A multi-region and multi-information thin-layer identification method for multi-origin rhubarb medicinal materials

Abstract

The present invention relates to a multi-region multi-information thin-layer identification method for multi-base rhubarb medicinal materials. The method is characterized in that the supernatant of the medicinal materials and the control medicinal materials subjected to methanol ultrasound is used as the test sample and control medicinal material solutions, and gradient developing agents of different polarities are used to identify the supernatant of the medicinal materials in six regions, two types of prefabricated silica gel GF and high-efficiency silica gel GF 254 On the thin layer plate, under different inspection conditions, a total of 2 exclusive distinguishing characteristic spots of medicinal rhubarb, 10 exclusive distinguishing characteristic spots of palm leaf rhubarb, and 8 exclusive distinguishing characteristic spots of tangut rhubarb were detected. These exclusive distinguishing characteristic spots are distributed in multiple areas, and are judged by multi-component exclusive spots with different polarities, which achieves multi-regional specificity complementarity and effectively improves the accuracy of the judgment results. The intuitive and beautiful color spots are easy to compare and judge between the origins, which is reported for the first time. It solves the difficult problems that are not easy to solve in fingerprints and molecular biology at a low cost and high efficiency, which is creative, practical and leading.

Description

Technical Field

[0001] The present invention relates to a multi-region multi-information thin-layer identification method for multi-origin rhubarb medicinal materials, and belongs to the field of thin-layer identification. That is, in the six regions of fat solubility, medium polarity and water solubility, in the form of thin-layer identification, a total of 2 exclusive distinguishing characteristic spots of medicinal rhubarb control medicinal materials are detected; 10 exclusive distinguishing characteristic spots of palm leaf rhubarb control medicinal materials; and 8 exclusive distinguishing characteristic spots of tangut rhubarb control medicinal materials are detected; these exclusive distinguishing characteristic points can make variety identification and identification of three types of original rhubarb medicinal materials in a simple, fast, multi-information and accurate manner. Background Art

[0002] The identification of multi-origin Chinese herbal medicines is still a very difficult problem. The reason is that if the identification is based on the pharmacognosy characteristics inherited for thousands of years, there must be a whole plant specimen as a control sample. If the herbal medicine is a whole herb, there must be a complete whole herb sample; if it is a rhizome herb, there must be a corresponding whole plant above ground; if it is a flower, fruit, or leaf, there must also be a corresponding whole plant above ground, and so on. Imagine a commercially available tuber, flower, fruit, or leaf, how to obtain the corresponding whole plant above ground, so pharmacognosy experts are helpless. Although modern high-performance liquid fingerprints can conduct multi-information qualitative research on multiple components of a certain herbal medicine from water-soluble to fat-soluble, the same-looking component peaks presented in the spectrum do not have beautiful and intuitive color presentation. Without skilled professionals, it is difficult to make comparative judgments, and the instrument is expensive. At present, no reports have been found that can identify multi-origin varieties. Even in the forefront of plant molecular biology, there are no reports of successful identification of polybasic species, and the high cost of determination is self-evident. How to easily, quickly, efficiently, low-cost, and accurately identify the polybasic medicinal materials sold on the market has become a very difficult problem, and there is no successful report yet.

[0003] The Chinese herbal medicine rhubarb is a variety included in the previous editions of the Chinese Pharmacopoeia. Under this variety, there are three original varieties, namely medicinal rhubarb, palm leaf rhubarb and tangut rhubarb. However, in all the test items such as its properties, identification, inspection and even content determination, the three original rhubarbs are not described as varieties, that is, the three original rhubarbs are the same and have no distinguishing characteristics. As a result, there is no distinction or identification basis for which original rhubarb medicinal material is available on the market.

[0004] As the requirements for research and development data of innovative Chinese medicines in my country continue to increase, multiple origin varieties are required, and the multiple origin medicinal materials in the prescription must have their origins clearly defined. Then, medicinal materials like rhubarb encounter the problem of how to determine the variety and conduct origin identification. For this reason, I have consulted a senior pharmacognosy expert, and the answer given is: it is necessary to provide whole plant samples of three rhubarb medicinal materials, and then compare them with their corresponding plant specimens, through the various characteristics of leaves and flowers, such as palmate rhubarb-the leaves are palmately half-cleft, each lobe has coarse teeth, the panicle inflorescence is terminal, the branches are attached to the stem, the flowers are small, purple-red or purple-red. And the microstructure, texture, cells and other parameters of the cross-section of the rhizome are compared to identify the origin variety. Imagine that it is impossible to obtain the corresponding above-ground part of rhubarb purchased from the medicinal materials market, so it is very difficult to identify the origin variety of rhubarb from the perspective of pharmacognosy. I have also consulted modern cutting-edge instrument detection methods and high-performance liquid fingerprints, but only one article was found. 【1】 "Study on HPLC fingerprints and content differences of rhubarb medicinal materials with different origins", although the gradient fingerprints of three kinds of rhubarb within 65 minutes were provided, it was difficult to judge their different components; the contents of free and bound aloe-emodin, rhein, rhein, chrysophanol and physophanol methyl ether were also given, but the same components and different contents could not identify their origin varieties, so the conclusion given by the authors of this paper is: Based on the results of this study, it can be seen that there are obvious differences in the components and contents of rhubarb medicinal materials with different origins, and the simple use of fingerprint similarity evaluation cannot effectively evaluate their internal differences. Therefore, when evaluating multi-origin medicinal materials, it is recommended to add indicators such as the peak range of the peak area per unit mass (A / W) of the main chromatographic peak, the number of common peaks, the number of non-common peaks and the sum of peak areas, and use statistical methods to perform appropriate principal component analysis and cluster analysis to comprehensively evaluate the batch differences of multi-origin medicinal materials.

[0005] It is clear from the conclusions of the above authors that high performance liquid chromatography fingerprinting cannot be used to identify the origin of multiple origin varieties.

[0006] However, the fingerprint of HPLC makes us think of the following: for example, several inexpensive thin layer plates can replace the expensive HPLC with complete functions; the gradient developing agent for thin layer identification can replace the gradient mobile phase of HPLC; the bright and intuitive spot characteristics of each component with different colors can replace the component peaks with the same appearance presented in the fingerprint; the standard reference medicinal materials with a price of tens of yuan per tube can replace the monomer reference materials with a price of hundreds or even thousands of yuan; the intuitive visual observation can replace the software that can only judge by similarity, etc., and the thin layer plates with high separation effect can be used to compare the color spots of various well-separated thin layer plates from fat-soluble components to water-soluble components. Perhaps, under various inspection conditions, we can find the differential exclusive spots of different original reference medicinal materials by comparing the composition and proportion of the optimal gradient developing agent. The workload of its exploration and research is very large. After success, the differential exclusive spots of each origin control medicinal material in multiple regions are summarized as a whole characteristic exclusive spot identification evaluation series. With this as the reference standard, the origin of the identified medicinal material can be accurately determined. Once the method is determined, the specific implementation is simple and fast, with low cost, high efficiency, beautiful and clear spots, and intuitive and easy to judge. Taking the above assumptions as the research goal, the multi-regional multi-information thin-layer identification method of multi-origin rhubarb medicinal materials was studied.

