Thin layer identification method for Xiebai, and method for distinguishing Xiebai from counterfeit products

Abstract

The present invention discloses a thin-layer identification method for Xiebai, a method for distinguishing Xiebai from counterfeit products, and the steps of preparing a test sample solution and a control medicinal material solution; spotting the samples on the same thin-layer plate, unfolding the sample, taking the sample out, drying the sample, spraying the sample with sulfuric acid ethanol solution, heating the sample until the spots are clearly colored, and inspecting the sample under sunlight and ultraviolet light; when a main spot or a fluorescent main spot of the same color appears in the chromatogram of the test sample at a position corresponding to the chromatogram of the control medicinal material, the test sample is Xiebai. The present invention solves the problem of inapplicability of the pharmacopoeia method due to the small amount of fat-soluble components in Xiebai formula granules; the sample is treated with a simple treatment method and inspected under sunlight and ultraviolet light at the same time, resulting in a large number of spots and good separation, and multiple components in the sample can be detected, greatly improving the specificity of the method; Xiebai and its counterfeit products, such as schizonepeta tenuifolia and shallot, can be easily distinguished, and has great practical value.

Description

Technical Field

[0001] The invention relates to a thin-layer identification method for Xiebai and a method for distinguishing Xiebai from counterfeit Xiebai, and belongs to the field of thin-layer identification of Xiebai. Background Art

[0002] Xiebai (Xiebai) is the dried bulb of Allium macrostemon Bge. or Allium chinense G. Don, both of the Liliaceae family. It is harvested in summer and autumn, cleaned, and its fibrous roots removed. It is then steamed or blanched thoroughly in boiling water, then sun-dried. Xiebai is primarily produced in Northeast China, Hebei, Jiangsu, and Hubei, but is also found throughout most of China. It contains a rich variety of medicinal ingredients, including saponins, volatile oils, acidic substances, and nitrogenous compounds.

[0003] Formulated Chinese medicine granules represent a breakthrough in the application of traditional Chinese medicine slices. They are typically made from slices of Chinese medicine through standardized extraction (usually water extraction), concentration, drying, and granulation. Their properties, flavor, meridian distribution, and efficacy remain essentially the same as the original slices. They also require no decoction, are easy to prepare, store, and consume, and are gradually gaining widespread application. The 2020 edition of the Chinese Pharmacopoeia, Volume 1, under the heading of Xiebai (Baibai), only includes a thin-layer identification (TLC) test for Xiebai. Currently, there are no corresponding quality control standards for Xiebai formula granules.

[0004] Most literature uses pharmacopoeial methods, using n-hexane as the extraction solvent, for thin-layer chromatography identification of Xiebai medicinal materials, decoction pieces, and granules. Some literature uses n-butanol as the extraction solvent, followed by multiple column purification to extract water-soluble components. However, after water extraction, the fat-soluble components of traditional Chinese medicine granules are relatively low. Direct application of the pharmacopoeial method can result in a small number of spots, unclear results, and poor resolution. The column separation process is complex, and while multiple column separations have been shown to effectively purify samples, the preparation process is cumbersome and requires high operator skills, making it unsuitable for practical application.

[0005] The patent application (A method for processing water-soluble components in Xiebai (Xiebai), a thin-layer chromatography identification method, and the application of a developing agent, application number: 2022106862707) uses n-butanol as the extraction solvent and then purifies the sample through multiple column passes to extract the water-soluble components. While the column pass process is complex and can effectively purify the sample, the preparation process is cumbersome and requires high operator skills, making it unsuitable for practical application.

[0006] Therefore, it is an important topic to establish a rapid, simple and low-cost thin layer identification method for Xiebai formula granules. Summary of the Invention

[0007] Purpose of the invention: The technical problem to be solved by the present invention is to provide a thin layer identification method for Xiebai and a method for distinguishing Xiebai from counterfeit products.

