CN113391019B - Thin-layer chromatography detection method for flos abelmoschi manihot medicinal material, extract or preparation - Google Patents
Thin-layer chromatography detection method for flos abelmoschi manihot medicinal material, extract or preparation Download PDFInfo
- Publication number
- CN113391019B CN113391019B CN202110501062.0A CN202110501062A CN113391019B CN 113391019 B CN113391019 B CN 113391019B CN 202110501062 A CN202110501062 A CN 202110501062A CN 113391019 B CN113391019 B CN 113391019B
- Authority
- CN
- China
- Prior art keywords
- solution
- extract
- preparation
- detection method
- acetone
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/94—Development
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/95—Detectors specially adapted therefor; Signal analysis
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention relates to the technical field of medicine quality control, in particular to a thin-layer chromatography detection method of an abelmoschus manihot medicinal material, an extract or a preparation, which comprises the following steps: respectively dripping any one of test solution and reference solution of flos Abelmoschi Manihot flower medicinal material, extract or preparation on the same thin layer plate, developing with toluene-acetone-methanol as developing agent, taking out, air drying, spraying color-developing agent until the color development of spots is clear, and inspecting under ultraviolet lamp 365 nm. Compared with the prior art, the method can simultaneously identify 4 components of rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin, can quickly perform qualitative and quantitative analysis on the sunset abelmoschus flower medicinal material, the extract and the preparation or the composition containing the 4 flavonoid components, and has the advantages of simple operation and good reproducibility.
Description
Technical Field
The invention belongs to the technical field of medicine quality control, and particularly relates to a thin-layer chromatography detection method of an abelmoschus manihot medicinal material, an extract or a preparation.
Background
Abelmoschus manihot (L.) Medicus, belonging to the genus Abelmoschus of the family Malvaceae, was originally recorded in Jiayou materia Medica, "spring seedlings leaves, which resemble Althaea manihot, but leaves of narrow and scarce, blossoming in late summer and pale yellow". Abelmoschus manihot is widely distributed and abundant in resources, and is recorded in compendium of materia Medica: its flower is sweet, cold, slippery and nontoxic in flavor, mainly used for treating stranguria with urination and hastening growth, and it is used for treating malignant sores, pus and non-healing after long-term use, and it is the essential herb for sores, eliminating deep-rooted carbuncle and swelling, soaking oil and applying soup for fire injury. Researches show that the total flavonoids of abelmoschus manihot mainly comprise the following 10 flavonols and glycoside compounds thereof: gossypetin-8-0-beta-D-glucuronide, gossypetin-3 ' -O-glucoside, myricetin-3 ' -O-glucoside, myricetin-3-0-glucoside, quercetin-3 ' -O-glucoside, isoquercitrin, hyperoside, and quercetin-3-locust glucoside.
The abelmoschus manihot capsule is prepared by extracting abelmoschus manihot, has the effects of clearing away heat and dampness, detoxifying and reducing swelling, is used for treating chronic nephritis, and has a remarkable clinical curative effect. The flavonoid component is the main active component, and only one component of quercetin is identified in 2020 edition Chinese pharmacopoeia and quality standard of abelmoschus esculentus capsules. In order to ensure the product quality, it is important to establish a method suitable for the rapid identification and content determination of various flavone components in extracts (including an abelmoschus manihot extract, a primary concentrated solution, a secondary concentrated solution and a dry extract) in various working procedures in the production process of abelmoschus manihot medicinal materials and abelmoschus manihot capsules and abelmoschus manihot preparations.
