Identification method of Dutong pills

Abstract

The invention provides an identification method for stomach pain pills, wherein the components of the stomach pain pills include 40 to 60 parts of cinnamon, 90 to 110 parts of costus root, 90 to 110 parts of dried ginger, 40 to 60 parts of magnolia bark, 40 to 60 parts of cardamom, 40 to 60 parts of amomum, 10 to 30 parts of longan, 10 to 30 parts of poppy shell, 90 to 110 parts of immature bitter orange and 40 to 60 parts of lindera; the identification method includes: using thin layer chromatography to detect one or more of the cinnamon, costus root, dried ginger and magnolia bark. The identification method is simple, fast and accurate, improves the accuracy and comprehensiveness of the quality control of the stomach pain pills, and is more conducive to the establishment and overall evaluation of the quality standard of the stomach pain pills.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine detection, in particular to an identification method for abdominal pain pills. Background Art

[0002] Dutong Pills are composed of 10 medicinal materials, including dried ginger, cardamom, cinnamon bark, immature bitter orange, costus root, lindera strychnifolia, magnolia bark, amomum villosum, longan and poppy shell. They have the effects of warming the middle and dispelling cold, regulating qi and relieving pain. The prescription of Dutong Pills has a long history of use. Under the guidance of traditional Chinese medicine theory, Dutong Pills are mainly used to treat epigastric pain, fullness, vomiting and heartburn caused by cold in the middle jiao and food accumulation. In modern medicine, Dutong Pills are mainly used for indigestion caused by food stagnation, gastric and duodenal ulcers, gastric neurosis, irritable bowel syndrome and other diseases and symptoms, and have a wide range of indications.

[0003] Dutong Pills are currently included in the "Pharmaceutical Standards of the Ministry of Health of the People's Republic of China" (Volume 6 of Chinese Medicine Formula Preparations), and only have routine inspection items under the pills such as appearance, properties, moisture, and weight difference. Dutong Pills have many medicinal flavors and complex and diverse chemical components, but there are no identification items in its quality standards. Without a definite quality standard, it is impossible to effectively control the quality of Dutong Pills, and it is impossible to truly reflect the relationship between the efficacy and chemical components of Dutong Pills. The quality of Chinese medicine is a reflection and characterization of the effectiveness and safety of Chinese medicine. It is an important basis for the clinical use of Chinese medicine and the effectiveness control of Chinese patent medicines. Therefore, an effective quality control method should be formulated to control the quality of this Chinese patent medicine.

[0004] The 2020 edition of the Chinese Pharmacopoeia includes TLC identification methods for single herbs: Magnolia officinalis, costusroot, cinnamon bark, and dried ginger. In addition, Zhao Li et al. discovered a method for identifying the Chinese herbal medicine cinnamon heart, and provided a method for TLC identification of cinnamon heart (a type of cinnamon bark). However, the detection of single herbal ingredients cannot conduct comprehensive quality control and evaluation of the Dutong Pills product. Other traditional methods provide methods for identifying one or more of dried ginger oil, costusroot oil, Magnolia officinalis extract, and Coptis chinensis extract in Huanghou Antidiarrhea Drops. Although Huanghou Antidiarrhea Drops have some of the same medicinal flavors as Dutong Pills, there are still significant differences in the overall prescription composition and preparation process, which makes this TLC detection method unsuitable for Dutong Pills. Summary of the invention

[0005] Based on this, the present invention provides an identification method for Duchu Pills, which makes the qualitative identification of Duchu Pills simpler, faster and more accurate by performing thin layer chromatography detection on one or more of the four medicinal materials of cinnamon, costus root, magnolia bark and dried ginger in Duchu Pills, thereby improving the accuracy and comprehensiveness of Duchu Pills quality control, and is more conducive to the establishment of Duchu Pills quality standards and overall evaluation.

[0006] The present invention is achieved through the following technical solutions.

[0007] A method for identifying abdominal pain pills, wherein the abdominal pain pills include, by weight, 40 to 60 parts of cinnamon bark, 90 to 110 parts of costusroot, 90 to 110 parts of dried ginger, 40 to 60 parts of magnolia bark, 40 to 60 parts of cardamom, 40 to 60 parts of amomum, 10 to 30 parts of piper longum, 10 to 30 parts of poppy shell, 90 to 110 parts of immature bitter orange and 40 to 60 parts of lindera striata;

[0008] The identification method comprises: using thin layer chromatography to detect one or more of the cinnamon bark, costusroot, dried ginger and magnolia bark.

