A method and device for quality identification of Chinese herbal medicine

Abstract

The application relates to the technical field of Chinese herbal medicine identification, and particularly discloses a Chinese herbal medicine quality identification method and device. The method comprises five steps of screening, tablet preparation, observation, reflux and color development comparison. Seven structures, including calcium oxalate cluster crystals, screw conduits, yellow block secretions, wood fibers, leaf epidermal cells, non-glandular hairs and pollen grains, are observed under a microscope and combined with color development comparison, so that the technical problem that the prior art identification method for malachra herb is only used for identifying saponin compounds contained in the malachra herb and is not accurate enough is solved.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine identification, in particular to a method and device for quality identification of traditional Chinese medicine. Background Art

[0002] Spergularia salina is a relatively common traditional Chinese medicine in China, which has the effects of cooling blood, detoxifying, killing insects and relieving itching. In terms of appearance, Spergularia salina is 5 to 20 cm long, with slender stems, cespitose growth and multiple branches; the surface of its stems is pale green to golden yellow, the texture is brittle, and the cross-section is light green to yellowish white. It is commonly used for treating lacquer itch, favus, eczema, erysipelas, scrofula, unknown swelling and toxin, venomous snake bite, nasosinusitis, dental caries pain, traumatic internal injury and other conditions.

[0003] Identifying medicinal materials is the first step in medication use. If the medicinal material cannot be correctly identified, misuse of other medicinal materials will inevitably lead to unpredictable consequences. When Spergularia salina is fresh, its characteristics are obvious and can be easily identified, and the quality of the traditional Chinese medicine can be judged according to its appearance characteristics. However, all traditional Chinese medicinal materials need to be dried. After drying, they turn yellow and brown, and the rods, stems and leaves will shrink and deform, making their characteristics difficult to identify. If the material is further chopped and ground into medicinal powder, its appearance characteristics completely disappear at this time, and only experienced veteran traditional Chinese medicine practitioners can distinguish it. It is difficult for ordinary traditional Chinese medicine practitioners to identify the medicinal material from its appearance and judge its quality.

[0004] In the prior art, the identification method for Spergularia salina is the acetic anhydride-concentrated sulfuric acid test. The process is usually as follows: taking coarse powder of Spergularia salina, adding water and hot-soaking (50-60°C) for 60 minutes, filtering, taking 2 mL of the filtrate placing in a small evaporating dish, evaporating to dryness in a water bath, adding 1 mL of glacial acetic acid dropwise to the residue to dissolve it, then adding 1 mL of acetic anhydride, filtering, adding 2 to 4 drops of sulfuric acid dropwise to the edge of the filtrate. If the solution turns red, it is identified as Spergularia salina.

[0005] Although the above method can qualitatively identify Spergularia salina, it only identifies the triterpenoid saponins contained in Spergularia salina to distinguish Spergularia salina from other medicinal materials. However, not only Spergularia salina contains saponin compounds in traditional Chinese medicinal materials, for example, traditional Chinese medicines such as Panax notoginseng, Ophiopogon japonicus and ginseng also contain saponin compounds. Of course, the possibility that other weeds also contain saponin compounds cannot be ruled out. Therefore, it cannot accurately identify Spergularia salina. Summary of the Invention

[0006] The purpose of the present invention is to provide a method and device for quality identification of traditional Chinese medicine, so as to solve the technical problem mentioned above that in the prior art, the identification method for Spergularia salina only identifies the saponin compounds contained therein and is not accurate enough.