The traditional high performance liquid chromatography has the advantages of high sensitivity, high accuracy and the like, and becomes one of important means for controlling the quality of traditional Chinese medicines, but the traditional high performance liquid chromatography has the defects of expensive instruments, long detection time and more toxic organic reagents, and brings certain influence on the environment. The thin layer chromatography is a chromatographic analysis technique which can rapidly perform the qualitative and quantitative analysis of the pharmaceutical ingredients by using a support coated on a support plate as a stationary phase and a proper solvent as a mobile phase. The flavonoids in the flower of abelmoschus manihot are complex in components, the parent nucleus structures are similar, and the chromatographic separation difficulty is high. In the prior art, the quality standard of abelmoschus manihot and the quality standard of abelmoschus manihot capsules, which are collected in the first part of 2020 edition of Chinese pharmacopoeia, only carry out thin-layer chromatography identification on quercetin; yanYu et al identified two flavonoid compounds rutin and quercetin by the same thin layer method in the study of thin layer chromatography identification of flavonoid compounds, but the identified ingredients are few and the inspection effect is not good; patent CN103487546B discloses a method for identifying flos abelmoschi manihot medicinal materials by using rutin, hyperoside, and quercetin as reference substances or rutin and hyperoside as reference substances and using glacial acetic acid as developing agent, wherein the number of identification components is small.
Therefore, the thin-layer chromatography identification and quantification method for multiple flavone components in the sunset abelmoschus flower medicinal material, extract and preparation has important significance.
Disclosure of Invention
In order to overcome the technical problems, the invention provides a thin-layer chromatography detection method for flavone in a flower of abelmoschus manihot medicinal material, an extract or a preparation, the method is simple and easy to operate, accurate identification results can be obtained, the reproducibility is good, and meanwhile, the rapid qualitative and quantitative analysis for detecting various flavone components at one time can be realized.
In order to realize the purpose, the technical scheme provided by the invention is as follows:
on the one hand, the invention provides a thin-layer chromatography detection method of an abelmoschus manihot medicinal material, an extract or a preparation, which is used for carrying out the thin-layer chromatography detection of the abelmoschus manihot medicinal material by taking toluene-acetone-methanol as a developing agent and polyamide as a thin-layer chromatography material.
Specifically, the developing solvent is a toluene-acetone-methanol solution with a volume ratio of 6-7.
Specifically, a polyamide film is used as the thin-layer sheet.
Specifically, an aluminum trichloride test solution was used as a color-developing agent, and the test piece was placed under an ultraviolet lamp at 365nm for inspection.
Specifically, the thin layer chromatography detection method comprises the following steps: respectively dropping test solution and reference solution of flos Abelmoschi Manihot flower medicinal material, extract or preparation on the same thin layer plate, developing with toluene-acetone-methanol as developing agent, taking out, air drying, spraying color developing agent until the spots are clearly developed, and inspecting under ultraviolet lamp 365 nm.
More specifically, the test solution is a test solution of a sunset abelmoschus herb, an extract test solution or a preparation test solution.
More specifically, the preparation method of the test solution comprises the following steps: taking an extract, an extract or a preparation of the sunset abelmoschus flower medicinal material, adding water for dissolving according to a material-liquid ratio of 1.5, adding ethyl acetate for extraction, and combining extracting solutions to obtain a test solution.
Further specifically, the preparation method of the test solution of the abelmoschus manihot medicinal material comprises the following steps: extracting flos Abelmoschi Manihot with water or organic solvent, concentrating the extractive solution to obtain extract, dissolving in water, extracting with ethyl acetate for 1-3 times, mixing extractive solutions, and processing into ethyl acetate solution containing 0.08g medicinal material per ml; the preparation method of the extract test solution or the preparation test solution comprises the following steps: dissolving the extract or preparation in water, extracting with ethyl acetate for 1-3 times, mixing extractive solutions, and processing into extract test solution or preparation test solution containing 0.08g medicinal material test sample per ml.
More specifically, the sunset abelmoschus flower extract or preparation is prepared from sunset abelmoschus flower medicinal materials under production conditions.
More specifically, the extract comprises a first concentrated solution obtained by primarily concentrating an abelmoschus manihot extract, a second concentrated solution obtained by further concentrating a first concentrated solution of abelmoschus manihot and a dry extract obtained by drying in the production process.
More specifically, the preparation method of the abelmoschus manihot flower extract comprises the following steps: taking flos Abelmoschi Manihot flower, heating and reflux-extracting with ethanol to obtain extractive solution, filtering, concentrating the filtrate to obtain primary concentrated solution, refrigerating to remove oil, concentrating the secondary concentrated solution to obtain secondary concentrated solution, and drying to obtain dry extract.