[0007] By comparing the separation effects of various thin-layer plates, the combination of components of different gradient developing agents, the discussion of the ratios between components, and the selection of accompanying color developers and inspection conditions, a multi-region and multi-information thin-layer identification method for multi-origin rhubarb medicinal materials was obtained. Summary of the invention

[0008] The present invention relates to a multi-region multi-information thin-layer identification method for multi-base rhubarb medicinal materials. The method is characterized in that: the supernatant of methanol ultrasonic medicinal materials and control medicinal materials is used as the test sample and control medicinal material solution, and gradient developing agents of different polarities are used to analyze the supernatant of methanol ultrasonic medicinal materials and control medicinal materials in 6 regions, high-efficiency and ordinary prefabricated silica gel GF 254 On the thin layer plate, under different inspection conditions, a total of 2 exclusive distinguishing characteristic spots of medicinal rhubarb control materials were detected; 10 exclusive distinguishing characteristic spots of palm leaf rhubarb control materials; 8 exclusive distinguishing characteristic spots of tangut rhubarb control materials were detected; these exclusive distinguishing characteristic spots are distributed in multiple regions of fat solubility, medium polarity and water solubility, and are judged by multi-component exclusive spots with different polarities, achieving multi-regional specificity complementarity and effectively improving the accuracy of the judgment results. The color spots with neat and consistent Rf values ​​and intuitive and beautiful colors are easy to compare and judge between origins, which is the first report. It solves the difficult problems of fingerprints and molecular biology in a simple, fast, low-cost and efficient way, which is creative, practical and leading.

[0009] The solution adopted by the present invention to solve the technical problem is:

[0010] (1) Differentiation of three types of rhubarb in the region of fat-soluble components 0.2 g of each of the control medicinal materials of medicinal rhubarb, palmate rhubarb, tangut rhubarb and commercially available medicinal rhubarb powder were taken, 2 ml of methanol was added, ultrasonically treated for 15 minutes, and centrifuged. The supernatant was used as the control medicinal material solution and the test sample solution respectively; 5 μl of each of the above four solutions was taken and spotted on the same prefabricated high-efficiency silica gel GF 254 On the thin layer plate, cyclohexane-ethyl acetate-formic acid with a volume ratio of 12:3:0.1 was used as the developing agent, developed, taken out, dried, and examined under a 365nm ultraviolet lamp. The medicinal rhubarb control material showed 6 fluorescent spots; the palm leaf rhubarb control material showed 7 fluorescent spots; the tangut rhubarb control material showed 5 fluorescent spots; the medicinal rhubarb control material in this area had an exclusive light blue-green fluorescent spot; the palm leaf rhubarb control material had 2 exclusive spots, one was blue-green and the other was pink; and the tangut rhubarb control material had no characteristic spots; in this way, the three original rhubarb control materials were easily distinguished by their exclusive spots; the spots presented by the test rhubarb medicinal material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb;

[0011] (2) Differentiation of the three types of rhubarb in the area of ​​partial fat-soluble components: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated silica gel GF 254 On the thin layer plate, cyclohexane-ethyl acetate-formic acid with a volume ratio of 7:4:0.15 was used as the developing solvent. After development, the plate was taken out, dried, and examined under a UV lamp at 254nm. The medicinal rhubarb control material showed 4 clear brown spots; the palm leaf rhubarb control material showed 6 clear brown spots; the tangut rhubarb control material showed 3 clear brown spots; in this area, the palm leaf rhubarb control material had an exclusive brown spot and a spot with a significantly higher content than other varieties; and the medicinal rhubarb control material had significantly more spots than the tangut rhubarb control material; in this way, the three original rhubarb control materials can also be distinguished from the number and content of spots; the number of spots presented by the test rhubarb material is The rhubarb was basically consistent with the medicinal rhubarb control medicinal material, so it was judged to be medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and examined under a 365nm ultraviolet lamp. The medicinal rhubarb control medicinal material showed 9 fluorescent spots of different colors; the palm leaf rhubarb control medicinal material showed 10 fluorescent spots of different colors; the tangut rhubarb control medicinal material showed 7 fluorescent spots of different colors; the palm leaf rhubarb control medicinal material in this area had 1 pink exclusive spot, while other rhubarb control medicinal materials did not have this spot; in this way, the three original rhubarb control medicinal materials can be distinguished from the number and color of spots; the spots presented by the test rhubarb medicinal material were consistent with the medicinal rhubarb control medicinal material, so it was judged to be medicinal rhubarb;

[0012] (3) Differentiation of the three types of rhubarb in the medium polarity component region: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated high-efficiency silica gel GF 254 On the thin layer plate, cyclohexane-ethyl acetate-formic acid with a volume ratio of 3:7:0.2 was used as the developing agent, developed, taken out, dried, and examined under a 365nm ultraviolet lamp. The medicinal rhubarb control material showed 3 fluorescent spots and 1 blue-purple spot; the palm leaf rhubarb control material showed 4 fluorescent spots and 1 blue-purple spot; the tangut rhubarb control material showed 3 fluorescent spots and 2 blue-purple spots; in this area, the palm leaf rhubarb control material had 1 pink fluorescent exclusive spot, and the gute rhubarb control material had 1 blue-purple exclusive spot; while the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials were easily distinguished; the spots presented by the test rhubarb medicinal material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb;

[0013] (4) Differentiation of the three types of rhubarb in the water-soluble component region: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated silica gel GF 254 On the thin layer plate, cyclohexane-ethyl acetate-methanol-formic acid with a volume ratio of 2:8:1:0.2 was used as the developing solvent, and the plate was developed, taken out, dried, and examined under a UV lamp at 254nm. The medicinal rhubarb control material showed 4 clearer brown spots; the palm leaf rhubarb control material showed 5 clearer brown spots; the tangut rhubarb control material showed 5 clearer brown spots; in this area, the palm leaf rhubarb control material and the tangut rhubarb control material each had an exclusive brown spot with a different Rf value, while the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials can be distinguished by whether they have exclusive spots; the spots presented by the test rhubarb material are basically the same as those of the medicinal rhubarb control material, It was judged to be medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and examined under ultraviolet light at 365nm. The medicinal rhubarb control material showed 8 fluorescent spots of different colors; the palm leaf rhubarb control material showed 9 fluorescent spots of different colors; the Tangut rhubarb control material showed 7 fluorescent spots of different colors and 1 light blue-purple spot; in this area, the palm leaf rhubarb control material had 1 pink exclusive spot, and the Tangut rhubarb control material had 1 light blue-purple exclusive spot; the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials could be distinguished by whether or not they had exclusive spots; the spots presented by the test rhubarb material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb;

