Thin-layer identification method of herba patriniae formula granules and application of thin-layer identification method

Abstract

The invention discloses a thin-layer identification method of herba patriniae formula granules and application thereof. The thin-layer identification method comprises the following steps: adding water into test sample powder for dissolving, shaking and extracting by using ethyl acetate, drying by distillation, and adding methanol into residues for dissolving to obtain a test sample solution; the method comprises the following steps: taking a dahurian patrinia herb reference medicinal material, heating, refluxing, filtering, concentrating, adding methanol, carrying out ultrasonic treatment, filtering, drying by distillation, adding water for dissolving, extracting by shaking with ethyl acetate, drying by distillation, and dissolving residues by adding methanol to obtain a reference medicinal material solution; spotting the sample on the same thin-layer plate, developing by using cyclohexane-ethyl acetate-methanol-formic acid as a developing solvent, and inspecting after developing; when spots with the same color appear at the positions corresponding to the chromatogram of the reference medicinal material in the chromatogram of the test sample, the test sample is the patrinia formula granules. According to the method, various components in the sample can be detected, and the specificity of the method is greatly improved; the patrinia scabiosaefolia and patrinia villosa And the patrinia and counterfeit products can be easily distinguished, so that the method has a relatively high practical value.

Description

Technical Field

[0001] The invention relates to a thin layer identification method for Patrinia patriniae formula granules and application thereof, belonging to the field of quality control of Patrinia patriniae formula granules. Background Art

[0002] Patrinia scabiosaefolia Fisch. or Patrinia villosa Juss. is a dried whole herb of Patriniaceae. It is harvested before the flowers bloom in summer, dried in the sun until half dry, tied into bundles, and then dried in the shade. Patrinia scabiosaefolia is mainly produced in Jiangsu, Zhejiang, Anhui, Hubei and other places, and has the functions of clearing away heat and detoxifying, removing blood stasis and draining pus. Studies have found that Patrinia scabiosaefolia contains a variety of effective ingredients such as terpenes, flavonoids, lignans, volatile oils, etc.

[0003] Chinese medicine formula granules are a breakthrough in the application of traditional Chinese medicine slices. They are usually made from slices of Chinese medicine, and are extracted (usually mainly with water), concentrated, dried and granulated. Their nature, flavor, meridians and efficacy are basically the same as the original slices. They also have the advantages of not requiring decoction, easy adjustment, storage and convenient use, and are gradually being widely used. The quality standards for Patrinia patriniae and Patrinia patriniae formula granules are not included in the first volume of the 2020 edition of the Chinese Pharmacopoeia.

[0004] In the prior art, most of the literature conducts thin-layer identification on the herb Patrinia chinensis, rather than water extracts such as formula granules; the literatures "An Improved Quality Detection Method for Longqing Capsules", "Research on Quality Standards for Yiren Liumiao Granules", and "Identification Methods for Patrinia chinensis in Traditional Chinese Medicine Compounds" conduct thin-layer identification on the herb Patrinia chinensis components in compound granule preparations. However, after water extraction, the Chinese medicine formula granules have fewer fat-soluble components, and the thin-layer identification method for medicinal materials may not be applicable to formula granules. Chinese medicine formula granules are single prescriptions, and the ingredients are quite different from those of compound granules. The method for compound granules may not be applicable to the thin-layer identification of formula granules. Moreover, there are fewer thin-layer chromatographic spots of Patrinia chinensis in the literature, and its specificity is not strong, and the identification effect is not ideal. Therefore, it is an important topic to establish a fast, simple, and low-cost thin-layer identification method for Patrinia chinensis formula granules. Summary of the invention

[0005] Purpose of the invention: The technical problem to be solved by the present invention is to provide a rapid, simple and low-cost thin layer identification method of Patrinia scabra formula granules and its application.

[0006] Technical solution: To solve the above technical problems, the present invention provides a thin layer identification method for Patrinia scabra formula granules, comprising the following steps:

[0007] (1) Preparation of test sample solution: Dissolve the test sample powder in water and shake with ethyl acetate to extract. Evaporate the ethyl acetate to dryness and dissolve the residue in methanol to prepare the test sample solution.

