Detection method of infantile cough syrup

Abstract

The invention belongs to the technical field of medicines, and particularly relates to an infantile cough relieving syrup detection method which comprises the following steps: identifying radix peucedani, platycodon grandiflorum, pericarpium citri reticulatae and cortex mori radicis in infantile cough relieving syrup by adopting a thin layer chromatography; and simultaneously detecting the fructus arctii and the radix scutellariae by adopting a high performance liquid chromatography. The detection method for the infantile cough relieving syrup has the advantages of being rapid in analysis, easy to operate, high in accuracy, wide in adaptability and the like, and establishment and overall evaluation of the quality standard of the infantile cough relieving syrup are facilitated.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to a detection method for pediatric cough syrup. Background Technology

[0002] Pediatric Cough Syrup contains 15 herbs: mulberry bark, mulberry leaf, bitter almond, burdock seed, trichosanthes fruit, angelica root, scutellaria root, platycodon root, medicated leaven (charred), malt (charred), hawthorn (charred), loquat leaf, tangerine peel, reed rhizome, and fritillaria bulb. Its main functions are to clear the lungs, relieve cough, and resolve phlegm. It is used for cough caused by wind-heat attacking the lungs, tracheitis, bronchitis, and the recovery period of pneumonia. This medicine is suitable for children from newborn to 12 years old.

[0003] Since its approval in December 1999, Xiaokewu Ning Syrup has adhered to this standard for drug quality control and release. However, due to the outdated nature of the quality standard, it only provides qualitative identification of burdock seed in the prescription; control methods for the other fourteen ingredients in the prescription have not been established. The existing standard can no longer accurately, scientifically, and comprehensively meet the requirements of drug quality control. Therefore, to better control the quality of Xiaokewu Ning Syrup, it is necessary to research new quality standards and testing methods. Summary of the Invention

[0004] The purpose of this invention is to provide a new method for quality testing of Pediatric Cough Syrup. By analyzing the components in the formulation, a new thin-layer chromatography method is added for the identification of multiple medicinal materials such as Peucedanum praeruptorum, Citrus reticulata peel, Platycodon grandiflorus and Morus alba root bark, thereby improving the accuracy and comprehensiveness of the quality control of Pediatric Cough Syrup.

[0005] This invention provides a thin-layer chromatography detection method for pediatric cough syrup, wherein the raw materials of the pediatric cough syrup include: mulberry bark, mulberry leaf, bitter almond, burdock seed, trichosanthes fruit, angelica root, scutellaria root, platycodon root, medicated leaven, malt, hawthorn, loquat leaf, tangerine peel, reed rhizome, and fritillaria bulb;

[0006] The detection method for the pediatric cough syrup includes at least one of the following: a thin-layer chromatography detection method for Peucedanum praeruptorum and a thin-layer chromatography detection method for Platycodon grandiflorus:

[0007] The thin-layer chromatography detection method for the aforementioned Peucedanum praeruptorum includes the following steps:

[0008] Pediatric cough syrup test solution A and Peucedanum praeruptorum reference solution were prepared separately and developed using a mixed solvent of petroleum ether, dichloromethane and ethyl acetate in a volume ratio of (3-5):(1-3):1 as the developing solvent for the first inspection.

[0009] The thin-layer chromatography detection method for Platycodon grandiflorus includes the following steps:

[0010] Pediatric cough syrup test solution B and Platycodon grandiflorus reference solution were prepared separately. They were developed using a mixed solvent of n-butanol, acetic acid and water in a volume ratio of (3-5):(1-3):(4-6). After color development, a second inspection was performed.

[0011] Furthermore, in the thin-layer chromatography detection method for Peucedanum praeruptorum, the volume ratio of petroleum ether, dichloromethane, and ethyl acetate is 4:2:1; in the thin-layer chromatography detection method for Platycodon grandiflorus, the volume ratio of n-butanol, acetic acid, and water is 4:2:5.

[0012] The novel thin-layer chromatography method for detecting Angelica dahurica provided by this invention has a higher resolution than the thin-layer chromatography method for Angelica dahurica in the 2020 edition of the Chinese Pharmacopoeia. In addition, the thin-layer chromatography method for detecting Platycodon grandiflorus has a simpler pretreatment method and uses more environmentally friendly solvents compared to the 2020 edition of the Chinese Pharmacopoeia, while still meeting the detection requirements.

[0013] In the thin-layer chromatography detection method of Peucedanum praeruptorum:

[0014] The preparation of the test solution A of the Pediatric Cough Syrup includes the following steps: mixing the Pediatric Cough Syrup with a non-polar or weakly polar aliphatic hydrocarbon solvent for extraction to obtain organic phase 1 and aqueous solution 1; concentrating and diluting the organic phase 1 to prepare the test solution A; further, the non-polar or weakly polar aliphatic hydrocarbon solvent is petroleum ether with a boiling point of 30-90℃.

[0015] The preparation of the Peucedanum reference solution includes the following steps: taking the Peucedanum praeruptorum A reference standard, dissolving it in a first organic solvent to obtain the Peucedanum praeruptorum reference solution; further, the first organic solvent is one or more of methanol, ethanol and ethyl acetate.

[0016] In the thin-layer chromatography detection method for Platycodon grandiflorus:

[0017] The preparation of the test solution B of the Pediatric Cough Syrup includes the following steps: the aqueous solution 1 is mixed with ethyl acetate for extraction to obtain organic phase 2 and aqueous solution 2; the aqueous solution 2 is mixed with water-saturated n-butanol for extraction to obtain organic phase 3; the organic phase 3 is washed with alkaline solution and / or water saturated with n-butanol; the solution is concentrated and diluted to prepare the test solution B.

[0018] The preparation of the Platycodon grandiflorus reference solution includes the following steps: taking Platycodon grandiflorus saponin D reference standard, dissolving it in a second organic solvent to obtain the Platycodon grandiflorus reference solution; further, the second organic solvent is one or more of methanol, ethanol and ethyl acetate.

[0019] In this invention, the detection method further includes a thin-layer chromatography detection method for dried tangerine peel, comprising the following steps:

[0020] The test solution C of Xiaokeshen Syrup and the reference solution of tangerine peel were prepared separately. The mixture of ethyl acetate, methanol and water in a volume ratio of (90-110):(15-20):(8-12) was used as the developing solvent. After color development, the solution was examined for the third time.

[0021] In the thin-layer chromatography detection method for dried tangerine peel:

[0022] The preparation of the test solution C of the Pediatric Cough Syrup includes the following steps: concentrating and diluting the organic phase 2 to prepare the test solution C;

[0023] The preparation of the tangerine peel reference solution includes the following steps: taking hesperidin reference standard, adding a third organic solvent to dissolve it to prepare a saturated solution, thereby obtaining the tangerine peel reference solution; further, the third organic solvent is one or more of methanol, ethanol and ethyl acetate.