[0014] (5) Differentiation of the three types of rhubarb in the water-soluble component region: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated high-efficiency silica gel GF254 On the thin layer plate, cyclohexane-ethyl acetate-methanol-formic acid with a volume ratio of 2:8:1:0.2 was used as the developing agent, and the plate was developed, taken out, dried, and examined under a UV lamp at 365nm. The medicinal rhubarb control material showed 6 fluorescent spots of different colors and 1 blue-purple spot; the palm leaf rhubarb control material showed 6 fluorescent spots of different colors and 2 blue-purple spots; the tangut rhubarb control material showed 3 fluorescent spots of different colors and 2 blue-purple spots; in this area, the palm leaf rhubarb and tangut rhubarb control materials each had an exclusive blue-purple spot, but the Rf values ​​of the two were different, and the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials can be distinguished from the Rf values ​​of the exclusive spots; the spots presented by the test rhubarb material are basically consistent with those of the medicinal rhubarb control material, and it can be judged that It is medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots are clearly colored, and examined under a 254nm ultraviolet lamp, the medicinal rhubarb control material showed 6 fluorescent spots of different colors and 1 blue-purple spot; the palm leaf rhubarb control material showed 6 fluorescent spots of different colors and 2 blue-purple spots; the tangut rhubarb control material showed 4 fluorescent spots of different colors and 2 light blue-purple spots; in this area, the palm leaf rhubarb control material had 1 blue-purple exclusive spot, and the tangut rhubarb control material had 1 light blue-purple exclusive spot, but the Rf values ​​of the two were different; the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials can be distinguished by whether there are exclusive spots; the spots presented by the test rhubarb medicinal material are consistent with those of the medicinal rhubarb control material, and it is judged to be medicinal rhubarb;

[0015] (6) Differentiation of the three types of rhubarb in the water-soluble component region: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated silica gel GF 254On the thin layer plate, chloroform-ethyl acetate-methanol-formic acid-glacial acetic acid with a volume ratio of 1:4:3:0.2:0.2 was used as the developing agent, and the plate was developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, heated at 105°C until the spots were clearly colored, and examined under a UV lamp at 365nm. The medicinal rhubarb control material showed 5 fluorescent spots of different colors, one of which was light yellow with a slight red; the palm leaf rhubarb control material showed 5 fluorescent spots of different colors, one of which was light yellow; the tangut rhubarb control material showed 5 fluorescent spots of different colors, one of which was light brown and one was a blue-purple spot; in this area, the medicinal rhubarb control material had 1 light yellow with a slight red exclusive spot, the palm leaf rhubarb control material had 1 light yellow exclusive spot, and the tangut rhubarb control material had 1 light brown exclusive fluorescent spot and 1 blue-purple exclusive spot. In this way, the three The original rhubarb control medicinal material can be distinguished by the color of the exclusive spots. The spots presented by the test rhubarb medicinal material are similar to those of the medicinal rhubarb control medicinal material, so it is judged to be medicinal rhubarb; the thin layer plate is then examined under a 254nm ultraviolet lamp, and the medicinal rhubarb control medicinal material presents 4 fluorescent spots of different colors and 2 brown spots; the palm-leaf rhubarb control medicinal material presents 4 fluorescent spots of different colors and 1 brown spot; the tangut rhubarb control medicinal material presents 2 fluorescent spots of different colors and 3 brown spots; in this area, the tangut rhubarb control medicinal material has two fewer fluorescent spots and one more brown exclusive spot, while the palm-leaf rhubarb control medicinal material has one less brown spot than the medicinal rhubarb control medicinal material. In this way, the three original rhubarb control medicinal materials can be distinguished by the number of brown spots; the spots presented by the test rhubarb medicinal material are consistent with the medicinal rhubarb control medicinal material, so it is judged to be medicinal rhubarb.

[0016] The principle of the present invention is as follows:

[0017] According to the chemical structure and properties of each effective ingredient of traditional Chinese medicine, following the principle of similar and compatible extraction, suitable extraction solvents are used to prepare test sample and reference medicinal material solutions simply and quickly. Then, using developing agents of different polarities for development, various chemical components will be well separated on their respective thin layer plates according to the different adsorption, desorption, re-adsorption and re-desorption abilities of different developing agents. As long as the thin layer plate has a good separation effect and the selected developing agent components and proportions are appropriate, a variety of original rhubarb reference medicinal materials can obtain different information spots with unique specificity of each variety. Using the multi-information spots presented by a variety of original rhubarb reference medicinal materials as a reference, compare the spot color, shape, Rf value, etc. of the accompanying rhubarb medicinal material to be identified, and identify the original variety with which original rhubarb reference medicinal material is consistent. Taking into account that a developing agent or a thin layer plate may have limited information due to regional limitations, resulting in deviations in the judgment results, a multi-region and multi-information thin layer differentiation and identification index from fat-soluble, medium-polar to water-soluble components was designed. If the origins of multi-region and multi-information identification are consistent, the results will be accurate and correct.

[0018] The innovative features and beneficial effects of the present invention are as follows:

[0019] 1. Using cyclohexane-ethyl acetate-formic acid in a volume ratio of 12:3:0.1 as the developing solvent, the same prefabricated high-efficiency silica gel GF 254 Using thin layer plates as carriers, the three original rhubarb control medicinal materials and the fat-soluble components of the rhubarb medicinal materials to be identified with the same sampling amount were developed and examined under ultraviolet light at 365nm. The medicinal rhubarb control medicinal materials showed 6 fluorescent spots; the palm leaf rhubarb control medicinal materials showed 7 fluorescent spots; the Tangut rhubarb control medicinal materials showed 5 fluorescent spots; in this area, the medicinal rhubarb control medicinal materials had an exclusive blue-green fluorescent spot at an Rf value of about 0.13; the palm leaf rhubarb control medicinal materials had 2 exclusive spots, one was blue-green at an Rf value of about 0.6, and the other was pink at an Rf value of about 0.15; and the Tangut rhubarb control medicinal materials had no characteristic spots; the remaining 5 fluorescent spots presented were the same for the three rhubarbs. In this way, using the three characteristic exclusive spots, the three original rhubarb control medicinal materials can be easily distinguished; the spots presented by the test rhubarb medicinal materials are consistent with the medicinal rhubarb control medicinal materials, and are judged to be medicinal rhubarb ( Figure 1 );

