the test sample chromatogram, the control sample, the single-ingredient medicine chromatogram, while the negative control sample does not show the fluorescent spots of the same color on the same positions, indicating that the Scutellaria baicalensis Georgi is contained.

[0199] Figure 6 The TLC chromatogram of the Gleditsia sinensis Lam. in the embodiment of the present application is shown in the figure. As shown in the figure, the fluorescent spots appear on the corresponding positions of the test sample chromatogram, the control sample, the single-ingredient medicine chromatogram, while the negative control sample does not show the fluorescent spots on the same positions, indicating that the Gleditsia sinensis Lam. is contained.

[0200] Figure 7 The TLC chromatogram of the Glycyrrhiza uralensis Fisch. in the embodiment of the present application is shown in the figure. As shown in the figure, the fluorescent spots of the same color appear on the corresponding positions of the test sample chromatogram, the glycyrrhizin control sample, the ammonium glycyrrhizate control sample, the Glycyrrhiza uralensis Fisch. control sample and the single-ingredient medicine chromatogram, while the negative control sample does not show the fluorescent spots of the same color on the same positions, indicating that the Glycyrrhiza uralensis Fisch. is contained.

[0201] The above description is only the preferred embodiments of the present application, and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A thin layer chromatography identification method of traditional Chinese medicine granules for treating sinusitis, which identifies six medicinal materials of Houttuynia cordata, Scutellaria baicalensis, Ligusticum chuanxiong, Gleditsia sinensis, Glycyrrhiza uralensis and Lonicera japonica in the traditional Chinese medicine granules for treating sinusitis, characterized in that, Comprising the following steps: Step 1: Preparation of test solution; Step 2: Preparation of control medicinal material solution; Step 3: Preparation of negative control sample; Step 4: Preparation of single herb sample; Step 5: Thin layer chromatography: the thin layer chromatography test of Houttuynia cordata, Scutellaria baicalensis, Chuanxiong Rhizoma, Gleditsia sinensis and Glycyrrhiza uralensis uses silica gel G thin layer plate, the thin layer chromatography test of Magnolia biondii uses silica gel H thin layer plate, Houttuynia cordata, Magnolia biondii, Gleditsia sinensis and Glycyrrhiza uralensis are sprayed with color developing agent, heated to develop color spots, and observed under 365 nm ultraviolet light; In the test sample chromatogram, if the same color fluorescent spots are shown at the corresponding positions of the control medicinal material chromatogram, single herb, and reference substance, while no same color fluorescent spots are shown at the corresponding positions of the negative control, it indicates that Magnolia biondii, Scutellaria baicalensis, Houttuynia cordata, Gleditsia sinensis, Glycyrrhiza uralensis and Chuanxiong Rhizoma are contained.

2. The thin layer chromatography identification method of the traditional Chinese medicine granules for treating sinusitis according to claim 1, characterized in that, The traditional Chinese medicine granules for treating sinusitis are prepared by the following method: The following weight fractions of raw materials are weighed: Houttuynia cordata 250 parts, Patrinia scabiosaefolia 250 parts, Gypsum fibrosum 250 parts, Scutellaria baicalensis 166.7 parts, Magnolia biondii 166.7 parts, Gleditsia sinensis 166.7 parts, Chuanxiong Rhizoma 100 parts, Angelica dahurica 100 parts, and Glycyrrhiza uralensis 50 parts; Distillation extraction: Houttuynia cordata, Patrinia scabiosaefolia and Magnolia biondii are extracted by water vapor distillation, and the residue, the water solution after distillation and the volatile oil are collected; Decoction concentration: the residue is mixed with Scutellaria baicalensis, Gypsum fibrosum, Gleditsia sinensis, Chuanxiong Rhizoma, Angelica dahurica and Glycyrrhiza uralensis to obtain decoction material, water is added to the decoction material and decoction is performed, decoction liquid is obtained, combined with the water solution after distillation, and evaporated and concentrated to obtain concentrated liquid; Purification: anhydrous ethanol is added to the concentrated liquid, and the ethanol is recovered after standing to obtain extract; Granule preparation: excipients are added to the extract, mixed uniformly, granulated and dried, and the volatile oil is sprayed, to obtain the traditional Chinese medicine granules for treating sinusitis.

