Method for testing medicine for treating sequela of apoplexy

Abstract

The invention discloses a detection method of a medicine for treating apoplexy sequela, and relates to the technical field of traditional Chinese medicinal material detection. The capsule disclosed by the invention comprises the following components: ginseng, deer-horn glue, tortoise shell, fructus lycii, turtle shell, leech, ligusticum wallichii, rheum officinale, hawthorn, earthworm, scorpio, bungarus parvus, gastrodia elata, stiff silkworm and borneol. During identification, microscopic identification and thin-layer chromatography identification are adopted, the gastrodia elata and the hawthorn are identified by the thin-layer chromatography, and the microscopic identification is used for identifying a capsule finished product; the aims of accurately detecting the ginseng and snake hemiplegia capsule and reducing the large influence on the detection result due to the change of different temperature and humidity in the environment are achieved.

Description

Technical Field

[0001] The invention relates to the technical field of Chinese medicinal material detection, in particular to a method for detecting drugs for treating sequelae of stroke. Background Art

[0002] Stroke, also known as cerebrovascular disease, includes ischemic stroke and hemorrhagic stroke. One of the characteristics of stroke is its high disability rate. Those patients who are saved by early treatment often have serious sequelae, the most common of which are complete and incomplete hemiplegia, hemiplegia, facial paralysis, slurred speech, cramps in hands and feet, numbness of limbs, edema of feet and knees, incontinence, pain in muscles and bones, etc. These sequelae greatly endanger the quality of life of patients.

[0003] There is no specific medicine for treating stroke sequelae in the current medical community. The efficacy of various drugs for treating stroke sequelae currently used in clinical practice is far from satisfactory, and it is very necessary to explore drugs that are easy to use and have better efficacy. Traditional Chinese medicine has its own characteristics and advantages in treating stroke sequelae, and some Chinese patent medicines with their own characteristics and certain efficacy have emerged. However, these Chinese patent medicines generally have the disadvantages of low efficacy and slow onset. The inventor has continuously conducted research and exploration on the treatment of stroke sequelae with traditional Chinese medicine in many years of medical practice, and has achieved technical achievements including the present invention.

[0004] Among them, Shenshe Hemiplegia Capsule is included in the State Food and Drug Administration standard WS-5018 (B-0018)-2004, and its quality standard can be found in the application document item 1. Its functions and indications are "replenishing qi and tonifying the kidney, promoting blood circulation and removing blood stasis, extinguishing wind and unblocking collaterals. It is used for sequelae of stroke, with symptoms of hemiplegia, crooked mouth and tongue, speech difficulties, dizziness, tinnitus, and soreness of waist and knees." For example, Chinese patent CN100344318C.

[0005] There are still inconveniences in the original testing process. In order to more effectively control the quality of drugs, the quality testing method needs to be optimized. Summary of the invention

[0006] The purpose of the present invention is to provide a method for testing drugs for treating sequelae of stroke, so as to achieve accurate detection of Shenshe Hemiplegia Capsules and reduce the influence of changes in environmental temperature and humidity on the detection results.

[0007] In order to achieve the above object, the present invention adopts the following technical means: A method for testing a drug for treating sequelae of stroke, wherein the capsule comprises the following ingredients: ginseng, deer antler glue, tortoise shell, wolfberry, turtle shell, leech, chuanxiong, rhubarb, hawthorn, earthworm, scorpion, golden snake, gastrodia, silkworm and borneol; Microscopic identification and thin layer chromatography identification are used for identification. The gastrodia elata and the hawthorn are identified by the thin layer chromatography, and the microscopic identification is used to identify the finished capsule product.

[0008] Preferably, in parts by weight, the ingredients include 45-50 parts of ginseng, 20-25 parts of antler glue, 33-38 parts of tortoise shell, 33-38 parts of wolfberry, 33-38 parts of turtle shell, 45-50 parts of leech, 33-38 parts of Chuanxiong, 13-18 parts of rhubarb, 25-30 parts of hawthorn, 25-30 parts of earthworm, 25-30 parts of scorpion, 45-50 parts of snakehead, 33-38 parts of gastrodia, 20-25 parts of bombyx batryticatus and 5-10 parts of borneol.

