Traditional Chinese medicine composition for invigorating qi and blood and activating meridians to stop pain

Abstract

The invention discloses a traditional Chinese medicine composition for invigorating qi and blood and activating meridians to stop pain, and belongs to the field of traditional Chinese medicine, the traditional Chinese medicine composition comprises 5 parts of ginseng, 17 parts of Poria cocos, 10 parts of bighead atractylodes rhizome, 3 parts of licorice, 10 parts of Chinese angelica, 5 parts of Ligusticum wallichii, 10 parts of white peony root, 15 parts of prepared rehmannia root, 17 parts of root of kudzu vine, 5 parts of Ligusticum wallichii, 5 parts of dahurian angelica root and 5 parts of corydalis tuber, and the prescription is composed of eight-treasure decoction, the root of kudzu vine, Ligusticum wallichii, dahurian angelica root and corydalis tuber. Rhizoma atractylodis macrocephalae and poria cocos strengthen the spleen and excrete dampness; the radix angelicae sinensis and the radix paeoniae alba are used for nourishing blood and harmonizing ying, and assisting the radix rehmanniae preparata to tonify yin blood, and are used as ministerial drugs, the rhizoma chuanxiong and the rhizoma corydalis are used for activating blood and promoting qi circulation, dispelling wind and relieving pain, the radix puerariae is used for resolving muscles and bringing down a fever, and promoting the production of body fluid to quench thirst, the radix ligustici sinensis is used for expelling wind and removing cold, and removing dampness and relieving pain, the radix angelicae is used for relieving exterior syndrome and dispelling wind, and opening orifices and relieving pain, and are used as adjuvant drugs; the traditional Chinese medicine composition disclosed by the invention is scientific in compatibility, proper in medicine flavor increase and decrease and synergistic in traditional Chinese medicinal materials, and typical cases prove that the traditional Chinese medicine composition disclosed by the invention has a remarkable curative effect on qi-blood deficiency type migraine.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for tonifying qi and blood and dredging collaterals to relieve pain. Background Art

[0002] Migraine is a common clinical neurological disease. A migraine epidemiology study in China in 2011 showed that the prevalence of this disease in the patient population was 9.3%. This disease often recurs easily, seriously affecting the patient's daily study, work, and quality of life. The main clinical manifestations are unilateral, moderate to severe pulsating headache, generally lasting for 4 to 72 hours, often accompanied by symptoms such as nausea, vomiting, photophobia, and phonophobia. In recent years, many scholars at home and abroad have investigated and found that migraine is closely related to genetic factors, hemodynamic factors, endocrine factors, sleep disorder factors, psychosocial factors, metabolic factors, mitochondrial dysfunction, drug factors, etc.

[0003] The "Clinical Practice Guidelines for Traditional Chinese Medicine in Migraine (Draft for Comment)" points out that 5 classic famous prescriptions including Chuanxiong Chatiao Powder, Chuanxiong Dingtong Decoction, Sanpian Decoction, Xuefu Zhuyu Decoction, and Tongqiao Huoxue Decoction, as well as 4 proprietary Chinese medicines, Zhengtian Pills, Tou Tongning Capsules, Tongtian Oral Liquid, and Yangxue Qingnao Granules (Pills), all have good preventive and therapeutic effects on migraine, and can reduce the attack frequency and degree of migraine in patients, and improve the quality of life. Most of the migraine patients in our hospital are of the type of qi and blood deficiency. At present, there are very few traditional Chinese medicine compositions for treating migraine of qi and blood deficiency type in clinical practice, which cannot meet the drug needs of patients with different syndromes.

[0004] The preparation of this formulation is improved for the above-mentioned existing defects. It is made entirely of traditional Chinese medicines, with a simple formula, using commonly used medicinal materials in traditional Chinese medicine, having small toxic and side effects, high safety, and strong applicability. Summary of the Invention

[0005] In order to solve the problems mentioned in the above background art, the present invention provides a traditional Chinese medicine composition for tonifying qi and blood and dredging collaterals to relieve pain.