[0007] In order to solve the above problems, the technical solution adopted by the present invention is as follows: a method and device for quality identification of traditional Chinese medicine, comprising the following steps: S1 Screening: The powder of *Lysimachia christinae* herb was screened through a No. 3 sieve, and the screened powder was used as the identification sample powder. S2 preparation: Take a portion of the sample powder from S1, place it in a small petri dish, add 10-15 ml of water and 10-15% glycerol, cover the small petri dish with filter paper, and then place the petri dish in an environment of 25-35℃ to allow the water to slowly evaporate to the concentration of pure glycerol. S3 observation: Use tweezers to take a small amount of powder from the small petri dish after the water in S2 has evaporated, spread it in the center of the slide, add a small drop of pure glycerin, cover with a coverslip and observe under a microscope; S4 reflux: Take 1-3g of sample powder from S1, add 15-45ml of ethanol, heat under reflux for 1-2 hours, filter, evaporate the filtrate, and dissolve the residue in 1-3ml of methanol to obtain a powder solution. S5 colorimetric comparison: Take 1-2 μL of S4 herbal powder solution and spot it on the polyamide film. Develop it with ethanol-water (7:3) as the developing solvent. After development, remove the polyamide film and let it dry. Spray the polyamide film with colorimetric agent and compare it with the standard chromatogram prepared in advance using standard *Lysimachia christinae* according to this method. It is essential to ensure that seven structures are observed in S3: calcium oxalate clusters, spiral vessels, yellow lumpy secretions, wood fibers, leaf epidermal cells, non-glandular hairs, and pollen grains. Furthermore, the color development position of the sample powder in S5 must correspond to that of the standard *Lysimachia christinae* chromatogram. Sample powder showing the same color spots is identified as *Lysimachia christinae* herbal powder.

[0008] The beneficial effects of this implementation plan are as follows: 1. In this application, the ethanol-soluble components of *Lysimachia christinae* are extracted using extraction. After extraction, thin-layer chromatography (TLC) is performed using a polyamide membrane. Since various organic components have different solubilities in the developing solvent, as the solvent flows along the polyamide membrane, components with high solubility in the solvent are less likely to be adsorbed by the polyamide membrane due to their high mixing capacity. These components flow along the polyamide membrane to the upper end, while components with low solubility precipitate easily from the solvent and are adsorbed by the polyamide membrane, thus being adsorbed at the lower end. Therefore, the various components of *Lysimachia christinae* can be separated on the polyamide membrane and displayed using different colorimetric reagents. Furthermore, the solubility of a particular colorimetric component among all components can be determined based on the colorimetric height, which is more accurate than the prior art that only performs qualitative analysis on one component.

[0009] 2. The identification method of this application also includes powder microscopic identification, which utilizes the fact that the powder is more transparent under glycerol to observe the cellular structure of *Rubus idaeus*. This is combined with thin-layer chromatography analysis to ensure that the identification of *Rubus idaeus* is accurate.

[0010] Furthermore, the heating reflux process includes a chassis, an electric heater installed on the chassis, a water tank on the side of the chassis, a water pump on the water tank to control water circulation, an extraction bottle, an extractor, and a condenser installed inside the chassis. The characteristic feature is that the chassis is also equipped with a temperature sensor that can collect temperature data inside the extraction bottle and a pressure sensor that can collect pressure data inside the condenser. Both the pressure sensor and the temperature sensor are connected to a host computer.

[0011] Furthermore, in step S5, a 3-5% aluminum trichloride ethanol solution is used as the color developer, and the solution is dried after spraying and compared under ultraviolet light.

[0012] Furthermore, the heating reflux temperature in S4 is controlled at 60°C.

[0013] Furthermore, the host computer includes a start-up control module, a data receiving module, and a data analysis module; the data receiving module is used to receive data collected by temperature sensors and pressure sensors; the data analysis module is used to perform rule calculations on the data collected by temperature sensors and pressure sensors; the start-up control module is used to control the speed of the electric heater and the water pump.