[0009] Furthermore, in parts by weight, the ingredients include 48 parts of ginseng, 22 parts of antler glue, 36 parts of tortoise shell, 36 parts of wolfberry, 36 parts of turtle shell, 48 parts of leech, 36 parts of Chuanxiong, 15 parts of rhubarb, 28 parts of hawthorn, 28 parts of earthworm, 28 parts of scorpion, 48 parts of snakehead, 36 parts of gastrodia, 22 parts of bombyx batryticatus and 8 parts of borneol.

[0010] Furthermore, the microscopic identification includes taking a capsule sample and observing it under a microscope: The muscle fibers are layered, colorless, slightly yellow or light brown, slightly curved, and arranged vertically and staggered; Body wall fragments are pale yellow to yellow, with reticular texture and round hair pits, and brown setae are seen; The body wall fragments are colorless and have very fine mycelium on the surface.

[0011] Furthermore, the thin layer chromatography identification of Gastrodia elata includes: S110. Sample preparation; S120. Preparation of control medicinal material solution; S130. Preparation of negative control samples; S140. Perform a thin layer chromatography test, which uses a silica gel G thin layer plate and ethyl acetate-methanol-water (9:1:0.2) as a developing agent. After spraying with 10% phosphomolybdic acid ethanol solution, it is heated at 105° C. until the spots are colored.

[0012] Furthermore, the step S110 includes: taking 5 samples, adding 40 ml of water-saturated n-butanol, ultrasonically treating for 30 minutes, filtering and evaporating the filtrate, dissolving the residue in 2 ml of water, and passing through a polyamide column, eluting with 50 ml of water, collecting the eluate, evaporating and dissolving the residue in 1 ml of methanol as a sample solution.

[0013] Furthermore, the filler of the polyamide column is 5 g and has a mesh size of 30-60, and the column is filled by a wet method.

[0014] Furthermore, the thin layer chromatography identification of hawthorn includes: S210. Sample preparation; S220. Preparation of control medicinal material solution; S230. Preparation of negative control samples; S240. Perform a thin layer chromatography test using a silica gel G thin layer plate, cyclohexane-acetone-ethyl acetate-formic acid (9:2:1:0.2) as a developing agent, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are colored, and inspect under daylight or ultraviolet light.

[0015] Furthermore, the step S210 includes: taking 10 samples, adding 50 ml of ethyl acetate, ultrasonically treating for 30 minutes, filtering, evaporating the filtrate to dryness, and adding 2 ml of ethyl acetate to the residue to dissolve it as a sample solution.

[0016] Furthermore, the wavelength of the ultraviolet lamp is 365nm.

[0017] The present invention has the following beneficial effects during application: For the identification of Gastrodia elata and Crataegus pinnatifida, under normal and low temperature conditions, the main spots of the sample chromatogram and the reference medicinal material chromatogram were clearly colored, with good separation, no tailing, and no background interference. The sample chromatogram showed spots of the same color at the corresponding positions of the reference medicinal material chromatogram, indicating that the decrease in temperature had no obvious effect on the thin-layer identification of Gastrodia elata and Crataegus pinnatifida, indicating that the thin-layer identification method has good durability under normal and low temperature conditions.

[0018] At the same time, for the identification of Gastrodia elata and Crataegus pinnatifida, under high humidity and low humidity conditions, the main spots of the sample chromatogram and the reference medicinal material chromatogram were clearly colored, with good separation, no tailing phenomenon, and no background interference. The sample chromatogram showed spots of the same color at the corresponding positions of the reference medicinal material chromatogram, indicating that humidity had no obvious effect on the thin-layer identification of Gastrodia elata and Crataegus pinnatifida, indicating that the thin-layer identification method has good durability to different humidity levels. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments.

[0020] Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features of the embodiments may be combined with each other.

[0022] Example 1 A method for testing a drug for treating sequelae of stroke, wherein the capsule comprises the following ingredients: ginseng, deer antler glue, tortoise shell, wolfberry, turtle shell, leech, chuanxiong, rhubarb, hawthorn, earthworm, scorpion, golden snake, gastrodia, silkworm and borneol; Microscopic identification and thin layer chromatography identification are used for identification. The gastrodia elata and the hawthorn are identified by the thin layer chromatography, and the microscopic identification is used to identify the finished capsule product.