[0006] In order to achieve the above object, the present invention adopts the following technical scheme: A traditional Chinese medicine composition for tonifying qi and blood and dredging collaterals to relieve pain is made from the following raw medicinal materials in parts by weight: 5 parts of ginseng, 17 parts of poria, 10 parts of atractylodes macrocephala, 3 parts of licorice, 10 parts of angelica sinensis, 5 parts of chuanxiong, 10 parts of white peony root, 15 parts of prepared rehmannia root, 17 parts of kudzu root, 5 parts of ligusticum sinense oliv, 5 parts of angelica dahurica, and 5 parts of Corydalis yanhusuo.

[0007] A preparation method of a traditional Chinese medicine composition for tonifying qi and blood and dredging collaterals to relieve pain includes the following steps: (1) Take twelve Chinese medicinal materials respectively, select and remove impurities, weigh and take, and wash. (2) Add 8 times the amount of water, soak for 1 hour, decoct for 2 hours, filter, and reserve the filtrate; add 6 times the amount of water to the residue, decoct for another 2 hours, combine the filtrate with the previous filtrate, and discard the residue. (3) Concentrate the filtrate to a relative density of 1.15 - 1.20 (measured at 60 °C) to obtain an extract. (4) Dry the extract at 80 ± 5 °C until the water content ≤ 5.0% to obtain a dry extract. (5) Crush the dry extract, add pharmaceutical excipients, and mix evenly. (6) Add ethanol for wet mixing, make into granules, dry, test the granules, and package to obtain. Preferably, the temperature for concentrating the filtrate is controlled at 60 °C.

[0008] Preferably, the traditional Chinese medicine composition is pretreated, decocted, granulated with starch, and dried. The weight ratio of the pretreated, decocted, and granulated is 1:4 - 8:1; the drying temperature is 80 ± 5 °C, the decoction is adding 12 or 8 times the amount of water, after boiling per minute, decoct for 30 - 60 minutes, and the starch is evenly mixed with 85% ethanol and granulated with 10% syrup.

[0009] On the other hand, the present invention also provides a traditional Chinese medicine granule with the efficacy of tonifying qi and blood and dredging collaterals to relieve pain, which is made from the traditional Chinese medicine composition and one or several of diluents, lubricants, disintegrants, flavoring agents, suspending agents, taste masking agents, preservatives, coloring agents, disintegrants, or sweeteners.

[0010] Preferably, the diluent is such as sucrose, starch, dextrin, lactose, or maltose; the lubricant is such as talc, magnesium stearate, microcrystalline silica, or calcium stearate; the disintegrant is such as dry starch, sodium carboxymethyl starch, or low-substituted hydroxypropyl cellulose; the taste masking agent is such as cyclodextrin, stevioside, or saccharin sodium; the coloring agent is such as pigment, iron oxide brown, iron oxide red, or sunset yellow; the sweetener is such as saccharin sodium, aspartame, sucralose, or steviol glycoside.

[0011] Preferably, the granulation temperature is 30 - 80 °C; the granulation time is 5 hours - 20 hours.

[0012] The present invention also provides a method for identifying ginseng in a Chinese medicine composition for replenishing qi and blood, unblocking meridians and relieving pain. The method comprises the following steps: taking 15g of the product, grinding it, adding 200ml of 90% methanol, heating and refluxing for extraction for 1 hour, filtering, concentrating the filtrate to dryness, adding 10ml of water to dissolve it, adding 100ml of water-saturated n-butanol, ultrasonically treating it for 30 minutes, adding 100ml of concentrated ammonia water for washing, discarding the alkali solution, taking the n-butanol solution, evaporating it to dryness, adding 10ml of water to dissolve the residue, adding it to the treated D101 macroporous resin, wherein the D101 macroporous resin has an inner diameter of 1.5cm and a length of 15cm, first eluting it with 50ml of water, then eluting it with 50ml of 20% ethanol, discarding the eluate, and then eluting it with 50ml of 80% ethanol, collecting the eluate, evaporating it Dried, add 1ml of methanol to dissolve the residue as the test solution, take another 2g of ginseng reference medicinal material, prepare the control medicinal material solution in the same way, then take appropriate amount of ginsenoside Rg1 and ginsenoside Re reference substances, add methanol to prepare a mixed solution containing 0.5mg of each in each ml, as the reference substance solution, test according to the thin layer chromatography method, take 5-20μl of each of the three solutions, spot them on the same silica gel G thin layer plate, use chloroform-ethyl acetate-methanol-water (15:40:22:10) placed below 10℃ as the developing agent, develop, take out, dry, spray with 10% ethanol sulfuric acid solution, heat at 105℃ until the spots are clearly colored, and in the test sample chromatogram, spots of the same color appear at the corresponding positions of the control medicinal material chromatogram and the reference substance chromatogram.