More specifically, the preparation method of the abelmoschus manihot flower extract comprises the following steps: taking abelmoschus manihot as a medicinal material, adding 95% ethanol according to a material-liquid ratio of 1-20, performing reflux extraction for 1-3 times, each time for 0.5-1.5h, filtering, combining filtrates to obtain an extracting solution, recovering ethanol to obtain a primary concentrated solution, concentrating the filtrate to a specific gravity of 1.10-1.30, storing and standing the concentrated solution, removing an oil layer of the cold-stored solution, concentrating again to obtain a secondary concentrated solution, and performing vacuum drying to obtain a dry extract.
More specifically, the flos abelmoschi manihot medicinal material, the extract or the preparation contains flavonoid components, wherein the flavonoid components are one or more of myricetin, quercetin, gossypetin-8-O-beta-D-glucuronide, hyperoside, isoquercitrin and rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin are preferred.
More specifically, the abelmoschus manihot preparation contains an abelmoschus manihot extract and is any one of granules, tablets, capsules, oral liquid or other pharmaceutically acceptable dosage forms.
More specifically, the preparation method of the reference solution comprises the following steps: taking rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin reference substances respectively, and adding an ethanol solution to prepare a mixed reference substance solution with the concentration ratio of the reference substances of 4-5.
Further specifically, the concentration ratio of rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin in the mixed reference solution is 4.5.
More specifically, the concentration of the ethanol solution is 50 to 100%, preferably 50 to 95%, and more preferably 65 to 75%.
More specifically, the sample application amount of the test solution is 1-3 μ L, and the sample application amount of the mixed control solution is 5-15 μ L.
More specifically, the sample amount of the test solution is 2 μ L, and the sample amount of the mixed control solution is 10 μ L.
In another aspect, the invention provides a thin-layer chromatography detection method for a composition containing rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin, wherein the thin-layer chromatography detection is carried out by taking toluene-acetone-methanol as a developing agent and polyamide as a thin-layer chromatography material.
Specifically, the volume ratio of the developing solvent toluene-acetone-methanol is (6-7).
Specifically, the detection method comprises the following steps: respectively dispensing the composition solution and the reference solution on the same thin layer plate, developing with toluene-acetone-methanol as developing agent, taking out, air drying, spraying color-developing agent until the spots are clearly developed, and inspecting under ultraviolet lamp 365 nm.
More specifically, the thin layer plate is a polyamide thin film, and the color developing agent is an aluminum trichloride test solution.
More specifically, the preparation method of the reference solution comprises the following steps: taking rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin reference substances respectively, and adding an ethanol solution to prepare a mixed reference substance solution with the concentration ratio of the reference substances of 4-5; the concentration ratio of rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin in the reference solution is 4.5; the concentration of the ethanol solution is 50-100%, preferably 50-95%, and more preferably 65-75%.
More specifically, the composition solution is applied in a spot amount of 0.5 to 6. Mu.L, preferably 2. Mu.L; the sample size of the mixed control solution is 5-15 μ L, preferably 10 μ L.
Compared with the prior art, the invention has the following beneficial effects:
the invention leads 4 flavonoid components in the extract or the preparation of the abelmoschus manihot medicinal material to be as follows: rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin are separated by thin-layer chromatography, so that 4 flavone components in the sunset abelmoschus flower medicinal material, the extract and the preparation can be rapidly qualitatively and quantitatively analyzed at one time.
Drawings
FIG. 1 is a view of thin layer chromatography in example 1, wherein 1 is a mixed control solution and 2 is a preparation sample.
FIG. 2 is a view of thin layer chromatography in example 2, wherein 1 is a mixed control solution and 2 is a preparation test sample.
FIG. 3 is a view of thin layer chromatography in example 3, wherein 1 is a mixed control solution and 2 is a preparation test sample.
