CN115184536A - Thin-layer chromatography detection method for safflower in Naoan dropping pills - Google Patents
Thin-layer chromatography detection method for safflower in Naoan dropping pills Download PDFInfo
- Publication number
- CN115184536A CN115184536A CN202210885788.3A CN202210885788A CN115184536A CN 115184536 A CN115184536 A CN 115184536A CN 202210885788 A CN202210885788 A CN 202210885788A CN 115184536 A CN115184536 A CN 115184536A
- Authority
- CN
- China
- Prior art keywords
- solution
- thin
- preparing
- acetone
- safflower
- 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.)
- Pending
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 60
- 244000020518 Carthamus tinctorius Species 0.000 title claims abstract description 40
- 235000003255 Carthamus tinctorius Nutrition 0.000 title claims abstract description 37
- 239000009999 naoan Substances 0.000 title claims abstract description 33
- 239000006187 pill Substances 0.000 title claims abstract description 32
- 238000004809 thin layer chromatography Methods 0.000 title claims abstract description 32
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 123
- 239000000243 solution Substances 0.000 claims abstract description 92
- 239000000047 product Substances 0.000 claims abstract description 62
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 44
- 238000012360 testing method Methods 0.000 claims abstract description 40
- 238000011161 development Methods 0.000 claims abstract description 33
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 29
- ZCHPKWUIAASXPV-UHFFFAOYSA-N acetic acid;methanol Chemical compound OC.CC(O)=O ZCHPKWUIAASXPV-UHFFFAOYSA-N 0.000 claims abstract description 27
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims abstract description 27
- 238000001704 evaporation Methods 0.000 claims abstract description 25
- 239000012085 test solution Substances 0.000 claims abstract description 24
- 238000001914 filtration Methods 0.000 claims abstract description 21
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 17
- 238000005507 spraying Methods 0.000 claims abstract description 17
- 238000001816 cooling Methods 0.000 claims abstract description 16
- 238000000227 grinding Methods 0.000 claims abstract description 16
- 239000013558 reference substance Substances 0.000 claims abstract description 16
- 238000007789 sealing Methods 0.000 claims abstract description 16
- 239000000706 filtrate Substances 0.000 claims abstract description 8
- 239000000843 powder Substances 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims description 78
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 51
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 claims description 31
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- 239000000523 sample Substances 0.000 claims description 29
- 239000012488 sample solution Substances 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 26
- 238000009210 therapy by ultrasound Methods 0.000 claims description 18
- 239000000463 material Substances 0.000 claims description 17
- 238000002360 preparation method Methods 0.000 claims description 16
- 229960000583 acetic acid Drugs 0.000 claims description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims description 15
- 239000012362 glacial acetic acid Substances 0.000 claims description 15
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 10
- 238000007689 inspection Methods 0.000 claims description 10
- 238000007605 air drying Methods 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- 238000011179 visual inspection Methods 0.000 claims description 6
- 238000010438 heat treatment Methods 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 4
- 239000013074 reference sample Substances 0.000 claims description 4
- 239000007921 spray Substances 0.000 claims description 4
- 238000004090 dissolution Methods 0.000 claims description 3
- 238000005470 impregnation Methods 0.000 claims description 2
- 229940079593 drug Drugs 0.000 claims 2
- 239000012088 reference solution Substances 0.000 abstract description 15
- 239000000126 substance Substances 0.000 abstract description 5
- 238000001035 drying Methods 0.000 abstract description 3
- KIWBPDUYBMNFTB-UHFFFAOYSA-N Ethyl hydrogen sulfate Chemical compound CCOS(O)(=O)=O KIWBPDUYBMNFTB-UHFFFAOYSA-N 0.000 abstract 1
- 241000628997 Flos Species 0.000 description 19
- 239000002904 solvent Substances 0.000 description 9
- 239000000203 mixture Substances 0.000 description 8
- 238000002791 soaking Methods 0.000 description 7
- 208000002193 Pain Diseases 0.000 description 4
- 238000004806 packaging method and process Methods 0.000 description 4
- 239000002775 capsule Substances 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 230000036407 pain Effects 0.000 description 3
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 206010000077 Abdominal mass Diseases 0.000 description 1
- 201000000736 Amenorrhea Diseases 0.000 description 1
- 206010001928 Amenorrhoea Diseases 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 241000208838 Asteraceae Species 0.000 description 1
- 208000005171 Dysmenorrhea Diseases 0.000 description 1
- 206010013935 Dysmenorrhoea Diseases 0.000 description 1
- 206010061876 Obstruction Diseases 0.000 description 1
- 235000003181 Panax pseudoginseng Nutrition 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 206010042674 Swelling Diseases 0.000 description 1
- 231100000540 amenorrhea Toxicity 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000000038 chest Anatomy 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229940040145 liniment Drugs 0.000 description 1
- 239000000865 liniment Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 230000008736 traumatic injury Effects 0.000 description 1
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
- G01N30/95—Detectors specially adapted therefor; Signal analysis
-
- 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
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)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention provides a thin-layer chromatography detection method for safflower in Naoan dropping pills, which comprises the steps of preparing a detection product, grinding the product, subpackaging filter paper bags, preventing powder leakage, adding 75% ethanol, sealing, performing ultrasonic filtration, evaporating filtrate to dryness, cooling, adding 85% acetone for dissolving to obtain a test solution, preparing 85% acetone to obtain a blank reference substance, sucking the two solutions, the test solution and the reference solution, respectively dropping the two solutions on the same silica gel G thin-layer plate, taking ethyl acetate-water-glacial acetic acid-methanol as a developing agent, developing, taking out, drying, spraying an ethanol sulfate solution, and placing the thin-layer chromatography detection product under an ultraviolet lamp (365 nm). According to the examination of different temperatures, according to the thin layer chromatography, the examination of low temperature (5 ℃) and high temperature (35 ℃) is carried out, and as a result, when the development temperature is low temperature (5 ℃) and high temperature (35 ℃), the same orange and yellow green spots are displayed on the corresponding positions of the chromatogram of the reference substance in the chromatogram of the test substance, the spots are clear, the resolution is good, and the Rf values are consistent.