[0013] The invention also provides a method for identifying pueraria root in a Chinese medicine composition for invigorating qi and blood, unblocking meridians and relieving pain. The method comprises the following steps: taking 10 g of the product, grinding it into powder, adding 40 ml of ethyl acetate, ultrasonically treating it for 30 minutes, filtering it, evaporating the filtrate to dryness, adding 30 ml of water and 30 ml of ether to the residue to dissolve it, placing it in a separating funnel to stand and separate the layers, taking the lower layer solution and evaporating it to dryness, and adding 1 ml of methanol to the residue to dissolve it as a test solution. A puerarin reference substance is separately taken, and methanol is added to prepare a solution containing 0.5 mg per 1 ml as a reference substance solution. According to a thin layer chromatography test, 5 to 10 μl of each of the two solutions are taken and spotted on the same silica gel G thin layer plate respectively, and chloroform-methanol-water (28:10:1) is used as a developing agent. The plate is developed, taken out, dried, and fumigated in ammonia vapor for 15 minutes. The plate is taken out and inspected under an ultraviolet lamp (365 nm). In the chromatogram of the test product, fluorescent spots of the same color are displayed at positions corresponding to the chromatogram of the reference substance.

[0014] Compared with the prior art, the present invention has the following beneficial effects: 1. The Chinese medicine composition of the present invention has a reasonable formula and has a significant therapeutic effect on headache, dizziness, palpitation, fatigue, chest tightness and shortness of breath caused by deficiency of both qi and blood. It can relieve the patient's discomfort in a short time and has a significant therapeutic effect.

[0015] 2. The composition of the present invention is prepared from all traditional Chinese medicines. The formula is simple, using commonly used medicinal materials in traditional Chinese medicine, with small toxic and side effects, high safety, and strong practicability.

[0016] 3. It is composed of Bazhen Decoction plus Pueraria lobata, Ligusticum sinense, Angelica dahurica, and Corydalis yanhusuo. In the formula, ginseng and rehmannia glutinosa are combined to supplement qi and nourish blood, and they are the monarch drugs together; Atractylodes macrocephala and Poria cocos strengthen the spleen and promote diuresis, assisting ginseng in strengthening the spleen and replenishing qi; Angelica sinensis and Paeonia lactiflora nourish blood and regulate qi and blood, assisting rehmannia glutinosa in tonifying yin and blood, and they are all ministerial drugs. Chuanxiong and Corydalis yanhusuo promote blood circulation and qi movement, dispel wind and relieve pain, Pueraria lobata relieves muscle heat and generates fluid to quench thirst, Ligusticum sinense dispels wind and cold, relieves pain due to dampness, and Angelica dahurica dispels wind from the superficies, opens orifices and relieves pain, and they are all assistant drugs. Licorice replenishes qi and harmonizes the middle energizer, regulating the actions of various drugs, and it is the guiding drug. The compatibility of the traditional Chinese medicine composition of the present invention is scientific, the addition and subtraction of medicinal flavors are appropriate, and the various Chinese medicinal materials synergistically enhance the efficacy. Typical cases have confirmed that the traditional Chinese medicine composition of the present invention has a significant curative effect on migraine of qi and blood deficiency type. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0018] Figure 1 : Process flow chart; Figure 2 : TLC chromatogram of Bushen Yangxue Zhitong Granules (ginsenoside); Figure 3 : TLC chromatogram of Bushen Yangxue Zhitong Granules (paeoniflorin); Figure 4 : TLC chromatogram of Bushen Yangxue Zhitong Granules (puerarin). DETAILED DESCRIPTION OF THE EMBODIMENTS

[0019] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the embodiments of the present invention. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts fall within the protection scope of the present invention.