[0008] Technical solution: To solve the above technical problems, the present invention provides a thin layer identification method for Xiebai, comprising the following steps:

[0009] (1) Prepare the test solution: take the test sample powder, add methanol, ultrasonically treat, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with a saturated n-butanol solution with water, combine the n-butanol solution, wash with ammonia test solution, discard the ammonia test solution, wash with a saturated n-butanol aqueous solution, discard the water solution, evaporate the n-butanol solution to dryness, dissolve the residue in methanol, and use it as the test solution;

[0010] (2) Preparation of Xiebai control solution: Take Xiebai control medicinal material, add water, heat to reflux, cool, filter, concentrate the filtrate to nearly dryness, add methanol to the residue, ultrasonicate, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with a saturated n-butanol solution with water, combine the n-butanol solution, wash with ammonia test solution, discard the ammonia test solution, wash with a saturated n-butanol aqueous solution, discard the water solution, evaporate the n-butanol solution to dryness, dissolve the residue in methanol, and use it as the control medicinal material solution;

[0011] (3) Spot the test sample solution and the Xiebai control solution on the same thin layer plate, develop with chloroform-ethyl acetate-methanol-water as the developing solvent, remove, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and inspect under sunlight and ultraviolet light respectively;

[0012] (4) When a main spot or fluorescent main spot of the same color appears in the chromatogram of the test sample at the corresponding position in the chromatogram of the control medicinal material, the test sample is Xiebai.

[0013] Wherein, the amount of the sample to be tested in step (3) is 5 to 15 μl.

[0014] Wherein, the spotting volume of the Xiebai control solution in step (3) is 5 to 15 μl.

[0015] Wherein, the temperature of the development in step (3) is 10-40°C.

[0016] Wherein, the humidity of the unfolding in step (3) is 18% to 88%.

[0017] The present invention also provides a method for distinguishing Xiebai from counterfeit products, comprising the following steps:

[0018] (1) Prepare the test sample solution and the control solution according to the method;

[0019] (2) Spot the test sample solution and the Xiebai control solution on the same thin layer plate, develop with chloroform-ethyl acetate-methanol-water as the developing solvent, remove, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and inspect under sunlight and ultraviolet light respectively;

[0020] (3) When the main spot or fluorescent main spot of the same color appears in the chromatogram of the test sample at the corresponding position of the chromatogram of the control medicinal material, the test sample is Xiebai; otherwise, it is a counterfeit product.

[0021] Wherein, the amount of the sample to be tested in step (2) is 5 to 15 μl.

[0022] Wherein, the spotting volume of the Xiebai control solution in step (2) is 5 to 15 μl.

[0023] Wherein, the temperature of the development in step (2) is 10-40°C.

[0024] Wherein, the humidity of the unfolding in step (2) is 18% to 88%.

[0025] Among them, the thin layer plate described in step (2) includes silica gel H thin layer plate, silica gel G thin layer plate, high-efficiency silica gel G thin layer plate, silica gel GF254 thin layer plate, silica gel HF254 thin layer plate, and high-efficiency silica gel H thin layer plate.

[0026] The article (Research on the Quality Standard of Gua Xie Jiang Zhi Pills, Zheng Ji, Wang Hui, Sun Zhiqiang, Dai Long, Liaoning Journal of Traditional Chinese Medicine, Vol. 39, No. 9, 2012) and the patent (A Thin Layer Identification Method for Xie Bai Samples, Application No.: 202110735282X) disclose sample processing methods and developer compositions that are quite different from those of this method. The components determined are also different from those of this method. The main components extracted using ethyl acetate as the extraction solvent are fat-soluble components, while this method mainly determines water-soluble components. Therefore, they are not of reference significance for this method.