[0020] 2. Using cyclohexane-ethyl acetate-formic acid with a volume ratio of 7:4:0.15 as the developing solvent, the same prefabricated silica gel GF 254Using thin layer plate as carrier, the fat-soluble components of three original rhubarb reference medicinal materials and rhubarb medicinal materials to be identified with the same sampling amount were developed and examined under ultraviolet light of 254nm. The medicinal rhubarb reference medicinal materials showed 4 clear brown spots; the palm leaf rhubarb reference medicinal materials showed 6 clear brown spots; the Tanggut rhubarb reference medicinal materials showed 3 clear brown spots; in this area, the palm leaf rhubarb reference medicinal materials had an exclusive brown spot at the Rf value of 0.55 and a spot at the Rf value of 0.35, which was significantly higher than other varieties; and the spot size and concentration of the medicinal rhubarb reference medicinal materials were significantly larger and denser than those of Tanggut rhubarb; in this way, the three original rhubarb reference medicinal materials can also be distinguished from the number and content of spots; the spots presented by the test rhubarb medicinal materials are basically consistent with those of the medicinal rhubarb reference medicinal materials, and are judged to be medicinal rhubarb ( Figure 2 ); sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and examined under ultraviolet light at 365nm. The medicinal rhubarb control material showed 10 fluorescent spots of different colors; the palm leaf rhubarb control material showed 10 fluorescent spots of different colors; the tangut rhubarb control material showed 8 fluorescent spots of different colors, which were also the spots shared by the other two rhubarbs; in this area, the palm leaf rhubarb control material had a pink exclusive spot at an Rf value of about 0.35, while the other rhubarbs had no such spot; in this way, the three original rhubarb control materials could be distinguished by the number and color of spots; the spots presented by the test rhubarb material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb ( Figure 3 );

[0021] 3. Using cyclohexane-ethyl acetate-formic acid with a volume ratio of 3:7:0.2 as the developing solvent, the same prefabricated high-efficiency silica gel GF 254 The thin layer plate was used as a carrier. The polar components of three original rhubarb reference medicinal materials and the rhubarb medicinal materials to be identified were developed and examined under ultraviolet light at 365nm. The medicinal rhubarb reference medicinal materials showed 3 fluorescent spots and 1 blue-purple spot; the palm leaf rhubarb reference medicinal materials showed 4 fluorescent spots and 1 blue-purple spot; the tangut rhubarb reference medicinal materials showed 3 fluorescent spots and 2 blue-purple spots; in this area, the palm leaf rhubarb reference medicinal materials had a pink fluorescent exclusive spot at an Rf value of about 0.7, and the tangut rhubarb reference medicinal materials had a blue-purple exclusive spot at an Rf value of about 0.3; while the medicinal rhubarb reference medicinal materials had no characteristic exclusive spots; in this way, the three original rhubarb reference medicinal materials could be distinguished by their respective exclusive spots; the spots presented by the test rhubarb medicinal materials were consistent with those of the medicinal rhubarb reference medicinal materials, and were judged to be medicinal rhubarb ( Figure 4 );

[0022] 4. Using cyclohexane-ethyl acetate-methanol-formic acid with a volume ratio of 2:8:1:0.2 as the developing solvent, the same prefabricated silica gel GF254 The thin layer plate was used as a carrier. The water-soluble components of the three original rhubarb reference medicinal materials and the rhubarb medicinal materials to be identified with the same sampling amount were developed and examined under ultraviolet light at 254nm. The medicinal rhubarb reference medicinal materials showed 4 clearer brown spots; the palm leaf rhubarb reference medicinal materials showed 5 clearer brown spots; the tangut rhubarb reference medicinal materials showed 5 clearer brown spots; in this area, the palm leaf rhubarb reference medicinal materials had an exclusive brown spot at the Rf value of about 0.5, and the tangut rhubarb reference medicinal materials had an exclusive brown spot at the Rf value of about 0.7, and the medicinal rhubarb reference medicinal materials had no characteristic exclusive spots; in this way, the three original rhubarb reference medicinal materials can be distinguished by whether they have exclusive spots; the spots presented by the test rhubarb medicinal materials are basically consistent with those of the medicinal rhubarb reference medicinal materials, and it is judged to be medicinal rhubarb ( Figure 5 ); sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and examined under ultraviolet light at 365nm. The medicinal rhubarb control material showed 8 fluorescent spots of different colors; the palm leaf rhubarb control material showed 9 fluorescent spots of different colors; the Tangut rhubarb control material showed 7 fluorescent spots of different colors and 1 light blue-purple spot; in this area, the palm leaf rhubarb control material had a pink exclusive spot at an Rf value of about 0.8, and the Tangut rhubarb control material had a light blue-purple exclusive spot at an Rf value of about 0.8; the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials could be distinguished by the presence or absence of exclusive spots; the spots presented by the test rhubarb material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb ( Figure 6 );

[0023] 5. Using cyclohexane-ethyl acetate-methanol-formic acid with a volume ratio of 2:8:1:0.2 as the developing solvent, the same prefabricated high-efficiency silica gel GF 254 Using thin layer plates as carriers, the water-soluble components of three original rhubarb reference materials and the rhubarb materials to be identified with the same sampling amount were developed and examined under ultraviolet light at 365nm. The medicinal rhubarb reference materials showed 6 fluorescent spots of different colors and 1 blue-purple spot; the palm leaf rhubarb reference materials showed 6 fluorescent spots of different colors and 2 blue-purple spots; the tangut rhubarb reference materials showed 3 fluorescent spots of different colors and 2 blue-purple spots; the palm leaf rhubarb reference materials in this area showed There is a half-moon-shaped exclusive blue-purple spot at the Rf value of about 0.45, while the Tangut rhubarb control medicinal material has an exclusive blue-purple spot at the Rf value of about 0.65. The two blue-purple spots have different Rf values ​​and shapes. The medicinal rhubarb control medicinal material has no characteristic exclusive spots. In this way, the three original rhubarb control medicinal materials can be distinguished by the presence or absence of exclusive spots and the Rf value of the spots; the spots presented by the test rhubarb medicinal material are basically the same as those of the medicinal rhubarb control medicinal material, and it is judged to be medicinal rhubarb ( Figure 7); sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and examined under ultraviolet light at 254nm. The medicinal rhubarb control material showed 6 fluorescent spots of different colors and 1 blue-purple spot; the palm leaf rhubarb control material showed 6 fluorescent spots of different colors and 2 light blue-purple spots; the tangut rhubarb control material showed 4 fluorescent spots of different colors and 2 dark blue-purple spots; in this area, the palm leaf rhubarb control material had a horizontally long oval blue-purple exclusive spot at an Rf value of about 0.45, and the tangut rhubarb control material had a quasi-circular blue-purple exclusive spot at an Rf value of about 0.65. The two blue-purple spots are actually Figure 7 The exclusive spots in the Figure 7 clearer, playing a role of further supplementation and strengthening; the medicinal rhubarb control medicinal material has no characteristic exclusive spots; in this way, the three original rhubarb control medicinal materials are further distinguished by the presence or absence of exclusive spots; the spots presented by the test rhubarb medicinal material are consistent with those of the medicinal rhubarb control medicinal material, and it is judged to be medicinal rhubarb ( Figure 8 ).