[0020] Unless otherwise specified, the raw materials used in the present invention are all conventional products purchased from the market.

[0021] Example 1: The ratio of the traditional Chinese medicine composition is as follows: 5 parts of ginseng, 17 parts of poria, 10 parts of atractylodes macrocephala, 3 parts of licorice, 10 parts of angelica sinensis, 5 parts of chuanxiong rhizome, 10 parts of white peony root, 15 parts of rehmannia glutinosa, 17 parts of kudzu root, 5 parts of ligusticum sinense, 5 parts of angelica dahurica and 5 parts of Corydalis yanhusuo. The weight ratio of ginseng, poria, atractylodes macrocephala, licorice, angelica sinensis, chuanxiong rhizome, white peony root, rehmannia glutinosa, kudzu root, ligusticum sinense, angelica dahurica and Corydalis yanhusuo is 5:17:10:3:10:5:10:15:17:5:5:5.

[0022] A preparation method of a traditional Chinese medicine composition for tonifying qi and blood and dredging collaterals to relieve pain, comprising the following steps: Pretreatment of medicinal materials: Take twelve traditional Chinese medicinal materials respectively, select and remove impurities, weigh, and wash.

[0023] Extraction: Add 8 times the amount of water, soak for 1 hour, decoct for 2 hours, filter, and reserve the filtrate; Add 6 times the amount of water to the medicinal residues, Decoct for another 2 hours, combine the filtrate with the previous filtrate, and discard the medicinal residues.

[0024] Concentration: Concentrate the filtrate to a relative density of 1.15 - 1.20 (measured at 60 °C) to obtain an extract.

[0025] Drying: Dry the extract at 80 ± 5 °C until the moisture content ≤ 5.0% to obtain a dry extract.

[0026] Pulverization: Pulverize the dry extract into 80-mesh powder to obtain dry extract powder.

[0027] Mixing: Weigh dextrin and mix it evenly with the dry extract powder.

[0028] Granulation: Prepare soft materials with 30% ethanol, granulate with a 16-mesh sieve, dry at 80 ± 5 °C, control the moisture content: 3% - 5%, and screen the granules with a 14-mesh sieve.

[0029] Sub-packaging: Debug the granule filling machine, sub-package, and control the device: 10 g / bag ± 5%.

[0030] Outer packaging: Put the instruction manual into the box together to obtain the product.

[0031] The medicinal excipients include but are not limited to one or several of flavoring agents, diluents, lubricants, dispersants, thickeners, preservatives, and colorants.

[0032] The traditional Chinese medicine preparation of the traditional Chinese medicine composition is an oral traditional Chinese medicine preparation. Preferably, the oral traditional Chinese medicine preparation is one of decoction, pill, tablet, mixture, capsule, granule, powder or ointment.

[0033] Example 2: A method for identifying the components of a traditional Chinese medicine composition, providing the following two component identifications: Identification method for ginseng in traditional Chinese medicine composition for supplementing qi and blood, dredging collaterals and relieving pain, characterized in that: take 15 g of this product, grind it finely, add 200 ml of 90% methanol, heat under reflux for 1 hour, filter, concentrate the filtrate to dryness, add 10 ml of water to dissolve, add 100 ml of water-saturated n-butanol, perform ultrasonic treatment for 30 minutes, add 100 ml of concentrated ammonia water for washing, discard the alkaline solution, separate the n-butanol solution, evaporate to dryness, add 10 ml of water to dissolve the residue, load it onto the pretreated D101 macroporous resin, the inner diameter of the D101 macroporous resin is 1.5 cm and its length is 15 cm, first elute with 50 ml of water, then elute with 50 ml of 20% ethanol, discard the eluate, continue to elute with 50 ml of 80% ethanol, collect the eluate, evaporate to dryness, add 1 ml of methanol to dissolve the residue, and use it as the test solution. Additionally, take 2 g of ginseng reference medicinal material, prepare the reference medicinal material solution in the same way. Then take appropriate amounts of ginsenoside Rg1 and ginsenoside Re reference substances, add methanol to make a mixed solution containing 0.5 mg of each per 1 ml, and use it as the reference substance solution. According to the thin-layer chromatography test, absorb 5 - 20 μl of each of the above three solutions, respectively spot them on the same silica gel G thin-layer plate, use the lower layer solution of chloroform - ethyl acetate - methanol - water (15:40:22:10) placed below 10 °C as the developing agent, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, heat at 105 °C until the spots are clearly visible. In the test solution chromatogram, at the positions corresponding to the reference medicinal material chromatogram and the reference substance chromatogram, there are spots showing the same color.

