CN118067911A - Thin-layer chromatography identification method for multiple components in Chinese patent medicine - Google Patents
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a thin-layer chromatography identification method for multiple components in a Chinese patent medicine, belonging to the technical field of detection of traditional Chinese medicines. Using ethyl acetate extract of Chinese medicinal materials of herba Paederiae, fructus crataegi, folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae as sample solution; taking herba Paederiae, fructus crataegi, folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae as reference materials, decocting with water slightly, filtering, concentrating the filtrate, shaking and extracting with ethyl acetate for 2-4 times, mixing ethyl acetate solutions, concentrating to obtain reference medicinal material solution, detecting the sample solution and reference material solution by thin layer chromatography, and determining herba Paederiae, fructus crataegi, folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae components. The invention only needs to prepare one sample solution, simultaneously identifies five medicinal materials, does not need expensive reference substances and complicated chromatographic column separation operation, shortens the identification time, reduces the cost, has strong specificity and good reproducibility, and provides a new idea for thin-layer chromatographic identification of the hawthorn inner gold preparation.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine detection, and particularly relates to a thin-layer chromatography identification method for multiple components in a Chinese patent medicine.
Background
The traditional Chinese medicine is the magnificent of China, the safety, the effectiveness and the quality controllability of the traditional Chinese medicine are basic requirements, and the quality standard is taken as the speaking right and is the pioneer leading factor of the development of the traditional Chinese medicine. Currently, traditional quality control modes of Chinese patent medicines have certain limitations: different medicines need to correspond to different sample processing methods and thin-layer chromatography conditions, and meanwhile, in order to avoid the interference of negative samples, multiple separation and purification means are often needed to remove impurity interference, so that the operation is complicated, and the detection workload is heavy.
The haw endojin preparation includes haw endojin oral liquid and haw endojin capsule, is a kind of medicine composition with natural plant and animal medicine as main medicinal components, and is a kind of medicine of Shenwei medicine company. The preparation is prepared from eight medicinal materials including hawthorn, tibetan calamus, shepherd's purse, fevervine, weeping forsythiae capsule, loquat leaf, cicada slough and chicken's gizzard-membrane, and has the effects of strengthening spleen and stomach, and removing food retention and resolving stagnation. Can be used for treating infantile malnutrition caused by food stagnation, inappetence, abdominal distention and pain, dyspepsia, and dysuria, and the existing standard of preparation is adopted in spleen and stomach partial volume of Chinese patent medicine standard assembly internal medicine. The original standard only carries out thin-layer identification on two medicines of the fevervine and the loquat leaf, but the feeding condition of three medicines of the hawthorn, the Tibetan calamus and the weeping forsythiae cannot be reflected.
The TLC identification method of the feverfew in the original standard shows that the color development of the characteristic spots is greatly influenced by temperature, weak characteristic spots appear at the corresponding positions of the sample and the reference medicinal material after long-time low-temperature color development, and interference spots exist near the characteristic spots after heating at 105 ℃ to influence judgment.
Disclosure of Invention
Aiming at the problems that the existing standard TLC detection method of the hawthorn inner gold preparation is not obvious in spots, has serious interference and single in existing standard detection components in the prior art, the invention provides a thin-layer chromatography identification method of multiple components in a Chinese patent medicine, and only one sample solution is needed to be prepared, so that five medicinal materials of fevervine, hawthorn, loquat leaf, sweelflag rhizome and fructus forsythiae can be identified simultaneously, the operation is simple, the detection efficiency is improved, and the detection cost is reduced.