3. The thin layer chromatography identification method of the traditional Chinese medicine granules for treating sinusitis according to claim 1, characterized in that, The specific method of thin layer chromatography identification of Houttuynia cordata is as follows: Step 1.1.1 Preparation of test solution: 3g of the traditional Chinese medicine granules for treating sinusitis is dissolved in 5mL of water, 2mL of which is taken, dissolved in 20mL of water, extracted with ethyl acetate, and the ethyl acetate liquid is combined, evaporated, and the residue is dissolved in 1mL of methanol as the test solution; Step 2.1.1 Preparation of control medicinal material solution: 5g of Houttuynia cordata control medicinal material is prepared into Houttuynia cordata control medicinal material solution according to the method of step 1.1.1; Step 3.1.1 Preparation of negative control sample: eight kinds of medicinal materials of Patrinia scabiosaefolia, Gypsum fibrosum, Scutellaria baicalensis, Magnolia biondii, Chuanxiong Rhizoma, Angelica dahurica, Gleditsia sinensis and Glycyrrhiza uralensis are prepared into extract according to the preparation method of the traditional Chinese medicine granules for treating sinusitis, 2mL of the extract is taken, and the Houttuynia cordata negative control sample is prepared according to the method of step 1.1.1; Step 4.1.1 Preparation of Houttuynia cordata single herb sample: Decoction concentration: 250g of Houttuynia cordata is soaked in water, distilled, and the residue, the water solution after distillation and the volatile oil are collected; The residue is added with water and decoction is performed, the decoction liquid is filtered and combined, and combined with the water solution after distillation, heated and concentrated to 3L to obtain concentrated liquid; Purification: the concentrated liquid is placed at room temperature, anhydrous ethanol is added to make the alcohol content reach 70%, stirred, and the supernatant is taken after standing, and the ethanol is recovered, concentrated to 500mL to obtain extract; Mix the extract and volatile oil, take 3 mL, and prepare the test sample solution of Houttuynia cordata according to the method of Step 1.1.1 to obtain the single-ingredient solution of Houttuynia cordata; Step 5.1.1 TLC: Take the bretschneideraneoside reference substance, add methanol to prepare a solution containing 1 mg per 1 mL as the bretschneideraneoside reference solution; Take 5 μL of the bretschneideraneoside reference solution, the test sample solution of Step 1.1.1, the Houttuynia cordata reference medicinal material solution of Step 2.1.1, the Houttuynia cordata negative reference substance solution of Step 3.1.1, and the Houttuynia cordata single-ingredient solution of Step 4.1.1 respectively, and spot them on the same silica gel G thin layer plate. Use a mixture of ethyl acetate, toluene, and formic acid (15:3:1 by volume) as the developing agent, and pre-saturate for 30 min. Develop, remove, and air dry. Spray with 10% sulfuric acid ethanol, heat at 105°C until the spots are clear, and observe under a 365 nm ultraviolet light. If the test sample chromatogram contains fluorescent spots of the same color as the reference medicinal material, the reference substance, and the single-ingredient solution at the corresponding positions, and the negative control does not contain the same fluorescent spots at the same positions, then the sample contains Houttuynia cordata.

4. The thin layer chromatography identification method of the traditional Chinese medicine granules for treating sinusitis according to claim 1, characterized in that, The specific method for identifying Scutellaria by TLC is as follows: Step 1.1.2 Preparation of test sample solution: Take 3 g of the Chinese medicine granules for treating rhinosinusitis, dissolve in 5 mL of water, take 2 mL, add 20 mL of methanol, and ultrasonically treat. Filter, evaporate the filtrate, dissolve the residue in 2 mL of methanol, and use as the test sample solution. Step 2.1.2 Preparation of reference medicinal material solution: Take 1 g of Scutellaria reference medicinal material, and prepare the Scutellaria reference medicinal material solution according to the method of Step 1.1.