[0024] 6. Using chloroform-ethyl acetate-methanol-formic acid-acetic acid in a volume ratio of 1:4:3:0.2:0.2 as the developing solvent, the same prefabricated silica gel GF 254 Using thin layer plate as carrier, the water-soluble components of three original rhubarb reference medicinal materials and rhubarb medicinal materials to be identified with the same sampling amount were developed, sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and examined under ultraviolet light at 365nm. The medicinal rhubarb reference medicinal material showed 5 fluorescent spots of different colors, one of which was light yellow with a slight red; the palm leaf rhubarb reference medicinal material showed 5 fluorescent spots of different colors, one of which was light yellow; the tangut rhubarb reference medicinal material showed 5 fluorescent spots of different colors, one of which was light brown and one was blue-purple. In this area, the medicinal rhubarb control material has a light yellow and slightly reddish exclusive spot at an Rf value of about 0.65, the palm leaf rhubarb control material has a light yellow exclusive spot at an Rf value of about 0.65, and the Tangut rhubarb control material has a light brown-red exclusive fluorescent spot at an Rf value of about 0.65 and a blue-purple exclusive spot at an Rf value of about 0.95. In this way, the three original rhubarb control materials can be distinguished from the colors of their exclusive spots. The spots presented by the test rhubarb medicinal material are close to those of the medicinal rhubarb control material, and it is judged to be medicinal rhubarb ( Fig. 9); then the thin layer plate was placed under ultraviolet light at 254nm for inspection, and the medicinal rhubarb control material showed 4 fluorescent spots of different colors and 2 brown spots; the palm leaf rhubarb control material showed 4 fluorescent spots of different colors and 1 brown spot; the Tangut rhubarb control material showed 2 fluorescent spots of different colors and 3 brown spots; in this area, the Tangut rhubarb control material had two fewer fluorescent spots, and one more brown exclusive spot at the Rf value of about 0.95, while the palm leaf rhubarb control material had one less brown spot at the Rf value of about 0.65. In this way, the three original rhubarb control materials can be distinguished by the number of brown spots; the spots presented by the test rhubarb material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb ( Fig.10 ).

[0025] 7. For the water-soluble components of the three basic rhubarb reference medicinal materials, two prefabricated silica gel GFs, ordinary and high-efficiency, were used. 254 Thin layer carrier, from the developed spectrum, it can be clearly seen that the spot chromatograms separated on the two carriers are different, and double characteristic spot confirmation is obtained, which enhances the accuracy of origin identification.

[0026] 8. Use gradient developing agents of different polarities to prepare two types of silica gel GF in 6 regions: high-efficiency and ordinary 254 On the thin layer plate, under different inspection conditions, a total of 2 unique distinguishing characteristic spots of medicinal rhubarb, 10 unique distinguishing characteristic spots of palm leaf rhubarb, and 8 unique distinguishing characteristic spots of tangut rhubarb were detected; these unique distinguishing characteristic spots are distributed in the full information area of ​​fat-soluble, medium-polar and water-soluble components, with the complementary characteristics of the characteristic spots and the function of multi-information identification. The spots have different colors, are intuitive and beautiful, and are easy to compare and judge between the origins. This is the first report. It solves the difficult problems of fingerprints and molecular biology in a simple, fast, low-cost and efficient way, which is creative, practical and leading. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is the thin layer TLC chart of the fat-soluble component area of ​​three original rhubarb reference medicinal materials and the medicinal materials to be identified under ultraviolet light at 365nm.

[0028] Figure 2 This is the thin layer TLC chart of three original rhubarb reference medicinal materials and the medicinal materials to be identified in the fat-soluble component area under ultraviolet light at 254nm.

[0029] Figure 3 This is the thin layer TLC diagram of three original rhubarb reference medicinal materials and medicinal materials to be identified in the fat-soluble component area under ultraviolet light at 365nm after sulfuric acid ethanol color development.

[0030] Figure 4The thin layer TLC images of three original rhubarb reference medicinal materials and medicinal materials to be identified in the medium polar component region under ultraviolet light at 365nm.

[0031] Figure 5 These are the thin layer TLC images of three original rhubarb reference medicinal materials and medicinal materials to be identified in the area of ​​water-soluble components under ultraviolet light at 254nm.

[0032] Figure 6 This is the thin layer TLC diagram of three original rhubarb reference medicinal materials and medicinal materials to be identified in the water-soluble component area under ultraviolet light at 365nm after sulfuric acid ethanol color development.

[0033] Figure 7 These are the thin layer TLC images of three original rhubarb reference medicinal materials and medicinal materials to be identified in the area of ​​water-soluble components under ultraviolet light at 365nm.

[0034] Figure 8 This is the thin layer TLC diagram of three original rhubarb reference medicinal materials and medicinal materials to be identified in the water-soluble component area under ultraviolet light at 254nm after sulfuric acid ethanol color development.

[0035] Fig. 9 This is the thin layer TLC diagram of three original rhubarb reference medicinal materials and medicinal materials to be identified in the water-soluble component area under ultraviolet light at 365nm after sulfuric acid ethanol color development.

[0036] Fig.10 This is the thin layer TLC diagram of three original rhubarb reference medicinal materials and medicinal materials to be identified in the water-soluble component area under ultraviolet light at 254nm after sulfuric acid ethanol color development.

[0037] Figure 1 Among them, 1 and 2 are reference materials for medicinal rhubarb; 3 and 4 are reference materials for palmate rhubarb; 5 and 6 are reference materials for tangut rhubarb; 7 and 8 are rhubarb materials to be identified.

[0038] Figure 2 , 3 These are TLC chromatograms of the same TLC plate under different inspection conditions, among which 1 and 2 are reference medicinal rhubarb; 3 and 4 are reference medicinal rhubarb; 5 and 6 are reference medicinal rhubarb; 7 and 8 are rhubarb medicinal materials to be identified.

[0039] Figure 4 Among them, 1 and 2 are reference materials for medicinal rhubarb; 3 and 4 are reference materials for palmate rhubarb; 5 and 6 are reference materials for tangut rhubarb; 7 and 8 are rhubarb materials to be identified.

