A preparation process of Fuzhiqing ointment medicinal composition

Abstract

The invention discloses a preparation process of a Fuzhiqing ointment pharmaceutical composition. The preparation process is as follows: extracting with alcohol from ginseng, phellodendron, and sophora flavescens, and decocting and extracting rhubarb, cinnabar root, wild chrysanthemum, Viola yedoensis, snow gall, parsley, Dioscorea bulbifera, turmeric, dandelion, and kudingcha, and removing impurities; the extract obtained by alcohol extraction and water extraction is mixed and concentrated into a clear paste, and polyethylene glycol 4000, polyethylene glycol 400, Tween-80 and glycerol base matrix, as well as auxiliary materials such as preservatives and transdermal absorption enhancers are added to prepare an ointment. Compared with the traditional process, the alkaloid content extracted by the process of the present invention is greatly improved, and the effect is remarkable; the ointment matrix has a good effective ingredient permeability, low production cost, and reduces the amount of medicinal materials, and the therapeutic effect is remarkable.

Description

Technical Field

[0001] The invention relates to the field of traditional Chinese medicines, and in particular to a preparation process of a Fuzhiqing ointment pharmaceutical composition. Background Art

[0002] Fuzhiqing Ointment is a product produced by Guizhou Green Sun Pharmaceutical Co., Ltd. It is one of the representative products of Guizhou Miao medicine brand (Drug Approval Number: National Medicine Standard Z20025745). It has the effects of clearing away heat and detoxifying, removing blood stasis and swelling, removing dampness and relieving itching. It is used for tinea pedis, tinea corporis, tinea cruris, maceration, internal hemorrhoids, external hemorrhoids, swelling, pain, bleeding and leucorrhea caused by damp-heat accumulation. Due to its significant clinical efficacy, this product is favored by patients and doctors. It has been sold for more than 100 million yuan for many consecutive years. It is one of the few Guizhou Miao medicine varieties with sales of more than 100 million yuan.

[0003] The prescription of Fuzhiqing ointment is composed of drugs such as jinguolan, rhubarb, phellodendron, cinnabar root, wild chrysanthemum, violet yedoensis, snow dandelion, sophora flavescens, borneol, parsley, dioscorea bulbifera, turmeric, dandelion, kudingcha and menthol. The original preparation process is: the 15 ingredients in the prescription, except borneol and menthol, the remaining 13 ingredients such as phellodendron, are decocted twice with water, the first time for 2 hours and the second time for 1.5 hours, the decoctions are combined, filtered, and the filtrate is concentrated to a clear ointment with a relative density of about 1.05 (60℃); sorbitol 60g, guar gum 4. 2g, 30g of beta-cyclodextrin, heat (80℃) to dissolve, and set aside; take another 10g of stearic acid, 80g of monoglyceride, 80g of octadecyl alcohol, 20g of polysorbate 80, 20g of paraffin oil, and 40g of Span 80, heat (80℃) to melt, add to the above solution, mix, emulsify for 15 minutes, keep warm at 80℃ for 30 minutes, when the temperature drops to 40℃, add borneol, menthol and 10g of laurocapram, 6g of benzoic acid, and 1.5g of ethylparaben, mix well, and make 1000g.

[0004] Fuzhiqing Ointment has been on the market for nearly 30 years. As a drug, according to Article 5 of the "Regulations on the Management of Post-Marketing Changes of Drugs (Trial)": "The holder shall take the initiative to conduct post-marketing research on drugs to achieve full life cycle management of drugs. Holders are encouraged to use new production technologies, new methods, new equipment, and new scientific and technological achievements to continuously improve and optimize production processes, continuously improve drug quality, and enhance drug safety, effectiveness, and quality controllability." With the continuous development of science and technology, the present invention team believes that it is necessary to make the following improvements to Fuzhiqing Ointment.

[0005] (1) In the original preparation process, except for borneol and menthol, the rest of the medicinal materials were extracted with water. Modern pharmacological studies have shown that the active ingredients of the prescription, such as Euphorbia miliacea, Phellodendron amurense and Sophora flavescens, that exert bactericidal and anti-inflammatory effects are all alkaloids, which are not very soluble in water. Therefore, the water extraction process has a low extraction rate of alkaloids, which inevitably leads to a waste of medicinal materials. It is necessary to optimize the extraction process.

[0006] (2) The original preparation process used a total of 12 excipients in the ointment base, which was too complicated and resulted in high costs. With the emergence of new materials and new processes, it is necessary to optimize the types and amounts of excipients. Summary of the invention

[0007] The purpose of the invention is to provide a preparation process of a Fuzhiqing ointment pharmaceutical composition.

