CN117849258A - Qualitative identification method for water-soluble components of safflower - Google Patents
Qualitative identification method for water-soluble components of safflower Download PDFInfo
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000004809 thin layer chromatography Methods 0.000 claims abstract description 10
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- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 39
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 27
- 239000000463 material Substances 0.000 claims description 27
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 13
- 238000001914 filtration Methods 0.000 claims description 13
- 239000003960 organic solvent Substances 0.000 claims description 10
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 8
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- 241000628997 Flos Species 0.000 description 2
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- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of detection of traditional Chinese medicine components, and particularly relates to a qualitative identification method of safflower water-soluble components. The qualitative identification method of the water-soluble components in the safflower adopts water as an extraction solvent to extract the water-soluble components in the safflower, and then adopts thin layer chromatography to identify, thereby effectively solving the technical problem that the safflower can not be effectively detected after being boiled in water in the prior art, and being suitable for the detection of classical prescription preparations containing the safflower.
Description
Technical Field
The invention belongs to the technical field of detection of traditional Chinese medicine components, and particularly relates to a qualitative identification method of safflower water-soluble components.
Background
The safflower alias safflower and thorn safflower are plants of the genus safflower of the family Compositae, and have the effects of activating blood circulation, removing blood stasis, relieving Yu Kang depression, resisting inflammation, easing pain, regulating menstruation, activating collaterals, improving digestion and the like. Safflower contains a large amount of flavonoids, and also contains volatile oil, phytosterol, polysaccharide, amino acid, fatty oil, etc. The main component of the volatile oil is alkyne mixture, and safflower polysaccharide consists of glucose, xylose, arabinose and galactose which are connected by beta-bond.
In the prior art, the identification of the components in the safflower is carried out by extracting the safflower with an organic solvent, and then the identification is carried out, the method is only suitable for identifying the fat-soluble components in the safflower, the water-soluble components can not be effectively detected, and the extracted components contain organic solvent residues. The identification method of safflower recorded in Chinese pharmacopoeia is as follows: adding 80% acetone solution 5mL into 0.5g of safflower powder, sealing, shaking for 15min, standing, collecting supernatant as sample solution, and collecting 0.5g of safflower control medicinal material to obtain control medicinal material solution. According to a thin layer chromatography test, 50 mu L of each of the two solutions is absorbed and respectively spotted on the same silica gel H thin layer plate, and the two solutions are spread by using ethyl acetate-formic acid-water-methanol (7:2:3:0.4) as a developing agent, taken out and dried.
In addition, patent publication No. CN103940929A discloses a detection method of a traditional Chinese medicine injection for treating cardiovascular and cerebrovascular diseases, the detection method comprises a thin layer identification method of red sage root and safflower medicinal materials, and the identification method of the safflower medicinal materials is as follows: taking 5ml of the injection, regulating the pH value to 2 by using dilute hydrochloric acid, shaking and extracting by using 10ml of ethyl acetate, separating ethyl acetate solution, evaporating to dryness, and adding 2ml of absolute ethyl alcohol into residues to dissolve the residues to obtain a sample solution. 0.5g of safflower reference medicine is added with 25mL of water, reflux is carried out for 2 hours, cooling is carried out, filtration is carried out, the pH value of the filtrate is regulated to 2 by dilute hydrochloric acid, the filtrate is extracted by shaking with 20mL of ethyl acetate, ethyl acetate liquid is separated and evaporated to dryness, and the residue is added with 1mL of absolute ethyl alcohol to be dissolved to be used as reference medicine solution. According to a thin layer chromatography test, absorbing 10 mu L of each of the test solution and the control medicinal solution under the identification (1), and respectively spotting on the same silica gel GF254 thin layer plate according to the volume ratio of 10:0.5:0.5 trichloromethane-acetone-98% formic acid as developing agent, developing, taking out, air drying, and inspecting under 254nm ultraviolet lamp, wherein spots with the same color appear on the corresponding position of the chromatogram of the sample to be tested. Although water is used as an extraction solvent, the method is used for identifying the injection, and is not a safflower medicinal material.
In summary, the prior art has the technical problem that the effective detection cannot be performed after the safflower is boiled.