[0023] In parts by weight, the ingredients include 48 parts of ginseng, 22 parts of antler glue, 36 parts of tortoise shell, 36 parts of wolfberry, 36 parts of turtle shell, 48 parts of leech, 36 parts of chuanxiong, 15 parts of rhubarb, 28 parts of hawthorn, 28 parts of earthworm, 28 parts of scorpion, 48 parts of snakehead, 36 parts of gastrodia, 22 parts of bombyx batryticatus and 8 parts of borneol.

[0024] The microscopic identification includes taking a capsule sample and observing it under a microscope: The muscle fibers are layered, colorless, slightly yellow or light brown, slightly curved, and arranged vertically and staggered; Body wall fragments are pale yellow to yellow, with reticular texture and round hair pits, and brown setae are seen; The body wall fragments are colorless and have very fine mycelium on the surface.

[0025] Furthermore, the thin layer chromatography identification of Gastrodia elata includes: S110. Sample preparation; S120. Preparation of control medicinal material solution; S130. Preparation of negative control samples; S140. Perform a thin layer chromatography test, which uses a silica gel G thin layer plate and ethyl acetate-methanol-water (9:1:0.2) as a developing agent. After spraying with 10% phosphomolybdic acid ethanol solution, it is heated at 105° C. until the spots are colored.

[0026] Furthermore, the step S110 includes: taking 5 samples, adding 40 ml of water-saturated n-butanol, ultrasonically treating for 30 minutes, filtering and evaporating the filtrate, dissolving the residue in 2 ml of water, and passing through a polyamide column, eluting with 50 ml of water, collecting the eluate, evaporating and dissolving the residue in 1 ml of methanol as a sample solution.

[0027] Furthermore, the filler of the polyamide column is 5 g and has a mesh size of 30-60, and the column is filled by a wet method.

[0028] Furthermore, the thin layer chromatography identification of hawthorn includes: S210. Sample preparation; S220. Preparation of control medicinal material solution; S230. Preparation of negative control samples; S240. Perform a thin layer chromatography test using a silica gel G thin layer plate, cyclohexane-acetone-ethyl acetate-formic acid (9:2:1:0.2) as a developing agent, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are colored, and inspect under daylight or ultraviolet light.

[0029] Furthermore, the step S210 includes: taking 10 samples, adding 50 ml of ethyl acetate, ultrasonically treating for 30 minutes, filtering, evaporating the filtrate to dryness, and adding 2 ml of ethyl acetate to the residue to dissolve it as a sample solution.

[0030] Furthermore, the wavelength of the ultraviolet lamp is 365nm.

[0031] Example 2 A method for testing a drug for treating sequelae of stroke, wherein the capsule comprises the following ingredients: ginseng, deer antler glue, tortoise shell, wolfberry, turtle shell, leech, chuanxiong, rhubarb, hawthorn, earthworm, scorpion, golden snake, gastrodia, silkworm and borneol; Microscopic identification and thin layer chromatography identification are used for identification. The gastrodia elata and the hawthorn are identified by the thin layer chromatography, and the microscopic identification is used to identify the finished capsule product.

[0032] Preferably, in parts by weight, the ingredients include 45 parts of ginseng, 20 parts of antler glue, 33 parts of tortoise shell, 33 parts of wolfberry, 33 parts of turtle shell, 45 parts of leech, 33 parts of Chuanxiong, 13 parts of rhubarb, 25 parts of hawthorn, 25 parts of earthworm, 25 parts of scorpion, 45 parts of snakehead, 33 parts of gastrodia, 20 parts of bombyx batryticatus and 5 parts of borneol.

[0033] Furthermore, the microscopic identification includes taking a capsule sample and observing it under a microscope: The muscle fibers are layered, colorless, slightly yellow or light brown, slightly curved, and arranged vertically and staggered; Body wall fragments are pale yellow to yellow, with reticular texture and round hair pits, and brown setae are seen; The body wall fragments are colorless and have very fine mycelium on the surface.

[0034] Furthermore, the thin layer chromatography identification of Gastrodia elata includes: S110. Sample preparation; S120. Preparation of control medicinal material solution; S130. Preparation of negative control samples; S140. Perform a thin layer chromatography test, which uses a silica gel G thin layer plate and ethyl acetate-methanol-water (9:1:0.2) as a developing agent. After spraying with 10% phosphomolybdic acid ethanol solution, it is heated at 105° C. until the spots are colored.