[0008] (2) Preparation of control medicinal material solution: Take Patrinia chinensis control medicinal material, add water and heat to reflux, filter, concentrate the filtrate to near dryness, add methanol to the residue, ultrasonically treat, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with ethyl acetate, evaporate the ethyl acetate to dryness, and dissolve the residue in methanol to prepare the control medicinal material solution;

[0009] (3) Take the control medicinal material solution and the test sample solution and spot them on the same thin layer plate, develop them with cyclohexane-ethyl acetate-methanol-formic acid as the developing solvent, take them out, dry them, spray them with sulfuric acid ethanol solution, heat them until the spots are clearly colored, and inspect and compare the thin layer chromatograms;

[0010] (4) When spots of the same color appear in the chromatogram of the test sample at the corresponding positions in the chromatogram of the control medicinal material, it means that the test sample is the formula granules of Patrinia scabra.

[0011] Among them, Patrinia salsa includes Patrinia leucoderma or Patrinia foetida.

[0012] Wherein, the development temperature in step (3) is 4 to 40°C.

[0013] Wherein, the humidity of the development in step (3) is 18-88%.

[0014] The present invention also provides application of the thin layer identification method in authenticity identification of Patrinia scabra formula granules.

[0015] The present invention also provides a method for distinguishing the herbaceous granules from the herbaceous granules and the herbaceous granules, comprising the following steps:

[0016] (1) Preparation of test sample solution: Dissolve the test sample powder in water and shake with ethyl acetate to extract. Evaporate the ethyl acetate to dryness and dissolve the residue in methanol to prepare the test sample solution.

[0017] (2) Preparation of control medicinal material solution: Take Patrinia chinensis control medicinal material, add water and heat to reflux, filter, concentrate the filtrate to near dryness, add methanol to the residue, ultrasonically treat, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with ethyl acetate, evaporate the ethyl acetate to dryness, and dissolve the residue in methanol to prepare the control medicinal material solution;

[0018] (3) Take the control medicinal material solution and the test sample solution and spot them on the same thin layer plate, develop them with cyclohexane-ethyl acetate-methanol-formic acid as the developing solvent, take them out, dry them, spray them with sulfuric acid ethanol solution, heat them until the spots are clearly colored, and inspect and compare the thin layer chromatograms;

[0019] (4) When spots of the same color appear in the chromatogram of the test sample at the corresponding positions in the chromatogram of the control medicinal material, it means that the test sample is the formula granules of Patrinia scabra.

[0020] Wherein, the development temperature in step (3) is 4 to 40°C.

[0021] Wherein, the humidity of the development in step (3) is 18-88%.

[0022] The present invention also provides a method for distinguishing the formula granules of Patrinia lutea and Patrinia odorata, comprising the following steps:

[0023] (1) Preparation of test sample solution: Dissolve the test sample powder in water and shake with ethyl acetate to extract. Evaporate the ethyl acetate to dryness and dissolve the residue in methanol to prepare the test sample solution.

[0024] (2) Preparation of control medicinal material solution: Take Patrinia chinensis control medicinal material, add water and heat to reflux, filter, concentrate the filtrate to near dryness, add methanol to the residue, ultrasonically treat, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with ethyl acetate, evaporate the ethyl acetate to dryness, and dissolve the residue in methanol to prepare the control medicinal material solution;

[0025] (3) Take the control medicinal material solution and the test sample solution and spot them on the same thin layer plate, develop them with cyclohexane-ethyl acetate-methanol-formic acid as the developing solvent, take them out, dry them, spray them with sulfuric acid ethanol solution, heat them until the spots are clearly colored, and inspect and compare the thin layer chromatograms;

[0026] (4) When the test sample chromatogram shows spots of the same color at the corresponding positions with the control medicinal material chromatogram, and there are no yellow-brown spots but blue spots at around Rf value 0.2, it means that the test sample is Baihua Baijiang formula granules; when the test sample chromatogram shows spots of the same color at the corresponding positions with the control medicinal material chromatogram, and there are yellow-brown spots but no blue spots at around Rf value 0.2, it means that the test sample is Huanghua Baijiang formula granules.

[0027] Wherein, the development temperature in step (3) is 4 to 40°C.

[0028] Wherein, the humidity of the development in step (3) is 18-88%.