[0024] The detection method also includes a thin-layer chromatography detection method for mulberry bark, comprising the following steps:

[0025] The test solution D of Xiaokeshen Syrup and the reference solution of mulberry bark were prepared separately. They were developed using a mixed solvent of butyl acetate, formic acid and water in a volume ratio of (6-8):(2-3):(2-3) as the developing solvent for the fourth inspection.

[0026] In the thin-layer chromatography detection method for mulberry bark:

[0027] The preparation of the test solution D of the Pediatric Cough Syrup includes the following steps: mixing the Pediatric Cough Syrup with ethyl acetate for extraction to obtain organic phase 4, washing the organic phase 4 with an alkaline solution, concentrating, diluting, and preparing the test solution D;

[0028] The preparation of the mulberry bark reference medicinal material solution includes the following steps: mulberry bark reference medicinal material is mixed with water for extraction to obtain an extract; the extract is mixed with ethyl acetate for extraction to obtain organic phase 5; organic phase 5 is washed with an alkaline solution, concentrated, diluted, and the reference medicinal material solution is prepared.

[0029] Furthermore, the color developing agent is at least one of vanillin sulfuric acid solution, vanillic acid sulfuric acid solution, and aluminum trichloride ethanol solution; the color developing temperature is 100℃-110℃.

[0030] In this invention, the detection method further includes: a high-performance liquid chromatography method for simultaneously detecting the contents of burdock seed and scutellaria baicalensis; comprising the following steps:

[0031] (1) Sample pretreatment: Take the pediatric cough syrup sample, add 50-70% methanol solution, extract by ultrasonication for 20-30 minutes, and filter to obtain the sample solution to be tested;

[0032] (2) Chromatographic analysis: The sample solution to be tested was analyzed and determined by high performance liquid chromatography;

[0033] Furthermore; the mobile phase was methanol (A) - 0.4% phosphoric acid aqueous solution (B), with a volume ratio of A:B = 39:61; the stationary phase was an octadecyl bonded silica column.

[0034] The beneficial effects of the technical solution of this invention:

[0035] 1) This invention identifies and tests multiple medicinal materials for Pediatric Cough Syrup, which more accurately, scientifically and comprehensively meets the requirements for drug quality control and can better control the quality of Pediatric Cough Syrup;

[0036] 2) The pre-processing procedures for identifying Peucedanum praeruptorum, Citrus reticulata peel, and Platycodon grandiflorus have been standardized and the operation simplified;

[0037] 3) The HPLC method can simultaneously determine the content of burdock seed and scutellaria baicalensis, which simplifies the operation and reduces costs;

[0038] 4) It avoids introducing the identification of multiple other medicinal materials that may affect the judgment, thus reducing misjudgment; it also eliminates the thin-layer chromatography identification of burdock seeds in the existing method, thus reducing the detection cost. Attached Figure Description

[0039] Figure 1 The image shows the thin-layer chromatogram of Peucedanum praeruptorum (A represents developing system 1; B represents developing system 2; C represents developing system 3).

[0040] Figure 2 These are thin-layer chromatograms of different batches of Angelica dahurica detected under developing system 1.

[0041] Figure 3 The chromatograms of tangerine peel in thin layer chromatography are shown below (left 1: UV light under development system 1; left 2: daylight under development system 1; right 1: daylight under development system 2; right 2: UV light under development system 2).

[0042] Figure 4 Thin-layer chromatograms of different batches of dried tangerine peel under developing system 1;

[0043] Figure 5 The image shows a thin-layer chromatogram of Platycodon grandiflorus (A represents developing system 1; B represents developing system 2; C represents developing system 3).

[0044] Figure 6 The image shows a thin-layer chromatogram of mulberry bark (development system 1 on the left; development system 2 on the right).

[0045] Figure 7 This is a thin-layer chromatogram of reed rhizome;

[0046] Figure 8This is a thin-layer chromatogram of loquat leaves;

[0047] Figure 9 This is a thin-layer chromatogram of bitter almonds;

[0048] Figure 10 This is a thin-layer chromatogram of Trichosanthes kirilowii. Detailed Implementation

[0049] The present invention will be further described in detail below with reference to specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of the disclosure of the present invention.

[0050] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0051] The term "preferred" in this invention refers to an embodiment of the invention that provides certain beneficial effects under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the description of one or more preferred embodiments does not imply that other embodiments are unavailable, nor is it intended to exclude other embodiments from the scope of the invention. That is, in this invention, "preferred" merely describes a more effective implementation or embodiment, but does not constitute a limitation on the scope of protection of the invention.

[0052] In this invention, terms such as "further" and "even further" are used for descriptive purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this invention.

[0053] In this invention, "at least one" means one or more, such as one, two or more. "Multiple" or "several" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0054] When a numerical range is disclosed in this invention, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Further, when the range refers to an integer, it includes every integer between the minimum and maximum values ​​of the range. Moreover, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0055] Unless otherwise specified, all steps of this invention may be performed sequentially or randomly. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or it may include steps (b) and (a) performed sequentially. For example, the method may also include step (c), indicating that step (c) may be added to the method in any order. For example, the method may include steps (a), (b), and (c), or it may include steps (a), (c), and (b), or it may include steps (c), (a), and (b), etc.

[0056] The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature.

[0057] In this invention, the terms "comprising," "including," "containing," "having," or other variations are intended to cover non-closed inclusion, and no distinction is made between these terms. The term "comprising" means that other steps and ingredients may be added without affecting the final result.

[0058] Pediatric Cough Syrup is made from 15 medicinal ingredients, including mulberry bark, mulberry leaf, bitter almond, burdock seed, trichosanthes fruit, angelica root, scutellaria root, platycodon root, styrax root, charred medicated leaven, malt, hawthorn, loquat leaf, tangerine peel, reed rhizome, and fritillaria bulb.

[0059] Thin-layer chromatography (TLC) involves coating a suitable stationary phase onto a glass plate, plastic, or aluminum substrate to form a uniform thin layer. After spotting and development, the ratio shift (Rf) is compared with the ratio shift (Rf) of a suitable reference chromatogram obtained using the same method. This method is used for drug identification, impurity detection, or content determination. As a crucial experimental technique for the rapid separation and qualitative analysis of small amounts of substances, TLC features short development time, strong separation capability, and high sensitivity, making it a vital analytical tool for the quality control of traditional Chinese medicine.

[0060] In this invention, unless otherwise specified, thin-layer chromatography plates are generally made of silicone, and silicone G plates are further used. To improve the accuracy and comprehensiveness of quality control for Pediatric Cough Syrup, this invention provides a thin-layer chromatography detection method for detecting four medicinal materials in Pediatric Cough Syrup: Peucedanum praeruptorum, Platycodon grandiflorus, Citrus reticulata peel, and Morus alba root bark.