The invention is realized by the following technical scheme:
A thin-layer chromatography identification method for multiple components in a Chinese patent medicine comprises the following steps:
(1) Preparation of test solution: comprises ethyl acetate extract of Chinese medicinal materials of herba Paederiae, fructus crataegi, folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae as sample solution;
(2) Preparation of control medicinal material solution: taking herba Paederiae, fructus crataegi, folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae as reference materials respectively, decocting with water slightly boiling, filtering, concentrating the filtrate, shaking and extracting with ethyl acetate for 2-4 times, mixing ethyl acetate solutions, concentrating to obtain reference medicinal material solution;
(3) Thin layer chromatography detection:
detecting the fevervine: the test solution and the herba Paederiae control medicinal material solution are respectively spotted on a thin layer plate, and the volume ratio is 9:2: developing 0.2 trichloromethane-methanol-concentrated ammonia solution as developing agent, taking out, air drying, inspecting under 365nm ultraviolet lamp, and detecting fluorescent spots of the same color at the position corresponding to herba Paederiae control medicinal material;
Detection of hawthorns and loquat leaves: the sample solution, the hawthorn control medicinal material solution and the loquat leaf control medicinal material solution are respectively spotted on a thin layer plate, and the volume ratio is 20:6:1, developing with cyclohexane-ethyl acetate-glacial acetic acid as developing agent, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until the color of spots is clear, inspecting under sunlight and 365nm ultraviolet lamp, and detecting spots and fluorescent spots of the same color at positions corresponding to fructus crataegi control medicinal material and folium Eriobotryae control medicinal material;
Detection of Tibetan calamus: the sample solution and the rhizoma acori graminei control medicinal material solution are respectively spotted on a thin layer plate, and the volume ratio is 9:1, developing with dichloromethane-ethyl acetate as developing agent, taking out, air drying, inspecting under 365nm ultraviolet lamp, and detecting fluorescent spots with the same color at the position corresponding to the rhizoma Acori Calami control medicinal material;
Detection of fructus forsythiae: the sample solution and the weeping forsythia control medicinal material solution are respectively spotted on a thin layer plate, and the volume ratio is 12:2.5:2: developing 0.2 trichloromethane-acetone-methanol-formic acid as developing agent, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until the color of spots is clear, and detecting spots of the same color at the positions corresponding to fructus forsythiae control medicinal materials.
Further, the Chinese patent medicine is a hawthorn inner gold preparation, and each milliliter of the test solution corresponds to 3-6 g of the Chinese patent medicine.
Further, the hawthorn inner gold preparation is hawthorn inner gold capsules or hawthorn inner gold oral liquid.
Further, the preparation method of the sample solution of the hawthorn endocyst capsule comprises the steps of adding methanol into the content of the hawthorn endocyst capsule for dissolving, carrying out ultrasonic treatment for 20-50 min, filtering, evaporating filtrate to dryness, adding water into residues for dissolving, extracting for 2-4 times by using ethyl acetate in a shaking way, combining ethyl acetate solutions, and concentrating to obtain the sample solution of the hawthorn endocyst capsule; the preparation method of the sample solution of the Hawthorn inner gold oral liquid comprises the steps of shaking and extracting the Hawthorn inner gold oral liquid with ethyl acetate for 2-4 times, combining the ethyl acetate solutions, and concentrating to obtain the sample solution of the Hawthorn inner gold oral liquid.
Further, the concentrated ammonia solution in the step (3) is ammonia water with the volume concentration of 25-28%.
Further, the thin layer plate in the step (3) is a silica gel G thin layer plate.
Further, the point sample amount in the step (3) is 3-10 mu L.
Further, 25 ml of water is added into each gram of the reference medicinal material in the step (2), and the micro-boiling decoction time is 20-40 min.
The beneficial effects obtained by the invention are as follows:
According to the thin-layer chromatography identification method provided by the invention, only one sample solution is needed to be prepared, so that five medicinal materials of fevervine, hawthorn, loquat leaf, tibetan calamus and fructus forsythiae in the Chinese patent medicine (hawthorn inner gold preparation) can be identified simultaneously, and expensive reference substances and complicated chromatographic column separation operations are not needed, so that the thin-layer identification time is shortened, the consumption of the expensive reference substances and organic reagents is reduced, and the detection cost is reduced; the method has the advantages of relatively simple operation, strong specificity and good repeatability, provides a new thought for thin-layer chromatography identification of the hawthorn inner gold preparation, can replace or supplement the existing standard content, is beneficial to quality control, inspection and detection of the hawthorn inner gold preparation, and enhances rationality and reliability.