Description
Technical Field
The invention relates to the field of safflower thin layers in Naoan dropping pills, in particular to a thin-layer chromatography detection method for safflower in Naoan dropping pills.
Background
The Carthami flos is dried flower of Carthamus Tinctorius of Compositae. Picking the flower when the flower turns yellow red in summer, and drying in the shade or in the sun. Mainly produced in Henan, hunan, sichuan, xinjiang, etc. Has the effects of promoting blood circulation, dredging channels, removing blood stasis and relieving pain. Can be used for treating amenorrhea, dysmenorrhea, lochiorrhea, abdominal mass, thoracic obstruction, cardialgia, abdominal pain due to blood stasis, pricking pain in chest and hypochondrium, traumatic injury, and swelling and pain due to pyocutaneous disease. The product can also be combined with other traditional Chinese medicines to prepare a plurality of traditional Chinese medicine compound varieties, such as Naoan dripping pills, naoan capsules, qili capsules, pseudo-ginseng vulnerary capsules, and pain-relieving liniment, wherein the Naoan dripping pills are unique varieties, although safflower is contained in the prescription, a thin layer safflower identification method is established in a few quality standards, and the operation is complicated or the interference components are easy to show.
The results of the identification of the safflower in the Naoan dropping pill by using the identification methods show that the methods can not be detected, have poor separation degree or have obvious interference components. At present, the quality control of Chinese herbal compound varieties is more and more strict at home and abroad, so that a stable, reliable and simple thin-layer chromatography which can accurately and effectively identify the safflower components of the Naoan dropping pills is urgently needed. Therefore, the quality control standard of the thin layer chromatography identification method of the safflower in the Naoan dropping pill is improved, and a thin layer chromatography detection method of the safflower in the Naoan dropping pill is provided.
Disclosure of Invention
The invention aims to: in order to solve the problems of the prior art, the invention provides the following technical scheme: a thin-layer chromatography detection method for Carthami flos in NAOAN dripping pill comprises preparing detection product, grinding the product, packaging with filter paper to prevent powder leakage, adding 75% ethanol, sealing, ultrasonic filtering, evaporating filtrate to dryness, cooling, and dissolving with 85% acetone to obtain test solution. Preparing 1g of safflower control medicinal material, and preparing a control solution by the same method;
and step two, respectively sucking 5 mul of each sample solution and the reference solution, respectively dropping the sample solution and the reference solution on the same silica gel G thin plate, developing by using ethyl acetate-water-glacial acetic acid-methanol as a developing agent, taking out, airing, spraying a sulfuric acid ethanol solution, and inspecting under an ultraviolet lamp (365 nm).
As a preferred technical scheme of the invention, the method also comprises a first detection product preparation step, wherein the first detection product preparation step comprises the steps of grinding the detection product, subpackaging filter paper bags, adding 75% ethanol into a conical flask, sealing the conical flask, carrying out ultrasonic treatment, filtering, evaporating to dryness, cooling, adding 85% acetone for dissolution, and taking the solution as a test solution. Preparing control solution from 1g of Carthami flos by the same method; developing agent formula I, ethyl acetate-water-glacial acetic acid-methanol; the first color development method is to spray sulfuric acid ethanol solution and to inspect the color under ultraviolet light (365 nm).
The preferable technical scheme of the invention comprises a second detection product preparation step, wherein the second detection product preparation step comprises grinding the detection product, subpackaging filter paper bags, adding 85% acetone for soaking, adding 75% ethanol, placing in a conical flask, sealing, carrying out ultrasonic treatment, filtering, drying by distillation, cooling, adding 85% acetone for dissolving, and taking the solution as a test solution. Preparing control solution from 1g of Carthami flos by the same method; developing solvent formula II, ethyl acetate-water-glacial acetic acid-methanol; and a second color development method, spraying sulfuric acid ethanol solution and placing under ultraviolet light (365 nm) for inspection.