[0014] Furthermore, an operation panel is provided on the front of the chassis. Attached Figure Description

[0015] Figure 1 This is a front view of the extraction device according to an embodiment of the present invention. Figure 2 This is a side view of the extraction device according to an embodiment of the present invention. Figure 3 This is a logic block diagram of the extraction equipment control and adjustment system according to an embodiment of the present invention. Figure 4 This is a microscopic structural diagram of the powder used in an embodiment of the present invention. Detailed Implementation

[0016] The following detailed description illustrates the specific implementation method: The reference numerals in the accompanying drawings include: water tank 1, water pump 11, inlet pipe 12, outlet pipe 13, control panel 2, display screen 21, operation button 22, mounting port 3, extraction bottle 31, extractor 32, condenser tube 33, fixing clamp 34, temperature sensor 35, chassis 4, electric heater 41, and pressure sensor 42.

[0017] Example: A method for quality identification of traditional Chinese medicine, comprising the following steps: S1 Screening: The powder of *Lysimachia christinae* herb was screened through a No. 3 sieve, and the screened powder was used as the identification sample powder. After being dried, *Lysimachia christinae* is processed into powder. However, the powders vary in size. To ensure better results during the preparation of glass slides and extraction for identification, and to facilitate observation, the *Lysimachia christinae* powder is sieved. Only the powder that can pass through a No. 3 sieve is used as the sample powder.

[0018] S2 preparation: Take a portion of the sample powder from S1, place it in a small petri dish, add 10-15 ml of water and 10-15% glycerol, cover the small petri dish with filter paper, and then place the petri dish in an environment of 25-35℃ to allow the water to slowly evaporate to the concentration of pure glycerol. Mix the powder, water, and glycerin. Use water to spread the powder and glycerin to wet it. Yellow lumpy secretions can be observed in the powder.

[0019] S3 observation: Take a small amount of powder from the small petri dish after the water in S2 has evaporated using tweezers, spread it in the center of a glass slide, add a small drop of pure glycerin, cover with a coverslip, and observe under a microscope; Adding glycerin is to ensure that the powder on the slide does not dry out during observation, which would reduce transparency and make observation difficult. Glycerin also enhances the transparency of the powder section and prevents the powder section from drying out.

[0020] The powder of *Lysimachia christinae* is yellowish-green. Under a microscope, calcium oxalate clusters can be observed, with a diameter of 40-96 μm and mostly short, blunt edges; spiral vessels are present, with a diameter of about 9-16 μm, and free vessels can reach a diameter of 50 μm; there are yellow, blocky secretions; wood fibers are bundled, square, slender, with slightly thick walls, and a diameter of about 11-21 μm; leaf epidermal cells are wavy, with transverse stomata in between; non-glandular hairs are easily seen, mostly 3-celled, with basal cells swollen into a trumpet shape and terminal cells triangular, the whole plant is mostly curved to a certain degree, with warty protrusions on the surface, about 110-200 μm long; pollen grains are spherical, with a diameter of 21-30 μm, with protrusions on the surface, and can reach a diameter of about 38 μm after imbibition. Only when seven structures are observed—calcium oxalate clusters, spiral vessels, yellow lumpy secretions, wood fibers, leaf epidermal cells, non-glandular hairs, and pollen grains—can the microscopic identification of *Rubus idaeus* powder be confirmed as successful. These seven structural features are as follows: Figure 4 As shown.

[0021] S4 reflux: Take 1-3g of sample powder from S1, add 15-45ml of ethanol, heat under reflux for 1-2 hours, filter, take the filtrate and evaporate it, add 1-3ml of methanol to dissolve the residue after evaporation, and dissolve it to obtain the test powder solution. Take 1g of the powder (passed through a No. 3 sieve), add 15ml of ethanol, heat under reflux for 1 hour, filter, evaporate the filtrate to dryness, dissolve the residue in 1ml of methanol to prepare the test solution. Separately, take 1g of *Lysimachia christinae* reference material. Extraction equipment used in the heating reflux process, such as Figure 1-3 As shown, with Figure 1 The direction described is as follows: It includes a chassis 4, which is a box-like structure with a mounting opening 3 in the middle, exposing the internal space. An electric heater 41 is located below the chassis 4. The electric heater 41 is a rectangular block with a recessed upper surface to accommodate an extraction bottle 31. An electric heating wire is placed inside the recess. Therefore, a horizontally positioned fixing clip 34 on the rear side of the chassis 4 can fix the extraction bottle 31, ensuring its lower spherical end is within the recess of the electric heater 41 without contacting it. An extractor 32 is connected to the upper end of the extraction bottle 31, communicating with the opening of the extraction bottle 31. A condenser tube 33, a spherical condenser tube, is fitted onto the upper end of the extractor 32. The extraction bottle 31, extractor 32, and condenser tube 33 are all existing chemical glassware, installed using the fixing clip 34 to ensure a sealed, airtight connection.