FIG. 4 is a positioning and detecting view of each reference chromatogram, wherein 1 is myricetin, 2 is quercetin, 3 is gossypetin-8-O-beta-D-glucuronide, 4 is hyperoside, 5 is isoquercitrin, 6 is rutin, and 7 is a mixed reference.
FIG. 5 is the thin layer chromatography test chart of example 4, in which the band 1 is the mixed control solution, and the other bands are 0.5. Mu.L, 1. Mu.L, 2. Mu.L, and 3. Mu.L of the preparation test solution in this order.
Fig. 6 is a thin-layer chromatography examination view of example 5, and an examination view of a sample solution of the sunset abelmoschus root medicinal material and each extract, wherein the chromatographic points from left to right are respectively: 1. mixing reference substance, 2. Abelmoschus manihot flower sample, 3. Extract (extractive solution) sample, 4. Extract (primary concentrated solution) sample, 5. Extract (secondary concentrated solution) sample, and 6. Extract (dry extract) sample.
FIG. 7 is a thin layer chromatography assay of example 6, in which 1 is a mixed control solution and 2 is a preparation test sample.
FIG. 8 is a thin layer chromatography examination of comparative example 1, in which 1. Granule preparation test article solution.
Detailed Description
The present invention will be described in further detail with reference to specific examples, which are not intended to limit the present invention, but to illustrate the present invention. The experimental methods used in the following examples are not specifically described, and the materials, reagents and the like used in the following examples are generally commercially available under the usual conditions without specific descriptions.
1.1 preparation of a test solution of abelmoschus manihot medicinal materials:
extracting 4g of a sunflower flower medicinal material by using 75-95% ethanol, concentrating or concentrating and drying an extracting solution, adding water into a concentrated solution or a dry extract until 10g of the water is uniformly stirred, adding ethyl acetate for extracting for 3 times, adding the ethyl acetate into a measuring flask with the volume constant of (3.
1.2 preparation of test solutions of the extracts
(1) Preparation method of flos Abelmoschi Manihot extract
Taking 4000g of abelmoschus manihot, performing reflux extraction for 2 times (1 h each time) by using 15 times (mass/volume ratio) of 95% ethanol, filtering, and combining filtrates to obtain an extracting solution; recovering ethanol to obtain primary concentrated solution; the filtrate was concentrated to a specific gravity of 1.20 to obtain a secondary concentrated solution. Standing the secondary concentrated solution at 0-4 deg.C for 24 hr, removing oil layer of the cold storage solution, concentrating, slowly adding into vacuum belt drier, drying, pulverizing, and packaging into clean double-layer plastic bag to obtain flos Abelmoschi Manihot flower extract dry extract.
(2) Preparation of extract test solution
Taking the extracting solution, the primary concentrated solution, the secondary concentrated solution and the dry extract which are equivalent to 4g of the flower of sunset abelmoschus and prepared in the step (1), respectively adding water to 10g, adding ethyl acetate for extraction for 3 times, adding the ethyl acetate in a ratio of (3.
1.3 preparation of test solutions of the preparations
(1) Preparation of abelmoschus manihot extract granules
Taking the dry extract of the abelmoschus manihot flowers prepared in the step 1.2 (1), crushing and sieving by a 80-mesh sieve, taking 500g of the extract, adding 550g of mannitol and 4g of magnesium stearate, and performing dry granulation to prepare granules.
(2) Preparation of test solution of flos Abelmoschi Manihot granule
Taking an abelmoschus manihot granule preparation which is equivalent to 4g of an abelmoschus manihot medicinal material, preparing into 10g of aqueous solution, uniformly stirring, adding ethyl acetate to extract for 3 times (3.
2.1 Single control solution preparation: taking appropriate amount of rutin, hyperoside, isoquercitin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin, respectively, adding appropriate amount of 70% ethanol to make into respective control solutions (with concentration of 0.225mg/mL, 0.570mg/mL, 1.330mg/mL, 0.877mg/mL, 1.315mg/mL, 1.080 mg/mL).