[0029] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: 1. It solves the problem that most of the existing literature is about the identification method of medicinal materials rather than the identification method of particles; 2. It solves the problem that some literatures identify particles but have few spots and the identification effect is not ideal; 3. A control medicinal material is used as a control substance, and the sample is treated with a simple treatment method and inspected under ultraviolet light (365nm). The number of spots is large and the separation is good, and multiple components in the sample can be detected, which greatly improves the specificity of the method; 4. This method can distinguish between the two basic sources of Patrinia jasminoides and Patrinia dahurica; 5. In addition, it can also easily distinguish Patrinia dahurica from counterfeit products such as Cyperus rotundus and Cyperus rotundus, which has a high practical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 TLC spectra of test solution and control medicinal material solution under different preparation conditions;

[0031] Figure 2 The TLC spectra of the test solution and the reference medicinal material solution at different sample amounts;

[0032] Figure 3 To investigate the specificity of the samples;

[0033] Figure 4 TLC spectra of test solution and control medicinal material solution at different temperatures;

[0034] Figure 5 The TLC spectrum of Patrinia scabra granules under different humidity conditions;

[0035] Figure 6 The TLC spectra of the Patrinia scabra granules prepared by different manufacturers are shown below;

[0036] Figure 7 The thin layer identification chromatograms of different batches of Patrinia scabra granules are shown;

[0037] Figure 8 These are the thin layer identification chromatograms of the formula granules of Patrinia scabra (yellow flower Patrinia scabra), the formula granules of Patrinia scabra (white flower Patrinia scabra), the formula granules of Cynanchum wilfordii, and the formula granules of Mutouhui. DETAILED DESCRIPTION

[0038] The technical solution of the present invention is further described below in conjunction with the accompanying drawings.

[0039] 1. Instruments and test drugs

[0040] Instruments: CAMAG TLC VISUALIZER, METTLERTOLEDO, KQ-250E ultrasonic cleaner (Kunshan Ultrasonic Electronics Co., Ltd.), silica gel G thin layer plate (Qingdao Ocean Chemical Co., Ltd., Qingdao Kangyexin Medicinal Silica Gel Desiccant Co., Ltd., Yantai Xincheng Silica Gel Material Co., Ltd.).

[0041] Reagents: methanol (Sinopharm Chemical Reagent Co., Ltd.), ethanol (Sinopharm Chemical Reagent Co., Ltd.), cyclohexane (Sinopharm Chemical Reagent Co., Ltd.), ethyl acetate (Sinopharm Chemical Reagent Co., Ltd.), formic acid (Sinopharm Chemical Reagent Co., Ltd.), sulfuric acid (Shanghai Lingfeng Chemical Reagent Co., Ltd.) were all of analytical grade, and water; the reference drug of Patrinia scabra (batch number: 121271-201201) was purchased from China Food and Drug Inspection Institutes. The formula granules of Patrinia salsa (batch numbers: 18041129, 18041139, 18041149), the formula granules of Patrinia salsa (yellow flower Patrinia salsa) (11803650, 11803651, 11803652), the formula granules of Patrinia salsa (white flower Patrinia salsa) (1806028, 1806029, 1806030), the formula granules of Cyperus rotundus (2407001, 2407025, 2407026), and the formula granules of Mutouhui (2206047, 2206050, 2206052) are provided by Jiangyin Tianjiang Pharmaceutical Co., Ltd.

[0042] 2. Experimental methods

[0043] 2.1 Thin layer identification conditions

[0044] A silica gel G thin layer plate was used with cyclohexane-ethyl acetate-methanol-formic acid (6:4:0.5:0.1) as the developing agent. The plate was developed, taken out, air-dried, sprayed with 10% sulfuric acid ethanol solution, heated at 105°C until the spots were clearly colored, and examined under ultraviolet light (365nm).

[0045] 2.2 Solution preparation

[0046] 2.2.1 Preparation of test solution

[0047] Method 1: Take 1g of Patrinia scabra granules, grind them into powder, add 25ml of methanol, treat with ultrasound for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of methanol to the residue to dissolve it, and use it as the test solution.

[0048] Method 2: Take 1g of Patrinia scabra granules, grind them into powder, add 20ml of water to dissolve them, shake and extract them with ethyl acetate twice, 20ml each time, combine the ethyl acetate solutions, evaporate them to dryness, add 1ml of methanol to dissolve the residue, and use it as the test solution.