[0035] Furthermore, the step S110 includes: taking 5 samples, adding 40 ml of water-saturated n-butanol, ultrasonically treating for 30 minutes, filtering and evaporating the filtrate, dissolving the residue in 2 ml of water, and passing through a polyamide column, eluting with 50 ml of water, collecting the eluate, evaporating and dissolving the residue in 1 ml of methanol as a sample solution.

[0036] Furthermore, the filler of the polyamide column is 5 g and has a mesh size of 30-60, and the column is filled by a wet method.

[0037] Furthermore, the thin layer chromatography identification of hawthorn includes: S210. Sample preparation; S220. Preparation of control medicinal material solution; S230. Preparation of negative control samples; S240. Perform a thin layer chromatography test using a silica gel G thin layer plate, cyclohexane-acetone-ethyl acetate-formic acid (9:2:1:0.2) as a developing agent, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are colored, and inspect under daylight or ultraviolet light.

[0038] Furthermore, the step S210 includes: taking 10 samples, adding 50 ml of ethyl acetate, ultrasonically treating for 30 minutes, filtering, evaporating the filtrate to dryness, and adding 2 ml of ethyl acetate to the residue to dissolve it as a sample solution.

[0039] Furthermore, the wavelength of the ultraviolet lamp is 365nm.

[0040] Example 3 A method for testing a drug for treating sequelae of stroke, wherein the capsule comprises the following ingredients: ginseng, deer antler glue, tortoise shell, wolfberry, turtle shell, leech, chuanxiong, rhubarb, hawthorn, earthworm, scorpion, golden snake, gastrodia, silkworm and borneol; Microscopic identification and thin layer chromatography identification are used for identification. The gastrodia elata and the hawthorn are identified by the thin layer chromatography, and the microscopic identification is used to identify the finished capsule product.

[0041] Preferably, in parts by weight, the ingredients include 50 parts of ginseng, 25 parts of deer antler glue, 38 parts of tortoise shell, 38 parts of wolfberry, 38 parts of turtle shell, 50 parts of leech, 38 parts of Chuanxiong, 18 parts of rhubarb, 30 parts of hawthorn, 30 parts of earthworm, 30 parts of scorpion, 50 parts of snakehead, 38 parts of gastrodia, 25 parts of bombyx batryticatus and 10 parts of borneol.

[0042] Furthermore, the microscopic identification includes taking a capsule sample and observing it under a microscope: The muscle fibers are layered, colorless, slightly yellow or light brown, slightly curved, and arranged vertically and staggered; Body wall fragments are pale yellow to yellow, with reticular texture and round hair pits, and brown setae are seen; The body wall fragments are colorless and have very fine mycelium on the surface.

[0043] Furthermore, the thin layer chromatography identification of Gastrodia elata includes: S110. Sample preparation; S120. Preparation of control medicinal material solution; S130. Preparation of negative control samples; S140. Perform a thin layer chromatography test, which uses a silica gel G thin layer plate and ethyl acetate-methanol-water (9:1:0.2) as a developing agent. After spraying with 10% phosphomolybdic acid ethanol solution, it is heated at 105° C. until the spots are colored.

[0044] Furthermore, the step S110 includes: taking 5 samples, adding 40 ml of water-saturated n-butanol, ultrasonically treating for 30 minutes, filtering and evaporating the filtrate, dissolving the residue in 2 ml of water, and passing through a polyamide column, eluting with 50 ml of water, collecting the eluate, evaporating and dissolving the residue in 1 ml of methanol as a sample solution.

[0045] Furthermore, the filler of the polyamide column is 5 g and has a mesh size of 30-60, and the column is filled by a wet method.

[0046] Furthermore, the thin layer chromatography identification of hawthorn includes: S210. Sample preparation; S220. Preparation of control medicinal material solution; S230. Preparation of negative control samples; S240. Perform a thin layer chromatography test using a silica gel G thin layer plate, cyclohexane-acetone-ethyl acetate-formic acid (9:2:1:0.2) as a developing agent, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are colored, and inspect under daylight or ultraviolet light.

[0047] Furthermore, the step S210 includes: taking 10 samples, adding 50 ml of ethyl acetate, ultrasonically treating for 30 minutes, filtering, evaporating the filtrate to dryness, and adding 2 ml of ethyl acetate to the residue to dissolve it as a sample solution.