[0022] A water tank 1 is located on the right side of the chassis 4. A water pump 11 is located at the bottom of the water tank 1 where it contacts the rear of the chassis. The water pump 11 is a three-speed adjustable water pump. The water pump 11 connects the water in the water tank to the water inlet pipe 12 located inside the rear of the chassis 4. One end of the water inlet pipe 12 is connected to the inlet of the condenser pipe 33, and the other end is connected to the bottom of the water tank 1. The water outlet pipe 13 of the condenser pipe 33 is connected to the water outlet pipe 13 inside the rear panel of the chassis. The water outlet pipe 13 pours the water in the condenser pipe 33 back into the upper part of the water tank 1, thus completing the water circulation. A pressure sensor is located at the position where the upper horizontal plate of the chassis 4 contacts the condenser pipe 33. The chassis 4 has an installation port for the condenser pipe 33, which contains a rubber gasket and a sealing ring. Therefore, when the condenser pipe 33 is inserted into the upper horizontal plate of the chassis 4, a seal is formed between the condenser pipe 33 and the chassis. The pressure sensor 42 is inserted into the condenser pipe 33. A temperature sensor 35 is located at the same height as the opening of the extraction bottle 31 on the rear panel of the chassis 4. The temperature sensor is a temperature probe type sensor, so the temperature sensor 35 can be inserted into the extraction bottle 31 for real-time temperature detection during use.

[0023] An operation panel 2 is provided at the front of the chassis 4. The operation panel 2 is equipped with a display screen 21 and operation buttons 22 to display the temperature and pressure during the extraction process.

[0024] This extraction equipment also includes a temperature and pressure control system, which comprises detection equipment, control equipment, and a host computer, such as... Figure 3 As shown, it includes a start-up control module, a data receiving module, and a data analysis module, which are electrically connected. The detection equipment includes the aforementioned temperature sensor 35 and pressure sensor 42. The control equipment includes a water pump 11 and an electric heater 41.

[0025] The data receiving module is used to receive data detected by the temperature sensor 35 and the pressure sensor 42 of the detection equipment, and to package and transmit it to the data analysis module.

[0026] The data analysis module processes the data from the data receiving module and then outputs it to the startup control module.

[0027] The start control module is used to control the opening and closing of the water pump 11 and the electric heater 41 and their speed settings.

[0028] Temperature sensor 35 collects data every 1 second, so the data receiving module receives data x1, x2, x3, x4, and x5 every 5 seconds and packages them. Pressure sensor 42 collects data every 2.5 seconds, so the data receiving module receives data y1 and y2 every 5 seconds and packages them. The data receiving module sends the packaged data to the data analysis module. The data analysis module calculates x0 by weighted averaging x1, x2, x3, x4, and x5. It compares x0 with the set value x. When x0 is greater than, less than, or equal to, the corresponding temperature outputs are -1, 0, and 1, respectively. It calculates y0 by averaging y1 and y2 and compares y0 with the set value y. When y0 is greater than, less than, or equal to, the corresponding air pressure outputs are 0, 1, and 2, respectively.