[0009] In one embodiment, detecting the cinnamon comprises the following steps:

[0010] Mixing the Dutong Pills with a first solvent to prepare a first test solution;

[0011] Mixing a cinnamaldehyde reference substance with the first solvent to prepare a first reference substance solution;

[0012] The first test solution and the first reference solution are respectively taken, spotted on a silica gel thin layer plate, placed in a first developing agent for development, taken out to dry, and sprayed with a first color developer;

[0013] Wherein, the first solvent is methanol; and the first developing agent is a mixture of petroleum ether and ethyl acetate.

[0014] In one embodiment, the volume ratio of petroleum ether to ethyl acetate in the first developing solvent is (17±0.5):(3±0.5).

[0015] In one embodiment, the first color developing agent is dinitrophenylhydrazine ethanol solution.

[0016] In one embodiment, the detection of the costusroot comprises the following steps:

[0017] Mixing the Dutong Pills with a second solvent to prepare a second test solution;

[0018] Mixing the reference substances of dehydrocostus lactone and costunolide with the second solvent to prepare a second reference substance solution;

[0019] The second test solution and the second reference solution are respectively taken, spotted on a silica gel thin layer plate, placed in a second developing agent for development, taken out and dried, then heated at 100°C to 110°C, and sprayed with a second color developer;

[0020] Wherein, the second developing agent is petroleum ether and ethyl acetate in a volume ratio of (17±0.5):(3±0.5).

[0021] In one embodiment, the second solvent is selected from one or more of ethyl acetate, methanol and ethanol; and / or

[0022] The second color developing agent is 10% sulfuric acid ethanol solution.

[0023] In one embodiment, the detection of the Magnolia officinalis comprises the following steps:

[0024] Mixing the Dutong Pills, activated carbon and a third solvent to prepare a third test solution;

[0025] Respectively mixing magnolol and honokiol reference substances with the third solvent to prepare a third reference substance solution;

[0026] The third test solution and the third reference solution are respectively taken, spotted on a silica gel thin layer plate, placed in a third developing agent for development, taken out and dried, and then examined at 254 nm;

[0027] Wherein, the third developing agent is a mixture of chloroform and methanol.

[0028] In one embodiment, in the third developing solvent, the volume ratio of chloroform to methanol is (17±0.5):(1±0.5); and / or

[0029] The third solvent is selected from one or more of methanol and ethanol.

[0030] In one embodiment, the detection of the dried ginger comprises the following steps:

[0031] Mixing the Dutong Pills with the fourth solvent to prepare a fourth test solution;

[0032] Mixing the dried ginger reference medicinal material with the fourth solvent to prepare a reference medicinal material solution;

[0033] The fourth test solution and the reference medicinal material solution are respectively taken, spotted on a silica gel thin layer plate, placed in a fourth developing agent for development, taken out and dried, then heated at 100° C. to 110° C., and sprayed with a third color developer;

[0034] Wherein, the fourth developing agent is a mixture of petroleum ether, chloroform and ethyl acetate; and the third color developing agent is a 10% sulfuric acid ethanol solution.

[0035] In one embodiment, in the fourth developing solvent, the volume ratio of petroleum ether, chloroform and ethyl acetate is (2±0.5):(1±0.5):(1±0.5); and / or

[0036] The fourth solvent is selected from one or more of methanol, ethyl acetate and ethanol.

[0037] Compared with the prior art, the identification method of the stomachache pills of the present invention has the following beneficial effects:

[0038] The present invention provides a quality control method for thin layer chromatography detection of one or more of the four medicinal materials of cinnamon, costusroot, magnolia bark and dried ginger in Dutong Pills. Furthermore, the present invention limits the selection of medicinal material extraction solvent and developing agent, and ultimately makes the qualitative identification of Dutong Pills more simple, rapid and reproducible, making up for the deficiency of Dutong Pills without current identification standards. The identification method established by the present invention has strong applicability, is more conducive to the establishment and overall evaluation of the quality standard of Dutong Pills, and not only ensures the product quality, but also improves the controllability of product production quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 The cinnamon thin layer identification chromatogram provided by the present invention; wherein 1 represents a cinnamon negative sample, 2 represents Z00001, 3 represents Z00002, 4 represents Z00003, and 5 represents a cinnamaldehyde reference substance;