[0008] The present invention is achieved through the following technical solutions:

[0009] The preparation process of the Fuzhiqing ointment pharmaceutical composition of the present invention comprises the following steps:

[0010] S1, according to the prescription ratio, take three medicinal materials, namely, Olea europaea, Phellodendron chinense, and Sophora flavescens, add 6 to 10 times of 40% to 80% ethanol, heat and extract 1 to 3 times, extract for 0.5 to 1.5 hours, filter, and recover the filtrate to obtain an ethanol extract for standby use;

[0011] S2, according to the prescription ratio, take 10 kinds of herbs including rhubarb, cinnabar root, wild chrysanthemum, Viola yedoensis, snow dandelion, Paris polyphylla, Dioscorea bulbifera, Curcuma longa, Sanguisorba officinalis, and Kudingcha, add water and boil for 1 to 3 times, each time for 0.5 to 1.5 hours, add 6 to 10 times of water each time, combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of 1.05 to 1.10 at 60°C;

[0012] S3, when the water-extracted paste is cooled to 50°C, add 1% to 3% chitosan in 1% acetic acid solution under stirring, stir at 30 to 50°C for 10 to 30 minutes, let stand for 24 hours, filter, and set aside the filtrate;

[0013] S4, combining the alcohol extract and the filtrate after impurity removal, and concentrating them into a clear paste with a 1:1 ratio of medicinal materials;

[0014] S5, add 40-80 parts of glycerin, 3 parts of ethylparaben, and 10 parts of laurocapram to the clear paste, mix well, add to a mixed matrix consisting of 200-600 parts of polyethylene glycol 4000 and 60-100 parts of polyethylene glycol 400, quickly stir at 70°C until the mixture is uniform, stop heating, add borneol and menthol when the temperature drops to 40°C, then add 12-20 parts of Tween-80, continue stirring until condensed, and make 1000 parts.

[0015] Preferably, step S1 of the present invention is: taking three medicinal materials of Oleracea chinensis, Phellodendron chinense and Sophora flavescens according to the prescription ratio, adding 6 times of ethanol, heating extraction twice, each time for 1 hour, filtering, and recovering the filtrate to obtain ethanol extract for standby use.

[0016] The concentration of ethanol in step S1 of the present invention is 40-80% ethanol.

[0017] Preferably, the concentration of ethanol in step S1 of the present invention is: 60% ethanol.

[0018] Preferably, step S2 of the present invention is: taking 10 medicinal materials including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, paridis, dioscorea bulbifera, turmeric, dandelion and kudingcha according to the prescription ratio, adding water and boiling for 2 times, each time for 1 hour, adding 8 times of water each time, combining the decoctions, filtering, and concentrating the filtrate to a clear paste with a relative density of 1.05 to 1.10 at 60°C.

[0019] Preferably, step S3 of the present invention is: when the water-extracted paste is cooled to 50° C., 2% chitosan solution in 1% acetic acid is added under stirring, stirred at 40° C. for 10 min, allowed to stand for 24 h, filtered, and the filtrate is used for later use.

[0020] Preferably, step S5 of the present invention is: add 60 parts of glycerol, 3 parts of ethylparaben, and 10 parts of laurocapram to the clear paste, mix well, add to a mixed matrix consisting of 400 parts of polyethylene glycol 4000 and 80 parts of polyethylene glycol 400, quickly stir at 70°C until the mixture is evenly mixed, stop heating, add borneol and menthol when the temperature drops to 40°C, then add 16 parts of Tween-80, continue stirring until condensed, and make 1000 parts.

[0021] The ratio of polyethylene glycol 4000, polyethylene glycol 400, Tween-80 and glycerol in step S5 of the present invention is: polyethylene glycol 4000: polyethylene glycol 400: Tween-80: glycerol = 100: 10-20: 2-4: 5-15.

[0022] Preferably, the ratio of polyethylene glycol 4000, polyethylene glycol 400, Tween-80 and glycerol in step S5 of the present invention is: polyethylene glycol 4000: polyethylene glycol 400: Tween-80: glycerol = 100:20:4:15.

[0023] The preparation method of the mixed matrix in step S5 is as follows: polyethylene glycol 4000 and polyethylene glycol 400 are taken according to the formula ratio, heated to melt at 70° C., and stirred at 70° C. for 15 minutes.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] 1. The extraction effect of effective ingredients is remarkable. After the three medicinal materials of Jin Guolan, Huangbai and Sophora flavescens in the prescription of Fuzhiqing ointment are extracted with ethanol, the total alkaloid content in the extract is measured to be 1.49 mg / ml, of which the content of Gulunbin is 0.067 mg / ml, the content of berberine hydrochloride is 0.65 mg / ml, and the content of oxymatrine is 0.37 mg / ml; while the total alkaloid content extracted by the prior art is only 0.52 mg / ml, of which the content of Gulunbin is 0.024 mg / ml, the content of berberine hydrochloride is 0.23 mg / ml, and the content of oxymatrine is 0.12 mg / ml. It can be seen that the extraction process of the present invention greatly increases the alkaloid content in the extract, and the extraction effect is remarkable.