2. Step 3.1.2 Preparation of negative reference sample: Take Houttuynia cordata, Herba Patullae, Gypsum Fibrosum, Magnolia, Chuanxiong, Baizhi, Radix Sophorae, and Radix Glycyrrhizae, and prepare the extract according to the preparation method of the Chinese medicine granules for treating rhinosinusitis. Take 2 mL of the extract, and prepare the Scutellaria negative reference substance solution according to the method of Step 1.1.

2. Step 4.1.2 Preparation of Scutellaria single-ingredient sample: Concentration by decoction: Take 166.7 g of Scutellaria, add water, and decoct. Filter and combine the decocted liquid, heat, and concentrate to 3 L to obtain the concentrated liquid. Purification: Let the concentrated liquid reach room temperature, add anhydrous ethanol to reach an alcohol content of 70%, stir, stand, take the supernatant, recover the ethanol, and concentrate to 500 mL to obtain the extract. Take 3 mL of the extract, and prepare the test sample solution according to the method of Step 1.1.2 to obtain the single-ingredient solution of Scutellaria. Step 5.1.2 TLC: Take the Scutellaria glycoside reference substance, add methanol to prepare a solution containing 1 mg per 1 mL as the Scutellaria glycoside reference solution. Take 3 μL of the Scutellaria glycoside reference solution, the test sample solution of Step 1.1.2, the Scutellaria reference medicinal material solution of Step 2.1.2, the Scutellaria negative reference substance solution of Step 3.1.2, and the Scutellaria single-ingredient solution of Step 4.1.2 respectively, and spot them on the same silica gel G thin layer plate. Use a mixture of ethyl acetate, acetone, formic acid, and water (5:3:1:1 by volume) as the developing agent, and pre-saturate for 30 min. Develop, remove, and air dry. Heat at 105°C until the spots are clear, and observe under a 365 nm ultraviolet light. If the same color fluorescent spots appear in the chromatogram of the test sample at the corresponding positions of the chromatograms of the control medicinal material, the control substance and the single-ingredient medicinal material, while no same color fluorescent spots appear at the same positions of the negative control, then the test sample contains Radix Paeoniae Alba.