Drawings
FIG. 1 is a thin-layer chromatography detection chart of herba Paederiae in the Hawthorn endocyst capsule of example 1, 1. Test article 22031702,2, test article 22031804,3, test article 21122701,4, herba Paederiae control medicinal material, 5, herba Paederiae negative control;
FIG. 2 is a chart of the chromatogram of haw and loquat She Baoceng in the capsule of Crataegus pinnatifida of example 1, (a) under sunlight, (b) under 365nm ultraviolet lamp; 1. sample 22031702,2, sample 22031804,3, sample 21122701,4, loquat leaf reference medicine, 5, haw reference medicine, 6, ursolic acid reference substance, 7, haw and loquat She Yinxing;
FIG. 3 is a thin layer chromatography detection chart of the Tibetan calamus in the Hawthorn endocardium capsule of example 1, a test article 22031702,2, a test article 22031804,3, a test article 21122701,4, a Tibetan calamus control medicinal material, and 5, a Tibetan calamus negative control;
FIG. 4 is a thin-layer chromatography detection chart of fructus forsythiae in the Hawthorn inner gold capsule of example 1, wherein the chart is a sample of 1, a sample of 22031702,2, a sample of 22031804,3, a sample of 21122701.4, a fructus forsythiae reference medicine, a fructus forsythiae glycoside reference substance, and a fructus forsythiae negative reference medicine;
FIG. 5 is a thin-layer chromatography detection chart of herba Paederiae in the capsule of Hawthorn fruit of comparative example 1, (a) at low temperature, (b) after heating at 105deg.C; 1. herba Paederiae control, 2, test piece 22031702,3, test piece 22031804,4, and test piece 21122701;
Fig. 6 is a chromatographic detection chart of loquat She Baoceng in the capsule of hawthorn endocardium of comparative example 2, 1, loquat leaf control medicine, 2, test piece 22031702,3, test piece 22031804,4, and test piece 21122701.
Detailed Description
In order to make the technical solution of the present application better understood, the following description of the technical solution of the present application will be made in a clear and complete manner, and other similar embodiments obtained by those skilled in the art without making any inventive effort on the basis of the embodiments of the present application shall fall within the scope of protection of the present application.
Hawthorn endocyst capsules in the following examples are manufactured by Shenwei pharmaceutical industry (Kunming) Co., ltd, and 10 batches of products are provided, wherein the production batch numbers are 22031701, 22031702, 22031803, 22031804, 21122701, 21122702, 21122803, 21122804, 21122905 and 21122906 respectively.
Example 1
(1) Preparation of test solution: adding 50ml of methanol into 4g of the content of the gold capsule in the hawthorn, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating the filtrate, adding 30ml of water into the residue to dissolve the residue, extracting for 3 times by shaking with ethyl acetate for 20ml each time, discarding water solution, combining ethyl acetate solutions, and concentrating to 1ml to obtain a sample solution;
(2) Preparation of control medicinal material solution: respectively taking 1g of herba Paederiae control medicinal material, 2g of fructus crataegi control medicinal material, 1g of folium Eriobotryae control medicinal material, 2g of rhizoma Acori Calami control medicinal material and 1g of fructus forsythiae control medicinal material, adding 50mL of water, boiling for 30min, filtering, concentrating the filtrate to 30mL, extracting with ethyl acetate for 3 times under shaking for 20mL each time, discarding water solution, mixing ethyl acetate solutions, concentrating to 0.5 mL, and collecting control medicinal material solution;
(3) Preparing a negative control medicinal material solution, namely respectively removing other medicinal materials of herba Paederiae, fructus crataegi, folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae in a prescription of the gold capsule in fructus crataegi, preparing negative control samples of herba Paederiae, fructus crataegi+folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae according to a prescription proportion, and respectively preparing a herba Paederiae negative control solution, a fructus crataegi and fructus Eriobotryae She Yinxing control solution, a rhizoma Acori Calami negative control solution and a fructus forsythiae negative control solution according to a preparation method of the sample solution in the step (1);
(4) Control solution: respectively taking appropriate amounts of ursolic acid and forsythin reference substances, and adding methanol to prepare a reference substance solution containing 0.5mg of ursolic acid per 1ml and a reference substance solution containing 1mg of forsythin per 1 ml;
(5) Thin layer chromatography detection:
Detecting the fevervine: respectively dotting 5 mu l of test solution (3 batches) and 5 mu l of fevervine control medicinal material solution and 5 mu l of fevervine negative control solution on a silica gel G thin-layer plate according to the volume ratio of 9:2:0.2 of chloroform-methanol-concentrated ammonia solution (25%) is used as a developing agent, and after developing, the solution is taken out, dried and inspected under an ultraviolet lamp of 365nm, the result is shown as a figure 1 (1. Sample 22031702,2. Sample 22031804,3. Sample 21122701.4. Fevervine control medicinal material, 5. Fevervine negative control), in the figure 1, fluorescent spots with the same color appear on the position corresponding to the fevervine control medicinal material in the chromatography of the sample, and the fevervine negative control is free of interference;