[0040] Figure 2 The chromatogram of the thin layer identification of costus root provided by the present invention; wherein 1 represents a negative sample of costus root, 2 represents Z00001, 3 represents Z00002, 4 represents Z00003, and 5 represents a mixed reference substance;

[0041] Figure 3 The thin layer identification chromatogram of Magnolia officinalis provided by the present invention; in the left figure, 1 represents a negative sample of Magnolia officinalis, 2 represents Z00001, 3 represents Z00002, 4 represents Z00003, and 5 represents a mixed reference substance; in the right figure, 1 represents X00009M, 2 represents X00008M, 3 represents a negative sample of Magnolia officinalis, 4 represents a mixed reference substance, 5 represents XS0001, 6 represents XS0002, 7 represents XS0003, and 8 represents X00007M;

[0042] Figure 4 The present invention provides a thin layer identification chromatogram of dried ginger; in the left figure, 1 represents a negative sample of dried ginger, 2 represents Z00001, 3 represents Z00002, 4 represents Z00003, and 5 represents a reference medicinal material of dried ginger; in the right figure, 1 represents X00007M, 2 represents X00008M, 3 represents X00009M, 4 represents a negative sample of dried ginger, 5 represents a reference medicinal material of dried ginger, 6 represents XS0001, 7 represents XS0002, 8 represents X00003M, and 9 represents a reference medicinal material of dried ginger. DETAILED DESCRIPTION

[0043] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0044] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the invention, the meaning of "multiple" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined. In the description of the present invention, the meaning of "several" is at least one, such as one, two, etc., unless otherwise clearly and specifically defined.

[0045] The words "preferably", "more preferably", etc. in the present invention refer to embodiments of the present invention that may provide certain beneficial effects in certain circumstances. However, other embodiments may also be preferred under the same circumstances or other circumstances. In addition, the description of one or more preferred embodiments does not imply that other embodiments are not applicable, nor is it intended to exclude other embodiments from the scope of the present invention.

[0046] When a numerical range is disclosed herein, the above range is considered to be continuous and includes the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be combined. In other words, unless otherwise indicated, all ranges disclosed herein should be understood to include any and all subranges included therein.

[0047] Unless otherwise specified, all percentages, fractions and ratios are calculated by the total mass of the composition of the present invention. Unless otherwise specified, all masses of the listed ingredients are given to the content of active substances, so they do not include solvents or by-products that may be contained in commercially available materials. The term "mass percentage content" herein can be represented by the symbol "%". Unless otherwise specified, all molecular weights in this article are weight average molecular weights expressed in Daltons. Unless otherwise specified, all formulations and tests in this article occur in an environment of 25°C. "Including", "including", "containing", "containing", "having" or other variants herein are intended to cover non-closed inclusions, and these terms are not distinguished. The term "comprising" means that other steps and ingredients that do not affect the final result can be added. The composition and method / process of the present invention include, consist of and are essentially composed of the necessary elements and restrictions described herein and any additional or optional ingredients, components, steps or restrictions described herein. The terms "efficacy", "performance", "effect" and "efficacy" are not distinguished between herein.

[0048] In addition, the drawings are not drawn in a 1:1 ratio, and the relative sizes of the elements are drawn in the drawings only as examples to facilitate understanding of the present invention, but are not necessarily drawn in true proportion. The proportions in the drawings do not constitute a limitation to the present invention.

[0049] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0050] The invention provides an identification method for abdominal pain pills. The abdominal pain pills include, by weight, 40 to 60 parts of cinnamon bark, 90 to 110 parts of costusroot, 90 to 110 parts of dried ginger, 40 to 60 parts of magnolia bark, 40 to 60 parts of cardamom, 40 to 60 parts of amomum, 10 to 30 parts of piper longum, 10 to 30 parts of poppy shell, 90 to 110 parts of immature bitter orange and 40 to 60 parts of lindera stalk;

[0051] The identification method includes: using thin layer chromatography to detect one or more of cinnamon bark, costus root, dried ginger and magnolia bark.