2.2 preparation of mixed control solutions: taking appropriate amount of rutin, hyperoside, isoquercitin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin reference solution respectively, mixing to obtain mixed reference solution (with concentration of 88.906 μ g/mL, 55.425 μ g/mL, 81.790 μ g/mL, 33.235 μ g/mL, 36.088 μ g/mL, 21.496 μ g/mL respectively).
Example 1
Taking 2 mu L of the test solution of the sunset abelmoschus flower granular preparation prepared in the pretreatment step 1.3 and 10 mu L of the mixed reference solution prepared in the pretreatment step 2.2, respectively dropping the test solution and the mixed reference solution on the same polyamide film, taking out the polyamide film, airing the polyamide film, spraying an aluminum trichloride color developing agent until spots are clearly developed, and placing the polyamide film under an ultraviolet lamp 365nm for inspection. The detection result is shown in fig. 1, and spots are clearly separated.
Example 2
Taking 2 mu L of the test solution of the sunset abelmoschus flower granular preparation prepared in the pretreatment step 1.3 and 10 mu L of the mixed reference solution prepared in the pretreatment step 2.2, respectively dropping the test solution and the mixed reference solution on the same polyamide film, taking out the polyamide film, airing the polyamide film, spraying an aluminum trichloride color developing agent until spots are clearly developed, and placing the polyamide film under an ultraviolet lamp 365nm for inspection. The detection result is shown in fig. 2, and the spots are clearly separated.
Example 3
Taking 2 mu L of the test solution of the abelmoschus manihot granule preparation prepared in the pretreatment step 1.3 and 10 mu L of the mixed reference solution prepared in the pretreatment step 2.2, respectively dropping the test solution and the mixed reference solution on the same polyamide film, developing by taking a toluene-acetone-methanol (3. The detection result is shown in fig. 3, and the spots are clearly separated.
Example 4 different dot amount test
1. Location in chromatogram of each control
And (3) respectively taking 6 mu L of the single reference substance solution prepared in the pretreatment step 2 and 10 mu L of the mixed reference substance solution, spotting the single reference substance solution and the mixed reference substance solution on a polyamide film, placing the polyamide film in a development cylinder, developing, taking out, airing, spraying an aluminum trichloride test solution, and observing under ultraviolet light (365 nm). The results are shown in FIG. 4.
2. Thin-layer identification of test sample of abelmoschus manihot preparation
Respectively dotting 0.5 muL, 1 muL, 2 muL and 3 muL of the test solution of the sunset abelmoschus flower preparation prepared in the pretreatment step 1.3 on the same polyamide film; 10 mu L of the mixed reference substance solution prepared in the pretreatment step 2.2 is respectively spotted on the same polyamide film, toluene-acetone-methanol (2.
Example 5 thin layer identification of sunset abelmoschus herb extract
Spotting 10 μ L of the mixed reference solution prepared in the pretreatment step 2.2 and 2 μ L of the abelmoschus manihot medicinal material and extract test solution prepared in the pretreatment steps 1.1 and 1.2 on the same polyamide film, developing with toluene-acetone-methanol (2. The results are shown in FIG. 6. As a result of inspection, spots of the same color were respectively shown on the chromatogram of the test sample at positions corresponding to those on the chromatogram of the control sample. The method is proved to have comprehensive and accurate detection results.
Example 6 thin layer identification of Abelmoschus manihot granule formulations
Taking abelmoschus manihot granules which are prepared in the pretreatment step 1.3 (1) and are equivalent to 4g of abelmoschus manihot medicinal materials, adding water to 10g of abelmoschus manihot granules, uniformly stirring, adding ethyl acetate to extract for 3 times (30mL, 10mL and 10mL), combining extracting solutions, and fixing the volume by using ethyl acetate to a 50mL measuring bottle to obtain a test solution. Taking appropriate amount of rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin as reference substances, respectively, adding 70% ethanol to obtain solutions containing 90 μ g, 30 μ g, 40 μ g and 20 μ g per 1mL, respectively, and making into mixed reference substance solution. Performing thin layer chromatography (China pharmacopoeia 2020 edition four-part general rule 0502) by respectively sucking 10 test L of mixed control solution and 2 mu L of test solution, spotting on the same polyamide film, and mixing with toluene: acetone: methanol (2. In the chromatogram of the test sample (as shown in FIG. 7), the fluorescence spots with the same color appear at the corresponding positions of the chromatogram of the mixed control sample, and have no substantial difference from any of the test samples in FIG. 6.