[0061] The thin-layer chromatography detection method for the aforementioned Peucedanum praeruptorum includes the following steps:

[0062] Pediatric cough syrup test solution A and Peucedanum praeruptorum reference solution were prepared separately and developed using a mixed solvent of petroleum ether, dichloromethane and ethyl acetate in a volume ratio of (3-5):(1-3):1 as the developing solvent for the first inspection.

[0063] The thin-layer chromatography detection method for Platycodon grandiflorus includes the following steps:

[0064] Pediatric cough syrup test solution B and Platycodon grandiflorus reference solution were prepared separately. They were developed using a mixed solvent of n-butanol, acetic acid and water in a volume ratio of (3-5):(1-3):(4-6). After color development, a second inspection was performed.

[0065] It can be understood that the volume ratio of the mixed solvent of petroleum ether, dichloromethane and ethyl acetate can be, but is not limited to, 4:1:1, 4:2:1, 4:3:1, 3:2:1, 5:2:1, etc.; preferably, the volume ratio of the mixed solvent of petroleum ether, dichloromethane and ethyl acetate is 4:2:1.

[0066] It can be understood that the volume ratio of the mixed solvent of n-butanol, acetic acid and water can be, but is not limited to, 4:1:5, 4:2:5, 4:3:5, 3:2:5, 5:2:5, 4:2:4, 4:2:6, etc., and preferably the volume ratio of the mixed solvent of n-butanol, acetic acid and water is 4:2:5.

[0067] Understandably, the color development step of the color developer includes: adding the color developer to a thin-layer plate and heating it at a certain temperature until the spots are clear.

[0068] In some specific embodiments, the color developing agent is at least one of vanillin sulfuric acid solution, vanillic acid sulfuric acid solution, and aluminum trichloride ethanol solution; the color developing temperature is 100℃-110℃.

[0069] This can be understood as meaning that one or more colorimetric reagents can be used; when multiple colorimetric reagents are used, they can be mixed or used sequentially, and the order is not limited. For example, vanillin sulfuric acid solution can be used first, followed by aluminum chloride ethanol solution, or aluminum chloride ethanol solution can be used first, followed by vanillic acid sulfuric acid solution. Preferably, vanillin sulfuric acid solution is used as the colorimetric reagent in the thin-layer chromatography identification of Platycodon grandiflorus.

[0070] Understandably, the temperature of the color development treatment includes, but is not limited to, 100℃, 102℃, 104℃, 105℃, 108℃, 110℃, etc.; preferably, the temperature of the color development treatment is 105℃.

[0071] In some embodiments, the thin-layer chromatography plate is a silica gel G thin-layer plate.

[0072] In some embodiments, the inspection light source is an ultraviolet lamp and / or a fluorescent lamp; preferably, the first inspection light source is a 365nm ultraviolet lamp; the second inspection light source is a fluorescent lamp; the third inspection light source is a fluorescent lamp and a 365nm ultraviolet lamp; and the fourth inspection light source is a 365nm ultraviolet lamp.

[0073] Understandably, the fluorescent lamps include, but are not limited to, conventional incandescent lamps.

[0074] In the thin-layer chromatography detection method of Peucedanum praeruptorum:

[0075] The preparation of the test solution A of the Pediatric Cough Syrup includes the following steps: mixing the Pediatric Cough Syrup with a non-polar or weakly polar aliphatic hydrocarbon solvent for extraction to obtain organic phase 1 and aqueous solution 1; concentrating and diluting the organic phase 1 to prepare the test solution A.

[0076] The preparation of the Peucedanum reference solution includes the following steps: take the Peucedanum praeruptorum A reference standard, add a first organic solvent to dissolve it, and the Peucedanum praeruptorum reference solution is obtained.

[0077] It can be understood that the nonpolar or weakly polar aliphatic hydrocarbon solvent includes, but is not limited to, petroleum ether, n-pentane, n-hexane, cyclohexane, n-heptane, etc.; preferably, the nonpolar or weakly polar aliphatic hydrocarbon solvent is petroleum ether.

[0078] It can be understood that the petroleum ether can be a petroleum ether with a boiling point of 30℃-60℃ or a petroleum ether with a boiling point of 60℃-90℃; preferably, the petroleum ether is a petroleum ether with a boiling point of 30℃-60℃.

[0079] This can be understood as the number of extractions being 1, 2, 3, 4, or 5 times; preferably, the number of extractions is 2 times.

[0080] Understandably, the concentration specifically involves evaporating the extract of the Pediatric Cough Syrup to dryness, obtaining a residue.

[0081] Understandably, the dilution specifically involves dissolving the concentrated residue in an organic solvent; the organic solvent is one or more of methanol, ethanol, and ethyl acetate.

[0082] In some embodiments, the thin-layer chromatography detection method for Peucedanum praeruptorum further includes preparing a reference herb solution and making an auxiliary determination by comparing the chromatograms of the test sample and the reference herb solution.

[0083] In the thin-layer chromatography detection method for Platycodon grandiflorus:

[0084] The preparation of the test solution B of the Pediatric Cough Syrup includes the following steps: the aqueous solution 1 is mixed with ethyl acetate for extraction to obtain organic phase 2 and aqueous solution 2; the aqueous solution 2 is mixed with water-saturated n-butanol for extraction to obtain organic phase 3; the organic phase 3 is washed with alkaline solution and / or water saturated with n-butanol; the solution is concentrated and diluted to prepare the test solution B.

[0085] The preparation of the Platycodon grandiflorus reference solution includes the following steps: take Platycodon grandiflorus saponin D reference standard, add a second organic solvent to dissolve it, and thus obtain the Platycodon grandiflorus reference solution.

[0086] In some embodiments, the extraction is performed by mixing the aqueous solution 2 with water-saturated n-butanol, and the number of extractions and the number of washes with the alkaline solution and / or water saturated with n-butanol are each 1-5 times independently.

[0087] Understandably, the washing with alkaline solution and / or water saturated with n-butanol is a sequential washing process, that is, washing with alkaline solution first, and then washing with water saturated with n-butanol.

[0088] Understandably, the number of times the aqueous solution 2 is extracted with water-saturated n-butanol can be 1, 2, 3, 4, or 5 times; preferably, the number of extractions is 2.

[0089] Understandably, the number of times the alkaline solution is used for washing can be 1, 2, 3, 4, or 5 times; preferably, the number of times the washing is 1.

[0090] Understandably, the number of times the water saturated with n-butanol is washed can be 1, 2, 3, 4, or 5 times; preferably, the number of times the washing is 1.