[0027] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: 1. It solves the problem that the pharmacopoeia method is not applicable due to the small amount of fat-soluble components in the Xiebai formula granules; 2. A control medicinal material is used as a control substance, and the sample is treated with a simple treatment method. It is inspected under sunlight and ultraviolet light (365nm) at the same time. The number of spots is large and the separation is good. Multiple components in the sample can be detected, which greatly improves the specificity of the method; this method can easily distinguish Xiebai and its counterfeit products, such as schizonepeta tenuifolia and shallot, and has great practical value; 3. The present invention uses n-butanol for shaking extraction and uses ammonia test solution for washing. While effectively extracting its water-soluble components, it removes the impurity components therein to the greatest extent; 4. The method of the present invention is simple to operate, eliminates the tedious column step, has low requirements for experimental personnel, and has high application value in actual production. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The TLC spectra of Xiebai formula granules prepared by different methods are as follows: A: TLC spectrum under sunlight, B: TLC spectrum under ultraviolet light;

[0029] Figure 2 The TLC spectra of Xiebai formula granules with different sample loadings are shown below: A: TLC spectra under sunlight, B: TLC spectra under UV light;

[0030] Figure 3 To investigate the specificity of Xiebai formula granules: A: TLC spectrum under sunlight, B: TLC spectrum under ultraviolet light;

[0031] Figure 4 The TLC spectra of Xiebai formula granules under different temperature conditions: A: TLC spectrum at 40℃ under sunlight, B: TLC spectrum at 40℃ under ultraviolet light; C: TLC spectrum at 10℃ under sunlight, D: TLC spectrum at 10℃ under ultraviolet light;

[0032] Figure 5 The TLC spectra of Xiebai formula granules under different humidity conditions are as follows: A: TLC spectrum under 18% humidity and sunlight, B: TLC spectrum under 18% humidity and ultraviolet light; C: TLC spectrum under 88% humidity and sunlight, D: TLC spectrum under 88% humidity and ultraviolet light;

[0033] Figure 6 The thin layer identification chromatograms of different batches of Xiebai formula granules: A: TLC spectrum under sunlight, B: TLC spectrum under ultraviolet light;

[0034] Figure 7 The thin-layer identification chromatograms of Xiebai formula granules, Mianzaoer formula granules, and Huoshang formula granules are shown: A: TLC spectrum under sunlight, B: TLC spectrum under ultraviolet light;

[0035] Figure 8 This is the thin layer identification chromatogram of Xiebai formula granules obtained by the pharmacopoeia method. DETAILED DESCRIPTION

[0036] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0037] Instruments and reagents:

[0038] Instruments: CAMAG TLC VISUALIZER, METTLERTOLEDO balance, KQ-250E ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.), silica gel H thin layer plate (Qingdao Ocean Chemical Co., Ltd.).

[0039] Reagents: Methanol (Sinopharm Chemical Reagent Co., Ltd.), n-butanol (Sinopharm Chemical Reagent Co., Ltd.), ammonia (Sinopharm Chemical Reagent Co., Ltd.), ethyl acetate (Sinopharm Chemical Reagent Co., Ltd.), chloroform (Shanghai Lingfeng Chemical Reagent Co., Ltd.), and sulfuric acid (Shanghai Lingfeng Chemical Reagent Co., Ltd.) were all analytical grade; water; Xiebai (Bulbus Allii) control material (batch number: 121130-202108) was purchased from the China Food and Drug Administration. Xiebai formula granules (batch numbers: 20090049, 20090059, 20090069), negative samples lacking Xiebai (excipients used in the granulation process), jujube herb formula granules, and shallot formula granules were provided by Jiangyin Tianjiang Pharmaceutical Co., Ltd. Xiebai formula granules and Xiebai control material all comply with the relevant requirements of the 2020 edition of the Chinese Pharmacopoeia.

[0040] Example 1 Investigation of the preparation methods of test solution and control medicinal material solution

[0041] 1. Thin layer identification conditions

[0042] Silica gel H thin layer plates were used with chloroform-ethyl acetate-methanol-water (15:40:22:10) as the developing agent. The plates were 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 sunlight and ultraviolet light (365nm).

[0043] 2. Solution Preparation

[0044] (1) Preparation of test solution

[0045] Method 1: Take 1 g of Xiebai formula granules (batch number: 20090049), grind it into powder, add 25 ml of methanol, ultrasonically treat it for 30 minutes, filter it, evaporate the filtrate to dryness, add 1 mL of methanol to dissolve the residue, and use it as the test solution.

[0046] Method 2: Take 1g of Xiebai formula granules (batch number: 20090049), grind it into powder, add 10ml of water to dissolve it, shake and extract it twice with a water-saturated n-butanol solution, 10ml each time, combine the n-butanol solution, evaporate to dryness, add 1ml of methanol to dissolve the residue, and use it as the test solution.

[0047] Method 3: Take 1 g of Xiebai formula granules (batch number: 20090049), grind it into powder, add 25 ml of methanol, ultrasonically treat it for 30 minutes, filter, evaporate the filtrate to dryness, add 10 ml of water to the residue to dissolve it, shake and extract it twice with a water-saturated n-butanol solution, 10 ml each time, combine the n-butanol solution, wash it with 20 ml of ammonia test solution, discard the ammonia test solution, and then wash it with 20 ml of an aqueous solution saturated with n-butanol, discard the water solution, evaporate the n-butanol solution to dryness, and add 0.5 ml of methanol to dissolve the residue as the test solution.

[0048] (2) Preparation of control medicinal material solution

[0049] Method 1: Take 2 g of Xiebai control medicinal material, add 25 ml of water, heat and reflux for 30 minutes, cool, filter, concentrate the filtrate to near dryness, add 25 ml of methanol to the residue, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 10 ml of water to the residue to dissolve it, shake and extract with a water-saturated n-butanol solution twice, 10 ml each time, combine the n-butanol solution, wash with 20 ml of ammonia test solution, discard the ammonia test solution, and then wash with 20 ml of an aqueous solution saturated with n-butanol, discard the water solution, evaporate the n-butanol solution to dryness, and add 0.5 ml of methanol to dissolve the residue as the control medicinal material solution.

[0050] Method 2: Take 2 g of Xiebai control medicinal material, add 25 ml of methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 10 ml of water to the residue to dissolve it, shake and extract twice with a water-saturated n-butanol solution, 10 ml each time, combine the n-butanol solution, wash with 20 ml of ammonia test solution, discard the ammonia test solution, and then wash with 20 ml of an aqueous solution saturated with n-butanol, discard the water solution, evaporate the n-butanol solution to dryness, and add 0.5 ml of methanol to dissolve the residue as the control medicinal material solution.

[0051] (3) Preparation of negative control solution

[0052] Take 1 g of the negative sample lacking Xiebai, grind it into powder, add 25 ml of methanol, ultrasonically treat it for 30 minutes, filter, evaporate the filtrate to dryness, add 10 ml of water to dissolve the residue, shake and extract twice with a water-saturated n-butanol solution, 10 ml each time, combine the n-butanol solution, wash with 20 ml of ammonia test solution, discard the ammonia test solution, wash with 20 ml of a saturated n-butanol aqueous solution, discard the water solution, evaporate the n-butanol solution to dryness, and add 0.5 ml of methanol to dissolve the residue as the negative control solution.

[0053] 3. Investigation of the preparation methods of test solution and control medicinal material solution

[0054] Take Xiebai formula granule sample (batch number: 20090049) and Xiebai control medicinal material according to the preparation method of the three test solution and the preparation method of the two control medicinal material solutions, and spot 10μl of the test solution and control medicinal material solution on the same silica gel H thin layer plate, develop at 28.5℃ and 55% humidity, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and examine under sunlight and ultraviolet light (365nm). Figure 1 As can be seen, the test solution prepared by method 3 has clear spots and good separation effect, so method 3 was selected as the sample processing method for this thin-layer identification. The Xiebai control medicinal material (method 1) corresponds more closely to the granular spots, so method 1 was selected as the control medicinal material processing method. Wherein, 1: test sample (method 1); 2: test sample (method 2); 3: test sample (method 3); S1: Xiebai control medicinal material (method 1); S2: Xiebai control medicinal material (method 2).

[0055] Example 2 Investigation of the influence of different spot amounts on the separation of spots

[0056] Take Xiebai formula granule sample (batch number: 20090049) and Xiebai control medicinal material according to the test solution method 3 and the control medicinal material solution preparation method 1 of Example 1, and spot them on the same silica gel H thin layer plate according to the thin layer chromatography conditions in Example 1 (the spot volume of the test solution is 2μl, 5μl, 10μl, and 15μl respectively; the spot volume of the control medicinal material solution is 2μl, 5μl, 10μl, and 15μl respectively), develop at 28.5℃ and 55% humidity, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and inspect under sunlight and ultraviolet light (365nm). Figure 2 It can be seen that when the sample volume of the test solution and the control medicinal material solution is 10 μl, the main spots at the corresponding positions in the test sample chromatogram and the control medicinal material chromatogram are clearly corresponding, without other interference. Therefore, the sample volume of the test solution and the control medicinal material solution is selected to be 10 μl. Among them, 1: 2 μl of test sample; 2: 5 μl of test sample; 3: 10 μl of test sample; 4: 15 μl of test sample; 5: 2 μl of control medicinal material; 6: 5 μl of control medicinal material; 7: 10 μl of control medicinal material; 8: 15 μl of control medicinal material.

[0057] Example 3 Sample specificity experiment

[0058] Take the Xiebai Formula Granules test solution (batch number: 20090049) prepared by method 3 of the test solution of Example 1, the negative control solution, and the control medicinal material solution prepared by method 1, and spot 10 μl of each on the same silica gel H thin layer plate according to the thin layer chromatography conditions in Example 1. Develop at 28.5°C and 55% humidity, remove, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clearly colored, and examine under sunlight and ultraviolet light (365nm). Figure 3 As can be seen, the chromatograms of the Xiebai Formula Granules test sample and the control herb exhibit the same main or fluorescent spots at the corresponding positions, and the negative control exhibits no interference, indicating that the TLC identification method has good specificity. Among them, 1-3: Xiebai Formula Granules (20090049); 4: Negative control; S: Xiebai control herb.

[0059] Example 4 Investigation of different temperatures

[0060] Take the Xiebai Formula Granules test solution (batch number: 20090049) prepared by method 3 of the test solution of Example 1 and the control medicinal material solution prepared by method 1 and spot 10 μl on the same silica gel H thin layer plate. According to the thin layer chromatography conditions in Example 1, develop at high temperature and low temperature conditions (55% humidity), remove, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clearly colored, and examine under sunlight and ultraviolet light (365nm). Figure 4 As can be seen, under different temperature conditions, the chromatograms of the Xiebai Formula Granules test sample and the control herb exhibited the same main spots or fluorescent main spots of the same color at the corresponding positions, and the separation effect was good. The experimental results show that temperature has no effect on the thin-layer chromatography identification of Xiebai Formula Granules, indicating that the thin-layer chromatography identification method has good durability at different temperatures.

[0061] Example 5 Investigation of different humidity

[0062] Take the Xiebai formula granules test solution (batch number: 20090049) prepared by method 3 of the test solution of Example 1 and the control medicinal material solution prepared by method 1 and spot 10 μl on the same silica gel H thin layer plate. According to the thin layer chromatography conditions in Example 1, develop under different humidity conditions (temperature is 28.5℃), remove, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and examine under sunlight and ultraviolet light (365nm). Figure 5 As can be seen, under different humidity conditions, the chromatograms of the Xiebai Formula Granules test sample and the control herb exhibited the same main or fluorescent spots at the corresponding positions, and the separation was good. The experimental results show that humidity has no effect on the TLC identification of Xiebai Formula Granules, indicating that the TLC identification method is robust under varying humidity conditions. Among them, 1-3: Xiebai Formula Granules (20090049); S: Xiebai control herb.

[0063] According to the above research results, the thin layer identification method of Xiebai formula granules is determined as follows:

[0064] Take 1g of Xiebai formula granules, grind it, add 25ml of methanol, ultrasonically treat it for 30 minutes, filter, evaporate the filtrate to dryness, add 10ml of water to dissolve the residue, shake and extract it twice with a water-saturated n-butanol solution, 10ml each time, combine the n-butanol solution, wash with 20ml of ammonia test solution, discard the ammonia test solution, wash with 20ml of an aqueous solution saturated with n-butanol, discard the water solution, evaporate the n-butanol solution to dryness, add 0.5ml of methanol to dissolve the residue, and use it as the test solution. Take another 2g of Xiebai control medicinal material, add 25ml of water, heat and reflux for 30 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 10ml of water to the residue to dissolve it, shake and extract with a water-saturated n-butanol solution twice, 10ml each time, combine the n-butanol solutions, wash with 20ml of ammonia test solution, discard the ammonia test solution, and then wash with 20ml of an aqueous solution saturated with n-butanol, discard the water solution, evaporate the n-butanol solution to dryness, add 0.5ml of methanol to dissolve the residue to prepare a control medicinal material solution. According to the thin-layer chromatography method (Chinese Pharmacopoeia 2020 edition, Part IV, General Chapter 0502), 10 μl of each of the test sample solution and the control herbal solution were spotted onto separate silica gel H thin-layer plates using chloroform-ethyl acetate-methanol-water (15:40:22:10) as the developing solvent. The plates were developed, removed, air-dried, and sprayed with 10% ethanolic sulfuric acid solution. The plates were heated at 105°C until the spots were clearly colored. The plates were then examined under sunlight and ultraviolet light (365 nm). In the chromatogram of the test sample, a main spot of the same color or a main fluorescent spot appeared at the corresponding position in the chromatogram of the control herbal substance.

[0065] Example 6 Thin layer chromatography identification of different batches of Xiebai formula granules

[0066] Take different batches of Xiebai formula granules (batch number: 20090049, 20090059, 20090069) and prepare the test solution and control medicinal material solution according to the preparation method of the test solution. Spot the sample on the same silica gel H thin layer plate, develop it, take it out, dry it, spray it with 10% sulfuric acid ethanol solution, heat it at 105℃ until the spots are clearly colored, and examine it under sunlight and ultraviolet light (365nm). Figure 6 As can be seen, the chromatogram of Xiebai Formula Granules and the chromatogram of the control herb show the same main spots or fluorescent main spots at the corresponding positions. Among them, 1: Xiebai Formula Granules (20090049); 2: Xiebai Formula Granules (20090059); 3: Xiebai Formula Granules (20090069); S: Xiebai control herb.

[0067] Example 7 Comparison of Xiebai and its counterfeit products

[0068] Take 1g each of Xiebai Formula Granules (Batch No. 20090049), Mianzaoer Formula Granules, and Shallot Formula Granules, grind them finely, add 25ml of methanol, sonicate for 30 minutes, filter, and evaporate the filtrate to dryness. Dissolve the residue in 10ml of water and shake and extract twice with a saturated n-butanol solution (10ml each time). Combine the n-butanol solutions, wash with 20ml of ammonia test solution, discard the ammonia test solution, and then wash with 20ml of a saturated n-butanol aqueous solution. Discard the water solution, evaporate the n-butanol solution to dryness, and dissolve the residue in 0.5ml of methanol to prepare the test solution. Separately, take 2g of Xiebai control medicinal material, add 25ml of water, heat and reflux for 30 minutes, cool, filter, and concentrate the filtrate to near dryness. Add 25ml of methanol to the residue and prepare the control medicinal material solution in the same manner. According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 edition Part IV General Chapter 0502), 10 μl of the test solution and the control medicinal material solution were taken and spotted on the same silica gel H thin layer plate, respectively, with chloroform-ethyl acetate-methanol-water (15:40:22:10) as the developing solvent, developed at 25.8°C and 55% humidity, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, heated at 105°C until the spots were clearly colored, and examined under sunlight and ultraviolet light (365nm). The results show that ( Figure 7 ), Xiebai Formula Granules and Mianzaoer Formula Granules showed significant differences under ultraviolet light (365nm), and Xiebai Formula Granules and Shallot Formula Granules showed significant differences under both ultraviolet light (365nm) and sunlight. Therefore, this thin-layer chromatography method can distinguish Xiebai Formula Granules from Mianzaoer Formula Granules and Shallot Formula Granules. Among them, 1: Xiebai Formula Granules (Batch No.: 20090049, 20090059, 20090069); 4-6: Mianzaoer Formula Granules (Batch No.: 2112005, 2112007, 2306031); 7-9: Shallot Formula Granules (Batch No.: 2306032, 2306033, 2306034); S: Xiebai control medicinal material.