Comparative example 1
Dripping 2 mu L of the test solution of the abelmoschus manihot flower granules prepared in the pretreatment step 1.3 on a polyamide film, and adding toluene: acetone: methanol: formic acid (0.5. The detection result is shown in fig. 8, the spot separation effect is poor, and the rutin strip is close to the upper edge of the thin-layer plate.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (19)
1. A thin-layer chromatography detection method for an abelmoschus manihot medicinal material, an extract or a preparation is characterized by comprising the following steps:
the method takes toluene-acetone-methanol as a developing solvent, wherein the developing solvent is a toluene-acetone-methanol solution with the volume ratio of 6-7;
the method takes polyamide as a thin-layer chromatography material;
the method takes an aluminum trichloride test solution as a color developing agent;
the preparation method of the reference substance solution in the method comprises the following steps: adding ethanol solution into rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin control substances to obtain control solution; the concentration of the ethanol solution is 50-100%;
the preparation method of the test solution in the method comprises the following steps: dissolving the extract, extract or preparation of flos Abelmoschi Manihot with water, extracting with ethyl acetate, and mixing extractive solutions to obtain test solution.
2. The detection method according to claim 1, characterized in that: the developing agent is a toluene-acetone-methanol solution with a volume ratio of 6-6.
3. The detection method according to claim 1, characterized in that: the developing solvent is a toluene-acetone-methanol solution with the volume ratio of 6-6.
4. The detection method according to claim 3, characterized in that: the developing solvent is a toluene-acetone-methanol solution with the volume ratio of 2.
5. The detection method according to any one of claims 1 to 4, wherein: respectively dropping any one of test solution and reference solution of flos Abelmoschi Manihot flower medicinal material, extract or preparation on the same thin layer plate, developing with toluene-acetone-methanol as developing agent, taking out, air drying, spraying color developing agent until the color development of spots is clear, and inspecting under ultraviolet lamp 365 nm.
6. The detection method according to claim 1, characterized in that: the concentration ratio of rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin in the reference solution is 4-5; the concentration of the ethanol solution is 50-95%.
7. The detection method according to claim 6, characterized in that: the concentration ratio of rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin in the reference solution is 4.5; the concentration of the ethanol solution is 65-75%.
8. The detection method according to claim 1, characterized in that: the test solution is an Abelmoschus manihot medicinal material test solution, an extract test solution or a preparation test solution;
the preparation method of the test solution comprises the following steps: taking an extract, an extract or a preparation of the sunset abelmoschus flower medicinal material, adding water for dissolving according to a material-liquid ratio of 1.5, adding ethyl acetate for extracting, and combining extracting solutions to obtain a test solution.
9. The detection method according to claim 8, characterized in that: the preparation method of the test solution of the abelmoschus manihot medicinal material comprises the following steps: extracting flos Abelmoschi Manihot with water or organic solvent, concentrating the extractive solution to obtain extract, dissolving in water, extracting with ethyl acetate for 1-3 times, mixing extractive solutions, and processing into ethyl acetate solution containing 0.08g medicinal material per ml; the preparation method of the extract test solution or the preparation test solution comprises the following steps: dissolving the extract or preparation in water, extracting with ethyl acetate for 1-3 times, mixing extractive solutions, and processing into extract test solution or preparation test solution containing 0.08g medicinal material test sample per ml.
10. The detection method according to claim 9, characterized in that: the extract comprises a first concentrated solution obtained by primarily concentrating an abelmoschus manihot flower extracting solution, a second concentrated solution obtained by further concentrating the first concentrated solution of the abelmoschus manihot flower and a dry extract obtained by drying in the production process;
the preparation method of the abelmoschus manihot extract comprises the following steps: heating and refluxing flos Abelmoschi Manihot flower with ethanol to obtain extractive solution, filtering, concentrating the filtrate to obtain primary concentrated solution, refrigerating to remove oil, concentrating the secondary concentrated solution to obtain secondary concentrated solution, and drying to obtain dry extract.