The preferable technical scheme of the invention also comprises a third detection product preparation step, wherein 25ml of acetone is added into the detection product of the third detection product preparation step, the impregnation is carried out, the acetone is removed, the evaporation is carried out at normal temperature, the heating is carried out in a hot water bath, the continuous operation is carried out twice, the water solution is evaporated to dryness, and 85% acetone is added to dissolve the acetone to be used as a test sample solution. Preparing a reference solution from 1g of safflower control medicinal material by the same method; developing solvent formula III, ethyl acetate-water-glacial acetic acid-methanol; and a third color development method, namely, using sulfuric acid ethanol solution to carry out visual inspection under ultraviolet light (254 nm).
The preferable technical scheme of the invention also comprises a fourth detection product preparation step, wherein the fourth detection product preparation step is to add methanol into the detection product, perform ultrasonic treatment, add hot water to dissolve the detection product, extract the detection product for 2 times by using ethyl acetate, combine ethyl acetate solutions and concentrate the ethyl acetate solutions to be used as a test solution. Preparing control solution from 1g of Carthami flos by the same method; the developing solvent is composed of chloroform-ethyl acetate-glacial acetic acid; and the fourth color development method is to use sulfuric acid ethanol solution to inspect under the sunlight.
As the preferable technical scheme, the method further comprises the steps of preparing a fifth detection product, preparing the fifth detection product, adding hot water to the fifth detection product to dissolve the fifth detection product, filtering, evaporating to dryness, adding 85% acetone, performing low-temperature ultrasonic treatment, and concentrating the mixture to obtain a test solution. Preparing 1g of safflower control medicinal material, and preparing a control solution by the same method; developing agent formula five, trichloromethane-ethyl acetate-glacial acetic acid; and a fifth color development method, and visual inspection under an ultraviolet lamp (254 nm).
As a preferred technical scheme of the invention, the method also comprises three special investigation steps, and 85% acetone is prepared to be used as a blank reference substance. Sucking the above two solutions, respectively dropping the sample solution and the reference solution on the same silica gel G thin layer plate, developing with ethyl acetate-water-glacial acetic acid-methanol as developing agent, taking out, air drying, spraying sulfuric acid ethanol solution, and inspecting under ultraviolet lamp (365 nm).
As a preferable technical scheme of the invention, the method also comprises a fourth step, wherein different temperatures are examined, low temperature (5 ℃) and high temperature (35 ℃) are examined according to thin layer chromatography, and when the development temperature is low temperature (5 ℃) and high temperature (35 ℃) as a result, in the chromatogram of the test sample, the same orange and yellow green spots are displayed at the corresponding positions of the chromatogram of the reference sample, the spots are clear, the separation degree is good, the Rf value is consistent, and the negative sample and the blank reference sample do not interfere. Showing that the development conditions of different temperatures have no obvious influence on the thin layer identification method.
The invention also comprises a detection instrument, a thin layer chromatography cylinder, a quantitative capillary, an ultrasonic cleaner, an electronic analytical balance and a silica gel G thin layer plate.
As a preferred technical scheme, the invention also comprises reagent reagents, finished Naoan dripping pills, negative samples of the Naoan dripping pills, safflower control medicinal materials, ethyl acetate, water, glacial acetic acid, methanol, acetone and 75% ethanol, wherein the reagents are analytically pure.
Compared with the prior art, the invention has the beneficial effects that:
in the scheme of the invention:
1. the preparation method comprises grinding the above materials, packaging with filter paper bag to prevent powder leakage, adding 75% ethanol, sealing, ultrasonic filtering, evaporating filtrate, cooling, and dissolving with 85% acetone to obtain test solution. Preparing control solution from 1g of Carthami flos by the same method; step two, respectively sucking 5 mul of each sample solution and the reference solution, respectively dropping the sample solution and the reference solution on the same silica gel G thin plate, developing by using ethyl acetate-water-glacial acetic acid-methanol as a developing agent, taking out, airing, spraying a sulfuric acid ethanol solution, and inspecting under an ultraviolet lamp (365 nm);
2. a blank control was prepared by formulating 85% acetone. Sucking the above two solutions, respectively dropping the sample solution and the reference solution on the same silica gel G thin layer plate, developing with ethyl acetate-water-glacial acetic acid-methanol as developing agent, taking out, air drying, spraying sulfuric acid ethanol solution, and inspecting under ultraviolet lamp (365 nm). And step four, examining different temperatures, examining low temperature (5 ℃) and high temperature (35 ℃) according to thin-layer chromatography, and when the development temperature is low temperature (5 ℃) and high temperature (35 ℃), displaying the same orange and yellow green spots on the corresponding positions of the chromatogram of the reference substance in the chromatogram of the test substance, wherein the spots are clear, the separation degree is good, and the Rf values are consistent.