Detection of hawthorns and loquat leaves: 10 mu l of sample solution (3 batches), 3 mu l of haw control medicinal material solution, 3 mu l of loquat leaf control medicinal material solution, 10 mu l of haw and loquat She Yinxing control solution and 3 mu l of ursolic acid control solution are respectively dotted on the silica gel G thin-layer plate, and the volume ratio is 20:6:1, developing the cyclohexane-ethyl acetate-glacial acetic acid serving as a developing agent, taking out the developing agent, airing the developing agent, spraying 10% sulfuric acid ethanol solution, heating the developing agent at 105 ℃ until spots are clear, and viewing the developing agent under sunlight and 365nm ultraviolet light, wherein the results are shown in a graph in which (a) under the sunlight, (b) under 365nm ultraviolet light, (1) a sample 22031702,2, a sample 22031804,3, a sample 21122701,4, a loquat leaf control medicinal material, 5, a haw control medicinal material, 6, a ursolic acid control, 7, haw and a loquat She Yinxing control), spots and fluorescent spots with the same color are shown in the positions corresponding to the loquat leaf control medicinal material, the haw control medicinal material and the ursolic acid control, and the loquat leaf and haw negative control have no interference, good separation effect and clear spots;
Detection of Tibetan calamus: the method comprises the steps of respectively dispensing 10 mu l of sample solution (3 batches) and 5 mu l of Tibetan calamus control medicinal material solution and 10 mu l of Tibetan calamus negative control medicinal material solution on a silica gel G thin-layer plate according to the volume ratio of 9:1, developing with dichloromethane-ethyl acetate as developing agent, taking out, air drying, and inspecting under 365nm ultraviolet lamp to obtain the result shown in figure 3 (1. Sample 22031702,2. Sample 22031804,3. Sample 21122701,4. Rhizoma Acori Calami control material, 5. Rhizoma Acori Calami negative control), wherein fluorescent spots with the same color are detected at the corresponding position of rhizoma Acori Calami control material in the sample chromatogram, and the rhizoma Acori Calami negative control has no interference;
Detection of fructus forsythiae: the method comprises the steps of respectively dotting 5 mu l of sample solution (3 batches) and 5 mu l of fructus forsythiae control medicinal material solution and 5 mu l of fructus forsythiae negative control medicinal material solution on a silica gel G thin-layer plate according to a volume ratio of 12:2.5:2:0.2 of trichloromethane-acetone-methanol-formic acid serving as a developing agent, taking out the developing agent after developing, airing the developing agent, spraying 10% sulfuric acid ethanol solution, heating the developing agent at 105 ℃ until spots develop clearly, wherein the result is shown in a graph in fig. 4 (1. A sample 22031702,2. A sample 22031804,3. A sample 21122701.4. A forsythia control medicinal material, 5. A forsythin control, 6. A forsythia negative control medicinal material), spots with the same color are detected at positions corresponding to the forsythia control medicinal material and the forsythia glycoside control medicinal material in the chromatogram of the sample, and the forsythia negative control is free from interference.
Comparative example 1
(1) Preparation of test solution: soaking fructus crataegi in 2g of gold capsule content in 30ml of water in water bath at 60deg.C for 30min, centrifuging, collecting supernatant, extracting with water saturated n-butanol for 3 times under shaking, 15ml each time, mixing n-butanol solutions, washing with n-butanol saturated water 30ml, collecting n-butanol solution, evaporating to dryness, dissolving residue in 2ml of methanol, adding into neutral alumina column (120 mesh, 3g, inner diameter 1cm, wet column loading, pre-washing with methanol), eluting with 80ml of methanol, collecting eluate, evaporating to dryness, dissolving residue in 1ml of methanol to obtain sample solution;
(2) Preparing a herba Paederiae control medicinal material solution: taking 3.5g of herba Paederiae control medicinal material, adding 100ml of water, decocting for 30 min at micro-boiling, filtering, concentrating the filtrate to 30ml, extracting with water saturated n-butanol for 3 times under shaking, 15ml each time, mixing n-butanol solutions, washing with water saturated n-butanol 30ml, taking n-butanol solution, evaporating to dryness, and adding 0.5ml of methanol into the residue to dissolve to obtain herba Paederiae control medicinal material solution;
(3) The 5 mu l of the test sample solution (3 batches) and the 5 mu l of the fevervine control medicinal material solution are respectively spotted on the silica gel G thin layer plate, and the volume ratio is 9:2:0.2 of chloroform-methanol-concentrated ammonia solution (25%) is used as a developing agent, after developing, the developing agent is taken out and dried, 10% sulfuric acid ethanol solution is sprayed, the developing agent is heated at 105 ℃ until spots develop clearly, and the result is shown in fig. 5 ((a) at low temperature, (b) after heating at 105 ℃,1. Chinese fevervine reference medicine, 2. Test article 22031702,3. Test article 22031804,4. Test article 21122701), as can be seen from fig. 5, the color development of the characteristic spots is greatly influenced by temperature under sunlight, weak characteristic spots appear at the corresponding positions of the test article and the reference medicine after long-time low-temperature color development, and interference spots exist near the characteristic spots after heating at 105 ℃, so that judgment is influenced.