[0049] Method 3: Take 1g of Patrinia scabra granules, grind them into powder, add 25ml of methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 20ml of water to the residue to dissolve it, shake and extract with ethyl acetate twice, 20ml each time, combine the ethyl acetate solutions, evaporate to dryness, add 1ml of methanol to the residue to dissolve it, and use it as the test solution.

[0050] 2.2.2 Preparation of reference medicinal material solution

[0051] Method 1: Take 2g of Patrinia salsa reference medicinal material, add 50ml of water, heat and reflux for 60 minutes, cool, filter, concentrate the filtrate to about 20ml, shake and extract with ethyl acetate twice, 20ml each time, combine the ethyl acetate liquid, evaporate to dryness, add 1ml of methanol to dissolve the residue, and use it as the reference medicinal material solution.

[0052] Method 2: Take 2g of Patrinia salsa reference medicinal material, add 50ml of water, heat and reflux for 60 minutes, cool, filter, concentrate the filtrate to near dryness, add 25ml of methanol to the residue, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 20ml of water to the residue to dissolve it, shake and extract with ethyl acetate twice, 20ml each time, combine the ethyl acetate solutions, evaporate to dryness, add 1ml of methanol to the residue to dissolve it, and use it as the reference medicinal material solution.

[0053] 2.2.3 Preparation of negative control solution

[0054] Take 1 g of the negative sample lacking Patrinia salsa and prepare a negative control solution using the same method as the test sample in "Method 2".

[0055] Example 1 Investigation of the preparation methods of the test solution and the control medicinal material solution

[0056] Take the test solution and control medicinal material solution prepared by the preparation methods of the three test solution and the two control medicinal material solutions of Patrinia patriniae formula granules (batch number: 18041139) and Patrinia patriniae control medicinal material (batch number: 121271-201201) according to the above three test solution preparation methods and two control medicinal material solution preparation methods, respectively spot on the same silica gel G thin layer plate, develop according to the above thin layer chromatography conditions, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365nm). Among them, the development temperature is 26.5°C and the humidity is 37%. Figure 1 1: Test sample method 1; 2: Test sample method 2; 3: Test sample method 3; S1: Patrinia scabra control medicinal material (method 1); S2: Patrinia scabra control medicinal material (method 2). Figure 1 As shown, the test solutions prepared by methods 2 and 3 have more abundant spots and better separation effects. Considering that method 2 is simpler to handle and the reagents used are less toxic, method 2 is selected as the sample processing method for this thin layer identification; the control medicinal material of Patrinia scabra (method 2) is more consistent with the granular spots, so method 2 is selected as the control medicinal material processing method.

[0057] Example 2 Investigation of different sample amounts

[0058] Take the Herba Patriniae Formula Granules (Batch No.: 18041139) and Herba Patriniae Control Medicinal Materials (Batch No.: 121271-201201) and prepare the test solution and the control medicinal material solution according to the preparation method of the test solution and the preparation method of the control medicinal material solution determined above. According to the thin layer chromatography conditions in the experimental method, spot the samples on the same silica gel G thin layer plate, develop them, take them out, dry them, spray them with 10% sulfuric acid ethanol solution, heat them at 105°C until the spots are clearly colored, and examine them under ultraviolet light (365nm). Among them, the development temperature is 26.5°C and the humidity is 37%. The results are as follows: Figure 2 As shown, Figure 2 1: test sample 1μl; 2: test sample 2μl; 3: test sample 5μl; 4: control drug 5μl; 5: control drug 10μl; 6: control drug 15μl. Figure 2 It can be seen that when the sample solution spotting volume is 2 μl and the control medicinal material solution spotting volume is 10 μl, the spots at the corresponding positions in the test sample chromatogram and the control medicinal material chromatogram clearly correspond to each other without other interference. Therefore, the sample solution spotting volume is selected as 2 μl and the control medicinal material solution spotting volume is selected as 10 μl.