Description of the drawings:
FIG. 1 shows a preparation method of a sample and a formula of a developing agent according to the present invention;
FIG. 2 is a diagram of the preparation method of different detection products and the thin layer chromatography identification method of the developer combination provided by the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the attached tables. It is clear that the described embodiment is a specific implementation of the invention and is not limited to all embodiments.
Thus, the following detailed description of the embodiments of the invention is not intended to limit the scope of the invention as claimed, but is merely representative of some embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that the embodiments and features and aspects of the embodiments of the present invention may be combined with each other without conflict, and it should be noted that like reference numerals and letters refer to like items in the following attached tables, and thus, once an item is defined in one attached table, it need not be further defined and explained in the following attached tables.
Example (b): 1, please refer to table 1-2, a thin-layer chromatography detection method for safflower in Naoan drop pills, comprising the steps of preparing the detection product, grinding the product, subpackaging with filter paper bags to prevent powder leakage, adding 75% ethanol, sealing, performing ultrasonic filtration, evaporating the filtrate to dryness, cooling, and adding 85% acetone to dissolve the product to obtain a test solution. Preparing control solution from 1g of Carthami flos by the same method;
and step two, respectively sucking 5 mul of each sample solution and the reference solution, respectively dropping the sample solution and the reference solution on the same silica gel G thin plate, developing by using ethyl acetate-water-glacial acetic acid-methanol as a developing agent, taking out, airing, spraying a sulfuric acid ethanol solution, and inspecting under an ultraviolet lamp (365 nm).
Preparing a test sample, grinding the test sample, subpackaging with filter paper bags, adding 75% ethanol, placing in a conical flask, sealing, performing ultrasonic treatment, filtering, evaporating to dryness, cooling, adding 85% acetone, and dissolving to obtain a test sample solution. Preparing 1g of safflower control medicinal material, and preparing a control solution by the same method; developing agent formula I, ethyl acetate-water-glacial acetic acid-methanol; the first color development method is to spray sulfuric acid ethanol solution and to inspect the color under ultraviolet light (365 nm).
And secondly, preparing a test sample, grinding the test sample, subpackaging the ground test sample into filter paper bags, adding 85% acetone for soaking, adding 75% ethanol, placing the mixture into a conical flask, sealing the conical flask, performing ultrasonic treatment, filtering, evaporating to dryness, cooling, adding 85% acetone for dissolving, and taking the mixture as a test sample solution. Preparing 1g of safflower control medicinal material, and preparing a control solution by the same method; developing agent formula II, ethyl acetate-water-glacial acetic acid-methanol; and a second color development method comprises spraying sulfuric acid ethanol solution and carrying out inspection under ultraviolet light (365 nm).
And thirdly, adding 25ml of acetone into the test sample, soaking, removing the acetone, volatilizing at normal temperature, heating in hot water bath, continuously operating twice, evaporating the water solution to dryness, and adding 85% acetone to dissolve the acetone to obtain a test sample solution. Preparing control solution from 1g of Carthami flos by the same method; developing agent formula III, ethyl acetate-water-glacial acetic acid-methanol; and a third color development method, namely, using sulfuric acid ethanol solution to carry out visual inspection under ultraviolet light (254 nm).
The fourth step of preparing the test sample is to take the test sample, add methanol, perform ultrasonic treatment, add hot water to dissolve, extract with ethyl acetate for 2 times, combine ethyl acetate solutions, and concentrate to obtain a test sample solution. Preparing control solution from 1g of Carthami flos by the same method; the developing solvent is composed of chloroform-ethyl acetate-glacial acetic acid; and the fourth color development method is to use sulfuric acid ethanol solution to inspect under the sunlight.
And fifthly, preparing the detection product, dissolving the detection product in hot water, filtering, evaporating to dryness, adding 85% acetone, performing low-temperature ultrasonic treatment, and concentrating to obtain a test solution. Preparing control solution from 1g of Carthami flos by the same method; developing agent formula five, trichloromethane-ethyl acetate-glacial acetic acid; and a fifth color development method, and visual inspection under an ultraviolet lamp (254 nm).
And step three, special investigation is carried out, and 85% acetone is prepared to be used as a blank reference substance. Sucking the above two solutions, respectively dropping the sample solution and the reference solution on the same silica gel G thin layer plate, developing with ethyl acetate-water-glacial acetic acid-methanol as developing agent, taking out, air drying, spraying sulfuric acid ethanol solution, and inspecting under ultraviolet lamp (365 nm).