[0069] Comparative Pharmacopoeia Method

[0070] Take 4g of this product, grind it into powder, add 20ml of n-hexane, ultrasonically treat it for 20 minutes, filter it, evaporate the filtrate, add 1ml of n-hexane to dissolve the residue, and use it as the test solution. Take another 4g of Xiebai as a control medicinal material, and prepare a control medicinal material solution in the same way. According to the thin layer chromatography method (General Rule 0502), take 10μl of each of the above two solutions, spot them on the same silica gel G thin layer plate, use n-hexane-ethyl acetate (10:1) as the developing agent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clearly colored, and inspect under ultraviolet light (365nm). The results show that ( Figure 8), if the pharmacopoeia method is used directly, there will be problems such as few spots, unclearness, and poor separation. Among them, 1: Xiebai Formula Granules (20090049); 2: Xiebai Formula Granules (20090059); 3: Xiebai Formula Granules (20090069); S: Xiebai control medicinal material.

Claims

1. A thin layer detection method for Xiebai formula granules, characterized in that: The following steps are involved: (1) Prepare the test solution: Take the powder of the Xiebai formula granules, add methanol, ultrasonicate, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with a saturated n-butanol solution with water, combine the n-butanol solution, wash with ammonia test solution, discard the ammonia test solution, wash with a saturated n-butanol aqueous solution, discard the water solution, evaporate the n-butanol solution to dryness, dissolve the residue in methanol, and use it as the test solution; (2) Preparation of Xiebai control solution: Take Xiebai control medicinal material, add water, heat to reflux, cool, filter, concentrate the filtrate to nearly dryness, add methanol to the residue, ultrasonicate, filter, evaporate the filtrate to dryness, dissolve the residue in water, shake and extract with a saturated n-butanol solution with water, combine the n-butanol solution, wash with ammonia test solution, discard the ammonia test solution, wash with a saturated n-butanol aqueous solution, discard the water solution, evaporate the n-butanol solution to dryness, dissolve the residue in methanol, and use it as the control medicinal material solution; (3) Spot the test sample solution and the Xiebai control solution on the same silica gel H thin layer plate, develop it with chloroform-ethyl acetate-methanol-water in a volume ratio of 15:40:22:10, remove it, dry it, spray it with 10% sulfuric acid ethanol solution, heat it at 105℃ until the spots are clearly colored, and inspect it under sunlight and ultraviolet light respectively; (4) In the chromatogram of the test sample, a main spot of the same color or a main fluorescent spot appears at the corresponding position in the chromatogram of the control medicinal material.

2. The method according to claim 1, characterized in that The amount of the sample to be tested in step (3) is 5~15µl.

3. The method according to claim 1, characterized in that The spotting volume of the Xiebai control solution in step (3) is 5~15µl.

4. The method according to claim 1, characterized in that The temperature of the development in step (3) is 10-40°C.

5. The method according to claim 1, characterized in that: The humidity of the unfolding described in step (3) is 18% to 88%.

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