Comparative example 2
(1) Preparation of test solution: taking 2g of the content of the gold capsule in the hawthorn, adding 30ml of water, soaking in a water bath at 60 ℃ for 30 minutes, centrifuging, taking supernatant, shaking and extracting with water saturated n-butanol for 3 times, 15ml each time, combining n-butanol solutions, washing with water saturated with n-butanol for 30ml hours, taking n-butanol solution, evaporating to dryness, and adding 1ml of methanol into residues to dissolve the residues to serve as a test solution;
(2) Preparing a loquat leaf control medicinal material solution: taking 3g of loquat leaf reference medicinal material, adding 100ml of water, decocting for 30 minutes with micro boiling, filtering, concentrating the filtrate to 30ml, shaking and extracting with water saturated n-butanol for 3 times, 15ml each time, combining n-butanol solutions, washing with water saturated n-butanol for 30ml, taking n-butanol solution, evaporating to dryness, and adding 1ml of methanol into residues to dissolve the residues to obtain a loquat leaf reference medicinal material solution;
(3) And (3) respectively dispensing 5 mu l of the sample solution (3 batches) and 5 mu l of the loquat leaf reference medicinal material solution on a silica gel G thin layer plate, unfolding by taking cyclohexane-ethyl acetate-glacial acetic acid with the volume ratio of 16:8:0.1 as a developing agent, taking out, airing, spraying 10% sulfuric acid ethanol solution, heating at 105 ℃ until spots are clear, and obtaining the results as shown in fig. 6 (1. Loquat leaf reference medicinal material, 2. Sample 22031702,3. Sample 22031804,4. Sample 21122701), wherein the spots of the sample chromatograph on the corresponding positions of the loquat leaf reference medicinal material are not obvious in sunlight, and the background interference is serious.
Comparative example 3
The preparation methods of the sample solution, the loquat leaf control medicinal material solution, the ursolic acid control solution and the loquat She Yinxing control solution are the same as those of the example 1; the volume ratio of the developing agent is 8:2:0.2:0.2 of dichloromethane-ethyl acetate, taking out the dot plate after expanding, airing, spraying 10% sulfuric acid ethanol solution, and heating at 105 ℃ until the color of the dot is clear; the results show that in the chromatogram of the test sample, spots with the same color appear at the positions corresponding to the loquat leaf control medicinal material and the ursolic acid control product, but the spots are weak, and the loquat She Yinxing control has interference.
Comparative example 4
The preparation methods of the sample solution, the loquat leaf control medicinal material solution, the ursolic acid control solution and the loquat She Yinxing control solution are the same as those of the example 1; the volume ratio of the developing agent is 8:2:0.2:0.2, spreading the dot board, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until the color of the dot is clear, and inspecting under 365nm ultraviolet lamp; the results show that no spots appear on the positions corresponding to the loquat leaf control medicinal material and the ursolic acid control in the chromatogram of the test sample, and the spot R f of the control is higher.