And step four, inspecting different temperatures, inspecting the low temperature (5 ℃) and the high temperature (35 ℃) according to the thin-layer chromatography, and when the development temperature is the low temperature (5 ℃) and the high temperature (35 ℃) as a result, displaying the same orange and yellow green spots on the corresponding positions of the chromatogram of the reference substance in the chromatogram of the test substance, wherein the spots are clear, the separation degree is good, the Rf value is consistent, and the negative sample and the blank reference substance are not interfered. Showing that the development conditions of different temperatures have no obvious influence on the thin layer identification method.
A detection instrument, a thin layer chromatography cylinder, a quantitative capillary tube, an ultrasonic cleaner, an electronic analytical balance and a silica gel G thin layer plate. Reagent reagents, namely a finished product of the Naoan dropping pill, a negative sample of the Naoan dropping pill, a safflower control medicinal material, ethyl acetate, water, glacial acetic acid, methanol, acetone and 75% ethanol, wherein the reagents are analytically pure.
The embodiment is as follows: 2, required instruments, a thin layer chromatography cylinder, a quantitative capillary, an ultrasonic cleaner, an electronic analytical balance and a silica gel G thin layer plate.
Reagent reagents, namely a finished product of the Naoan dropping pill, a negative sample of the Naoan dropping pill, a safflower control medicinal material, ethyl acetate, water, glacial acetic acid, methanol, acetone and 75% ethanol, wherein the reagents are analytically pure.
The thin layer chromatography identification method comprises respectively dropping 5 μ l of sample and reference solution on the same silica gel G thin plate, developing with ethyl acetate-water-glacial acetic acid-methanol as developing agent, taking out, air drying, spraying sulfuric acid ethanol solution, and inspecting under ultraviolet lamp (365 nm).
The preparation method of the test substance comprises grinding the product, packaging with filter paper bag to prevent powder leakage, adding 75% ethanol, sealing, ultrasonic filtering, evaporating filtrate, cooling, and dissolving with 85% acetone to obtain test solution. Preparing 1g of safflower control medicinal material, and preparing a control solution by the same method.
Referring to table 1-2, test article preparation one, grinding test article, packaging with filter paper, adding 75% ethanol, placing into a conical flask, sealing, ultrasonic treating, filtering, evaporating, cooling, and dissolving with 85% acetone to obtain test solution. Preparing 1g of safflower control medicinal material, and preparing a control solution by the same method; developing agent formula I, ethyl acetate-water-glacial acetic acid-methanol; the first color development method is that sulfuric acid ethanol solution is sprayed to be inspected under ultraviolet light (365 nm);
and secondly, preparing a test sample, grinding the test sample, subpackaging the ground test sample into filter paper bags, adding 85% acetone for soaking, adding 75% ethanol, placing the mixture into a conical flask, sealing the conical flask, performing ultrasonic treatment, filtering, evaporating to dryness, cooling, adding 85% acetone for dissolving, and taking the mixture as a test sample solution. Preparing control solution from 1g of Carthami flos by the same method; developing solvent formula II, ethyl acetate-water-glacial acetic acid-methanol; secondly, spraying sulfuric acid ethanol solution and carrying out inspection under ultraviolet light (365 nm);
and thirdly, adding 25ml of acetone into the detection product, soaking, removing the acetone, volatilizing the acetone at normal temperature, heating in hot water bath, continuously operating twice, evaporating the water solution to dryness, and dissolving the acetone with the concentration of 85% to obtain a test solution. Preparing 1g of safflower control medicinal material, and preparing a control solution by the same method; developing solvent formula III, ethyl acetate-water-glacial acetic acid-methanol; a third color development method, using sulfuric acid ethanol solution to carry out inspection under ultraviolet light (254 nm);
and fourthly, adding methanol into the detection product, performing ultrasonic treatment, adding hot water to dissolve the detection product, extracting the detection product for 2 times by using ethyl acetate, combining ethyl acetate solutions, and concentrating the ethyl acetate solutions to obtain a test solution. Preparing control solution from 1g of Carthami flos by the same method; developing agent formula IV, trichloromethane-ethyl acetate-glacial acetic acid; viewing the obtained product in a sulfuric acid ethanol solution in sunlight;
and fifthly, preparing the detection product, dissolving the detection product in hot water, filtering, evaporating to dryness, adding 85% acetone, performing low-temperature ultrasonic treatment, and concentrating to obtain a test solution. Preparing control solution from 1g of Carthami flos by the same method; developing agent formula five, trichloromethane-ethyl acetate-glacial acetic acid; a fifth color development method, placing under an ultraviolet lamp (254 nm) for inspection;
the working principle is as follows: the using process of the invention comprises the steps of preparing a detection product, grinding the product, subpackaging filter paper bags to prevent powder leakage, adding 75% ethanol, sealing, carrying out ultrasonic filtration, evaporating filtrate to dryness, cooling, and adding 85% acetone to dissolve the mixture to be used as a test solution. Preparing control solution from 1g of Carthami flos by the same method; and step two, respectively sucking 5 mul of each sample solution and the reference solution, respectively dropping the sample solution and the reference solution on the same silica gel G thin plate, developing by using ethyl acetate-water-glacial acetic acid-methanol as a developing agent, taking out, airing, spraying a sulfuric acid ethanol solution, and inspecting under an ultraviolet lamp (365 nm).
Preparing a test sample, grinding the test sample, subpackaging with filter paper bags, adding 75% ethanol, placing into a conical flask, sealing, performing ultrasonic treatment, filtering, evaporating to dryness, cooling, adding 85% acetone, and dissolving to obtain a test sample solution. Preparing control solution from 1g of Carthami flos by the same method; developing agent formula I, ethyl acetate-water-glacial acetic acid-methanol; the first color development method is to spray sulfuric acid ethanol solution and to inspect the color under ultraviolet light (365 nm). And secondly, preparing a test sample, grinding the test sample, subpackaging the ground test sample into filter paper bags, adding 85% acetone for soaking, adding 75% ethanol, placing the mixture into a conical flask, sealing the conical flask, performing ultrasonic treatment, filtering, evaporating to dryness, cooling, adding 85% acetone for dissolving, and taking the mixture as a test sample solution. Preparing control solution from 1g of Carthami flos by the same method; developing solvent formula II, ethyl acetate-water-glacial acetic acid-methanol; and a second color development method comprises spraying sulfuric acid ethanol solution and carrying out inspection under ultraviolet light (365 nm). And thirdly, adding 25ml of acetone into the test sample, soaking, removing the acetone, volatilizing at normal temperature, heating in hot water bath, continuously operating twice, evaporating the water solution to dryness, and adding 85% acetone to dissolve the acetone to obtain a test sample solution. Preparing control solution from 1g of Carthami flos by the same method; developing solvent formula III, ethyl acetate-water-glacial acetic acid-methanol; and a third color development method, namely, using sulfuric acid ethanol solution to carry out inspection under ultraviolet light (254 nm). Preparation of test sample four the test sample is dissolved in hot water by adding methanol, ultrasonic treating, extracting with ethyl acetate for 2 times, mixing ethyl acetate solutions, and concentrating to obtain test sample solution. Preparing control solution from 1g of Carthami flos by the same method; the developing solvent is composed of chloroform-ethyl acetate-glacial acetic acid; and the fourth color development method is to use sulfuric acid ethanol solution to inspect under the sunlight. And fifthly, preparing the detection product, dissolving the detection product in hot water, filtering, evaporating to dryness, adding 85% acetone, performing low-temperature ultrasonic treatment, and concentrating to obtain a test solution. Preparing control solution from 1g of Carthami flos by the same method; developing agent formula five, trichloromethane-ethyl acetate-glacial acetic acid; and a fifth color development method, and visual inspection under an ultraviolet lamp (254 nm).
And step three, special investigation is carried out, and 85% acetone is prepared to be used as a blank reference substance. Sucking the above two solutions, respectively dropping the sample solution and the reference solution on the same silica gel G thin layer plate, developing with ethyl acetate-water-glacial acetic acid-methanol as developing agent, taking out, air drying, spraying sulfuric acid ethanol solution, and inspecting under ultraviolet lamp (365 nm). And step four, inspecting different temperatures, inspecting low temperature (5 ℃) and high temperature (35 ℃) according to thin-layer chromatography, and when the development temperature of the result is low temperature (5 ℃) and high temperature (35 ℃), displaying the same orange and yellow green spots on the corresponding positions of the chromatogram of the reference substance in the chromatogram of the test substance, wherein the spots are clear, the separation degree is good, the Rf value is consistent, and the negative sample and the blank reference substance are not interfered. Showing that the development conditions of different temperatures have no obvious influence on the thin layer identification method. A detection instrument, a thin layer chromatography cylinder, a quantitative capillary tube, an ultrasonic cleaner, an electronic analytical balance and a silica gel G thin layer plate. Reagent reagents, namely a finished product of the Naoan dropping pill, a negative sample of the Naoan dropping pill, a safflower control medicinal material, ethyl acetate, water, glacial acetic acid, methanol, acetone and 75% ethanol, wherein the reagents are analytically pure.
The above embodiments are only used for illustrating the invention and not for limiting the technical solutions described in the invention, and although the present invention has been described in detail in the present specification with reference to the above embodiments, the present invention is not limited to the above embodiments, and therefore, any modification or equivalent replacement of the present invention is made; all such modifications and variations are intended to be included herein within the scope of this disclosure and the appended claims.
Claims (10)
1. A thin-layer chromatography detection method for safflower in Naoan dropping pills is characterized by comprising the following steps:
step one, preparing a detection product, grinding the detection product, subpackaging filter paper bags to prevent powder leakage, adding 75% ethanol, sealing, performing ultrasonic filtration, evaporating filtrate to dryness, cooling, adding 85% acetone to dissolve the filtrate to obtain a test solution, taking 1g of a safflower control medicinal material, and preparing the control solution by the same method;
and step two, respectively sucking 5 mu l of each sample solution and 5 mu l of each reference substance solution, respectively dropping the sample solution and the reference substance solution on the same silica gel G thin plate, taking out the sample solution and the reference substance solution by taking ethyl acetate-water-glacial acetic acid-methanol as developing agents, airing the sample solution, spraying sulfuric acid ethanol solution, and placing the sample solution and the reference substance solution under an ultraviolet lamp (365 nm) for inspection.