[0052] In a specific example, the method for identifying abdominal pain pills includes the following steps:

[0053] Take Dutong Pills, perform pretreatment, add solvent for extraction, filter the extract, evaporate the filtrate to dryness, add solvent to dissolve the residue, and obtain a test solution;

[0054] Take the control medicinal material and prepare the control medicinal material solution according to the extraction method of the test solution;

[0055] Take the reference substance and dissolve it in the solvent to prepare the reference substance solution;

[0056] Separately aspirate the test sample solution, reference substance and / or reference medicinal material solution and spot them on the same thin layer plate, then place the spotted thin layer plate in a developing agent, develop it, take it out, dry it, develop the color, and inspect it.

[0057] In a specific example, detecting cinnamon includes the following steps:

[0058] Mixing the Dutong Pills with a first solvent to prepare a first test solution;

[0059] Mixing a cinnamaldehyde reference substance with a first solvent to prepare a first reference substance solution;

[0060] Take the first test solution and the first reference solution respectively, spot them on a silica gel thin layer plate, place them in the first developing agent for development, take them out to dry, and spray the first color developing agent;

[0061] The first solvent is methanol; and the first developing agent is a mixture of petroleum ether and ethyl acetate.

[0062] In a specific example, in the first developing solvent, the volume ratio of petroleum ether to ethyl acetate is (17±0.5):(3±0.5).

[0063] Preferably, in the first developing solvent, the volume ratio of petroleum ether to ethyl acetate is (17±0.3):(3±0.3). More preferably, in the first developing solvent, the volume ratio of petroleum ether to ethyl acetate is 17:3.

[0064] In a more specific example, the boiling range specification of petroleum ether in the first developing agent is 60°C to 90°C.

[0065] In a specific example, the first color developing agent is a dinitrophenylhydrazine ethanol solution.

[0066] In a specific example, preparing the first test solution includes the following steps:

[0067] The Dutong Pills and the first solvent are mixed, ultrasonically extracted for 30 min±10 min, filtered, the filtrate is taken, evaporated to dryness, and then the first solvent is added.

[0068] In a specific example, the silica gel thin layer plate used for detecting cinnamon is a silica gel G thin layer plate.

[0069] In a specific example, the detection of costus root includes the following steps:

[0070] Mixing the Dutong Pills with a second solvent to prepare a second test solution;

[0071] Mixing the reference substances of dehydrocostus lactone and costunolide with the second solvent to prepare a second reference substance solution;

[0072] Take the second test solution and the second reference solution respectively, spot them on the silica gel thin layer plate, place them in the second developer for development, take them out and dry them, then heat them at 100℃~110℃, and spray the second developer;

[0073] The second developing solvent is petroleum ether and ethyl acetate in a volume ratio of (17±0.5):(3±0.5).

[0074] Preferably, in the second developing solvent, the volume ratio of petroleum ether to ethyl acetate is (17±0.3):(3±0.3). More preferably, in the second developing solvent, the volume ratio of petroleum ether to ethyl acetate is 17:3.

[0075] In a more specific example, the boiling range specification of petroleum ether in the second developing solvent is 60°C to 90°C.

[0076] In a specific example, the second solvent is selected from one or more of ethyl acetate, methanol and ethanol.

[0077] In a specific example, the second developer is a 10% sulfuric acid ethanol solution.

[0078] In a specific example, preparing the second test solution includes the following steps:

[0079] Mix the Dutong Pills with the second solvent, perform ultrasonic extraction for 30 min±10 min, filter, take the filtrate, evaporate to dryness, and then add ethyl acetate.

[0080] In a specific example, the silica gel thin layer plate used for costus root detection is a 0.1% CMC-Na-G thin layer plate.

[0081] In a specific example, testing Magnolia officinalis includes the following steps:

[0082] Mixing the Dutong Pills, activated carbon and a third solvent to prepare a third test solution;

[0083] Mixing magnolol and honokiol reference substances with a third solvent to prepare a third reference substance solution;

[0084] Take the third test solution and the third reference solution respectively, spot them on the silica gel thin layer plate, place them in the third developing agent for development, take them out and dry them, and then examine them at 254nm;

[0085] Wherein, the third developing solvent is a mixture of chloroform and methanol.

[0086] In a specific example, in the third developing solvent, the volume ratio of chloroform to methanol is (17±0.5):(1±0.5).

[0087] Preferably, in the third developing solvent, the volume ratio of chloroform to methanol is (17±0.3):(1±0.3). More preferably, in the third developing solvent, the volume ratio of chloroform to methanol is 17:1.

[0088] In a specific example, the third solvent is selected from one or more of methanol and ethanol.

[0089] In a specific example, preparing the third test solution includes the following steps:

[0090] Mix the abdominal pain pills with activated carbon, add a third solvent, perform ultrasonic extraction for 30 min ± 10 min, filter, take the filtrate, evaporate to dryness, and then add the third solvent.

[0091] In a specific example, the silica gel thin layer plate used for the detection of Magnolia officinalis is GF 254 Thin layer board.

[0092] In a specific example, testing dried ginger includes the following steps:

[0093] Mixing the Dutong Pills with the fourth solvent to prepare a fourth test solution;

[0094] Mixing the dried ginger reference medicinal material with the fourth solvent to prepare a reference medicinal material solution;

[0095] Take the fourth test sample solution and the control medicinal material solution respectively, spot them on the silica gel thin layer plate, place them in the fourth developing agent for development, take them out and dry them, then heat them at 100℃~110℃, and spray the third color developing agent;

[0096] The fourth developing agent is a mixture of petroleum ether, chloroform and ethyl acetate; and the third color developing agent is a 10% sulfuric acid ethanol solution.

[0097] In a specific example, in the fourth developing solvent, the volume ratio of petroleum ether, chloroform and ethyl acetate is (2±0.5):(1±0.5):(1±0.5).

[0098] Preferably, in the fourth developing solvent, the volume ratio of petroleum ether, chloroform and ethyl acetate is (2±0.3):(1±0.3):(1±0.3). More preferably, in the fourth developing solvent, the volume ratio of petroleum ether, chloroform and ethyl acetate is 2:1:1.

[0099] In a more specific example, the boiling range specification of petroleum ether in the fourth developing agent is 60°C to 90°C.

[0100] In a specific example, the fourth solvent is selected from one or more of methanol, ethyl acetate and ethanol.

[0101] In a specific example, preparing the fourth test solution includes the following steps:

[0102] The Dutong Pills were mixed with the fourth solvent, subjected to ultrasonic extraction for 30 min±10 min, filtered, the filtrate was collected, evaporated to dryness, and then the fourth solvent was added.

[0103] In a specific example, preparing the control medicinal material solution includes the following steps:

[0104] The dried ginger control medicinal material was mixed with the fourth solvent, subjected to ultrasonic extraction for 30 min±10 min, filtered, the filtrate was collected, evaporated to dryness, and then the fourth solvent was added.

[0105] In a specific example, the silica gel thin layer plate used for the detection of dried ginger is a 0.1% CMC-Na-G thin layer plate.

[0106] The identification method of the stomachache pill of the present invention is further described in detail below in conjunction with specific examples. The raw materials used in the following examples, unless otherwise specified, are all commercially available products.

[0107] Example 1

[0108] This embodiment provides a method for thin layer chromatography identification of Dutong Pills, which is as follows:

[0109] (1) Thin layer identification of cinnamon:

[0110] Take 6g of Dutong Pills (samples with batch numbers Z00001, Z00002, and Z00003 are the first, second, and third batches of domestically produced preparations produced in 2019, respectively), grind them into powder, add 10ml of methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and add 1ml of methanol to the residue to dissolve it as the test solution. Prepare the negative solution without cinnamon in the same way.

[0111] Take another cinnamaldehyde reference substance and add methanol to make a solution containing 1 μl per 1 ml as the reference substance solution.

[0112] 4 μl of the test solution and 2 μl of the reference solution were respectively spotted on the same silica gel G thin layer plate, and petroleum ether (60-90°C)-ethyl acetate (17:3) was used as the developing solvent. Saturation was performed for 10 minutes, and the plate was developed. The plate was taken out, dried, and sprayed with dinitrophenylhydrazine ethanol reagent.

[0113] In the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatogram of the reference sample, such as Figure 1 shown.

[0114] (2) Thin layer identification of costus root:

[0115] Take 6g of Dutong Pills, grind them into powder, add 10ml of ethyl acetate, treat with ultrasound for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of methanol to dissolve the residue, and use it as the test solution. Prepare the negative solution without costusroot in the same way.

[0116] Separately, take dehydrocostus lactone reference substance and costunolide reference substance, add ethyl acetate to prepare solutions containing 1 mg of dehydrocostus lactone and costunolide per 1 ml, as mixed reference substance solutions.

[0117] 2 μl of each of the above two solutions were taken and spotted on the same silica gel 0.1% CMC-Na-G thin layer plate, and petroleum ether (60-90°C)-ethyl acetate (17:3) was used as the developing agent. After development, the plate was taken out, dried, heated at 105°C for 2 minutes, sprayed with 10% ethanolic sulfuric acid solution, and heated at 105°C until the spots were clearly colored.

[0118] In the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatogram of the reference sample, such as Figure 2 shown.

[0119] (3) Thin layer identification of Magnolia officinalis:

[0120] Take 6g of Dutong Pills, grind them into powder, add 3g of activated carbon, mix well, add 15ml of methanol, extract by ultrasonic for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of methanol to dissolve the residue, and use it as the test solution. Prepare the negative solution without Magnolia officinalis in the same way.

[0121] Separately, magnolol and honokiol reference substances were taken, and methanol was added to prepare a mixed reference substance solution containing 1 mg of magnolol and honokiol per 1 ml.

[0122] Pipette 2 μl of each of the above two solutions and spot them on the same silica gel GF 254 On the thin layer plate, chloroform-methanol (17:1) was used as the developing solvent, saturated for 20 minutes, developed, taken out, dried, and examined at 254 nm.

[0123] In the chromatogram of the test sample, a quenching spot of the same color appears at the corresponding position in the chromatogram of the reference sample, such as Figure 3 shown.

[0124] (4) Thin layer identification of dried ginger:

[0125] Take 6g of Dutong Pills, grind them into powder, add 10ml of methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of methanol to dissolve the residue, and use it as the test solution. Prepare the negative solution without dried ginger in the same way.

[0126] Take 1 g of dried ginger reference medicinal powder and extract it according to the extraction method of the test sample as the reference medicinal solution.

[0127] 2 μl of each of the above two solutions were taken and spotted on the same silica gel 0.1% CMC-Na-G thin layer plate, and petroleum ether (60-90°C)-chloroform-ethyl acetate (2:1:1) was used as the developing solvent. After development, the plate was taken out, dried, heated at 105°C for 1 minute, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored.

[0128] In the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatogram of the control medicinal material, such as Figure 4 shown.

[0129] Example 2

[0130] This embodiment provides a method for thin layer chromatography identification of Dutong Pills, which is as follows:

[0131] (1) TLC identification of cinnamon bark, compared with Example 1, the extraction solvent was changed:

[0132] Take 6g of Dutong Pills, grind them into powder, add 10ml of ethanol, treat with ultrasound for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of ethanol to the residue to dissolve it, and use it as the test solution.

[0133] Take cinnamaldehyde reference substance and add ethanol to make a solution containing 1 μl per 1 ml as the reference substance solution.

[0134] Pipette 4 μl of the test solution and 2 μl of the cinnamaldehyde reference solution, spot them separately on the same silica gel G thin layer plate, use petroleum ether (60-90℃)-ethyl acetate (17:3) as the developing solvent, saturate for 10 minutes, develop, take out, dry, and spray with dinitrophenylhydrazine ethanol reagent.

[0135] In the chromatogram of the test sample, spots of the same color appear at the corresponding positions of the chromatogram of the reference sample. However, compared with the thin layer identification of cinnamon in Example 1, the color of the spots is lighter and the effect is inferior to that of Example 1. Figure 1 shown.

[0136] (2) Thin layer identification of costusroot, compared with Example 1, the volume ratio of the developing agent was changed:

[0137] Take 6 g of Dutong Pills, grind them into powder, add 10 ml of ethyl acetate, treat with ultrasound for 30 minutes, filter, evaporate the filtrate to dryness, add 1 ml of methanol to dissolve the residue, and use it as the test solution.

[0138] Take dehydrocostus lactone reference substance and costunolide reference substance separately, add ethyl acetate to prepare solutions containing 1 mg per 1 ml, as reference substance solutions.

[0139] Take 2 μl of each of the above two solutions and spot them on the same silica gel 0.1% CMC-Na-G thin layer plate, use petroleum ether (60-90℃)-ethyl acetate (18:2) as the developing solvent, develop, take out, dry, heat at 105℃ for 2 minutes, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly colored. In the chromatogram of the test sample, spots of the same color appear at the corresponding position of the chromatogram of the reference sample.

[0140] In the chromatogram of the test sample, spots of the same color appeared at the corresponding positions of the chromatogram of the reference sample. However, compared with the thin layer identification of costusroot in Example 1, the separation effect of the spots of the reference sample was inferior to that of Example 1. Figure 2 shown.