Disclosure of Invention
In order to solve the technical problems, the invention provides a qualitative identification method of water-soluble components of safflower. The qualitative identification method of the water-soluble components of the safflower adopts water as an extraction solvent to extract the water-soluble components in the safflower, and then adopts thin layer chromatography to identify, thereby effectively solving the technical problem that the safflower can not be effectively detected after being boiled in water in the prior art, and being suitable for the detection of the compound preparation containing the safflower.
In order to achieve the above object, the technical scheme of the present invention is as follows:
a qualitative identification method of water-soluble components of safflower comprises the following steps:
s1, preparing a sample solution: adding water into safflower, heating and refluxing for extraction, filtering, adding organic solvent into filtrate for extraction, merging extract solutions, volatilizing to obtain residues, and adding absolute ethanol into the residues for dissolution to obtain a sample solution;
s2, preparing a medicinal material control solution: adding water into safflower control medicinal materials, heating and refluxing for extraction, filtering, adding an organic solvent into filtrate for extraction, merging extract solutions, volatilizing to obtain residues, and adding absolute ethanol into the residues for dissolution to obtain medicinal material control solution;
s3, identifying by thin layer chromatography: respectively sucking the sample solution prepared in the step S1 and the medicinal material control solution prepared in the step S2, spotting on the same silica gel thin layer plate, spreading in a spreading agent, taking out, air drying, and inspecting under an ultraviolet lamp.
Further, the mass ratio of the safflower to the water in the step S1 and the step S2 is 1:20-80.
Further, the reflux extraction temperature in the step S1 and the step S2 is 80-100 ℃, and the reflux extraction time is 1-3h.
Further, after the filtration in the step S1 and the step S2, the filtrate was cooled to room temperature.
Further, the organic solvent in the step S1 and the step S2 is diethyl ether, the dosage of the organic solvent is 0.2-5 times of the volume of the filtrate, the extraction mode is shaking extraction, and the extraction times are 1-5 times.
Further, the dosage of the absolute ethyl alcohol in the step S1 and the step S2 is 1-6 times of the mass of the safflower.
Further, the volatilizing in the step S1 and the step S2 may be replaced by evaporating.
Further, the sucking amount of the sample solution and the medicinal material control solution in the step S3 is 10-30 mu L.
Further, the silica gel thin layer plate in the step S3 is a high-efficiency silica gel G thin layer plate or a high-efficiency silica gel GF254 thin layer plate.
Further, the developing agent in the step S3 is a mixture of cyclohexane, dichloromethane and ethyl acetate.
Further, the mass ratio of cyclohexane, dichloromethane and ethyl acetate in the developing agent is 2-6:1:1.
Further, the wavelength of the ultraviolet lamp in the step S3 is 245nm or 365nm.
Compared with the prior art, the qualitative identification method of the safflower water-soluble component provided by the invention has the following technical advantages:
(1) The qualitative identification method of the water-soluble components of the safflower can effectively detect the safflower decoction, and is suitable for the compound preparation containing the safflower;
(2) The qualitative identification method of the safflower water-soluble components avoids the occurrence of the front edges of various solvents in the unfolding process, ensures the rounding of spots in the unfolding process and improves the identification accuracy;
(3) The qualitative identification method of the safflower water-soluble component does not need to add a color developing agent for color development reaction in a thin layer chromatography test;
(4) The qualitative identification method of the safflower water-soluble component provided by the invention is simple and convenient to operate, has quick and accurate separation effect and good reproducibility, and provides a basis for quality control of the compound preparation containing the safflower component.
Drawings
FIG. 1 is a view of example 1;
FIG. 2 is a view of example 2;
FIG. 3 is a view of example 3;
fig. 4 is a view of comparative example 1;
FIG. 5 is a view of comparative example 2;
FIG. 6 is a view of comparative example 3;
FIG. 7 is a view of comparative example 4;
in the figure, 1 is the chromatographic band of the medicinal material control solution, and 2 is the chromatographic band of the test sample solution.
Detailed Description
The present invention is further described below by way of specific embodiments, but the present invention is not limited to the following examples. Various modifications may be made by those skilled in the art in light of the basic idea of the invention, but are within the scope of the invention without departing from the basic idea of the invention.