11. The detection method according to claim 10, characterized in that: the flos Abelmoschi Manihot medicinal material, extract or preparation contains flavonoids, wherein the flavonoids are one or more of myricetin, quercetin, gossypetin-8-O-beta-D-glucuronide, hyperoside, isoquercitrin or rutin;
the abelmoschus manihot preparation contains abelmoschus manihot extract and is any one of pharmaceutically acceptable dosage forms.
12. The detection method according to claim 11, characterized in that: the flavonoid components comprise rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin.
13. A thin-layer chromatography detection method of a composition containing rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin is characterized in that: toluene-acetone-methanol is taken as a developing agent, and polyamide is taken as a thin-layer chromatography material; the developing agent is a toluene-acetone-methanol solution with the volume ratio of 6-7;
taking an aluminum trichloride test solution as a color developing agent;
the preparation method of the reference substance solution in the method comprises the following steps: adding ethanol solution into rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin control substances to obtain control solution; the concentration of the ethanol solution is 50-100%.
14. The detection method according to claim 13, characterized in that: the developing solvent is a toluene-acetone-methanol solution with the volume ratio of 6-6.
15. The detection method according to claim 13, characterized in that: the developing agent is a toluene-acetone-methanol solution with a volume ratio of 6-6.
16. The detection method according to claim 15, characterized in that: the developing agent is a toluene-acetone-methanol solution with a volume ratio of 2.
17. The detection method according to claim 13, characterized in that: the detection method comprises the following steps: respectively dropping the composition solution and the reference solution on the same thin layer plate, developing with toluene-acetone-methanol as developing agent, taking out, air drying, spraying color-developing agent until the spots are clearly developed, and inspecting under 365nm ultraviolet lamp.
18. The detection method according to claim 13, characterized in that: the concentration ratio of rutin, gossypetin-8-O-beta-D-glucuronide, myricetin and quercetin in the reference solution is 4.5; the concentration of the ethanol solution is 50-95%.
19. The detection method according to claim 18, characterized in that: the concentration of the ethanol solution is 65-75%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110501062.0A CN113391019B (en) | 2021-05-08 | 2021-05-08 | Thin-layer chromatography detection method for flos abelmoschi manihot medicinal material, extract or preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110501062.0A CN113391019B (en) | 2021-05-08 | 2021-05-08 | Thin-layer chromatography detection method for flos abelmoschi manihot medicinal material, extract or preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113391019A CN113391019A (en) | 2021-09-14 |
| CN113391019B true CN113391019B (en) | 2022-11-15 |
Family
ID=77616823
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110501062.0A Active CN113391019B (en) | 2021-05-08 | 2021-05-08 | Thin-layer chromatography detection method for flos abelmoschi manihot medicinal material, extract or preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113391019B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103487546A (en) * | 2013-09-03 | 2014-01-01 | 王志敏 | Identification method of flower of sunset abelmoschus |
| CN107519220A (en) * | 2017-09-15 | 2017-12-29 | 江苏苏中药业集团股份有限公司 | The extracting method of flavone compound in a kind of sunset abelmoschus flower |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102600219B (en) * | 2012-03-21 | 2013-02-13 | 江苏苏中药业集团股份有限公司 | Total flavone extract of abelmoschus manihot and preparing method of total flavone extract |
-
2021
- 2021-05-08 CN CN202110501062.0A patent/CN113391019B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103487546A (en) * | 2013-09-03 | 2014-01-01 | 王志敏 | Identification method of flower of sunset abelmoschus |