[0091] Understandably, the alkaline solution includes, but is not limited to, sodium hydroxide solution, potassium hydroxide solution, and ammonia solution; preferably, the alkaline solution is an ammonia solution.

[0092] Understandably, the concentration specifically involves evaporating the organic phase layer to dryness, yielding a residue.

[0093] Understandably, the dilution specifically involves dissolving the concentrated residue in an organic solvent; the organic solvent is one or more of methanol, ethanol, and ethyl acetate.

[0094] In some specific embodiments, the thin-layer chromatography detection method for Platycodon grandiflorus also includes preparing a reference medicinal material solution of Platycodon grandiflorus and making auxiliary determination by comparing the chromatograms of the test sample and the reference medicinal material solution.

[0095] The detection method also includes a thin-layer chromatography detection method for dried tangerine peel, comprising the following steps:

[0096] The test solution C of Xiaokeshen Syrup and the reference solution of tangerine peel were prepared separately. The mixture of ethyl acetate, methanol and water in a volume ratio of (90-110):(15-20):(8-12) was used as the developing solvent. After color development, the solution was examined for the third time.

[0097] It can be understood that the volume ratio of the mixed solvent of ethyl acetate, methanol and water can be, but is not limited to, 100:17:8, 100:17:9, 100:17:10, 100:17:11, 100:17:12, 90:17:10, 110:17:10, 100:15:10, 100:16:10, 100:18:10, 100:19:10, 100:20:10, etc., and preferably the volume ratio of the mixed solvent of ethyl acetate, methanol and water is 100:17:10.

[0098] In some embodiments, the preparation of the pediatric cough syrup test solution C includes the following steps: concentrating and diluting the organic phase 2 to prepare the test solution C;

[0099] The preparation of the tangerine peel reference solution includes the following steps: take hesperidin reference standard, add a third organic solvent to dissolve it to prepare a saturated solution, and thus obtain the tangerine peel reference solution.

[0100] Understandably, the concentration specifically involves evaporating the organic phase layer to dryness, yielding a residue.

[0101] Understandably, the dilution specifically involves dissolving the concentrated residue in an organic solvent; the organic solvent is one or more of methanol, ethanol, and ethyl acetate.

[0102] In some embodiments, the thin-layer chromatography detection method for tangerine peel further includes preparing a tangerine peel reference solution and making an auxiliary determination by comparing the chromatograms of the test sample and the reference solution.

[0103] In some implementation methods, for ease of operation and cost savings, the preparation of the test solution in the chromatography detection method for Peucedanum praeruptorum, Citrus reticulata peel, and Platycodon grandiflorus can be carried out in the following order:

[0104] (1) The pediatric cough syrup was mixed with a non-polar or weakly polar aliphatic hydrocarbon solvent for extraction to obtain organic phase 1 and aqueous solution 1. The organic phase 1 was concentrated and diluted to prepare the test solution A; it was used for the thin-layer chromatography detection of Peucedanum praeruptorum.

[0105] (2) The aqueous solution 1 is mixed with ethyl acetate for extraction to obtain organic phase 2 and aqueous solution 2. The organic phase 2 is concentrated and diluted to prepare the test solution C; used for thin-layer chromatography detection of dried tangerine peel.

[0106] (3) The aqueous solution 2 and water-saturated n-butanol are mixed for extraction to obtain organic phase 3. The organic phase 3 is washed with alkaline solution and / or water saturated with n-butanol, concentrated, diluted and prepared to prepare the test solution B; used for thin-layer chromatography detection of Platycodon grandiflorus.

[0107] The detection method also includes a thin-layer chromatography detection method for mulberry bark, comprising the following steps:

[0108] The test solution D of Xiaokeshen Syrup and the reference solution of mulberry bark were prepared separately. They were developed using a mixed solvent of butyl acetate, formic acid and water in a volume ratio of (6-8):(2-3):(2-3) as the developing solvent for the fourth inspection.

[0109] In some embodiments, the preparation of the pediatric cough syrup test solution D includes the following steps: mixing the pediatric cough syrup with ethyl acetate for extraction to obtain organic phase 4, washing the organic phase 4 with an alkaline solution, concentrating, diluting, and preparing the test solution D;

[0110] The preparation of the mulberry bark reference medicinal material solution includes the following steps: mulberry bark reference medicinal material is mixed with water for extraction to obtain an extract; the extract is mixed with ethyl acetate for extraction to obtain organic phase 5; organic phase 5 is washed with an alkaline solution, concentrated, diluted, and the reference medicinal material solution is prepared.

[0111] In some specific embodiments, the subsequent processing method of the extract in the preparation of the mulberry bark reference medicinal material solution is the same as that in the preparation method of the test solution D.

[0112] This can be understood as meaning that the pH should be controlled to be acidic during the preparation of the test solution D and the mulberry bark reference medicinal material solution; preferably, the pH is 1-2.

[0113] This can be understood as the number of times the ethyl acetate is extracted, which can be 1, 2, 3, 4, or 5 times; preferably, the number of extractions is 2.

[0114] Understandably, the alkaline solution includes, but is not limited to, sodium hydroxide solution, potassium hydroxide solution, and ammonia solution; preferably, the alkaline solution is an ammonia solution.

[0115] Understandably, the concentration specifically involves evaporating the organic phase layer to dryness, yielding a residue.

[0116] Understandably, the dilution specifically involves dissolving the concentrated residue in an organic solvent; the organic solvent is one or more of methanol, ethanol, and ethyl acetate.

[0117] Understandably, the extraction method of mulberry bark reference material by mixing it with water includes, but is not limited to, decoction extraction, reflux extraction, etc.; preferably, the extraction is decoction extraction.

[0118] In some specific embodiments, the decoction extraction time is 10-40 minutes.

[0119] Understandably, the simmering time includes, but is not limited to, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, and 40 minutes; preferably, the simmering time is 20 minutes.

[0120] In some embodiments, the first solvent, the second solvent, and the third solvent are all selected from one or more of methanol, ethanol, and ethyl acetate; preferably, the first solvent, the second solvent, and the third solvent are methanol.

[0121] The above-mentioned thin-layer chromatography method can identify the four medicinal materials in the Pediatric Cough Syrup: Peucedanum praeruptorum, Platycodon grandiflorus, Citrus reticulata peel, and Morus alba root bark. The corresponding thin-layer chromatographic spots are clear, which improves the accuracy and comprehensiveness of the quality control of the Pediatric Cough Syrup and is more conducive to the establishment and overall evaluation of the quality standard of the Pediatric Cough Syrup.