[0059] Example 3 Sample specificity experiment

[0060] Take the Patrinia patriniae formula granules (batch number: 18041139) and Patrinia patriniae control medicinal materials (batch number: 121271-201201) according to the preparation method of the test solution and the preparation method of the control medicinal material solution determined above to prepare the test solution, the control medicinal material solution and the negative control solution, and spot them on the same silica gel G thin layer plate according to the thin layer chromatography conditions in the experimental method, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365nm). Among them, the development temperature is 26.5°C and the humidity is 37%. Figure 3 1-3: Herba Patriniae Formula Granules (18041139); 4: Negative control; S: Herba Patriniae control medicinal material. Figure 3 It can be seen that the fluorescent spots of the same color appear at the corresponding positions of the chromatogram of the test sample of the Herba Patriniae formula granules and the chromatogram of the control medicinal material, and there is no interference from the negative control, indicating that the thin-layer identification method has good specificity.

[0061] Example 4 Investigation of different temperatures

[0062] Take the Patrinia patriniae formula granules (batch number: 18041139) and Patrinia patriniae control medicinal materials (batch number: 121271-201201) according to the preparation method of the test solution and the preparation method of the control medicinal material solution determined above, spot on the same silica gel G thin layer plate according to the thin layer chromatography conditions in the experimental method, develop at high temperature 40°C and low temperature 4°C, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365nm). Among them, the development humidity is 37%. Figure 4 1-3 in: Patrinia scabra formula granules (18041139); S: Patrinia scabra control medicinal material. Figure 4 It can be seen that under different temperature conditions, the same color fluorescent spots appear at the corresponding positions of the chromatogram of the test sample and the control medicinal material of the Herba Patriniae Formula Granules, and the separation effect is good. The experimental results show that temperature has no effect on the thin layer identification of the Herba Patriniae Formula Granules, indicating that the thin layer identification method has good durability at different temperatures.

[0063] Example 5 Investigation of different humidity

[0064] Take the Patrinia patriniae formula granules (batch number: 18041139) and Patrinia patriniae control medicinal materials (batch number: 121271-201201) according to the preparation method of the test solution and the preparation method of the control medicinal material solution determined above, spot on the same silica gel G thin layer plate according to the thin layer chromatography conditions in the experimental method, develop under different humidity conditions (18%, 88%), take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365nm). Among them, the development temperature is 26.5°C. Figure 5 1-3 in: Patrinia scabra formula granules (18041139); S: Patrinia scabra control medicinal material. Figure 5 It can be seen that under different humidity conditions, the same color fluorescent spots appear at the corresponding positions of the chromatogram of the test sample and the control medicinal material of the Herba Patriniae Formula Granules, and the separation effect is good. The experimental results show that humidity has no effect on the thin layer identification of the Herba Patriniae Formula Granules, indicating that the thin layer identification method has good durability under different humidity.

[0065] Example 6 Investigation of thin-layer plates from different manufacturers

[0066] Take the Herba Patriniae Formula Granules (Batch No.: 18041139) and Herba Patriniae Control Medicinal Materials (Batch No.: 121271-201201) and prepare the test solution and the control medicinal material solution according to the preparation method of the test solution and the preparation method of the control medicinal material solution determined above, spot them on the silica gel G thin layer plates of different manufacturers, and develop them at 26.5°C and 37% humidity according to the thin layer chromatography conditions in the experimental method, take them out, dry them, spray them with 10% sulfuric acid ethanol solution, heat them at 105°C until the spots are clearly colored, and examine them under ultraviolet light (365nm). The results are as follows Figure 6 As shown, 1 to 3 in the figure: Patrinia scabra formula granules (18041139); S: Patrinia scabra control medicinal material. As can be seen from Figure 6, under the conditions of thin layer plates from different manufacturers, the chromatograms of the Patrinia scabra formula granules test sample and the control medicinal material chromatograms show spots of the same color at the corresponding positions, and the separation effect is good. The experimental results show that the thin layer plate manufacturer has no significant effect on the thin layer identification of Patrinia scabra formula granules, indicating that the thin layer identification method has good durability for thin layer plates from different manufacturers.