[0048] Furthermore, the wavelength of the ultraviolet lamp is 365nm.

[0049] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for testing drugs for treating sequelae of stroke, characterized in that: The capsules include the following ingredients: ginseng, deer antler glue, tortoise shell, wolfberry, turtle shell, leech, chuanxiong, rhubarb, hawthorn, earthworm, scorpion, golden snake, gastrodia, silkworm and borneol; Microscopic identification and thin layer chromatography identification are used for identification. The gastrodia elata and the hawthorn are identified by the thin layer chromatography, and the microscopic identification is used to identify the finished capsule product.

2. A method for testing a drug for treating sequelae of stroke according to claim 1, characterized in that: In parts by weight, the invention comprises 45-50 parts of ginseng, 20-25 parts of antler glue, 33-38 parts of tortoise shell, 33-38 parts of wolfberry, 33-38 parts of turtle shell, 45-50 parts of leech, 33-38 parts of chuanxiong, 13-18 parts of rhubarb, 25-30 parts of hawthorn, 25-30 parts of earthworm, 25-30 parts of scorpion, 45-50 parts of snakehead, 33-38 parts of gastrodia, 20-25 parts of bombyx batryticatus and 5-10 parts of borneol.

3. A method for testing drugs for treating sequelae of stroke according to claim 2, characterized in that: In parts by weight, the ingredients include 48 parts of ginseng, 22 parts of antler glue, 36 parts of tortoise shell, 36 parts of wolfberry, 36 parts of turtle shell, 48 parts of leech, 36 parts of chuanxiong, 15 parts of rhubarb, 28 parts of hawthorn, 28 parts of earthworm, 28 parts of scorpion, 48 parts of snakehead, 36 parts of gastrodia, 22 parts of bombyx batryticatus and 8 parts of borneol.

4. The method for testing a drug for treating sequelae of stroke according to claim 1, characterized in that: The microscopic identification includes taking a capsule sample and observing it under a microscope: The muscle fibers are layered, colorless, slightly yellow or light brown, slightly curved, and arranged vertically and staggered; Body wall fragments are pale yellow to yellow, with reticular texture and round hair pits, and brown setae are seen; The body wall fragments are colorless and have very fine mycelium on the surface.

5. The method for testing a drug for treating sequelae of stroke according to claim 1, characterized in that: The thin layer chromatography identification of the Gastrodia elata comprises: S110. Sample preparation; S120. Preparation of control medicinal material solution; S130. Preparation of negative control samples; S140. Perform a thin layer chromatography test, which uses a silica gel G thin layer plate and ethyl acetate-methanol-water (9:1:0.2) as a developing agent. After spraying with 10% phosphomolybdic acid ethanol solution, it is heated at 105° C. until the spots are colored.

6. A method for testing drugs for treating sequelae of stroke according to claim 5, characterized in that: The step S110 comprises: taking 5 samples, adding 40 ml of saturated n-butanol, ultrasonically treating for 30 min, filtering and evaporating the filtrate, dissolving the residue in 2 ml of water, passing the residue through a polyamide column, eluting with 50 ml of water, collecting the eluate, evaporating the residue, and dissolving the residue in 1 ml of methanol as a sample solution.

7. A method for testing drugs for treating sequelae of stroke according to claim 6, characterized in that: The filler of the polyamide column is 5 g and has a mesh size of 30-60, and the column is filled by a wet method.

8. The method for testing a drug for treating sequelae of stroke according to claim 1, characterized in that: The thin layer chromatography identification of the hawthorn comprises: S210. Sample preparation; S220. Preparation of control medicinal material solution; S230. Preparation of negative control samples; S240. Perform a thin layer chromatography test using a silica gel G thin layer plate, cyclohexane-acetone-ethyl acetate-formic acid (9:2:1:0.2) as a developing agent, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are colored, and inspect under daylight or ultraviolet light.

9. A method for testing drugs for treating sequelae of stroke according to claim 8, characterized in that: The step S210 comprises: taking 10 portions of the sample, adding 50 ml of ethyl acetate, ultrasonically treating for 30 minutes, filtering, evaporating the filtrate to dryness, and adding 2 ml of ethyl acetate to dissolve the residue to obtain a sample solution.

10. The method for testing drugs for treating sequelae of stroke according to claim 8, characterized in that: The wavelength of the ultraviolet lamp is 365nm.