[0034] Identification method for pueraria lobata in traditional Chinese medicine composition for supplementing qi and blood, dredging collaterals and relieving pain, characterized in that: take 10 g of this product, grind it finely, add 40 ml of ethyl acetate, perform ultrasonic treatment for 30 minutes, filter, evaporate the filtrate to dryness, add 30 ml of water and 30 ml of ether to dissolve the residue, place it in a separating funnel and let it stand for layering, separate the lower layer solution and evaporate to dryness, add 1 ml of methanol to dissolve the residue, and use it as the test solution. Additionally, take puerarin reference substance, add methanol to make a solution containing 0.5 mg per 1 ml, and use it as the reference substance solution. According to the thin-layer chromatography test, absorb 5 - 10 μl of each of the above two solutions, respectively spot them on the same silica gel G thin-layer plate, use chloroform - methanol - water (28:10:1) as the developing agent, develop, take out, dry in the air, expose it to ammonia vapor for 15 minutes, take out, and examine it under an ultraviolet lamp (365 nm). In the test solution chromatogram, at the position corresponding to the reference substance chromatogram, there are fluorescent spots showing the same color.

[0035] As mentioned above, it is only the preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, according to the technical solution and inventive concept of the present invention, makes equivalent substitutions or changes, and all should be covered within the protection scope of the present invention.

Claims

1. A Chinese medicine composition for replenishing qi and blood, dredge meridians and relieve pain, characterized in that: The Chinese medicine composition is prepared from the following Chinese medicine components: ginseng, tuckahoe, atractylodes macrocephala, liquorice, angelica, ligusticum chuanxiong, white peony root, rehmannia root, kudzu root, ligusticum ligusticum, angelica dahurica and yanhusuo.

2. The Chinese medicine composition according to claim 1, characterized in that: The weight ratio of ginseng, Poria, Atractylodes macrocephala, licorice, Chinese angelica, Ligusticum chuanxiong, white peony root, Rehmannia root, Pueraria root, Ligusticum chuanxiong, Angelica dahurica and Corydalis yanhusuo is 5:17:10:3:10:5:10:15:17:5:5:

5.

3. The Chinese medicine composition according to claim 2, characterized in that: The Chinese medicine composition is prepared from the following raw materials in parts by weight: 5 parts of ginseng, 17 parts of Poria cocos, 10 parts of Atractylodes macrocephala, 3 parts of Licorice, 10 parts of Angelica sinensis, 5 parts of Chuanxiong, 10 parts of White Peony Root, 15 parts of Rehmannia glutinosa, 17 parts of Pueraria lobata, 5 parts of Ligusticum chuanxiong, 5 parts of Angelica dahurica and 5 parts of Corydalis yanhusuo.

4. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The preparation method is: (1) Take twelve Chinese medicinal materials respectively, select and remove impurities, weigh and wash; (2) Add 8 times the amount of water, soak for 1 hour, decoct for 2 hours, filter, and reserve the filtrate; add 6 times the amount of water to the residue, decoct for another 2 hours, combine the filtrate with the above filtrate, and discard the residue; (3) Concentrating the filtrate to a relative density of 1.15 to 1.20 (thermal measurement at 60°C) to obtain an extract; (4) Drying the extract at 80±5°C until the moisture content is ≤5.0% to obtain a dry extract; (5) crushing the dry paste, adding pharmaceutical excipients, and mixing evenly; (6) adding ethanol for wet mixing, forming granules, drying, testing the granules, and packaging.