[0040] Figure 5 , 6These are TLC chromatograms of the same TLC plate under different inspection conditions, among which 1 and 2 are reference medicinal rhubarb; 3 and 4 are reference medicinal rhubarb; 5 and 6 are reference medicinal rhubarb; 7 and 8 are rhubarb medicinal materials to be identified.

[0041] Figure 7 , 8 These are TLC chromatograms of the same TLC plate under different inspection conditions, among which 1 and 5 are rhubarb medicinal materials to be identified; 2. medicinal rhubarb reference medicinal material; 3. palm leaf rhubarb reference medicinal material; 4. tangut rhubarb reference medicinal material.

[0042] Fig. 9 , 10 These are TLC chromatograms of the same TLC plate under different inspection conditions, among which 1 and 2 are reference medicinal rhubarb; 3 and 4 are reference medicinal rhubarb; 5 and 6 are reference medicinal rhubarb; 7 and 8 are rhubarb medicinal materials to be identified.

[0043] Specific implementation methods of the present invention

[0044] (1) Differentiation of three types of rhubarb in the region of fat-soluble components. Medicinal rhubarb, palm leaf rhubarb, and tangut rhubarb were used as controls. 0.2 g of medicinal rhubarb and commercially available medicinal rhubarb powder were added with 2 ml of methanol, ultrasonically treated for 15 minutes, and centrifuged. The supernatants were used as the control medicinal solution and the test solution respectively. 5 μl of each of the above four solutions were taken and spotted on the same prefabricated high-efficiency silica gel GF 254 On the thin layer plate, cyclohexane-ethyl acetate-formic acid with a volume ratio of 12:3:0.1 was used as the developing agent, developed, taken out, dried, and examined under a 365nm ultraviolet lamp. The medicinal rhubarb control material showed 6 fluorescent spots; the palm leaf rhubarb control material showed 7 fluorescent spots; the tangut rhubarb control material showed 5 fluorescent spots; the medicinal rhubarb control material in this area had an exclusive light blue-green fluorescent spot; the palm leaf rhubarb control material had 2 exclusive spots, one was blue-green and the other was pink; and the tangut rhubarb control material had no characteristic spots; in this way, the three original rhubarb control materials were easily distinguished by their exclusive spots; the spots presented by the test rhubarb medicinal material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb;

[0045] (2) Differentiation of the three types of rhubarb in the area of ​​partial fat-soluble components: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated silica gel GF 254On the thin layer plate, cyclohexane-ethyl acetate-formic acid with a volume ratio of 7:4:0.15 was used as the developing solvent. After development, the plate was taken out, dried, and examined under a UV lamp at 254nm. The medicinal rhubarb control material showed 4 clear brown spots; the palm leaf rhubarb control material showed 6 clear brown spots; the tangut rhubarb control material showed 3 clear brown spots; in this area, the palm leaf rhubarb control material had an exclusive brown spot and a spot with a significantly higher content than other varieties; and the medicinal rhubarb control material had significantly more spots than the tangut rhubarb control material; in this way, the three original rhubarb control materials can also be distinguished from the number and content of spots; the number of spots presented by the test rhubarb material is The rhubarb was basically consistent with the medicinal rhubarb control medicinal material, so it was judged to be medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and examined under a 365nm ultraviolet lamp. The medicinal rhubarb control medicinal material showed 9 fluorescent spots of different colors; the palm leaf rhubarb control medicinal material showed 10 fluorescent spots of different colors; the tangut rhubarb control medicinal material showed 7 fluorescent spots of different colors; the palm leaf rhubarb control medicinal material in this area had 1 pink exclusive spot, while other rhubarb control medicinal materials did not have this spot; in this way, the three original rhubarb control medicinal materials can be distinguished from the number and color of spots; the spots presented by the test rhubarb medicinal material were consistent with the medicinal rhubarb control medicinal material, so it was judged to be medicinal rhubarb;

[0046] (3) Differentiation of the three types of rhubarb in the medium polarity component region: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated high-efficiency silica gel GF 254 On the thin layer plate, cyclohexane-ethyl acetate-formic acid with a volume ratio of 3:7:0.2 was used as the developing agent, developed, taken out, dried, and examined under a 365nm ultraviolet lamp. The medicinal rhubarb control material showed 3 fluorescent spots and 1 blue-purple spot; the palm leaf rhubarb control material showed 4 fluorescent spots and 1 blue-purple spot; the tangut rhubarb control material showed 3 fluorescent spots and 2 blue-purple spots; in this area, the palm leaf rhubarb control material had 1 pink fluorescent exclusive spot, and the gute rhubarb control material had 1 blue-purple exclusive spot; while the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials were easily distinguished; the spots presented by the test rhubarb medicinal material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb;

[0047] (4) Differentiation of the three types of rhubarb in the water-soluble component region: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated silica gel GF 254On the thin layer plate, cyclohexane-ethyl acetate-methanol-formic acid with a volume ratio of 2:8:1:0.2 was used as the developing solvent, and the plate was developed, taken out, dried, and examined under a UV lamp at 254nm. The medicinal rhubarb control material showed 4 clearer brown spots; the palm leaf rhubarb control material showed 5 clearer brown spots; the tangut rhubarb control material showed 5 clearer brown spots; in this area, the palm leaf rhubarb control material and the tangut rhubarb control material each had an exclusive brown spot with a different Rf value, while the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials can be distinguished by whether they have exclusive spots; the spots presented by the test rhubarb material are basically the same as those of the medicinal rhubarb control material, It was judged to be medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and examined under ultraviolet light at 365nm. The medicinal rhubarb control material showed 8 fluorescent spots of different colors; the palm leaf rhubarb control material showed 9 fluorescent spots of different colors; the Tangut rhubarb control material showed 7 fluorescent spots of different colors and 1 light blue-purple spot; in this area, the palm leaf rhubarb control material had 1 pink exclusive spot, and the Tangut rhubarb control material had 1 light blue-purple exclusive spot; the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials could be distinguished by whether or not they had exclusive spots; the spots presented by the test rhubarb material were consistent with those of the medicinal rhubarb control material, and it was judged to be medicinal rhubarb;