5. The thin layer chromatography identification method of the traditional Chinese medicine granules for treating sinusitis according to claim 1, characterized in that, The specific method for identifying the Gastrodia elata by thin layer chromatography is as follows: Step 1.1.3 Preparation of test sample solution: 3 g of the Chinese medicine granules for treating rhinosinusitis was dissolved in 5 mL of water, 1 mL of which was taken and added with 10 mL of ethyl acetate, and then shaken, left to stand, and taken to obtain the ethyl acetate layer solution. The ethyl acetate was removed, and the obtained product was dissolved in 1 mL of ethyl acetate to serve as the test sample solution; Step 2.1.3 Preparation of control medicinal material solution: 0.5 g of Gastrodia elata control medicinal material was prepared into a Gastrodia elata control medicinal material solution according to the method for preparing the test sample solution in step 1.1.3; Step 3.1.3 Preparation of negative control sample: 10 mL of extract was prepared from eight kinds of medicinal materials, i.e., Houttuynia cordata, Patrinia scabiosaefolia, Gypsum fibrosum, Radix Paeoniae Alba, Magnolia biondii, Sophora Vaciensis, Angelica dahurica and Glycyrrhiza uralensis, according to the method for preparing the test sample solution in step 1.1.3, to serve as the Gastrodia elata negative control substance solution; Step 4.1.3 Preparation of Gastrodia elata single-ingredient sample: Concentration by decoction: 100 g of Gastrodia elata was added with water and subjected to decoction, and the decocted liquid was filtered and combined. The combined decocted liquid was concentrated to 3 L by heating to obtain a concentrated liquid; Purification: the concentrated liquid was left to room temperature, and anhydrous ethanol was added to make the alcohol content reach 70%. After stirring, the supernatant was taken, and the ethanol was recovered. The supernatant was concentrated to 500 mL to obtain an extract; 3 mL of the extract was prepared into a Gastrodia elata single-ingredient solution according to the method for preparing the test sample solution in step 1.1.3; Step 5.1.3 Thin layer chromatography: 1 mg of ferulic acid was dissolved in 1 mL of methanol to serve as a ferulic acid control substance solution; 6 μL of the ferulic acid standard solution was taken, and 10 μL of each of the test sample solution in step 1.1.3, the Gastrodia elata control medicinal material solution in step 2.1.3, the Gastrodia elata negative control substance solution in step 3.1.3 and the Gastrodia elata single-ingredient solution in step 4.1.3 was taken and spotted on the same silica gel G thin layer plate. A mixture of toluene, ethyl acetate and formic acid at a volume ratio of 5:3:0.5 was used as a developing agent, and the thin layer plate was pre-saturated for 30 min, developed, taken out, dried, heated at 105 ℃ until the spots were clear, and observed under a 365 nm ultraviolet light. If the same color fluorescent spots appear in the chromatogram of the test sample at the corresponding positions of the chromatograms of the control medicinal material, the control substance and the single-ingredient medicinal material, while no same color fluorescent spots appear at the same positions of the negative control, then the test sample contains Gastrodia elata.

6. The thin layer chromatography identification method of the traditional Chinese medicine granules for treating sinusitis according to claim 1, characterized in that, The specific method for identifying the Sophora Vaciensis by thin layer chromatography is as follows: Step 1.1.4 Preparation of test sample solution: 3 g of the Chinese medicine granules for treating rhinosinusitis was dissolved in 5 mL of water, 2 mL of which was taken and added with 10 mL of methanol, and then subjected to ultrasonic treatment for 30 min. The filtrate was evaporated to dryness, the residue was dissolved in 20 mL of water, and then extracted with ethyl acetate. The ethyl acetate liquid was combined, evaporated to dryness, and the residue was dissolved in 3 mL of methanol to serve as the test sample solution; Step 2.1.4 Preparation of the control medicinal material solution: 2 g of the control Gleditsia sinensis L. medicinal material was added with 50 mL of water and boiled for 45 min, and then filtered. The filtrate was evaporated to dryness, and the residue was added with 20 mL of water to dissolve, and then extracted with ethyl acetate for 2 times, 10 mL each time. The ethyl acetate liquid was combined and evaporated to dryness. The residue was added with 2 mL of methanol to dissolve, and served as the Gleditsia sinensis L. control medicinal material solution; Step 3.1.4 Preparation of the negative control sample: Gynura japonica, Patrinia, Gypsum fibrosum, Scutellaria, Magnolia, Chuanxiong, Baizhi and licorice were taken out, and then the eight kinds of medicinal materials were prepared into extracts according to the preparation method of the Chinese medicine granules for treating sinusitis. 2 mL of each extract was taken out, and then the Gleditsia sinensis L. negative control sample solution was prepared according to the preparation method of the test sample solution in step 1.1.4; Step 4.1.4 Preparation of the Gleditsia sinensis L. single medicinal material sample: Concentration by decoction: 166.7 g of Gleditsia sinensis L. was taken out, and then water was added and decocted. The decocted liquid was filtered and combined, and then concentrated to 3 L by heating to obtain a concentrated liquid; Purification: the concentrated liquid was placed at room temperature, and then anhydrous ethanol was added to reach an alcohol content of 70%. After stirring, the supernatant was taken out, and then the ethanol was recovered. The supernatant was concentrated to 500 mL to obtain an extract; 3 mL of the extract was taken out, and then the test sample solution was prepared according to the preparation method of the test sample solution in step 1.1.4 to obtain the Chuanxiong single medicinal material solution; Step 5.1.4 Thin layer chromatography: the taxifolin control sample was added with methanol to prepare a solution containing 1 mg per 1 mL, and then served as the taxifolin control sample solution; 5 μL of the taxifolin control sample solution, the test sample solution in step 1.1.4, the Gleditsia sinensis L. control medicinal material solution in step 2.1.4, the Gleditsia sinensis L. negative control sample solution in step 3.1.4 and the Gleditsia sinensis L. single medicinal material solution in step 4.1.4 was taken out respectively, and then spotted on the same silica gel G thin layer plate. Petroleum ether: ethyl acetate: formic acid mixed liquid (10:10:0.5) was used as the developing agent, and then pre-saturation was performed for 30 min. After development, the thin layer plate was taken out, dried, sprayed with 10% sulfuric acid ethanol, and then heated at 105°C until the spots were clear. The thin layer plate was observed under a 365 nm ultraviolet light. If fluorescent spots were observed in the test sample chromatogram at the corresponding positions of the control sample, the control medicinal material and the single medicinal material chromatogram, and no fluorescent spots were observed at the same position of the negative control, then the Gleditsia sinensis L. was contained.