[0122] The detection method further includes: a high-performance liquid chromatography method for simultaneously detecting the content of burdock seed and scutellaria baicalensis, comprising the following steps:

[0123] (1) Sample pretreatment: Take the pediatric cough syrup sample, add 50-70% methanol solution, extract by ultrasonication for 20-30 minutes, and filter to obtain the sample solution to be tested;

[0124] (2) Chromatographic analysis: The sample solution to be tested was analyzed and determined by high performance liquid chromatography.

[0125] Understandably, the concentration of the methanol solution can be 50%, 55%, 60%, 70%, etc., and preferably, the concentration of the methanol solution is 50%.

[0126] Understandably, the filtration includes, but is not limited to, conventional filtration methods, such as filter paper filtration and filter membrane filtration.

[0127] Understandably, octadecyl-bonded silica columns are generally chosen, such as: Waters X-Bridge Cl8 4.6x250 mm 5μm column, Agilent Zorbax SB C18 4.6x250 mm 5μm column, and SHIMADZU Shim-pack GIST 4.6x250 mm 5μm column.

[0128] In some specific embodiments, the mobile phase is methanol (A) - 0.4% aqueous phosphoric acid solution (B), with a volume ratio of A:B = 39:61.

[0129] The above-mentioned high-performance liquid chromatography method can simultaneously detect the content of burdock seed and scutellaria baicalensis, avoiding the introduction of other medicinal materials that may affect the identification and reducing misjudgment; it also eliminates the need for thin-layer chromatography identification of burdock seed in existing methods, thus reducing detection costs.

[0130] The following description, in conjunction with specific embodiments and comparative examples, further illustrates the point:

[0131] Example 1: Preparation of Pediatric Cough Syrup

[0132] Prescription: Mulberry bark 45g, Mulberry leaf 90g, Bitter almond 90g, Burdock seed 90g, Trichosanthes fruit 90g, Peucedanum root 90g, Scutellaria root 90g, Platycodon root 90g, Medicated leaven (charred) 30g, Malt (charred) 30g, Hawthorn fruit (charred) 30g, Loquat leaf 90g, Tangerine peel 30g, Reed rhizome 90g, Fritillaria thunbergii bulb 45g

[0133] Preparation: Take the above fifteen ingredients, extract almond water from almonds, and decoct the remaining fourteen ingredients, including mulberry bark, twice with water, the first time for 2 hours and the second time for 1 hour. Filter, combine the filtrates, let stand for 48 hours, take the supernatant and concentrate to an appropriate amount, add 400g of sucrose and an appropriate amount of preservative, boil to dissolve, add almond water and mix well, let stand, take the supernatant, add flavoring and other ingredients while stirring, add water to 1000mL, stir well, and the product is ready.

[0134] Example 2: Identification of Peucedanum praeruptorum by Thin-Layer Chromatography

[0135] Take 25 mL of Pediatric Cough Syrup and extract twice with petroleum ether (30-60℃), 25 mL each time. Combine the petroleum ether extracts and keep the aqueous solution (1). Dry the extract and dissolve the residue in 1 mL of methanol to prepare the test solution. Take 1 g of Peucedanum praeruptorum reference material and soak it in 20 mL of petroleum ether (30-60℃) for 1 hour. Filter the solution, dry the filtrate, and dissolve the residue in 1 mL of methanol to prepare the reference material solution. Take Peucedanum praeruptorum glycoside reference standard and prepare a solution containing 1 mg per mL with methanol to prepare the reference standard solution. Apply 5 μL of each of the above three solutions to the same silica gel G thin-layer plate. Develop the plate using petroleum ether (30-60℃)-dichloromethane-ethyl acetate (4:2:1) as the developing solvent (developing system 1). Remove the plate, air dry, and examine under ultraviolet light (365 nm). Replace the developing solvent with n-hexane-ethyl acetate (8:4) (developing system 2); replace the developing solvent with petroleum ether (60-90℃)-ethyl acetate (3:1) (developing system 3).

[0136] Test results as follows Figure 1As shown, the test sample and the reference sample show the same color spots at the same positions. The negative sample shows no interference. The spots of different batches of test samples are consistent. The test sample spectrum and the reference material of Peucedanum praeruptorum can only correspond to one spot. Figure 1 A (Development System 1) has regular spots and better clarity than [the previous one]. Figure 1 B (Deployment System 2), 1C (Deployment System 3); Figure 1 C has poor separation.

[0137] The system was used to test 15 batches of samples. The results are as follows: Figure 2 As shown, spots of the same color appear at the corresponding positions of the chromatograms of the angelica reference material. Negative samples show no interference. Spots from different batches of test samples are consistent. The chromatograms of the test samples correspond to only one spot of the angelica reference material, indicating that the use of the angelica reference material may lead to misjudgment.

[0138] Example 3: Thin-layer chromatographic identification of dried tangerine peel

[0139] Take the aqueous solution from Example 2 (Aqueous Solution 1), extract twice with ethyl acetate, 25 mL each time, combine the ethyl acetate extracts (Aqueous Solution 2), evaporate to dryness, dissolve the residue in 1 mL of methanol, and use as the test solution. Separately, take 1 g of dried tangerine peel reference material, add 20 mL of methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1 mL of methanol, and use as the reference material solution. Also, take hesperidin reference standard, add methanol to prepare a saturated solution, and use as the reference solution. Apply 10 μL of each of the above three solutions to the same silica gel G thin-layer plate, develop using ethyl acetate-methanol-water (100:17:10) as the developing solvent (developing system 1), remove, air dry, spray with 1% aluminum trichloride ethanol solution, heat at 105°C for 5 minutes, and examine under 365 nm ultraviolet light; then spray with 5% vanillic acid sulfuric acid solution, heat until the spots are clearly visible, and examine under sunlight. Alternatively, the developing solvent was replaced with the upper layer solution of toluene-ethyl acetate-formic acid-water (20:10:1:1) (developing system 2).

[0140] Test results as follows Figure 3 As shown, the test sample and the reference sample show the same spots at the same positions. The negative sample has no interference. The spots of different batches of test samples are consistent. The test sample and the reference medicinal material spectrum can only correspond to one spot. The reference spot in the left spectrum is centered and the Rf value is higher than that in the right spectrum. Moreover, the color development effect of the left spectrum (development system 1) under ultraviolet light and sunlight is better than that of the right spectrum (development system 2).

[0141] The system was used to test 15 batches of samples. The results are as follows: Figure 4As shown, spots of the same color appear at the corresponding positions of the hesperidin reference chromatogram. Negative samples show no interference, and spots from different batches of test samples are consistent. The test sample and the reference medicinal material chromatogram can only correspond to one spot, indicating that the use of tangerine peel as a reference medicinal material may lead to misjudgment.