| CN107519220A (en) * | 2017-09-15 | 2017-12-29 | 江苏苏中药业集团股份有限公司 | The extracting method of flavone compound in a kind of sunset abelmoschus flower |
Non-Patent Citations (4)
| Title |
|---|
| 复方黄葵颗粒的薄层鉴别研究;贺婕等;《山西医科大学学报》;20170326(第03期);全文 * |
| 芪葵颗粒定性定量方法研究;李爽等;《药物分析杂志》;20120930(第09期);全文 * |
| 薄层色谱法分析苦豆子提取物中生物碱及黄酮类成分;李雪俐等;《新疆医科大学学报》;20161115(第11期);全文 * |
| 黄蜀葵花药材黄酮类成分的微乳薄层色谱定性鉴别研究;李琳等;《中国新药杂志》;20090315(第05期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113391019A (en) | 2021-09-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112666268B (en) | Method for identifying various components and measuring content of traditional Chinese medicine composition | |
| CN104013710B (en) | A kind of Cape jasmine cypress composition and its detection method | |
| CN109613166B (en) | Quality detection method of 'Jihui Tongbiang' capsule | |
| CN108279278B (en) | Method for separating flavonoid components and application thereof | |
| JP3386796B2 (en) | Quality determination method for plants of the genus Salicaceae and / or extracts thereof | |
| CN102078403B (en) | Detection method of Chinese traditional medicine preparation of oral liquid for treating infant cough with lung heat | |
| CN113109495B (en) | Quality detection method of jujube kernel nerve-soothing capsules based on thin-layer chromatography | |
| CN101244240B (en) | Quantitative and qualitative analysis method for four turmeric soup preparations | |
| CN100363029C (en) | Quality control method of cbinese medicinal preparation | |
| CN113391019B (en) | Thin-layer chromatography detection method for flos abelmoschi manihot medicinal material, extract or preparation | |
| CN100369614C (en) | Quality control method of compound gallblader freeflow solid preparation | |
| CN101690752A (en) | Quality control method of medicine composition | |
| CN108037200B (en) | Quality detection method of kidney nourishing and tranquilizing pills | |
| CN108037234B (en) | Quality detection method of abrus herb hepatitis granules | |
| CN115728409A (en) | Qualitative and quantitative detection method for Xiasangju granules | |
| CN115575541A (en) | Method for simultaneously determining multiple active ingredients in Yinhuang Erchen mixture | |
| CN104111295A (en) | Method for controlling quality of Chinese herbal preparation | |
| CN111830187B (en) | Rapid thin-layer identification method for multiple medicinal flavors in bupleurum tenuifolia granule finished product | |
| CN100998617B (en) | Quality control method for Xindakang preparation | |
| CN108802260B (en) | Thin-layer identification method for quality control of lotus leaf medicinal materials, decoction pieces and formula granules | |
| CN101879198B (en) | Method for identification of Xindakang preparation | |
| CN110907562A (en) | Quality detection method of stomach harmonizing and detoxifying capsules | |
| CN116087403B (en) | Thin layer identification method for folium Apocyni Veneti and flos Chrysanthemi in compound herba Apocyni Veneti granule | |
| CN100515445C (en) | Quality control method for traditional Chinese medicine preparation for treating generalized anxiety | |
| CN100565202C (en) | The detection method of medicine containing swertia psedochinensis and milk thistle silybum marianum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: No.1 Suzhong Road, Jiangyan District, Taizhou City, Jiangsu Province 225500 Applicant after: Suzhong Pharmaceutical Group Co.,Ltd. Applicant after: Jiangsu Suzhong Pharmaceutical Research Institute Co.,Ltd. Address before: 225500 No. 1 Suzhong Road, Taizhou City, Jiangsu Province Applicant before: JIANGSU SUZHONG PHARMACEUTICAL GROUP Co.,Ltd. Applicant before: Jiangsu Suzhong Pharmaceutical Research Institute Co.,Ltd. |
|
| CB02 | Change of applicant information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Ming Xi Inventor after: Tang Haitao Inventor after: Zhou Zhaoling Inventor after: Wang Dianguang Inventor after: Ge Haitao Inventor after: Xu Jie Inventor before: Tang Haitao Inventor before: Zhou Zhaoling Inventor before: Wang Dianguang Inventor before: Ge Haitao Inventor before: Xu Jie |
|
| CB03 | Change of inventor or designer information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |