A thin layer identification method of scutellaria baicalensis and its preparation

Abstract

The invention discloses a thin layer identification method of tongcao and its preparation, the method comprising: preparing a control medicinal material solution, a sample solution of a decoction piece, a formula granule and a standard decoction sample solution, taking the control medicinal material solution and the sample solution and spotting them on the same thin layer plate respectively, developing them with a developing agent and then spraying a color developer for color development, obtaining a thin layer chromatogram to complete the identification operation. The method of the invention is simple and fast to operate, has good chromatographic spot separation, low detection cost, safety, environmental protection, strong specificity, and can be used to distinguish the authenticity of tongcao and the distinction of different origins.

Description

Technical Field

[0001] The invention relates to a thin layer identification method, in particular to a thin layer identification method for smilax china and its preparation. Background Art

[0002] Small Tongcao is the dried stem pith of Stachyurus himalaicus Hook.f.et Thoms., Stachyurus chinensis Franch., or Helwingia japonica (Thunb.) Dietr. of the Cornaceae family. The stems are cut in autumn, cut into sections, and the pith is taken out while fresh, straightened, and dried. Small Tongcao has the effects of clearing heat and promoting diuresis, ventilating and promoting lactation. Through pharmacological experiments, it has been found that different varieties of Small Tongcao have good diuretic, anti-inflammatory and antipyretic effects. It is mainly used clinically for urinary tract infection, urine retention or oliguria, fever thirst, yellow and red urine, milk obstruction, etc.

[0003] The 2020 edition of the Chinese Pharmacopoeia does not include the TLC identification item of Xiaotongcao. Most of the existing TLC identification methods for Xiaotongcao and its preparations use chloroform as the developing agent, and the chromatographic information is not rich enough. Literature: Identification method of Xiaotongcao and its counterfeits [J]. Strait Pharmacy, 2006, 18(1): 104-105. This method is only used to distinguish Xiaotongcao and its counterfeits. The chromatographic information is single and there is only one distinguishing spot. Literature: Identification of Xiaotongcao and its counterfeits [J]. Journal of Traditional Chinese Medicine and Pharmacy, 2005, 11(2): 57-58. There are only microscopic and property identification, as well as ultraviolet spectrum absorption peaks. This method is not accurate. No accurate and effective TLC identification method has been reported in the existing literature, making it difficult to effectively distinguish Xiaotongcao and its counterfeits and different origins. Summary of the invention

[0004] Purpose of the invention: The present invention aims to provide a thin layer identification method of Herba Smilacis Glabrae and its preparations which has rich chromatographic spot information and can be used to distinguish counterfeit products and Herba Smilacis Glabrae of different origins.

[0005] Technical solution: The thin layer identification method of the herb and its preparation of the present invention comprises the following steps:

[0006] (1) Prepare control medicinal material solutions, decoction sample solutions, formula granules and standard decoction sample solutions;

[0007] (2) The control medicinal material solution and the test sample solution are spotted on the same thin layer plate, developed with a developing agent, and then sprayed with a color developer for color development. The plate is heated until the spots are clearly colored to obtain a daylight thin layer chromatogram, thereby completing the identification operation. The developing agent is a mixed solution of dichloromethane, methanol and water, wherein the volume ratio of dichloromethane:methanol:water is 18:2 to 2.5:1.

[0008] Furthermore, the preparation method of the control medicinal material solution and the decoction piece test solution in step (1) comprises the following steps: taking the Xiaotongcao control medicinal material and the Xiaotongcao decoction piece, respectively, decocting them with water or heating them under reflux, filtering them, ultrasonically treating them with ethanol solution, filtering them, evaporating the solution to dryness, dissolving the residue with alcohol solvent, and obtaining the control medicinal material solution and the decoction piece test solution; the preparation method of the formula granules and the standard decoction test solution comprises the following steps: taking the Xiaotongcao formula granules and the standard decoction samples, ultrasonically treating them with ethanol solution, filtering them, evaporating the solution to dryness, dissolving the residue with alcohol solvent, and obtaining the formula granules and the standard decoction test solution.

[0009] Furthermore, when the control medicinal material of xiaotongcao and the xiaotongcao decoction pieces are decocted with water, the mass volume of the control medicinal material, the xiaotongcao decoction pieces and water is 1:50g / mL~150g / mL, the decoction or heating reflux treatment time is 2h~4h, and the ultrasonic treatment power is 250-300W, the frequency is 40kHz, and the time is 15min~60min.

[0010] Furthermore, when the Xiaotongcao formula granules and the standard decoction are dissolved in ethanol, the mass volume of the Xiaotongcao formula granules, the standard decoction and ethanol is 1:10g / mL~20g / mL, the power of ultrasonic treatment is 250-300W, the frequency is 40kHz, and the time is 15min~60min.

[0011] Furthermore, in step (2), the spotting amount of the control medicinal material solution and the test solution is 5 μl to 20 μl; the color developer is a 10% sulfuric acid ethanol test solution, which is heated at 105°C for several minutes after spraying the color developer and inspected under white light; the thin layer plate is a silica gel G thin layer plate; on the thin layer chromatogram, a main spot of the same color is displayed in the chromatogram of the test solution at a position corresponding to the chromatogram of the control medicinal material solution.

[0012] Principle analysis: Based on the chemical structure and properties of each active ingredient of traditional Chinese medicine, following the principle of similar and compatible extraction, suitable extraction solvents are used to prepare test and control medicinal material solutions simply and quickly. Then, suitable developing agents are used for development. Various chemical components will be well separated on the thin layer plate according to the different adsorption, desorption, re-adsorption and re-desorption abilities of the selected developing agents. With the help of various active ingredients with similar polarity, different color spots are presented on the same thin layer plate under different inspection conditions, and multi-information thin layer chromatograms are obtained.

[0013] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: the chromatographic spots on the thin-layer chromatogram obtained by the method are rich in information, can be used to effectively distinguish fake smilax china and their different origins, and the operation is simple and fast, does not destroy the chemical composition of smilax china, and presents the prototype components of smilax china to the greatest extent. The chromatographic spots have good separation, and the use of toxic reagents such as chloroform is avoided. The detection cost is low, safe, and environmentally friendly, and it is more conducive to quality supervision. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a thin layer chromatogram of the Xiaotongcao formula granules in Example 1 of the present invention;

[0015] Figure 2 is a thin layer chromatogram of the standard decoction of Herba Lysimachiae in Example 1 of the present invention;

[0016] Figure 3 It is a thin layer chromatogram of the Herba Lysimachiae in Example 1 of the present invention;

[0017] Figure 4 is a thin layer chromatogram of different sample amounts in Example 2 of the present invention;

[0018] Figure 5 is a thin layer chromatogram at different temperatures in Example 2 of the present invention;

[0019] Figure 6 is a thin layer chromatogram of different humidity in Example 2 of the present invention;

[0020] Figure 7 It is a thin layer chromatogram of the small tongcao herb in comparative example 1 of the present invention;

[0021] Figure 8 It is a thin layer chromatogram of the Xiaotongcao formula granules in Comparative Example 2 of the present invention;

[0022] Fig. 9 It is a thin layer chromatogram of the Xiaotongcao formula granules in Comparative Example 2 of the present invention;

[0023] Fig.10 This is a thin layer chromatogram comparing the standard decoction of the small tongcao and its counterfeit in Comparative Example 3 of the present invention;

[0024] Fig.11 It is a thin layer chromatogram comparing the standard decoction of the small tongcao and its different batches of counterfeit products in comparative example 3 of the present invention;

[0025] Fig.12 It is a thin layer chromatogram comparing the small tongcao and its different base standard decoctions in comparative example 3 of the present invention. DETAILED DESCRIPTION

[0026] The technical solution of the present invention is further described below in conjunction with the accompanying drawings.

[0027] The present invention adopts the following instruments and reagents:

[0028] Instruments: CAMAG TLC VISUALIZER, 1 / 10,000 balance (Mettler-Toledo, Switzerland), KQ-250E ultrasonic cleaner (Kunshan Ultrasonic Electronics Co., Ltd.), electronic temperature-controlled electric furnace (Jiangyin Baoli Scientific Research Instrument Co., Ltd.), GKC temperature-controlled water bath (Nantong Huatai Experimental Instrument Co., Ltd.), silica gel G thin layer plate (Qingdao Ocean Chemical Plant).

[0029] Reagents: dichloromethane (Sinopharm Chemical Reagent Co., Ltd.), chloroform (Sinopharm Chemical Reagent Co., Ltd.), formic acid (Sinopharm Chemical Reagent Co., Ltd.), methanol (Sinopharm Chemical Reagent Co., Ltd.), and ethanol (Sinopharm Chemical Reagent Co., Ltd.) were all of analytical grade; water was ultrapure water.

[0030] The control medicinal material of Xiaotongcao (Chinese flag flower) (batch number: YC121906019, working control).

[0031] Xiaotongcao formula granules (batch numbers: 19120139, 19120149, 19120159), Xiaotongcao standard decoction (batch numbers: DG1908018, DG1908020, DG121905065), and Xiaotongcao medicinal materials (batch numbers: YC1908018, YC1908020, YC121905065) were prepared by Jiangyin Tianjiang Pharmaceutical Co., Ltd.

[0032] Standard decoction of Juncus effusus (batch number: DG1905088, DG1905099, DG2102011); Standard decoction of Hydrangea chinensis (batch number: DG1805019, DG1805020, DG1805021); Standard decoction of Rhizoma Anemarrhenae (batch number: DG2302006, DG2302007, DG2302008); Standard decoction of Rhizoma Cibotii (batch number: DG2201001, DG2201002, DG2201003)

[0033] Example 1

[0034] Preparation of test solution of formula granules or standard decoction: Take 2g of Xiaotongcao formula granules or Xiaotongcao standard decoction, grind it into powder, add 20ml of ethanol, ultrasonically treat it for 30 minutes, filter it, evaporate the filtrate to dryness, add 1ml of ethanol to the residue to dissolve it, and use it as the test solution.

[0035] Preparation of test solution of medicinal materials or decoction pieces: take 2g of Xiaotongcao medicinal materials or Xiaotongcao decoction pieces, add 200ml of water, heat and reflux for 3 hours, cool, filter, evaporate the filtrate to dryness, add 10ml of ethanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of ethanol to dissolve the residue, and use it as the test solution.

[0036] Preparation of control medicinal material solution: Take 2g of Chinese cyperus rotundus (Chinese flag flower) control medicinal material, add 200ml of water, heat and reflux for 3 hours, cool, filter, evaporate the filtrate to dryness, add 10ml of ethanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of ethanol to the residue to dissolve it to prepare a control medicinal material solution.

[0037] Thin layer identification steps: Pipette 10 μl of the test solution and the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, use dichloromethane-methanol-water (volume ratio 18:2:1) as the developing agent, develop, take out, dry, spray with 10% sulfuric acid ethanol test solution, heat at 105℃ for several minutes, and inspect under white light.

[0038] Evaluation requirements: In the chromatogram of the test sample, a main spot of the same color appears at the corresponding position in the chromatogram of the control medicinal material.

[0039] Result analysis: Figure 1 1 to 3 correspond to the chromatograms of the test samples of the granules of the herb Xiaotongcao with batch numbers 19120139, 19120149, and 19120159, respectively, and S is the chromatogram of the reference herb Xiaotongcao. Figure 2 1 to 3 correspond to the chromatograms of the test samples of the standard decoction of Smilax glabra with batch numbers DG1908018, DG1908020, and DG121905065, respectively, and S is the chromatogram of the reference medicinal material of Smilax glabra. Figure 3 1 to 3 correspond to the chromatograms of the test samples of the herb Xiaotongcao with batch numbers YC1908018, YC1908020, and YC121905065, respectively, and S is the chromatogram of the herb Xiaotongcao reference.

[0040] Example 2

[0041] The durability verification of the thin layer identification in Example 1 specifically includes:

[0042] (1) Investigation of different sample quantities:

[0043] Take Xiaotongcao formula granules (batch number: 19120149) and Xiaotongcao control medicinal materials, prepare test solution and control medicinal material solution respectively according to the method described in Example 1, and take 5μL, 8μL, 10μL, 12μL of Xiaotongcao formula granules test solution and 5μL, 8μL, 10μL, 12μL of Xiaotongcao control medicinal material solution respectively, spot on the same silica gel G thin layer plate, use dichloromethane-methanol-water (volume ratio 18:2:1) as the developing agent, develop, take out, dry, spray with 10% sulfuric acid ethanol test solution, heat at 105℃ for several minutes, and inspect under white light. The results are shown in Figure 4 , Figure 4 1 to 4 correspond to the chromatograms of 5 μL, 8 μL, 10 μL, and 12 μL of the test sample of the Xiaotongcao formula granules, and 5 to 8 correspond to the chromatograms of 5 μL, 8 μL, 10 μL, and 12 μL of the Xiaotongcao reference medicinal material. Figure 4 It can be seen that when the sample volume is 5μL~12μL, the chromatogram of the Xiaotongcao formula granules and the chromatogram of the control medicinal materials show clear spots at corresponding positions without other interference.

[0044] (2) Investigation at different temperatures:

[0045] Take Xiaotongcao formula granules (batch number: 19120149) and Xiaotongcao control medicinal materials, prepare test solution and control medicinal material solution respectively according to the method described in Example 1, and take 10μL of Xiaotongcao formula granules test solution and 10μL of Xiaotongcao control medicinal material solution respectively, spot on the same silica gel G thin layer plate, use dichloromethane-methanol-water (volume ratio 18:2:1) as the developing agent, develop at low temperature (T:4℃, RH:25%), room temperature (T:22.8℃, RH:25%) and high temperature (T:35℃, RH:25%), take out, dry, spray with 10% sulfuric acid ethanol test solution, heat at 105℃ for several minutes, and inspect under white light. Results are shown in Figure 5 , Figure 5 1 to 3 correspond to the chromatograms of the test sample of the Xiaotongcao formula granules, and S corresponds to the chromatogram of the Xiaotongcao reference medicinal material. Figure 5 It can be seen that under different temperature conditions, the main spots of the same color appear at the corresponding positions of the chromatograms of the test and control medicinal materials of the Xiaotongcao formula granules, and the separation effect is good. The experimental results show that temperature has little effect on the thin layer identification of Xiaotongcao formula granules, indicating that the thin layer identification method has good durability at different temperatures.

[0046] (3) Investigation of different humidity levels:

[0047] Take Xiaotongcao formula granules (batch number: 19120149) and Xiaotongcao control medicinal materials, prepare test solution and control medicinal material solution respectively according to the method described in Example 1, and take 10μL of Xiaotongcao formula granules test solution and 10μL of Xiaotongcao control medicinal material solution respectively, spot on the same silica gel G thin layer plate, use dichloromethane-methanol-water (volume ratio 18:2:1) as the developing agent, develop under different humidity conditions (RH: 18%, 25%, 88%), take out, dry, spray with 10% sulfuric acid ethanol test solution, heat at 105℃ for several minutes, and inspect under white light. The results are shown in Figure 6 , Figure 6 1 to 3 correspond to the chromatograms of the test sample of the Xiaotongcao formula granules, and S corresponds to the chromatogram of the Xiaotongcao reference medicinal material. Figure 6 It can be seen that under different humidity conditions, the chromatograms of the test and control herbal medicines of the Xiaotongcao formula granules show the same main spots of color at the corresponding positions, and the separation effect is good. The experimental results show that humidity has little effect on the thin layer identification of the Xiaotongcao formula granules, indicating that the thin layer identification method has good durability under different humidity.

[0048] Comparative Example 1

[0049] This comparative example adopts the method of the literature (Identification of Xiaotongcao and its counterfeit products [J]. Strait Pharmacy, 2006, 18 (1)) to perform thin layer chromatography on Xiaotongcao formula granules. The specific process is as follows:

[0050] Preparation of test solution: Take 3g of Herba Cynanchum wilfordii, crush it slightly, add 70% methanol to reflux and extract for 1h, cool, filter, evaporate the filtrate to dryness, add 1ml of methanol to dissolve it, and use it as the test solution.

[0051] Preparation of control medicinal material solution: Take 3 g of Chinese gentian (Chinese flag flower) medicinal material, add 70% methanol to reflux and extract for 1 hour, cool, filter, evaporate the filtrate to dryness, add 1 ml of methanol to dissolve, and use as the control medicinal material solution.

[0052] Thin layer identification steps: Pipette 5μl of the test solution and spot it on the same silica gel G thin layer plate, use chloroform-methanol (volume ratio 4:1) as the developing agent, develop, take out, dry, spray with 10% phosphomolybdic acid ethanol solution, heat at 100℃ until the spots are clearly colored, and inspect under white light.

[0053] Evaluation requirements: In the test sample chromatogram, a main spot of the same color appears at the corresponding position in the reference sample chromatogram.

[0054] Result analysis: Figure 7 In the chromatograms of the test sample and the reference medicinal material, the spots are blurred and the chromatographic information is not rich enough.

[0055] Comparative Example 2

[0056] This comparative example adopts different ratios of developing agents: chloroform-methanol-water-formic acid (volume ratio 18:2:1:0.05) and dichloromethane-methanol-water-formic acid (volume ratio 18:1:2:0.05) for development, and compares the preparation methods of the test sample and the control medicinal material solution under the present invention and the method, and the developing agent for the detection of the Xiaotongcao formula granules. The specific process is as follows:

[0057] 1: Take 2g of this product, grind it into powder, add 20ml of ethanol, ultrasonically treat it for 30 minutes, filter it, evaporate it to dryness, add 1ml of ethanol to the residue to dissolve it, and use it as the test solution.

[0058] 2: Take 2g of this product, grind it into powder, add 20ml of methanol, ultrasonically treat it for 30 minutes, filter it, evaporate it to dryness, add 1ml of ethanol to the residue to dissolve it, and use it as the test solution.

[0059] 3: Take 2g of this product, grind it into powder, add 20ml of water to dissolve it, shake and extract it twice with ethyl acetate, 25ml each time, combine the ethyl acetate liquid, evaporate to dryness, add 1ml of ethanol to dissolve the residue, and use it as the test solution.

[0060] S: Take 2g of the control medicinal material of Smilax glabra, add 200ml of water, boil for 3 hours, filter, evaporate to dryness, add 10ml of ethanol to the residue, ultrasonically treat for 30 minutes, filter, evaporate to dryness, add 1ml of ethanol to the residue to dissolve it, and use it as the control medicinal material solution.

[0061] 10 μL of each of the three test sample solutions and one control medicinal material solution was taken and spotted on a thin layer plate, and developed with silica gel G thin layer plate, chloroform-methanol-water-formic acid (volume ratio of 18:2:1:0.05) and silica gel G thin layer plate, dichloromethane-methanol-water-formic acid (volume ratio of 18:1:2:0.05), respectively. After development, take out, dry, spray with 10% sulfuric acid ethanol test solution, heat at 105°C for several minutes, and inspect under white light.

[0062] Result analysis: Figure 8-Figure 9 1 to 3 are the chromatograms of the test products of the granules of the herb Xiaotongcao prepared by the three preparation methods, and S is the chromatogram of the reference herb Xiaotongcao. Figure 7 The thin layer plate was a common silica gel G thin layer plate, chloroform-methanol-water-formic acid (volume ratio 18:2:1:0.05), Fig. 9 The thin layer plate was a common silica gel G thin layer plate, dichloromethane-methanol-water-formic acid (volume ratio 18:1:2:0.05). When the developing agent chloroform-methanol-water-formic acid (volume ratio 18:2:1:0.05) was used, the spots showed obvious stratification and poor separation. When the method 2 was used to prepare the sample of the Xiaotongcao formula granule, the sample spot information was less, indicating that the sample preparation method was not suitable for the preparation of Xiaotongcao.

[0063] Comparative Example 3

[0064] This comparative example uses the thin layer chromatography identification method in the Xiaotongcao formula granule standard published by Guangdong Province, Shanghai City, etc. to distinguish the Xiaotongcao standard soup and its counterfeit products, Juncus effusus, Hydrangea cylindrica, and Tongcao different original green pods by thin layer chromatography. The specific process is as follows:

[0065] Preparation of test solution: Take 2 g of Xiaotongcao and its counterfeits and different base original standard soups, treat them by ultrasonic for 30 minutes, filter, evaporate the filtrate to dryness, add 1 ml of ethanol to dissolve the residue, and use it as the test solution.

[0066] Preparation of control medicinal material solution: Take 1 g of Chinese gentian herb, add 50 ml of ethanol, and prepare the control medicinal material solution in the same way.

[0067] Thin layer identification steps: Pipette 10μl of the test solution and 5μl of the control medicinal material solution, respectively spot them on the same silica gel G thin layer plate, use dichloromethane-ethyl acetate (volume ratio 5:1) as the developing agent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ for several minutes, and inspect under ultraviolet light (365nm).

[0068] Evaluation requirements: In the test sample chromatogram, a main spot of the same color appears at the corresponding position in the reference sample chromatogram.

[0069] Result analysis: Fig.10 1 to 3 are three kinds of fake standard soups of Juncus effusus, Hydrangea chinensis, and Smilax glabra, 4 is a standard soup of different origins of Qingjiaoye, 5 is Smilax glabra (Chinese flag flower), and S is a control herb of Smilax glabra. The difference between the fakes and Smilax glabra is obvious in the comparison of chromatograms. However, there is almost no difference in the comparison of chromatograms of different origins and Smilax glabra, indicating that this method is difficult to distinguish different origins of Smilax glabra. Fig.11 The difference between different batches of counterfeit products and Xiaotongcao Biaotang is obvious. Fig.12 The method of the present invention is different from the Xiaotongcao standard soup, and there are obvious blue spots to distinguish them. This shows that the present invention can not only distinguish the fake Xiaotongcao, but also clearly distinguish Xiaotongcao and its different origins.

[0070] Comprehensive analysis: The thin-layer identification method of the herb and its preparation provided by the present invention is simpler and faster in the preparation of the sample, does not require steps such as ethyl acetate extraction, avoids the use of toxic reagents such as chloroform, overcomes the defect of the prior art that the chromatographic information is lacking when the herb is thin-layer identified, and is safer and more environmentally friendly. The thin-layer identification method of the present invention has more abundant chromatographic spots, reflects the information of the original chemical components in the sample, and overcomes the defect of the existing thin-layer identification method that the chromatographic spot information is less. The thin-layer identification method of the present invention adopts "the main spot of the same color in the chromatogram of the sample at the corresponding position of the chromatogram of the control medicinal material" for quality evaluation, and there are more spots corresponding to the control medicinal material in the sample. The thin-layer plate of the present invention only needs an ordinary silica gel G thin-layer plate, and does not need to use an expensive high-efficiency silica gel G thin-layer plate, and can achieve the effect of rich spot information and good separation. The present invention can clearly distinguish the herb and its counterfeit, and can also distinguish the herb and its different origins, and has practical applications in distinguishing authenticity and distinguishing different origins.

Claims

1. A thin layer identification method for a scutellaria baicalensis preparation, characterized in that: The steps include: (1) preparing control medicinal material solution, test solution of Herba Schizonepetae decoction pieces, Herba Schizonepetae formula granules and Herba Schizonepetae standard decoction test solution; (2) Take the control medicinal material solution and the test sample solution and spot them on the same thin layer plate respectively, develop them with a developing agent and then spray them with a color developing agent to obtain a thin layer chromatogram to complete the identification operation. The developing agent is a mixed solution of dichloromethane, methanol and water, wherein dichloromethane: The volume ratio of methanol: water is 18:2 to 2.5:1; The preparation method of the control medicinal material solution and the test solution of the herb slices of xiaotongcao in step (1) comprises the following steps: taking the herb control medicinal material and the herb slices of xiaotongcao, respectively, boiling them in water or heating them under reflux, filtering them, ultrasonically treating them with an ethanol solution, filtering them, evaporating the solution to dryness, dissolving the residue with an alcohol solvent, and obtaining the control medicinal material solution and the test solution of the herb slices of xiaotongcao; the preparation method of the herb formula granules and the herb standard decoction test solution comprises the following steps: taking the herb formula granules and the herb standard decoction samples, ultrasonically treating them with an ethanol solution, filtering them, evaporating the solution to dryness, dissolving the residue with an alcohol solvent, and obtaining the herb formula granules and the herb standard decoction test solution; The thin layer plate in step (2) is a silica gel G thin layer plate.

2. The thin layer identification method of the Xiaotongcao preparation according to claim 1 is characterized in that: The mass volume ratio of the reference medicinal material or the decoction pieces of the Herba Smilacis Glabrae to water is 1:50 g / mL to 150 g / mL.

3. The thin layer identification method of the Xiaotongcao preparation according to claim 2 is characterized in that: The mass volume ratio of the xiaotongcao formula granules or standard decoction to ethanol is 1:10g / mL to 20g / mL.

4. The thin layer identification method of the Xiaotongcao preparation according to claim 2 is characterized in that: In the preparation process of control medicinal material solution and decoction sample solution, the decoction or heating reflux treatment time is 2h~4h, the ultrasonic treatment power is 250-300W, the frequency is 40kHz, and the time is 15min~60min; in the preparation process of formula granules and standard decoction sample solution, the ultrasonic treatment power is 250-300W, the frequency is 40kHz, and the time is 15min~60min.

5. The thin layer identification method of the Xiaotongcao preparation according to claim 1 is characterized in that: In step (2), the spotting volume of the control medicinal material solution and the test sample solution is 5 μl to 20 μl.

6. The thin layer identification method of the Xiaotongcao preparation according to claim 1 is characterized in that: In step (2), the color developer is 10% sulfuric acid ethanol test solution. After spraying the color developer, heat it and inspect it under white light.

7. The thin layer identification method of the Xiaotongcao preparation according to claim 1 is characterized in that: In step (2), on the thin layer chromatogram, a main spot of the same color appears in the chromatogram of the test solution at a position corresponding to that in the chromatogram of the control medicinal material solution.

Images