Example 1, qualitative identification method of Water-soluble component of safflower
The qualitative identification method of the safflower water-soluble component comprises the following steps:
s1, preparing a sample solution: reflux extracting Carthami flos 0.5g with 10g water at 80deg.C for 3 hr, filtering, cooling the filtrate, shaking and extracting with diethyl ether for 1 time, wherein the diethyl ether amount is 0.2 times of the volume of the filtrate, mixing the extractive solutions, volatilizing to obtain residue, and dissolving the residue with 0.5g anhydrous ethanol to obtain sample solution;
s2, preparing a medicinal material control solution: adding 10g of water into 0.5g of safflower control medicinal material, heating and refluxing at 80 ℃ for extraction for 3 hours, filtering while the safflower control medicinal material is hot, cooling the filtrate, shaking and extracting the filtrate for 1 time by using diethyl ether, wherein the consumption of the diethyl ether is 0.2 times of the volume of the filtrate, merging the extracting solutions, volatilizing the extracting solutions to obtain residues, and adding 0.5g of absolute ethyl alcohol into the residues for dissolution to obtain medicinal material control solution;
s3, thin layer chromatography test (general rule 0502): respectively sucking 10 μl of the sample solution prepared in step S1 and the medicinal material control solution prepared in step S2, spotting on the same high-efficiency silica gel G thin layer plate, spreading in a spreading agent composed of cyclohexane, dichloromethane and ethyl acetate at a mass ratio of 2:1:1, taking out, air drying, and inspecting under ultraviolet light with wavelength of 365nm.
Example 2 qualitative identification method of Water-soluble component of safflower
The qualitative identification method of the safflower water-soluble component comprises the following steps:
s1, preparing a sample solution: adding 37.5g of water into 0.5g of safflower, heating and refluxing for extraction for 1h at 100 ℃, filtering while the safflower is still hot, cooling the filtrate, shaking and extracting for 5 times by using diethyl ether, wherein the use amount of the diethyl ether is 5 times of the volume of the filtrate, merging the extracting solutions, evaporating to dryness to obtain residues, and adding 3g of absolute ethyl alcohol into the residues for dissolution to obtain a sample solution;
s2, preparing a medicinal material control solution: adding 37.5g of water into 0.5g of safflower reference medicine, heating and refluxing for extraction for 1h at 100 ℃, filtering while the safflower reference medicine is hot, cooling the filtrate, shaking and extracting for 5 times by using diethyl ether, wherein the use amount of the diethyl ether is 5 times of the volume of the filtrate, merging the extracting solutions, evaporating to dryness to obtain residues, and adding 3g of absolute ethyl alcohol into the residues for dissolution to obtain medicine reference solution;
s3, thin layer chromatography test (general rule 0502): respectively sucking 30 mu L of the sample solution prepared in the step S1 and the medicinal material control solution prepared in the step S2, spotting on a thin layer plate of the same high-efficiency silica gel G thin layer plate, placing in a developing agent composed of cyclohexane, dichloromethane and ethyl acetate according to the mass ratio of 6:1:1, taking out, airing, and placing under an ultraviolet lamp with the wavelength of 365nm for inspection.
Example 3 qualitative identification method of Water-soluble component of safflower
The qualitative identification method of the safflower water-soluble component comprises the following steps:
s1, preparing a sample solution: reflux extracting Carthami flos 0.5g with 40g water at 90deg.C for 2 hr, filtering, cooling the filtrate, shaking and extracting with diethyl ether for 4 times, wherein the diethyl ether amount is 3 times of the volume of the filtrate, mixing the extractive solutions, volatilizing to obtain residue, and dissolving the residue with 2.5g anhydrous ethanol to obtain sample solution;
s2, preparing a medicinal material control solution: adding 40g of water into 0.5g of safflower control medicine, heating and refluxing at 90 ℃ for 2 hours, filtering while the mixture is hot, cooling the filtrate, shaking and extracting the filtrate for 4 times by using diethyl ether, wherein the use amount of the diethyl ether is 3 times of the volume of the filtrate, merging the extracting solutions, volatilizing the extracting solutions to obtain residues, and adding 2.5g of absolute ethyl alcohol into the residues for dissolving to obtain medicine control solution;
s3, thin layer chromatography test (general rule 0502): respectively sucking 20 mu L of the sample solution prepared in the step S1 and the medicinal material control solution prepared in the step S2, spotting on the same high-efficiency silica gel G thin-layer plate, placing in a developing agent consisting of cyclohexane, dichloromethane and ethyl acetate according to the mass ratio of 4:1:1, taking out, airing, and placing under an ultraviolet lamp with the wavelength of 365nm for inspection.