[0067] According to the above research results, the thin layer identification method of Patrinia scabra formula granules is determined as follows:

[0068] Take 1g of Patrinia scabra granules, grind them into powder, add 20ml of water to dissolve them, shake and extract them with ethyl acetate twice, 20ml each time, combine the ethyl acetate solution, evaporate to dryness, add 1ml of methanol to the residue to dissolve it, and use it as the test solution. Take another 2g of Patrinia scabra control medicinal material, add 50ml of water, heat and reflux for 60 minutes, cool, filter, concentrate the filtrate to near dryness, add 25ml of methanol to the residue, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 20ml of water to the residue to dissolve it, and prepare the control medicinal material solution in the same way. According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 Edition Part IV General Rules 0502), 2 μl of the test solution and 10 μl of the control medicinal material solution were taken and spotted on the same silica gel G thin layer plate, and cyclohexane-ethyl acetate-methanol-formic acid (6:4:0.5:0.1) was used as the developing agent. After development, 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 ultraviolet light (365nm). In the chromatogram of the test sample, fluorescent spots of the same color appeared at the corresponding position of the chromatogram of the control medicinal material.

[0069] Example 7 Thin layer identification of different batches of Patrinia scabra granules

[0070] Take different batches of Patrinia scabra granules according to the above test solution preparation method to prepare the test solution and the control drug solution, respectively, spot on the same silica gel G thin layer plate, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105 ° C until the spots are clearly colored, and examine under ultraviolet light (365nm). The results are as follows Figure 7As shown in the figure, 1: Patrinia scabra formula granules (18041129); 2: Patrinia scabra formula granules (18041139); 3: Patrinia scabra formula granules (18041149); S: Patrinia scabra control medicinal material. Figure 7 It can be seen that the chromatogram of the Herba Patriniae formula granules and the chromatogram of the control medicinal material show fluorescent spots of the same color at corresponding positions.

[0071] Example 8 Comparison of Patrinia odorata, Patrinia odorata and counterfeit products

[0072] Take 1g of each of the formula granules of Patrinia chinensis (yellow flower Patrinia chinensis), the formula granules of Patrinia chinensis (white flower Patrinia chinensis), the formula granules of Cyperus rotundus, and the formula granules of Mutouhui, grind them into powder, add 20ml of water to dissolve, shake and extract with ethyl acetate twice, 20ml each time, combine the ethyl acetate solution, evaporate to dryness, add 1ml of methanol to the residue to dissolve, and use it as the test solution. Take another 2g of the control medicinal material of Patrinia chinensis, add 50ml of water, heat and reflux for 60 minutes, cool, filter, concentrate the filtrate to near dryness, add 25ml of methanol to the residue, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 20ml of water to the residue to dissolve, and prepare the control medicinal material solution in the same way. According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 Edition Part IV General Rules 0502), according to the method of Example 7, 2 μl of the test solution and 10 μl of the control drug solution were taken and spotted on the same silica gel G thin layer plate, and cyclohexane-ethyl acetate-methanol-formic acid (6:4:0.5:0.1) was used as the developing agent. After development, 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 ultraviolet light (365nm). The results are as follows Figure 8 As shown in the figure, 1-3: Patrinia scabra (yellow Patrinia scabra) formula granules (11803650, 11803651, 11803652); 4-6: Patrinia scabra (white Patrinia scabra) formula granules (1806028, 1806029, 1806030); 7-9: Cyperus rotundus formula granules (2407001, 2407025, 2407026); 10-12: Mutouhui formula granules (2206047, 2206050, 2206052); S: Patrinia scabra control medicinal material. Figure 8 It can be seen that compared with the formula granules of Patrinia scabra (yellow flower Patrinia scabra), the formula granules of Patrinia scabra (white flower Patrinia scabra) lack yellow-brown spots at around Rf value 0.2 and have more blue spots. Therefore, the thin-layer method can identify the formula granules of Patrinia scabra (yellow flower Patrinia scabra) from the formula granules of Patrinia scabra (white flower Patrinia scabra); the number and position of spots on the formula granules of Cyperus rotundus, the formula granules of Mutouhui and the formula granules of Patrinia scabra are quite different and can be easily distinguished. Therefore, the thin-layer method can identify the formula granules of Patrinia scabra from the formula granules of Cyperus rotundus and the formula granules of Mutouhui.