7. The thin layer chromatography identification method of the traditional Chinese medicine granules for treating sinusitis according to claim 1, characterized in that, The specific method of the thin layer chromatography identification of the licorice is as follows: Step 1.1.5 Preparation of the test sample solution: 3 g of the Chinese medicine granules for treating sinusitis was added with 60 mL of water to completely dissolve, and then the supernatant was taken out. The supernatant was extracted with 40 mL of diethyl ether for 1 time, and then the diethyl ether liquid was discarded. The lower layer was extracted with n-butanol, and then the n-butanol liquid was combined and washed with water. The water liquid was discarded, and then the n-butanol liquid was evaporated to dryness. The residue was added with 5 mL of methanol to completely dissolve, and then served as the test sample solution; Step 2.1.5 Preparation of the control medicinal material solution: 3 g of the control licorice medicinal material was added with 60 mL of water to completely dissolve, and then the supernatant was taken out. The supernatant was extracted with 40 mL of diethyl ether for 1 time, and then the diethyl ether liquid was discarded. The lower layer was extracted with n-butanol, and then the n-butanol liquid was combined and washed with water. The water liquid was discarded, and then the n-butanol liquid was evaporated to dryness. The residue was added with 5 mL of methanol to completely dissolve, and then served as the licorice control medicinal material solution; Step 3.1.5 Preparation of negative control sample: Take the eight medicinal materials of Houttuynia cordata, Patrinia, Gypsum, Scutellaria, Magnolia, Sophora, Chuanxiong and Baishiyang, and prepare the extract according to the preparation method of the Chinese medicine granules for treating sinusitis; take 3 mL of the extract, and prepare the licorice negative control solution according to the preparation method of the test solution in step 1.1.5; Step 4.1.5 Preparation of licorice single herb sample: Decoction concentration: Take 50g of licorice, add water and decoct, filter and combine the decoction liquid, heat and concentrate to 3L to obtain the concentrated liquid; Purification: Let the concentrated liquid reach room temperature, add anhydrous ethanol to make the alcohol content reach 70%, stir evenly, stand, take the supernatant, recover the ethanol, and concentrate to 500mL to obtain the extract; Take 3mL of the extract, and prepare the licorice single herb solution according to the preparation method of the test solution in step 1.1.5; Step 5.1.5 Thin layer chromatography: Take the glycyrrhizin reference substance and ammonium glycyrrhizinate reference substance, add methanol to prepare a solution containing 2mg per 1mL as the glycyrrhizin reference substance solution and the ammonium glycyrrhizinate reference substance solution; Take 5μL of the glycyrrhizin reference substance solution, the ammonium glycyrrhizinate reference substance solution, the test solution in step 1.1.5, the licorice control medicinal material solution in step 2.1.5, the licorice negative control solution in step 3.1.5 and the licorice single herb solution in step 4.1.5 respectively, and point them on the same silica gel G thin layer plate; use a mixture of ethyl acetate, formic acid, glacial acetic acid and water in a volume ratio of 15:1:1:2 as the developing agent, presaturate for 30min, develop, take out, air dry, spray with 10% sulfuric acid ethanol, heat at 105℃ until the spots are clear, and observe under a 365nm ultraviolet light; if the same colored fluorescent spots appear in the corresponding positions of the glycyrrhizin reference substance, the ammonium glycyrrhizinate reference substance, the licorice control medicinal material and the licorice single herb in the test chromatogram, and the same colored fluorescent spots do not appear in the same positions of the negative control, then it contains licorice.