2. The method for detecting thin-layer chromatography of safflower in Naoan dripping pill according to claim 1, characterized in that it further comprises a first step of preparing a detection product, wherein the first step of preparing a detection product is grinding the detection product, subpackaging with a filter paper bag, adding 75% ethanol into a conical flask, sealing, performing ultrasonic treatment, filtering, evaporating to dryness, cooling, adding 85% acetone for dissolution to obtain a test solution, and preparing 1g of a safflower control drug to obtain a control solution in the same manner; developing agent formula I, ethyl acetate-water-glacial acetic acid-methanol; the first color development method is to spray sulfuric acid ethanol solution and to detect under ultraviolet light (365 nm).
3. The method for detecting thin-layer chromatography of safflower in Naoan dripping pills according to claim 2, characterized in that, the method further comprises a second preparation step of detection products, wherein the second preparation step of detection products is to grind the detection products, divide the ground detection products into filter paper bags, add 85% acetone for impregnation, add 75% ethanol into conical flasks, seal plugs, carry out ultrasonic treatment, filter, evaporate to dryness, cool the products, add 85% acetone for dissolution, use the solution as a test solution, take 1g of safflower control medicine, and prepare the control solution in the same way; developing agent formula II, ethyl acetate-water-glacial acetic acid-methanol; and a second color development method comprises spraying sulfuric acid ethanol solution and carrying out inspection under ultraviolet light (365 nm).
4. The method for detecting thin-layer chromatography of safflower in Naoan dripping pill according to claim 3, characterized in that, the method further comprises the third step of preparing a third detection product, wherein the third step of preparing the detection product comprises adding 25ml of acetone into the detection product, immersing, removing the acetone, volatilizing the acetone at normal temperature, heating in hot water in a water bath, continuously operating twice, evaporating the water solution, adding 85% acetone to dissolve the acetone to obtain a test solution, preparing 1g of safflower control drug, and preparing the control solution in the same way; developing agent formula III, ethyl acetate-water-glacial acetic acid-methanol; and a third color development method, namely, using sulfuric acid ethanol solution to carry out inspection under ultraviolet light (254 nm).
5. The method for detecting thin-layer chromatography of safflower in Naoan dripping pill according to claim 4, characterized in that, the method further comprises preparing a fourth detection product, adding methanol to the fourth detection product, performing ultrasonic treatment, dissolving with hot water, extracting with ethyl acetate for 2 times, combining ethyl acetate solutions, concentrating to obtain a test solution, taking 1g of safflower control material, and preparing a control solution by the same method; developing agent formula IV, trichloromethane-ethyl acetate-glacial acetic acid; and the fourth color development method is to use sulfuric acid ethanol solution to inspect under the sunlight.
6. The method of claim 5, further comprising preparing the test product as five, dissolving the test product in hot water, filtering, evaporating, adding 85% acetone, performing low temperature ultrasonic treatment, concentrating to obtain a test solution, collecting 1g of a control material, and preparing a control solution by the same method; developing agent formula five, trichloromethane-ethyl acetate-glacial acetic acid; and a fifth color development method, and visual inspection under an ultraviolet lamp (254 nm).
7. The method for detecting safflower carthamus tinctorius thin layer chromatography in Naoan dripping pills according to claim 6, further comprising three special attribute studies, preparing 85% acetone as a blank reference substance, respectively dropping the two solutions and the test solution and the reference substance solution on the same silica gel G thin layer plate, developing with ethyl acetate-water-glacial acetic acid-methanol as developing agent, taking out, air drying, spraying sulfuric acid ethanol solution, and inspecting under ultraviolet lamp (365 nm).
8. The method for detecting the thin-layer chromatography of the safflower in the Naoan dropping pill according to claim 7, which is characterized by further comprising a step four of examining different temperatures, wherein according to the thin-layer chromatography, the examination is carried out at a low temperature (5 ℃) and a high temperature (35 ℃), and when the development temperature is the low temperature (5 ℃) and the high temperature (35 ℃), the same orange and yellow green spots appear on the chromatogram of the test sample at the corresponding positions with the chromatogram of the reference sample, the spots are clear, the separation degree is good, the Rf value is consistent, the negative sample and the blank reference sample are free of interference, which indicates that the development conditions at different temperatures have no obvious influence on the thin-layer identification method.
9. The method for detecting safflower carthamus thin-layer chromatography in Naoan dropping pills according to claim 8, which is characterized by further comprising a detection instrument, a thin-layer chromatography cylinder, a quantitative capillary tube, an ultrasonic cleaner, an electronic analytical balance and a silica gel G thin-layer plate.