Claims

1. A thin layer identification method for Patrinia scabra granules, characterized in that: The following steps are involved: (1) Preparation of test sample solution: Dissolve the test sample powder in water and shake with ethyl acetate to extract. Evaporate the ethyl acetate to dryness and dissolve the residue in methanol to prepare the test sample solution. (2) Preparation of control medicinal material solution: Take Patrinia chinensis control medicinal material, add water and heat to reflux, filter, concentrate the filtrate to near dryness, add methanol to the residue, ultrasonically treat, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with ethyl acetate, evaporate the ethyl acetate to dryness, and dissolve the residue in methanol to prepare the control medicinal material solution; (3) Take the control medicinal material solution and the test sample solution and spot them on the same thin layer plate, develop them with cyclohexane-ethyl acetate-methanol-formic acid as the developing solvent, take them out, dry them, spray them with sulfuric acid ethanol solution, heat them until the spots are clearly colored, and inspect and compare the thin layer chromatograms; (4) When spots of the same color appear in the chromatogram of the test sample at the corresponding positions in the chromatogram of the control medicinal material, it means that the test sample is the formula granules of Patrinia scabra.

2. The thin layer identification method according to claim 1, characterized in that: The development temperature in step (3) is 4 to 40°C.

3. The thin layer identification method according to claim 1, characterized in that: The humidity of the development in step (3) is 18-88%.

4. Use of the thin layer identification method according to any one of claims 1 to 3 in authenticity identification of Patrinia scabra formula granules.

5. A method for distinguishing Patrinia scabra granules from Cynanchum indica granules and Cynanchum scabra granules, characterized in that: The following steps are involved: (1) Preparation of test sample solution: Dissolve the test sample powder in water and shake with ethyl acetate to extract. Evaporate the ethyl acetate to dryness and dissolve the residue in methanol to prepare the test sample solution. (2) Preparation of control medicinal material solution: Take Patrinia chinensis control medicinal material, add water and heat to reflux, filter, concentrate the filtrate to near dryness, add methanol to the residue, ultrasonically treat, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with ethyl acetate, evaporate the ethyl acetate to dryness, and dissolve the residue in methanol to prepare the control medicinal material solution; (3) Take the control medicinal material solution and the test sample solution and spot them on the same thin layer plate, develop them with cyclohexane-ethyl acetate-methanol-formic acid as the developing solvent, take them out, dry them, spray them with sulfuric acid ethanol solution, heat them until the spots are clearly colored, and inspect and compare the thin layer chromatograms; (4) When spots of the same color appear in the chromatogram of the test sample at the corresponding positions in the chromatogram of the control medicinal material, it means that the test sample is the formula granules of Patrinia scabra.

6. The method according to claim 5, characterized in that: The development temperature in step (3) is 4 to 40°C.

7. The method according to claim 5, characterized in that: The humidity of the development in step (3) is 18-88%.

8. A method for distinguishing the formula granules of Patrinia lutea and Patrinia odorata, characterized in that: The following steps are involved: (1) Preparation of test sample solution: Dissolve the test sample powder in water and shake with ethyl acetate to extract. Evaporate the ethyl acetate to dryness and dissolve the residue in methanol to prepare the test sample solution. (2) Preparation of control medicinal material solution: Take Patrinia chinensis control medicinal material, add water and heat to reflux, filter, concentrate the filtrate to near dryness, add methanol to the residue, ultrasonically treat, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with ethyl acetate, evaporate the ethyl acetate to dryness, and dissolve the residue in methanol to prepare the control medicinal material solution; (3) Take the control medicinal material solution and the test sample solution and spot them on the same thin layer plate, develop them with cyclohexane-ethyl acetate-methanol-formic acid as the developing solvent, take them out, dry them, spray them with sulfuric acid ethanol solution, heat them until the spots are clearly colored, and inspect and compare the thin layer chromatograms; (4) When the test sample chromatogram shows spots of the same color at the corresponding positions with the control medicinal material chromatogram, and there are no yellow-brown spots but blue spots at around Rf value 0.2, it means that the test sample is Baihua Baijiang formula granules; when the test sample chromatogram shows spots of the same color at the corresponding positions with the control medicinal material chromatogram, and there are yellow-brown spots but no blue spots at around Rf value 0.2, it means that the test sample is Huanghua Baijiang formula granules.

9. The method according to claim 8, characterized in that: The development temperature in step (3) is 4 to 40°C.

10. The method according to claim 8, characterized in that: The humidity of the development in step (3) is 18-88%.

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