Comparative example 5
(1) Sample solution preparation: taking 4g of the content of the gold capsule in the hawthorn, adding 50ml of methanol, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating the filtrate to dryness, adding 30ml of water into the residue to dissolve, extracting for 3 times by shaking with diethyl ether, each time 20ml of water is removed, merging diethyl ether solutions, evaporating to dryness, and adding 1ml of methanol into the residue to dissolve to obtain a sample solution;
(2) Preparation of a Tibetan calamus control medicinal material solution: 2g of a Tibetan calamus control medicinal material is taken, 50ml of water is added, the mixture is boiled for 30 minutes, filtered, the filtrate is concentrated to 30ml, the mixture is extracted for 3 times by shaking with diethyl ether, 20ml of water solution is removed each time, the diethyl ether solution is combined and evaporated to dryness, and 1ml of methanol is added into residues to dissolve the residues to obtain a Tibetan calamus control medicinal material solution;
(3) Preparation of a negative control solution of rhizoma Acori Calami: taking other medicinal herbs except for the Tibetan calamus in the prescription, preparing a negative control sample of the Tibetan calamus according to the prescription proportion, adding 50ml of water, decocting for 30 minutes with micro boiling, filtering, concentrating the filtrate to about 30ml, shaking and extracting for 3 times with diethyl ether for 20ml each time, discarding water solution, merging diethyl ether solution, evaporating to dryness, and adding 1ml of methanol into residues to dissolve the residues to serve as a Tibetan calamus negative control solution;
(4) The method comprises the steps of respectively dispensing 5 μl of sample solution, 5 μl of Tibetan calamus control medicinal material solution and 10 μl of Tibetan calamus negative control solution on a silica gel G thin layer plate, wherein the volume ratio is 4:1 (60-90 ℃) and ethyl acetate as developing agent, taking out, airing, and inspecting under 254nm ultraviolet lamp, wherein spots with the same color are not found on the corresponding positions of the sample chromatogram and the rhizoma acori graminei contrast medicinal material.
Comparative example 6
(1) Sample solution preparation: taking 4g of the content of the gold capsule in the hawthorn, adding 50ml of methanol, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating the filtrate to dryness, adding 30ml of water into the residue to dissolve, extracting for 3 times by shaking with diethyl ether, each time 20ml of water is removed, merging diethyl ether solutions, evaporating to dryness, and adding 1ml of methanol into the residue to dissolve to obtain a sample solution;
(2) Preparing a fructus forsythiae control medicinal material solution: taking 1g of fructus forsythiae reference medicine, adding 50ml of water, decocting for 30 minutes with micro boiling, filtering, concentrating the filtrate to 30ml, shaking and extracting 3 times with diethyl ether, 20ml each time, discarding water solution, combining diethyl ether solution, evaporating to dryness, and adding 1ml of methanol into residues to dissolve the residues to obtain a fructus forsythiae reference medicine solution;
(3) Preparing a forsythin reference substance solution: adding methanol into the forsythin reference substance to prepare a solution containing 1mg per 1ml, and taking the solution as the forsythin reference substance solution;
(4) Preparation of fructus forsythiae negative control solution: taking other medicinal herbs except fructus forsythiae in the prescription, preparing a negative control sample of fructus forsythiae deficiency according to the prescription proportion, adding 50ml of water, decocting for 30 minutes with micro boiling, filtering, concentrating the filtrate to about 30ml, shaking and extracting 3 times with diethyl ether for 20ml each time, discarding water solution, combining diethyl ether solutions, evaporating to dryness, and adding 1ml of methanol into residues to dissolve the residues to serve as a fructus forsythiae negative control solution;
(5) Respectively dotting 5 μl of sample solution, 5 μl of fructus forsythiae control medicinal material solution, 5 μl of fructus forsythiae glycoside control solution and 5 μl of fructus forsythiae negative control solution on the silica gel G thin-layer plate, wherein the volume ratio is 12:2.5:2: developing 0.2 of chloroform-acetone-methanol-98% formic acid as developing agent, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until the color of spots is clear, and collecting spots with the same color at the positions corresponding to the fructus forsythiae control material and the phillyrin control material in the chromatogram of the sample.
Comparative example 7
Preparation of a sample solution, preparation of a fructus forsythiae control medicinal material solution, preparation of a fructus forsythiae glycoside control substance solution, preparation of a fructus forsythiae negative control solution and the same comparison example 6, wherein 5 microliters of the sample solution, 5 microliters of the fructus forsythiae control medicinal material solution, 5 microliters of the fructus forsythiae control substance solution and 5 microliters of the fructus forsythiae negative control solution are respectively dotted on a silica gel G thin layer plate, and the volume ratio is 15:10:0.25 cyclohexane-ethyl formate-formic acid is taken as developing agent, taken out after developing, dried in the air, sprayed with 10% sulfuric acid ethanol solution, heated at 105 ℃ until spots develop clearly, spots with the same color appear at the positions corresponding to the weeping Forsythia control medicinal material in the chromatogram of the test sample, but the weeping Forsythia negative control has interference.