10. The method for detecting safflower carthamus thin-layer chromatography in Naoan dropping pills according to claim 9, which is characterized by further comprising reagent reagents, finished Naoan dropping pills, naoan dropping pill negative samples, safflower control medicinal materials, ethyl acetate, water, glacial acetic acid, methanol, acetone and 75% ethanol, wherein the reagents are analytically pure.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210885788.3A CN115184536A (en) | 2022-07-26 | 2022-07-26 | Thin-layer chromatography detection method for safflower in Naoan dropping pills |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210885788.3A CN115184536A (en) | 2022-07-26 | 2022-07-26 | Thin-layer chromatography detection method for safflower in Naoan dropping pills |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115184536A true CN115184536A (en) | 2022-10-14 |
Family
ID=83521302
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210885788.3A Pending CN115184536A (en) | 2022-07-26 | 2022-07-26 | Thin-layer chromatography detection method for safflower in Naoan dropping pills |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115184536A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101732387A (en) * | 2009-12-28 | 2010-06-16 | 辽源誉隆亚东药业有限责任公司 | Detection method of Naoan capsules |
-
2022
- 2022-07-26 CN CN202210885788.3A patent/CN115184536A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101732387A (en) * | 2009-12-28 | 2010-06-16 | 辽源誉隆亚东药业有限责任公司 | Detection method of Naoan capsules |
Non-Patent Citations (3)
| Title |
|---|
| 任红兵: "痛经灵颗粒质量标准研究", 现代中药研究与实践, vol. 25, no. 2, pages 76 - 79 * |
| 李清林 等: "补肾活血颗粒中红花、附子和甘草的薄层鉴别研究", 中华中医药学刊, vol. 31, no. 4, pages 817 - 818 * |
| 黄衍民 等: "吸收度法控制丹红注射液质量研究", 时珍国医国药, vol. 10, no. 10, pages 754 - 755 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107843677B (en) | Radix paeoniae rubra control extract and preparation method and application thereof | |
| CN106822203A (en) | A kind of levisticum particle and preparation method thereof and method of quality control | |
| CN109142588A (en) | A kind of LianZhixiaoyan Capsule HPLC characteristic spectrum and its construction method and application | |
| CN113759034A (en) | Establishing method of ophiopogon japonicus soup substance standard | |
| CN108088715B (en) | Moutan bark reference extract and preparation method and application thereof | |
| CN113109495B (en) | Quality detection method of jujube kernel nerve-soothing capsules based on thin-layer chromatography | |
| CN112858497B (en) | Method for constructing characteristic spectrum of rhizoma Menispermi standard decoction and detecting quality of rhizoma Menispermi standard decoction | |
| CN112946132B (en) | Chinese rose medicinal material fingerprint spectrum, construction method thereof and Chinese rose medicinal material quality detection method | |
| CN115184536A (en) | Thin-layer chromatography detection method for safflower in Naoan dropping pills | |
| CN106596759B (en) | A kind of HPLC analysis method of Ramulus et folium taxi cuspidatae extract and its preparation | |
| CN110927322A (en) | Detection method of agastache rugosus healthy energy mixture | |
| CN113484429B (en) | Method for establishing standard of peach pit qi-bearing soup material | |
| CN112083097B (en) | Thin-layer identification method for simultaneously identifying ferulic acid, calycosin glucoside and hesperidin | |
| CN114814068B (en) | Efficient thin-layer identification method for abrus herb and abrus herb | |
| CN118225962B (en) | Thin-layer identification method for rhizoma Curcumae, rhizoma anemarrhenae and rhizoma Ligustici Chuanxiong in traditional Chinese medicine compound containing herba Epimedii and application thereof | |
| CN112748213B (en) | Thin-layer chromatography identification method for single needle | |
| CN112630324B (en) | Method for constructing and detecting characteristic spectrums of platycladi seed medicinal materials, decoction pieces, standard decoction and formula granules | |
| CN110763642B (en) | Detection method of traditional Chinese medicine preparation | |
| CN116858982A (en) | Application of developing agent in thin layer identification method and thin layer identification method | |
| Li et al. | Thin-layer chromatographic quantification of three alkaloid compounds in Sophora alopecuroides and TLC—bioautography for screening antioxidant components | |
| CN107607669A (en) | A kind of quality determining method of safflower and safflower granule | |
| CN114487254B (en) | Quality control method for fried rice sprouts | |
| CN115166129A (en) | Thin-layer chromatography detection method for angelica sinensis in Naoan dropping pills | |
| CN117288882A (en) | Quality control method of compound pseudo-ginseng stomach-ache capsule | |
| CN118624801A (en) | Pseudo-ginseng saponin F in Jinchai capsule11Thin layer identification method of (2) |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20221014 |
|
| RJ01 | Rejection of invention patent application after publication |