[0142] Example 4: Thin-layer chromatographic identification of Platycodon grandiflorus

[0143] Take the aqueous solution (aqueous solution 2) from Example 3, and extract twice with 25 mL of water-saturated n-butanol each time. Combine the n-butanol extracts, extract once with 25 mL of ammonia solution, discard the ammonia solution, and then extract once with 25 mL of water saturated with n-butanol, discard the aqueous solution, evaporate the n-butanol solution to dryness, and dissolve the residue in 1 mL of methanol to prepare the test solution. Separately, take 1 g of Platycodon grandiflorus reference material, add 50 mL of water and decoct for 1 hour, filter, add 20 mL of water-saturated n-butanol to the filtrate, and prepare the reference material solution using the same method. Also, take Platycodon grandiflorus saponin D reference standard, add methanol to prepare a solution containing 1 mg per 1 mL, and use this as the reference solution. Take 5-10 μL of each of the above three solutions and spot them separately on the same silica gel G thin-layer plate. Use n-butanol-acetic acid-water (4:2:5) as the developing solvent (developing system 1), develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, heat at 105℃ for 5 minutes, and examine under sunlight. Change the developing solvent to n-butanol-methanol-acetic acid (5:1:1) (developing system 2); change the developing solvent to chloroform-ethyl ether (2:1) (developing system 3).

[0144] Test results as follows Figure 5 As shown, the test sample, the reference sample, and the reference medicinal material show the same spots in the corresponding positions. The negative sample shows no interference, and the spots of different batches of test samples are consistent. In spectrum A (development system 1), the spots are clear and round with good separation, and the overall identification effect is significantly better than that of spectrum B (development system 2) and spectrum C (development system 3). Spectrum C has poor separation and there is spot overlap.

[0145] Example 5: Thin-layer chromatographic identification of mulberry bark

[0146] Take 25 mL of Pediatric Cough Syrup, adjust the pH to 1-2 with dilute hydrochloric acid, extract twice with ethyl acetate (25 mL each time), combine the ethyl acetate extracts, wash twice with 10% sodium carbonate solution (15 mL each time), separate the ethyl acetate extract, evaporate to dryness, dissolve the residue in 1 mL of methanol, and use as the test solution. Separately, take 1 g of mulberry bark reference material, add 50 mL of water and decoct for 20 minutes, cool, centrifuge, take the supernatant, adjust the pH to 1-2 with dilute hydrochloric acid, and prepare the reference material solution using the same method. Apply 5-10 μL of each of the above two solutions to the same silica gel G thin-layer plate, using the upper layer of butyl acetate-formic acid-water (7:2.5:2.5) as the developing solvent (developing system 1), develop, and set the development distance to at least 12 cm. Remove, air dry, and examine under ultraviolet light (365 nm). The developing solvent (developing system 2) was replaced with dichloromethane-methanol (6:1).

[0147] Test results as follows Figure 6 As shown, in the left-hand spectrum (development system 1), the test sample and the control medicinal material show the same spots at corresponding positions, the negative sample has no interference, and the spots of different batches of test samples are consistent, with clear spots and good separation; in the right-hand spectrum (development system 2), the spots are blurry and the separation is poor.

[0148] Example 6: Simultaneous determination of the contents of burdock seed and scutellaria baicalensis by HPLC

[0149] 1) Instruments and reagents: Shimadzu LC-20AT HPLC, Shimadzu LC-40 HPLC, Agilent 1200 HPLC; Waters X-Bridge C18 4.6x250 mm 5μm column, Agilent Zorbax SB C18 4.6x250 mm 5μm column, SHIMADZU Shim-pack GIST C 18 4.6x250 mm 5μm column. Arctiin reference standard (National Institutes for Food and Drug Control 110819-201821, content calculated as 95.0%, no pretreatment required); Baicalin reference standard (National Institutes for Food and Drug Control 110715-201821, content calculated as 95.4%, no pretreatment required); Methanol was used as the mobile phase (chromatographic grade), and ultrapure water was used as the mobile phase; all other reagents were analytical grade.

[0150] 2) Flowability selection: Methanol-0.4% phosphoric acid (39:61) was used as the mobile phase.

[0151] 3) Selection of detection wavelength: Take the online DAD spectrum of the reference standard and the sample. Since the absorption curves of the two are consistent, the detection wavelength is set to 280nm.

[0152] 4) Investigation of extraction solvent:

[0153] Take 1 mL of Pediatric Cough Syrup (batch number 190202), shake well, and accurately measure 6 portions. Place each portion in a 25 mL volumetric flask and add 15 mL each of 30% methanol, 50% methanol, 70% methanol, 30% ethanol, 50% ethanol, and 70% ethanol. Sonicate the solutions (250 W, 40 kHz) for 20 minutes. Remove each flask, cool, and dilute to the mark with 30% methanol, 50% methanol, 70% methanol, 30% ethanol, 50% ethanol, and 70% ethanol. Shake well and filter through a 0.45 μm microporous membrane. Collect the filtrate. Inject 10 μL of each of the above test solutions into a liquid chromatograph, record the chromatograms, and calculate the peak areas (see Table 1). The results show that 50% methanol and 70% methanol have relatively high extraction efficiency, and the ratio of 50% methanol to the mobile phase is closer, making them the optimal choices.

[0154] Table 1. Investigation of extraction solvents

[0155]

[0156] 5) Examination of extraction methods:

[0157] Take pediatric cough syrup (batch number 190202), shake well, and accurately measure 1 mL of each sample into a 25 mL volumetric flask. Add 15 mL of 50% methanol. Sonicate the first sample (250 W, 40 kHz) for 20 minutes, remove, and cool. Shake the other sample for 5 minutes, then dilute to the mark with 50% methanol, shake well, and filter through a 0.45 μm microporous membrane. Collect the filtrate. Inject 10 μL of each of the above test solutions into a liquid chromatograph, record the chromatograms, and calculate the peak areas (results shown in Table 2). The results show that sonication is superior to shaking.

[0158] Table 2. Examination of Extraction Methods

[0159]

[0160] 6) Examination of extraction time:

[0161] Take 1 mL of Pediatric Cough Syrup (batch number 190202), shake well, and accurately measure three parallel aliquots. Place each aliquot in a 25 mL volumetric flask, add 15 mL of 50% methanol, and sonicate the three aliquots sequentially (power 250 W, frequency 40 kHz) for 10, 20, and 30 minutes. Remove, cool, shake well, and filter through a microporous membrane (0.45 μm). Collect the filtrate. Inject 10 μL of each of the above test solutions into a liquid chromatograph, record the chromatograms, and calculate the peak areas (results shown in Table 3). The results show that there is no significant difference between sonication for 30 minutes and 20 minutes.