Claims (8)
1. A thin-layer chromatography identification method for multiple components in a Chinese patent medicine is characterized by comprising the following steps:
(1) Preparation of test solution: comprises ethyl acetate extract of Chinese medicinal materials of herba Paederiae, fructus crataegi, folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae as sample solution;
(2) Preparation of control medicinal material solution: taking herba Paederiae, fructus crataegi, folium Eriobotryae, rhizoma Acori Calami and fructus forsythiae as reference materials respectively, decocting with water slightly boiling, filtering, concentrating the filtrate, shaking and extracting with ethyl acetate for 2-4 times, mixing ethyl acetate solutions, concentrating to obtain reference medicinal material solution;
(3) Thin layer chromatography detection:
detecting the fevervine: the test solution and the herba Paederiae control medicinal material solution are respectively spotted on a thin layer plate, and the volume ratio is 9:2: developing 0.2 trichloromethane-methanol-concentrated ammonia solution as developing agent, taking out, air drying, inspecting under 365nm ultraviolet lamp, and detecting fluorescent spots of the same color at the position corresponding to herba Paederiae control medicinal material;
Detection of hawthorns and loquat leaves: the sample solution, the hawthorn control medicinal material solution and the loquat leaf control medicinal material solution are respectively spotted on a thin layer plate, and the volume ratio is 20:6:1, developing with cyclohexane-ethyl acetate-glacial acetic acid as developing agent, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until the color of spots is clear, inspecting under sunlight and 365nm ultraviolet lamp, and detecting spots and fluorescent spots of the same color at positions corresponding to fructus crataegi control medicinal material and folium Eriobotryae control medicinal material;
Detection of Tibetan calamus: the sample solution and the rhizoma acori graminei control medicinal material solution are respectively spotted on a thin layer plate, and the volume ratio is 9:1, developing with dichloromethane-ethyl acetate as developing agent, taking out, air drying, inspecting under 365nm ultraviolet lamp, and detecting fluorescent spots with the same color at the position corresponding to the rhizoma Acori Calami control medicinal material;
Detection of fructus forsythiae: the sample solution and the weeping forsythia control medicinal material solution are respectively spotted on a thin layer plate, and the volume ratio is 12:2.5:2: developing 0.2 trichloromethane-acetone-methanol-formic acid as developing agent, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until the color of spots is clear, and detecting spots of the same color at the positions corresponding to fructus forsythiae control medicinal materials.
2. The thin-layer chromatography identification method for multiple components in a Chinese patent medicine according to claim 1, wherein the Chinese patent medicine is a hawthorn inner gold preparation, and each milliliter of test solution corresponds to 3-6 g of Chinese patent medicine.
3. The method for identifying the components of the Chinese patent medicine by thin-layer chromatography according to claim 1, wherein the hawthorn endo-gold preparation is hawthorn endo-gold capsule or hawthorn endo-gold oral liquid.
4. The method for identifying the components in the Chinese patent medicine by thin-layer chromatography according to claim 3, wherein the preparation method of the sample solution of the hawthorn inner gold capsule is characterized in that methanol is added into the content of the hawthorn inner gold capsule for dissolving, ultrasonic treatment is carried out for 20-50 min, filtration is carried out, filtrate is evaporated to dryness, residues are dissolved in water, ethyl acetate is used for shaking and extracting for 2-4 times, ethyl acetate liquid is combined, and concentrated to obtain the sample solution of the hawthorn inner gold capsule; the preparation method of the sample solution of the Hawthorn inner gold oral liquid comprises the steps of shaking and extracting the Hawthorn inner gold oral liquid with ethyl acetate for 2-4 times, combining the ethyl acetate solutions, and concentrating to obtain the sample solution of the Hawthorn inner gold oral liquid.
5. The thin-layer chromatography identification method for multiple components in a Chinese patent medicine according to claim 1, wherein the concentrated ammonia solution in the step (3) is ammonia water with the volume concentration of 25-28%.
6. The method for identifying the components in the Chinese patent medicine by thin-layer chromatography according to claim 1, wherein the thin-layer plate in the step (3) is a silica gel G thin-layer plate.
7. The method for identifying the components in the Chinese patent medicine by thin-layer chromatography according to claim 1, wherein the point sample amount in the step (3) is 3-10 mu L.
8. The method for identifying multiple components in Chinese patent medicine by thin layer chromatography according to claim 1, wherein 25 ml of water is added into each gram of reference medicinal material in the step (2), and the micro-boiling decoction time is 20-40 min.
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