[0048] (5) Differentiation of the three types of rhubarb in the water-soluble component region: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated high-efficiency silica gel GF 254On the thin layer plate, cyclohexane-ethyl acetate-methanol-formic acid with a volume ratio of 2:8:1:0.2 was used as the developing agent, and the plate was developed, taken out, dried, and examined under a UV lamp at 365nm. The medicinal rhubarb control material showed 6 fluorescent spots of different colors and 1 blue-purple spot; the palm leaf rhubarb control material showed 6 fluorescent spots of different colors and 2 blue-purple spots; the tangut rhubarb control material showed 3 fluorescent spots of different colors and 2 blue-purple spots; in this area, the palm leaf rhubarb and tangut rhubarb control materials each had an exclusive blue-purple spot, but the Rf values ​​of the two were different, and the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials can be distinguished from the Rf values ​​of the exclusive spots; the spots presented by the test rhubarb material are basically consistent with those of the medicinal rhubarb control material, and it can be judged that It is medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots are clearly colored, and examined under a 254nm ultraviolet lamp, the medicinal rhubarb control material showed 6 fluorescent spots of different colors and 1 blue-purple spot; the palm leaf rhubarb control material showed 6 fluorescent spots of different colors and 2 blue-purple spots; the tangut rhubarb control material showed 4 fluorescent spots of different colors and 2 light blue-purple spots; in this area, the palm leaf rhubarb control material had 1 blue-purple exclusive spot, and the tangut rhubarb control material had 1 light blue-purple exclusive spot, but the Rf values ​​of the two were different; the medicinal rhubarb control material had no characteristic exclusive spots; in this way, the three original rhubarb control materials can be distinguished by whether there are exclusive spots; the spots presented by the test rhubarb medicinal material are consistent with those of the medicinal rhubarb control material, and it is judged to be medicinal rhubarb;

[0049] (6) Differentiation of the three types of rhubarb in the water-soluble component region: 5 μl of the test solution and the control medicinal material solution under item (1) were respectively applied to the same prefabricated silica gel GF 254On the thin layer plate, chloroform-ethyl acetate-methanol-formic acid-glacial acetic acid with a volume ratio of 1:4:3:0.2:0.2 was used as the developing agent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, heated at 105°C until the spots were clearly colored, and examined under a UV lamp at 365nm. The medicinal rhubarb control medicinal material showed 5 fluorescent spots of different colors. One of them was light yellow with a slight red; the palm leaf rhubarb control medicinal material showed 5 fluorescent spots of different colors, one of which was light yellow; the tangut rhubarb control medicinal material showed 5 fluorescent spots of different colors, one of which was light brown and one was a blue-purple spot; in this area, the medicinal rhubarb control medicinal material had 1 light yellow with a slight red exclusive spot, the palm leaf rhubarb control medicinal material had 1 light yellow exclusive spot, and the tangut rhubarb control medicinal material had 1 light brown exclusive fluorescent spot and 1 blue-purple exclusive spot. In this way, the three The original rhubarb control medicinal material can be distinguished by the color of the exclusive spots. The spots presented by the test rhubarb medicinal material are similar to those of the medicinal rhubarb control medicinal material, so it is judged to be medicinal rhubarb; the thin layer plate is then examined under a 254nm ultraviolet lamp, and the medicinal rhubarb control medicinal material presents 4 fluorescent spots of different colors and 2 brown spots; the palm-leaf rhubarb control medicinal material presents 4 fluorescent spots of different colors and 1 brown spot; the tangut rhubarb control medicinal material presents 2 fluorescent spots of different colors and 3 brown spots; in this area, the tangut rhubarb control medicinal material has two fewer fluorescent spots and one more brown exclusive spot, while the palm-leaf rhubarb control medicinal material has one less brown spot than the medicinal rhubarb control medicinal material. In this way, the three original rhubarb control medicinal materials can be distinguished by the number of brown spots; the spots presented by the test rhubarb medicinal material are consistent with the medicinal rhubarb control medicinal material, so it is judged to be medicinal rhubarb.

[0050] References

[0051] 【1】Yuan Zhenhai, Wang Fang, Liang Dalian, et al., Study on HPLC fingerprint and content difference of different origin rhubarb medicinal materials, Food and Drug, 2021, 23(2):147-153.