[0029]

[0030] The sum of the temperature output and the air pressure output can be -1, 0, 1, or 2. When the sum is -1, the control module is activated, adjusting water pump 1 to level 1 and electric heater 41 to heating mode. When the sum is 0, the control module is activated, adjusting water pump 1 to level 1 and electric heater 41 to heating mode. When the sum is 1, water pump 1 is adjusted to level 2 and electric heater 41 to heat preservation mode. When the sum is 2, water pump 1 is adjusted to level 3 and electric heater 41 is turned off.

[0031] S5 colorimetric comparison: Take 1-2 μL of S4 herbal powder solution and spot it on the polyamide film. Develop it with ethanol-water (7:3) as the developing solvent. After development, remove the polyamide film and let it dry. Spray the polyamide film with colorimetric agent and compare it with the standard chromatogram prepared in advance using standard *Lysimachia christinae* according to this method. Take 2 μl of the above solution and spot it onto the same polyamide film. Use ethanol-water (7:3) as the developing solvent to develop the film, remove it, and let it air dry.

[0032] The colorimetric agent can be sprayed onto the dried polyamide film using a 5% aluminum trichloride ethanol solution. After the polyamide film is dried with hot air, it is examined under a UV lamp (365nm) and compared with the standard chromatogram of *Lysimachia christinae* prepared according to this method. Only those showing the same color fluorescent main spot at the same position can be confirmed as having passed the chromatographic identification.

[0033] Seven structures were observed in S3: calcium oxalate clusters, spiral vessels, yellow lumpy secretions, wood fibers, leaf epidermal cells, non-glandular hairs, and pollen grains. In S5, the sample powder and the standard *Rubus parvifolius* chromatogram showed the same color spots at the corresponding positions, and the sample powder was identified as *Rubus parvifolius* medicinal powder.

[0034] The above descriptions are merely embodiments of the present invention, and common knowledge regarding specific structures and characteristics in the solutions is not described in detail here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the structure of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the implementation of the present invention or the practicality of the patent.

Claims

1. A method for quality identification of traditional Chinese medicine, characterized in that: Includes the following steps: S1 Screening: The powder of *Lysimachia christinae* herb was screened through a No. 3 sieve, and the screened powder was used as the identification sample powder. S2 preparation: Take a portion of the sample powder from S1, place it in a small petri dish, add 10-15ml of water and 10-15% glycerol, cover the petri dish with filter paper, and then place the petri dish in an environment of 25-35℃ to allow the water to slowly evaporate until the glycerol concentration reaches pure glycerol. S3 observation: Use tweezers to take a small amount of powder from the small petri dish after the water in S2 has evaporated, spread it in the center of the slide, add a drop of pure glycerin, cover with a coverslip and observe under a microscope; S4 reflux: Take 1-3g of sample powder from S1, add 15-45ml of ethanol, heat under reflux for 1-2 hours, filter, take the filtrate and evaporate it, add 1-3ml of methanol to dissolve the residue after evaporation, and dissolve it to obtain the test powder solution. S5 colorimetric comparison: Take 1-2 μL of S4 herbal powder solution and spot it on a polyamide film. Develop the film using ethanol-water (volume ratio 7:3) as the developing solvent. After development, remove the polyamide film and air dry it. Spray it with 3-5% aluminum chloride ethanol solution, dry it with hot air, and examine it under a 365nm ultraviolet lamp. Compare it with the standard chromatogram of *Lysimachia christinae* prepared in advance using the same method. It is essential to ensure that seven structures are observed in S3: calcium oxalate clusters, spiral vessels, yellow lumpy secretions, wood fibers, leaf epidermal cells, non-glandular hairs, and pollen grains. Furthermore, the sample powder in S5 must show fluorescent main spots of the same color at the corresponding positions as the reference medicinal material chromatograms to be identified as *Lysimachia christinae* herbal powder.

2. The method for quality identification of traditional Chinese medicines according to claim 1, characterized in that: The heating reflux temperature in S4 is controlled at 60°C.

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