[0141] (3) Thin layer identification of Magnolia officinalis, compared with Example 1, the type of developing agent was changed and the developer was sprayed:

[0142] Take 6g of Dutong Pills (samples with batch numbers X00009M, X00008M, and X00007M are the ninth, eighth, and seventh batches of domestic sales products produced in 2017 that need to be frozen; samples with batch numbers XS0001, XS0002, and XS0003 are the first, second, and third batches of trial production products produced in 2017), grind them into powder, add 3g of activated carbon, mix well, add 15ml of methanol, extract by ultrasonication for 30 minutes, filter, evaporate the filtrate, add 1ml of methanol to dissolve the residue, and use it as the test solution.

[0143] Take magnolol and honokiol reference substances separately, add methanol to prepare reference substance solutions containing 1 mg per 1 ml.

[0144] 2 μl of each of the above two solutions were taken and spotted on the same silica gel G thin layer plate, and petroleum ether (60-90°C)-ethyl acetate (17:8) was used as the developing agent. After development, the plate was taken out, dried, sprayed with 1% vanillin sulfuric acid solution, and heated at 105°C until the spots were clearly colored.

[0145] In the sunlight, it can be seen that the test sample chromatogram has spots of the same color at the corresponding positions of the reference sample chromatogram. However, compared with the thin layer identification of Magnolia officinalis in Example 1, the use of non-fluorescent silica gel G thin layer plate, changing the type of developing agent and spraying with color developer, the impurity spots are more and more obvious, the separation effect of the reference sample spots and the impurity spots is inferior to that of Example 1, and R f Value = 0.72 is larger, such as Figure 3 shown.

[0146] (4) TLC identification of dried ginger: Compared with Example 1, the types of developing agent and developer were changed:

[0147] Take 6g of Dutong Pills (of which the samples with batch number X00003M are the third batch of domestic sales products produced in 2017 that need to be frozen), grind them into powder, add 10ml of ethanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate, and add 1ml of ethanol to the residue to dissolve it as the test solution.

[0148] Take another 1g of dried ginger reference medicinal material powder and extract it according to the extraction method of the test sample as the reference medicinal material solution.

[0149] Pipette 2 μl of each of the above two solutions and spot them on the same silica gel G thin layer plate, use cyclohexane-ethyl acetate-dichloromethane (5:4:2) as the developing solvent, develop, take out, dry, heat at 105°C for 1 minute, spray with 1% vanillin sulfuric acid solution, and heat at 105°C until the spots are clearly colored.

[0150] In the sunlight, the test sample chromatogram showed spots of the same color at the corresponding positions of the control medicinal material chromatogram. However, compared with the thin layer identification of dried ginger in Example 1, only one spot in the control medicinal material showed obvious color, and the other spots did not show obvious color. The test sample failed to compare better with the control medicinal material, and the effect was inferior to that of Example 1. Figure 4 shown.

[0151] Example 3

[0152] This example provides a method for thin layer chromatography identification of Dutong Pills, specifically thin layer chromatography identification of costusroot. Compared with Example 1, the extraction solvent is changed, as follows:

[0153] Take 6 g of Dutong Pills, grind them into powder, add 10 ml of ethyl acetate, treat with ultrasound for 30 minutes, filter, evaporate the filtrate to dryness, add 1 ml of methanol to dissolve the residue, and use it as the test solution.

[0154] Take dehydrocostus lactone reference substance and costunolide reference substance separately, add ethyl acetate to prepare solutions containing 1 mg per 1 ml, as reference substance solutions.

[0155] Pipette 2 μl of each of the above two solutions and spot them on the same silica gel G thin layer plate, use cyclohexane-ethyl formate-formic acid (15:5:1) as the developing agent, develop, take out, dry, heat at 105°C for 2 minutes, spray with 1% vanillin sulfuric acid solution, and heat at 105°C until the spots are clearly colored.

[0156] In the chromatogram of the test sample, spots of the same color appeared at the corresponding positions of the chromatogram of the reference sample. However, compared with the thin layer identification of costus root in Example 1, the reference sample had negative interference, and compared with Example 1 (R f =0.5), its R f Value = 0.2 is too small, and the separation effect is inferior to that of Example 1. Figure 2 shown.