[0162] Table 3. Examination of Extraction Time

[0163]

[0164] 7) Investigation of different chromatographic columns and chromatographs:

[0165] Shimadzu LC-20AT HPLC system, Shimadzu LC-40 HPLC system, Agilent 1200 HPLC system; Waters X-Bridge Cl8 4.6x250 mm 5μm column, Agilent Zorbax SB C18 4.6x250 mm 5μm column, SHIMADZU Shim-pack GIST 4.6x250 mm 5μm column.

[0166] The content determination method described above was applied to samples from the same batch (batch numbers 200601, 190302, and 190202). The results are shown in Tables 4 and 5. The results show that the method of the present invention is applicable to different chromatographic columns and chromatographs.

[0167] Table 4. Results of determination (arbuscular inosin) using different chromatographic columns and chromatograms.

[0168]

[0169] Table 5. Results of determination using different chromatographic columns and chromatograms (baicalin)

[0170]

[0171] 8) Sample determination:

[0172] According to the proposed content determination method, the contents of arctiin and baicalin in 15 batches of children's cough syrup were determined, and the results are shown in Table 6.

[0173] Table 6 Results of Sample Content Determination

[0174]

[0175] Comparative Example 1: Thin-layer Chromatographic Identification of Reed Rhizome

[0176] Take 20 mL of pediatric cough syrup, extract twice with 30 mL of chloroform each time, combine the chloroform extracts, evaporate to dryness, dissolve the residue in 1 mL of chloroform to obtain the test solution. Separately, take reed rhizome as a reference material, add 10 mL of chloroform, sonicate for 30 minutes, filter, and use the filtrate as the reference material solution. Take a negative sample lacking reed rhizome and prepare a negative sample according to the test solution. Apply 5-10 μL of each of the above three solutions to the same silica gel G thin-layer plate, develop using the upper layer of petroleum ether (60-90℃)-ethyl formate-formic acid (15:5:1) as the developing solvent, remove, air dry, and spray with phosphomolybdic acid test solution.

[0177] The results are as follows Figure 7 As shown, the test sample did not show fluorescent spots of the same color at the corresponding positions as the reference medicinal material, therefore, thin-layer chromatography of reed rhizome was not used for identification.

[0178] Comparative Example 2: Thin-layer chromatographic identification of loquat leaves

[0179] Take 20 mL of Pediatric Cough Syrup and extract twice with petroleum ether (30-60℃), 20 mL each time. Discard the petroleum ether solution. Extract the aqueous layer twice with ethyl acetate, 20 mL each time. Discard the ethyl acetate solution. Extract the aqueous layer twice with water-saturated n-butanol, 20 mL each time. Combine the n-butanol solutions and extract once with 20 mL of ammonia solution. Discard the ammonia solution. Extract once with 20 mL of water-saturated n-butanol. Discard the aqueous solution. Evaporate the n-butanol solution to dryness. Dissolve the residue in 1 mL of methanol to obtain the test solution. Separately, take 1 g of loquat leaf control material, add 20 mL of water, and decoct for 1 hour. Add 20 mL of water-saturated n-butanol to the aqueous solution and prepare the control material solution using the same method. Take a negative sample lacking loquat leaves and prepare a negative sample solution according to the test solution. Take 5-10 μL of each of the above three solutions and spot them separately on the same silica gel G thin-layer plate. Develop the plate using cyclohexane-ethyl acetate-glacial acetic acid (8:4:0.1) as the developing solvent. Remove the plate, air dry it, spray it with 5% vanillin sulfuric acid solution, heat it at 105°C for 5 minutes, and examine it under sunlight.

[0180] The results are as follows Figure 8 As shown, the test sample did not show spots of the same color at the corresponding positions as the reference medicinal material, therefore, thin-layer chromatography of loquat leaves was not used for identification.

[0181] Comparative Example 3: Thin-layer Chromatographic Identification of Bitter Almonds

[0182] Take 20 mL of Pediatric Cough Syrup and extract twice with petroleum ether (30-60℃), 20 mL each time. Discard the petroleum ether solution. Extract the aqueous layer twice with ethyl acetate, 20 mL each time. Discard the ethyl acetate solution. Extract the aqueous layer twice with water-saturated n-butanol, 20 mL each time. Combine the n-butanol solutions and extract twice with 20 mL of ammonia solution. Discard the ammonia solution. Wash twice with acetic acid solution (3→10), 20 mL each time. Combine the acetic acid solutions, evaporate to dryness, and dissolve the residue in 1 mL of methanol to obtain the test solution. Separately, take amygdalin reference standard and prepare a solution containing 1 mg per mL with methanol as the reference solution. Take a negative sample lacking bitter apricot kernels and prepare a negative sample according to the test solution. Take 10 μL of each of the above three solutions and spot them separately on the same silica gel G thin-layer plate. Use the lower layer solution of chloroform-ethyl acetate-methanol-water (15:40:22:10) that has been placed below 10℃ for 12 hours as the developing solvent. Develop, remove, air dry, and spray with phosphatidyl-aluminate-sulfuric acid solution (take 2 g of phosphatidyl-aluminate, add 20 mL of water to dissolve, then slowly add 30 mL of sulfuric acid and mix well). Heat at 105℃ until the spots are clearly visible and examine under sunlight.

[0183] The results are as follows Figure 9 As shown, the test sample did not show spots of the same color at the corresponding positions as the reference sample, therefore, bitter almond thin-layer chromatography was not used for identification.

[0184] Comparative Example 4: Thin-layer Chromatographic Identification of Trichosanthes kirilowii

[0185] Take 20 mL of pediatric cough syrup and extract twice with petroleum ether (30-60℃), 20 mL each time. Discard the petroleum ether solution. Extract the aqueous layer twice with ethyl acetate, 20 mL each time. Discard the ethyl acetate solution. Extract the aqueous layer twice with water-saturated n-butanol, 20 mL each time. Combine the n-butanol solutions and extract once with 20 mL of ammonia solution. Discard the ammonia solution. Extract once with 20 mL of water-saturated n-butanol. Discard the aqueous solution. Evaporate the n-butanol solution to dryness. Dissolve the residue in 1 mL of methanol to obtain the test solution. Separately, take 1 g of Trichosanthes kirilowii reference material, add 20 mL of water, and decoct for 1 hour. Add 20 mL of water-saturated n-butanol to the aqueous solution and prepare the reference material solution using the same method. Take a negative sample lacking Trichosanthes kirilowii and prepare the negative solution according to the test solution. Take 5-10 μL of each of the above three solutions and spot them separately on the same silica gel G thin-layer plate. Develop the plate using cyclohexane-ethyl acetate-glacial acetic acid (8:4:0.1) as the developing solvent. Remove the plate, air dry it, spray it with 5% vanillin sulfuric acid solution, heat it at 105°C for 5 minutes, and examine it under sunlight.