8. The method according to claim 1, wherein the method is characterized by the following steps: 1) applying the sample to the plate; 2) developing the plate with the developing solvent; 3) observing the plate under UV light; and 4) identifying the sample by comparing the developed plate with the standard plate. The specific method of thin layer chromatography identification of the Magnolia is: Step 1.1.6 Preparation of test solution: Take 3g of the Chinese medicine granules for treating sinusitis, dissolve in 5mL of water, take 3mL, add 20mL of chloroform, ultrasonic treatment, cool, extract and evaporate to dryness, add 5mL of methanol to the residue to dissolve, and take it as the test solution; Step 2.1.6 Preparation of control medicinal material solution: Take 2g of Magnolia control medicinal material, and prepare the Magnolia control medicinal material solution according to the preparation method of the test solution in step 1.1.6; Step 3.1.6 Preparation of negative control sample: Take the eight medicinal materials of Houttuynia cordata, Patrinia, Gypsum, Scutellaria, Sophora, Chuanxiong, Baishiyang and licorice, and prepare the extract according to the preparation method of the Chinese medicine granules for treating sinusitis; take 3mL of the extract, and prepare the Magnolia negative control sample according to the preparation method of the test solution in step 1.1.6; Step 4.1.6 Preparation of Magnolia single herb sample: Decoction concentration: Take 166.7g of Magnolia, soak in water, distill, collect the residue, the water solution after distillation and the volatile oil; Add water to the residue and decoct, filter and combine the decoction liquid, and combine with the water solution after distillation, heat and concentrate to 3L to obtain the concentrated liquid; Purification: the concentrated solution was placed at room temperature, anhydrous ethanol was added to reach 70% alcohol content, stirred, and left to stand for 24 h. The supernatant was recovered, the ethanol was recovered, and the solution was concentrated to 500 mL to obtain the extract; The extract was mixed with the volatile oil, and 3 mL was taken to prepare the Magnolia denudata single-herb solution according to the method of step 1.1.6; Step 5.1.6 thin layer chromatography: the magnolin reference substance was taken, methanol was added to prepare a solution containing 1 mg per 1 mL, which was used as the magnolin reference substance solution; 5 μL of the Magnolia denudata reference substance solution, the test solution of step 1.1.6, the Magnolia denudata reference material solution of step 2.1.6, the Magnolia denudata negative reference substance solution of step 3.1.6, and the Magnolia denudata single-herb solution of step 4.1.6 were taken respectively and spotted on the same silica gel H thin layer plate. A chloroform: diethyl ether mixture (volume ratio 12:1) was used as the developing agent, and the plate was pre-saturated for 30 min, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol, heated at 105°C until the spots were clear, and observed under white light; in the test sample chromatogram, if the same color fluorescent spots appeared at the corresponding positions of the magnolin reference substance and the Magnolia denudata reference material and single-herb chromatograms, but not in the negative control, then the sample contained Magnolia denudata.

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