Comparative example 1,
This comparative example is similar to example 3, and differs from example 3 in that: methanol was used instead of cyclohexane in the developing solvent in this comparative example.
Comparative example 2,
This comparative example is similar to example 3, and differs from example 3 in that: toluene was used instead of methylene chloride in the developing agent in this comparative example.
Comparative example 3,
This comparative example is similar to example 3, and differs from example 3 in that: acetone was used instead of ethyl acetate in the developing solvent in this comparative example.
Comparative example 4,
This comparative example is similar to example 3, and differs from example 3 in that: the developing agent in this comparative example consists of cyclohexane, methylene chloride and ethyl acetate in a mass ratio of 1:3:4.
Test results: as can be seen from fig. 1, 2 and 3, spots of the same color and main fluorescent spots are respectively displayed on the positions corresponding to the chromatogram of the medicinal material control solution in the chromatogram of the sample solution, the spots are round and uniform, the accuracy of the identification process is improved, and meanwhile, in the embodiment, the cleaned chromatogram can be obtained by only once sample application, once spreading and once inspection, so that the once success rate of identifying the water-soluble components of the safflower is remarkably improved.
As can be seen from fig. 4, 5, 6 and 7, compared with the chromatogram of the medicinal material control solution, the spots of the chromatogram of the sample solution are blurred and incomplete, and the positions of the spots do not correspond, which indicates that the identification result of the water-soluble components of safflower is affected by changing the formulation components and the amounts of the developing agent in comparative examples 1 to 4.
The above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.
Claims (10)
1. The qualitative identification method of the water-soluble component of the safflower is characterized by comprising the following steps of:
s1, preparing a sample solution: adding water into safflower, heating and refluxing for extraction, filtering, adding organic solvent into filtrate for extraction, merging extract solutions, volatilizing to obtain residues, and adding absolute ethanol into the residues for dissolution to obtain a sample solution;
s2, preparing a medicinal material control solution: adding water into safflower control medicinal materials, heating and refluxing for extraction, filtering, adding an organic solvent into filtrate for extraction, merging extract solutions, volatilizing to obtain residues, and adding absolute ethanol into the residues for dissolution to obtain medicinal material control solution;
s3, identifying by thin layer chromatography: respectively sucking the sample solution prepared in the step S1 and the medicinal material control solution prepared in the step S2, spotting on the same silica gel thin layer plate, spreading in a spreading agent, taking out, air drying, and inspecting under an ultraviolet lamp.
2. The method for qualitative identification of water-soluble components of safflower according to claim 1, wherein the mass ratio of safflower to water in step S1 and step S2 is 1:20-80.
3. The qualitative identification method of water-soluble components of safflower according to claim 1, wherein the reflux extraction temperature in step S1 and step S2 is 80-100 ℃ and the reflux extraction time is 1-3h.
4. The method for qualitative identification of water-soluble components of safflower according to claim 1, wherein the filtrate is cooled to room temperature after the filtration in step S1 and step S2, and the evaporation may be replaced by evaporation.
5. The qualitative identification method of water-soluble components of safflower according to claim 1, wherein the organic solvent in step S1 and step S2 is diethyl ether, the amount of the organic solvent is 0.2-5 times the volume of the filtrate, the extraction is performed by shaking, and the number of times of extraction is 1-5 times.
6. The method for qualitatively identifying water-soluble components of safflower according to claim 1, wherein the absolute ethanol is used in an amount of 1 to 6 times the mass of safflower in step S1 and step S2.
7. The method for qualitative identification of water-soluble components of safflower according to claim 1, wherein the amount of the sample solution and the medicinal material control solution sucked in step S3 is 10 to 30 μl.
8. The method for qualitative identification of water-soluble components of safflower according to claim 1, wherein the developing agent in step S3 is a mixture of cyclohexane, methylene chloride and ethyl acetate.
9. The method for qualitatively identifying water-soluble components of safflower according to claim 8, wherein the mass ratio of cyclohexane, dichloromethane and ethyl acetate in the developing agent is 2-6:1:1.
10. The method for qualitatively identifying water-soluble components of safflower according to claim 1, wherein the ultraviolet light in step S3 has a wavelength of 245nm or 365nm, and the silica gel thin layer plate is a high-efficiency silica gel G thin layer plate or a high-efficiency silica gel GF254 thin layer plate.
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