[0157] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0158] The above-described embodiments only express several implementation methods of the present invention, which are convenient for understanding the technical solutions of the present invention in detail, but they cannot be understood as limiting the scope of protection of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided by the present invention are all within the protection scope of the claims attached to the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the contents of the attached claims, and the description and drawings can be used to interpret the contents of the claims.

Claims

1. A method for identifying abdominal pain pills, characterized in that: In parts by weight, the components of the abdominal pain pill include 40 to 60 parts of cinnamon bark, 90 to 110 parts of costus root, 90 to 110 parts of dried ginger, 40 to 60 parts of magnolia bark, 40 to 60 parts of cardamom, 40 to 60 parts of amomum villosum, 10 to 30 parts of piper longum, 10 to 30 parts of poppy shell, 90 to 110 parts of immature bitter orange and 40 to 60 parts of lindera stalk; The identification method comprises: using thin layer chromatography to detect the cinnamon bark, costusroot, dried ginger and magnolia bark; Detecting the cinnamon bark comprises the following steps: Mixing the Dutong Pills with a first solvent to prepare a first test solution; Mixing a cinnamaldehyde reference substance with the first solvent to prepare a first reference substance solution; The first test solution and the first reference solution are respectively taken, spotted on a silica gel thin layer plate, placed in a first developing agent for development, taken out to dry, and sprayed with a first color developer; Wherein, the first solvent is methanol; the first developing agent is a mixture of petroleum ether and ethyl acetate; Detecting the woodruff comprises the following steps: Mixing the Dutong Pills with a second solvent to prepare a second test solution; Mixing the reference substances of dehydrocostus lactone and costunolide with the second solvent to prepare a second reference substance solution; The second test solution and the second reference solution are respectively taken, spotted on a silica gel thin layer plate, placed in a second developing agent for development, taken out and dried, then heated at 100°C to 110°C, and sprayed with a second color developer; Wherein, the second developing agent is petroleum ether and ethyl acetate in a volume ratio of (17±0.5):(3±0.5).

2. The method for identifying the stomachache pills according to claim 1, characterized in that: In the first developing solvent, the volume ratio of petroleum ether to ethyl acetate is (17±0.5):(3±0.5).

3. The identification method of the stomachache pill according to claim 2, characterized in that: The first color developing agent is dinitrophenylhydrazine ethanol solution.

4. The method for identifying the stomachache pills according to claim 1, characterized in that: The second solvent is selected from one or more of ethyl acetate, methanol and ethanol.

5. The method for identifying the stomachache pills according to claim 1, characterized in that: The second color developing agent is 10% sulfuric acid ethanol solution.

6. The method for identifying the stomachache pills according to claim 1, characterized in that: Detecting the Magnolia officinalis comprises the following steps: Mixing the Dutong Pills, activated carbon and a third solvent to prepare a third test solution; Mixing the reference substance of magnolol and honokiol with the third solvent to prepare a third reference substance solution; The third test solution and the third reference solution are respectively taken, spotted on a silica gel thin layer plate, placed in a third developing agent for development, taken out and dried, and then examined at 254 nm; Wherein, the third developing agent is a mixture of chloroform and methanol.

7. The method for identifying the stomachache pills according to claim 6, characterized in that: In the third developing solvent, the volume ratio of chloroform to methanol is (17±0.5):(1±0.5).

8. The method for identifying the stomachache pills according to claim 6, characterized in that: In the third developing agent, the third solvent is selected from one or more of methanol and ethanol.

9. The method for identifying the stomachache pills according to claim 1, characterized in that: Detecting the dried ginger comprises the following steps: Mixing the Dutong Pills with the fourth solvent to prepare a fourth test solution; Mixing the dried ginger reference medicinal material with the fourth solvent to prepare a reference medicinal material solution; The fourth test solution and the reference medicinal material solution are respectively taken, spotted on a silica gel thin layer plate, placed in a fourth developing agent for development, taken out and dried, then heated at 100° C. to 110° C., and sprayed with a third color developer; Wherein, the fourth developing agent is a mixture of petroleum ether, chloroform and ethyl acetate; and the third color developing agent is 10% sulfuric acid ethanol solution.

10. The method for identifying the abdominal pain pills according to claim 9, characterized in that: In the fourth developing solvent, the volume ratio of petroleum ether, chloroform and ethyl acetate is (2±0.5):(1±0.5):(1±0.5); and / or The fourth solvent is selected from one or more of methanol, ethyl acetate and ethanol.

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