[0026] 2. The ointment matrix has a good permeability of active ingredients, and the production cost is greatly reduced. The original preparation process ointment matrix used a total of 12 excipients. After the improvement of the present invention, the ointment matrix only uses a total of 6 excipients, which greatly reduces the production cost. At the same time, the new ointment matrix has a permeability of 71.88%, which is much higher than the permeability of 43.75% of the original matrix.

[0027] 3. Reduce the amount of medicinal materials used, and the treatment effect is significant. The present invention has subjected the prepared product to pharmacological experiments, and the results show that the new process group has the same treatment effect as the original process group in the mouse hemorrhoid model when the dosage is only 80% of that of the original process group. This shows that the present invention can reduce the amount of medicinal materials used and save medicinal material resources.

[0028] 4. The preparation process of the present invention is stable and feasible. Through single factor experimental investigation, the results are as follows: (1) Alcohol extraction: Add 6 times 60% ethanol to the three medicinal materials of Jin Guolan, Huangbai and Sophora flavescens, heat and extract twice, extract for 1 hour, and the effect of extracting total alkaloids is the best; (2) Purification and impurity removal: When the water-extracted clear paste is cooled to 50°C, add 2% chitosan 1% acetic acid solution under stirring, stir at 40°C for 10 minutes, and stand for 24 hours, the impurity removal rate is the best; (3) Matrix: According to the ratio of polyethylene glycol 4000: polyethylene glycol 400: Tween-80: glycerol = 100:20:4:15, the molding effect is the best; The samples were prepared under the optimal extraction conditions obtained by the above investigation, and the verification test showed that the indicators of the prepared Fuzhiqing ointment were good, and the process was stable, indicating that the process of the present invention is reasonable and feasible. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 :Chromatogram of the content of columbin extracted by water extraction

[0030] Figure 2 :Chromatogram of berberine hydrochloride content measured by water extraction

[0031] Figure 3 :Chromatogram of oxymatrine content measured by water extraction

[0032] Figure 4 :Chromatogram of the content of Gulumbin extracted by alcohol extraction

[0033] Figure 5 :Chromatogram of berberine hydrochloride content measured by alcohol extraction

[0034] Figure 6 :Chromatogram of oxymatrine content measured by alcohol extraction

[0035] Figure 7 :Chromatogram of the content of Gulumbin extracted by enzymatic hydrolysis

[0036] Figure 8 :Chromatogram of berberine hydrochloride content measured by enzymatic extraction

[0037] Fig. 9 :Chromatogram of oxymatrine content measured by enzymatic extraction

[0038] Fig.10 :Chromatogram of the content of gulombin extracted by ultrasonic extraction method

[0039] Fig.11 :Chromatogram of berberine hydrochloride content measured by ultrasonic extraction method

[0040] Fig.12 :Chromatogram of oxymatrine content measured by ultrasonic extraction

[0041] Fig.13 :Chromatogram of gulombin content measured by 40% ethanol extraction

[0042] Fig.14 :Chromatogram of berberine hydrochloride content measured by 40% ethanol extraction

[0043] Fig.15 :Chromatogram of oxymatrine content measured by 40% ethanol extraction

[0044] Fig.16 :Chromatogram of gulombin content measured by 60% ethanol extraction

[0045] Fig.17 :Chromatogram of berberine hydrochloride content measured by 60% ethanol extraction

[0046] Fig.18 :Chromatogram of oxymatrine content measured by 60% ethanol extraction

[0047] Fig.19 :Chromatogram of gulombin content measured by 80% ethanol extraction

[0048] Fig. 20 :Chromatogram of berberine hydrochloride content measured by 80% ethanol extraction

[0049] Fig.21:Chromatogram of oxymatrine content measured by 80% ethanol extraction

[0050] Fig. 22 :6 times the amount of 60% ethanol extraction to measure the chromatogram of the content of Gulumbin

[0051] Fig.23 :6 times the amount of 60% ethanol extraction to measure the berberine hydrochloride content chromatogram

[0052] Fig.24 :6 times the amount of 60% ethanol extraction to measure the oxymatrine content chromatogram

[0053] Fig.25 :Chromatogram of gulombin content measured by 8 times 60% ethanol extraction

[0054] Fig.26 :Chromatogram of berberine hydrochloride content measured by 8 times 60% ethanol extraction

[0055] Fig. 27 :Chromatogram of oxymatrine content measured by 8 times 60% ethanol extraction

[0056] Fig.28 :Chromatogram of the content of Gulumbin extracted with 10 times the amount of 60% ethanol

[0057] Fig.29 :Chromatogram of berberine hydrochloride content measured by 10 times 60% ethanol extraction

[0058] Fig.30 :10 times the amount of 60% ethanol extraction to measure the oxymatrine content chromatogram DETAILED DESCRIPTION

[0059] The technical solution of the present invention is further described in detail below through specific embodiments.