[0186] The results are as follows Figure 10As shown, the test sample did not show spots of the same color at the corresponding positions as the reference medicinal material, therefore, thin-layer chromatography of Trichosanthes kirilowii was not used for identification.

[0187] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for detecting pediatric cough syrup, characterized in that: The ingredients of the Pediatric Cough Syrup include: mulberry bark, mulberry leaf, bitter almond, burdock seed, trichosanthes fruit, angelica root, scutellaria root, platycodon root, medicated leaven, malt, hawthorn, loquat leaf, dried tangerine peel, reed rhizome, and fritillaria bulb; The detection method for the pediatric cough syrup includes at least one of the following: a thin-layer chromatography detection method for Peucedanum praeruptorum and a thin-layer chromatography detection method for Platycodon grandiflorus: The thin-layer chromatography detection method for the aforementioned Peucedanum praeruptorum includes the following steps: Pediatric cough syrup test solution A and Peucedanum praeruptorum reference solution were prepared separately and developed using a mixed solvent of petroleum ether, dichloromethane and ethyl acetate in a volume ratio of (3-5):(1-3):1 as the developing solvent for the first inspection. The thin-layer chromatography detection method for Platycodon grandiflorus includes the following steps: Pediatric cough syrup test solution B and Platycodon grandiflorus reference solution were prepared separately. They were developed using a mixed solvent of n-butanol, acetic acid and water in a volume ratio of (3-5):(1-3):(4-6). After color development, a second inspection was performed.

2. The method for detecting pediatric cough syrup according to claim 1, characterized in that: In the thin-layer chromatography detection method for Peucedanum praeruptorum, the volume ratio of petroleum ether, dichloromethane, and ethyl acetate is 4:2:1; in the thin-layer chromatography detection method for Platycodon grandiflorus, the volume ratio of n-butanol, acetic acid, and water is 4:2:

5.

3. The method for detecting pediatric cough syrup according to claim 1, characterized in that: In the thin-layer chromatography detection method of Peucedanum praeruptorum: The preparation of the test solution A of the Pediatric Cough Syrup includes the following steps: mixing the Pediatric Cough Syrup with a non-polar or weakly polar aliphatic hydrocarbon solvent for extraction to obtain organic phase 1 and aqueous solution 1; concentrating and diluting the organic phase 1 to prepare the test solution A; further, the non-polar or weakly polar aliphatic hydrocarbon solvent is petroleum ether with a boiling point of 30-90℃; The preparation of the Peucedanum reference solution includes the following steps: taking the Peucedanum praeruptorum A reference standard, dissolving it in a first organic solvent to obtain the Peucedanum praeruptorum reference solution; further, the first organic solvent is one or more of methanol, ethanol and ethyl acetate; In the thin-layer chromatography detection method for Platycodon grandiflorus: The preparation of the test solution B of the Pediatric Cough Syrup includes the following steps: Aqueous solution 1 is mixed with ethyl acetate for extraction to obtain organic phase 2 and aqueous solution 2; Aqueous solution 2 is mixed with water-saturated n-butanol for extraction to obtain organic phase 3; Organic phase 3 is washed with alkaline solution and / or water saturated with n-butanol; Concentrated; Diluted; and the test solution B is prepared. The preparation of the Platycodon grandiflorus reference solution includes the following steps: taking Platycodon grandiflorus saponin D reference standard, dissolving it in a second organic solvent to obtain the Platycodon grandiflorus reference solution; further, the second organic solvent is one or more of methanol, ethanol and ethyl acetate.

4. The method for detecting pediatric cough syrup according to claim 1, characterized in that: The detection method also includes a thin-layer chromatography detection method for dried tangerine peel, comprising the following steps: The test solution C of Xiaokeshen Syrup and the reference solution of tangerine peel were prepared separately. The mixture of ethyl acetate, methanol and water in a volume ratio of (90-110):(15-20):(8-12) was used as the developing solvent. After color development, the solution was examined for the third time.

5. The method for detecting pediatric cough syrup according to claim 4, characterized in that: In the thin-layer chromatography detection method for dried tangerine peel: The preparation of the test solution C of the Pediatric Cough Syrup includes the following steps: mixing the Pediatric Cough Syrup with a non-polar or weakly polar aliphatic hydrocarbon solvent for extraction to obtain organic phase 1 and aqueous solution 1; mixing the aqueous solution 1 with ethyl acetate for extraction to obtain organic phase 2; concentrating and diluting the organic phase 2 to prepare the test solution C. The preparation of the tangerine peel reference solution includes the following steps: taking hesperidin reference standard, adding a third organic solvent to dissolve it to prepare a saturated solution, thereby obtaining the tangerine peel reference solution; further, the third organic solvent is one or more of methanol, ethanol and ethyl acetate.

6. The method for detecting pediatric cough syrup according to claim 1, characterized in that: The detection method also includes a thin-layer chromatography detection method for mulberry bark, comprising the following steps: The test solution D of Xiaokeshen Syrup and the reference solution of mulberry bark were prepared separately. They were developed using a mixed solvent of butyl acetate, formic acid and water in a volume ratio of (6-8):(2-3):(2-3) as the developing solvent for the fourth inspection.

7. The method for detecting pediatric cough syrup according to claim 6, characterized in that: The preparation of the test solution D of the Pediatric Cough Syrup includes the following steps: mixing the Pediatric Cough Syrup with ethyl acetate for extraction to obtain organic phase 4, washing the organic phase 4 with an alkaline solution, concentrating, diluting, and preparing the test solution D; The preparation of the mulberry bark reference medicinal material solution includes the following steps: mulberry bark reference medicinal material is mixed with water for extraction to obtain an extract; the extract is mixed with ethyl acetate for extraction to obtain organic phase 5; organic phase 5 is washed with an alkaline solution, concentrated, diluted, and the reference medicinal material solution is prepared.

8. The method for detecting pediatric cough syrup according to claim 1 or 4, characterized in that: The colorimetric agent is at least one of vanillin sulfuric acid solution, vanillic acid sulfuric acid solution, and aluminum trichloride ethanol solution; the colorimetric treatment temperature is 100℃-110℃.

9. The method for detecting pediatric cough syrup according to claim 1, characterized in that: The detection method further includes: a high-performance liquid chromatography method for simultaneously detecting the content of burdock seed and scutellaria baicalensis, comprising the following steps: (1) Sample pretreatment: Take the pediatric cough syrup sample, add 50-70% methanol solution, extract by ultrasonication for 20-30 minutes, and filter to obtain the sample solution to be tested; (2) Chromatographic analysis: The sample solution to be tested was analyzed and determined by high performance liquid chromatography; Further; mobile phase: methanol (A) - 0.4% phosphoric acid aqueous solution (B), volume ratio A:B = 39:61; solid Phase fixation: octadecyl bonded silica column.

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