5. A Chinese medicine granule with the effects of nourishing qi and blood, unblocking meridians and relieving pain, characterized in that: The Chinese medicine granules are prepared from the Chinese medicine composition according to any one of claims 1 to 3 and one or more of a diluent, a lubricant, a disintegrant, a flavoring agent, a suspending agent, a taste masking agent, a preservative, a colorant, a disintegrant or a sweetener.

6. The Chinese medicine granules according to claim 5, characterized in that: Diluents such as sucrose, starch, dextrin, lactose or maltose; lubricants such as talc, magnesium stearate, micronized silica or calcium stearate; disintegrants such as dry starch, sodium carboxymethyl starch or low-substituted hydroxypropyl cellulose; masking agents such as cyclodextrin, stevioside or saccharin sodium; colorants such as pigments, iron oxide brown, iron oxide red or sunset yellow; sweeteners such as saccharin sodium, aspartame, sucralose or steviol glycosides.

7. The Chinese medicine granules according to claim 5, characterized in that: The granulation temperature is 30-80° C.; the granulation time is 5 hours to 20 hours.

8. The method for preparing the Chinese medicine composition according to claim 4, characterized in that: The Chinese medicine composition is pre-treated, decocted, granulated with starch, and dried, and the weight ratio of the pre-treatment, decoction and granulation is 1:4-8:1; the drying temperature is 80±5°C, the decoction is adding 12 or 8 times the amount of water, boiling every minute, and then decocting for 30-60 minutes, the starch is evenly mixed with 85% ethanol, and 10% syrup is added for granulation.

9. A method for identifying ginseng in a Chinese medicine composition for replenishing qi and blood, unblocking meridians and relieving pain, characterized in that: Take 15g of this product, grind it, add 200ml of 90% methanol, heat and reflux to extract for 1 hour, filter, concentrate the filtrate to dryness, add 10ml of water to dissolve it, add 100ml of water-saturated n-butanol, ultrasonically treat it for 30 minutes, wash it with 100ml of concentrated ammonia water, discard the alkali solution, take the n-butanol solution, evaporate it to dryness, add 10ml of water to dissolve the residue, add it to the treated D101 macroporous resin, the D101 macroporous resin has an inner diameter of 1.5cm and a length of 15cm, first elute it with 50ml of water, then elute it with 50ml of 20% ethanol, discard the eluent, continue to elute it with 50ml of 80% ethanol, collect the eluent, evaporate it to dryness, add 1ml of methanol to dissolve the residue as the test solution, and another Take 2g of ginseng reference medicinal material and prepare a reference medicinal material solution in the same way. Then take appropriate amounts of ginsenoside Rg1 and ginsenoside Re reference substances, add methanol to prepare a mixed solution containing 0.5mg of each in 1ml as the reference substance solution. According to the thin layer chromatography test, take 5-20μl of each of the three solutions and spot them on the same silica gel G thin layer plate. Use chloroform-ethyl acetate-methanol-water (15:40:22:10) placed below 10℃ as the developing agent. Develop, take out, dry, spray with 10% ethanolic sulfuric acid solution, heat at 105℃ until the spots are clearly colored, and in the test sample chromatogram, spots of the same color appear at the corresponding positions of the reference medicinal material chromatogram and the reference substance chromatogram.

10. A method for identifying Pueraria root in a Chinese medicine composition for replenishing qi and blood, unblocking meridians and relieving pain, characterized in that: Take 10g of this product, grind it, add 40ml of ethyl acetate, ultrasonically treat it for 30 minutes, filter it, evaporate the filtrate, dissolve the residue in 30ml of water and ether, place it in a separatory funnel and let it stand for stratification, take the lower layer solution and evaporate it, dissolve the residue in 1ml of methanol as the test solution, take another puerarin reference substance, add methanol to make a solution containing 0.5mg per 1ml as the reference substance solution, test it according to the thin layer chromatography method, take 5~10μl of each of the above two solutions, and spot them on the same silica gel G thin layer plate, use chloroform-methanol-water (28:10:1) as the developing agent, develop it, take it out, dry it, fumigate it in ammonia vapor for 15 minutes, take it out, and examine it under ultraviolet light (365nm). In the chromatogram of the test substance, at the corresponding position of the chromatogram of the reference substance, a fluorescent spot of the same color appears.

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