Claims

1. A multi-region multi-information thin-layer identification method for multi-origin rhubarb medicinal materials, characterized by: (1) Take 0.2 g of each of the medicinal rhubarb, palm leaf rhubarb, tangut rhubarb control medicinal materials and commercial medicinal rhubarb powder, add 2 ml of methanol, ultrasonically treat for 15 minutes, centrifuge, and use the supernatant as the respective control medicinal material solution and test sample solution; (2) Differentiation of the three types of rhubarb in the region of partial fat-soluble components 5 μl of the test solution and the control solution under item (1) were respectively spotted on the same prefabricated silica gel GF254 thin layer plate, and cyclohexane-ethyl acetate-formic acid with a volume ratio of 7:4:0.15 was used as the developing solvent. The plate was developed, taken out, dried, and examined under a UV lamp at 254 nm. The medicinal rhubarb control drug showed 4 clear brown spots; the palm leaf rhubarb control drug showed 6 clear brown spots; and the tangut rhubarb control drug showed 3 clear brown spots. In this region, the palm leaf rhubarb control drug had one exclusive brown spot and one spot with a significantly higher content than other varieties; and the medicinal rhubarb control drug had significantly more spots than the tangut rhubarb control drug. In this way, the three types of rhubarb control drugs were The number and content of spots can also be distinguished; the number of spots presented by the test rhubarb medicinal material is basically consistent with that of the medicinal rhubarb control medicinal material, so it is judged to be medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots are clearly colored, and inspected under a 365nm ultraviolet lamp, the medicinal rhubarb control medicinal material showed 9 fluorescent spots of different colors; the palm leaf rhubarb control medicinal material showed 10 fluorescent spots of different colors; the tangut rhubarb control medicinal material showed 7 fluorescent spots of different colors; the palm leaf rhubarb control medicinal material in this area has 1 pink exclusive spot, while other rhubarb control medicinal materials do not have this spot; in this way, the three original rhubarb control medicinal materials can be distinguished from the number and color of spots; the spots presented by the test rhubarb medicinal material are consistent with those of the medicinal rhubarb control medicinal material, so it is judged to be medicinal rhubarb; (3) Differentiation of three kinds of rhubarb in the region of medium polarity components 5 μl of the test solution and control solution under item (1) were respectively spotted on the same prefabricated high-efficiency silica gel GF254 thin layer plate, and cyclohexane-ethyl acetate-formic acid with a volume ratio of 3:7:0.2 was used as the developing solvent. The plate was developed, taken out, dried, and examined under a UV lamp at 365 nm. The control rhubarb showed 3 fluorescent spots and 1 blue-purple spot; the palm leaf rhubarb showed 4 fluorescent spots and 1 blue-purple spot; the tangut rhubarb showed 3 fluorescent spots and 2 blue-purple spots; in this region, the palm leaf rhubarb control had 1 pink fluorescent spot, and the tangut rhubarb control had 1 blue-purple spot; while the rhubarb control had no characteristic spots. In this way, the three kinds of rhubarb control were easily distinguished. The spots presented by the test rhubarb were consistent with those of the rhubarb control, and it was judged to be rhubarb. (4) Differentiation of the three types of rhubarb in the region of partially water-soluble components 5 μl of the test solution and the control solution under item (1) were respectively spotted on the same prefabricated silica gel GF254 thin layer plate, and cyclohexane-ethyl acetate-methanol-formic acid with a volume ratio of 2:8:1:0.2 was used as the developing solvent. The plate was developed, taken out, dried, and examined under a UV lamp at 254 nm. The control rhubarb showed 4 clear brown spots; the palm leaf rhubarb showed 5 clear brown spots; and the tangut rhubarb showed 5 clear brown spots. In this region, the palm leaf rhubarb control medicine and the tangut rhubarb control medicine each had an exclusive brown spot with a different Rf value, while the medicinal rhubarb control medicine had no characteristic exclusive spot. In this way, the three types of rhubarb control medicines can be distinguished by the presence or absence of exclusive spots. points; the spots presented by the test rhubarb medicinal material are basically consistent with those of the medicinal rhubarb control medicinal material, and it is judged to be medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots are clearly colored, and examined under a UV lamp at 365nm, the medicinal rhubarb control medicinal material presented 8 fluorescent spots of different colors; the palm leaf rhubarb control medicinal material presented 9 fluorescent spots of different colors; the Tanggut rhubarb control medicinal material presented 7 fluorescent spots of different colors and 1 light blue-purple spot; in this area, the palm leaf rhubarb control medicinal material has 1 pink exclusive spot, and the Tanggut rhubarb control medicinal material has 1 light blue-purple exclusive spot; the medicinal rhubarb control medicinal material has no characteristic exclusive spots; in this way, the three original rhubarb control medicinal materials can be distinguished by whether there are exclusive spots; the spots presented by the test rhubarb medicinal material are consistent with the medicinal rhubarb control medicinal material, and it is judged to be medicinal rhubarb; (5) Differentiation of three kinds of rhubarb in the area of ​​partial water-soluble components 5 μl of the test solution and the control solution under item (1) were respectively spotted on the same prefabricated high-efficiency silica gel GF254 thin layer plate, and cyclohexane-ethyl acetate-methanol-formic acid with a volume ratio of 2:8:1:0.2 was used as the developing solvent. The plate was developed, taken out, dried, and examined under a UV lamp at 365 nm. The control rhubarb showed 6 fluorescent spots of different colors and 1 blue-purple spot; the palm leaf rhubarb showed 6 fluorescent spots of different colors and 2 blue-purple spots; the tangut rhubarb showed 3 fluorescent spots of different colors and 2 blue-purple spots. In this area, the palm leaf rhubarb and tangut rhubarb control materials each had an exclusive blue-purple spot, but the Rf values ​​of the two were different. The medicinal rhubarb control material had no characteristic exclusive spot. In this way, the three kinds of rhubarb control materials can be distinguished from the Rf values ​​of the exclusive spots. ; The spots presented by the test rhubarb medicinal material are basically consistent with those of the medicinal rhubarb control medicinal material, and it is judged to be medicinal rhubarb; sprayed with 10% sulfuric acid ethanol solution, heated at 105℃ until the spots are clearly colored, and examined under a UV lamp at 254nm, the medicinal rhubarb control medicinal material presented 6 fluorescent spots of different colors and 1 blue-purple spot; the palm leaf rhubarb control medicinal material presented 6 fluorescent spots of different colors and 2 blue-purple spots; the tangut rhubarb control medicinal material presented 4 fluorescent spots of different colors and 2 light blue-purple spots; in this area, the palm leaf rhubarb control medicinal material has 1 blue-purple exclusive spot, and the tangut rhubarb control medicinal material has 1 light blue-purple exclusive spot, but the Rf values ​​of the two are different; the medicinal rhubarb control medicinal material has no characteristic exclusive spots; in this way, the three original rhubarb control medicinal materials can be distinguished by whether there are exclusive spots; the spots presented by the test rhubarb medicinal material are consistent with the medicinal rhubarb control medicinal material, and it is judged to be medicinal rhubarb; (6) Differentiation of the three types of rhubarb in the water-soluble component region: 5 μl of the test solution and the control drug solution under item (1) were respectively spotted on the same prefabricated silica gel GF254 thin layer plate, and chloroform-ethyl acetate-methanol-formic acid-glacial acetic acid in a volume ratio of 1:4:3:0.2:0.2 was used as the developing agent. The plate was taken out, dried, sprayed with 10% sulfuric acid ethanol solution, heated at 105°C until the spots were clearly colored, and examined under a UV lamp at 365 nm. The rhubarb control material showed 5 fluorescent spots of different colors, one of which was light yellow with a slight red; the palm leaf rhubarb control material showed 5 fluorescent spots of different colors, one of which was light yellow; the tangut rhubarb control material showed 5 fluorescent spots of different colors, one of which was light brown and one was blue-purple; the medicinal rhubarb control material in this area had one exclusive spot of light yellow with a slight red, the palm leaf rhubarb control material had one exclusive spot of light yellow, and the tangut rhubarb control material There is 1 light brown-red exclusive fluorescent spot and 1 bluish-purple exclusive spot. In this way, the three original rhubarb control materials can be distinguished by the color of the exclusive spots. The spots presented by the test rhubarb medicinal material are close to those of the medicinal rhubarb control medicinal material, so it is judged to be medicinal rhubarb; then the thin layer plate is placed under ultraviolet light at 254nm for inspection, and the medicinal rhubarb control medicinal material presents 4 fluorescent spots of different colors and 2 brown spots; the palm leaf rhubarb control medicinal material presents 4 fluorescent spots of different colors and 1 brown spot; the Tangut rhubarb control medicinal material presents 2 fluorescent spots of different colors and 3 brown spots; in this area, the Tangut rhubarb control medicinal material has two fewer fluorescent spots and one more brown exclusive spot, and the palm leaf rhubarb control medicinal material has one less brown spot than the medicinal rhubarb control medicinal material. In this way, the three original rhubarb control medicinal materials can be distinguished by the number of brown spots; the spots presented by the test rhubarb medicinal material are consistent with the medicinal rhubarb control medicinal material, so it is judged to be medicinal rhubarb.

2. The multi-region multi-information thin-layer identification method for multi-origin rhubarb medicinal material according to claim 1, characterized in that: The commercially available medicinal rhubarb was used as the verification object of the method. Through the comparison of multi-region and multi-component information maps, it was found that the maps were basically consistent with the maps of the medicinal control drugs, further verifying that this method can accurately identify the three original rhubarb drugs.

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