[0060] Example 1 Preparation process of a Fuzhiqing ointment pharmaceutical composition

[0061] (1) Obtaining medicinal materials

[0062] According to the prescription ratio, take 50g of Olea europaea, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Chrysanthemum indica, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Ilex kudingcha, 16g of Borneolum, and 3g of Menthol.

[0063] (2) Ethanol extraction

[0064] Take three medicinal materials, namely, Olea europaea, Phellodendron chinense and Sophora flavescens, add 6 times 60% ethanol, heat and extract twice, each time for 1 hour, filter, and recover the filtrate to obtain ethanol extract for later use.

[0065] (3) Water extraction

[0066] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and kudingcha tea, and boil them twice for 1 hour each time. Add 8 times more water each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of 1.05 (60℃).

[0067] (4) Removal of impurities

[0068] When the water-extracted paste is cooled to 50°C, add 2% chitosan in 1% acetic acid solution under stirring, stir at 40°C for 10 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0069] (5) Concentration

[0070] Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0071] (6) Preparation molding

[0072] Add 60g of glycerin, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the mixed matrix prepared in advance, stir quickly at 70°C until the mixture is evenly mixed, stop heating, add borneol and menthol when the temperature drops to 40°C, then add 16g of Tween-80, continue stirring until condensed, and make 1000g.

[0073] The mixed matrix is ​​prepared as follows: 400 g of polyethylene glycol 4000 and 80 g of polyethylene glycol 400 are heated to melt at 70° C., and stirred at 70° C. for 15 min.

[0074] Example 2 Preparation process of a Fuzhiqing ointment pharmaceutical composition

[0075] (1) Obtaining medicinal materials

[0076] According to the prescription ratio, take 50g of Olea europaea, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Chrysanthemum indica, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Ilex kudingcha, 16g of Borneolum, and 3g of Menthol.

[0077] (2) Ethanol extraction

[0078] Take three medicinal materials, namely, Olea europaea, Phellodendron chinense and Sophora flavescens, add 8 times of 40% ethanol, heat and extract 3 times, each time for 0.5 hour, filter, and recover the filtrate to obtain ethanol extract for later use.

[0079] (3) Water extraction

[0080] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and bitter melon tea, and boil them for 3 times, each time for 1.5 hours. Add 10 times the amount of water each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60℃).

[0081] (4) Removal of impurities

[0082] When the water-extracted paste is cooled to 50°C, add 1% chitosan in 1% acetic acid solution under stirring, stir at 30°C for 20 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0083] (5) Concentration

[0084] Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0085] (6) Preparation molding

[0086] Add 40g of glycerol, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the mixed matrix prepared in advance, stir quickly at 70°C until the mixture is evenly mixed, stop heating, add borneol and menthol when the temperature drops to 40°C, then add 12g of Tween-80, continue stirring until condensed, and make 1000g.

[0087] The mixed matrix is ​​prepared as follows: 400 g of polyethylene glycol 4000 and 60 g of polyethylene glycol 400 are heated to melt at 70° C., and stirred at 70° C. for 15 min.

[0088] Example 3 Preparation process of a Fuzhiqing ointment pharmaceutical composition

[0089] (1) Obtaining medicinal materials

[0090] According to the prescription ratio, take 50g of Olea europaea, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Chrysanthemum indica, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Ilex kudingcha, 16g of Borneolum, and 3g of Menthol.

[0091] (2) Ethanol extraction

[0092] Take three medicinal materials, namely, Olea europaea, Phellodendron chinense and Sophora flavescens, add 10 times 80% ethanol, heat and extract once, extract for 1.5 hours, filter, recover the filtrate to obtain ethanol extract, and set aside.

[0093] (3) Water extraction

[0094] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and kudingcha, and boil them once for 0.5 h. Add 6 times more water, combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60°C).

[0095] (4) Removal of impurities

[0096] When the water-extracted paste is cooled to 50°C, add 3% chitosan in 1% acetic acid solution under stirring, stir at 50°C for 30 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0097] (5) Concentration

[0098] Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0099] (6) Preparation molding

[0100] Add 75g of glycerin, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the mixed matrix prepared in advance, stir quickly at 70°C until the mixture is evenly mixed, stop heating, add borneol and menthol when the temperature drops to about 40°C, then add 20g of Tween-80, continue stirring until condensed, and make 1000g.

[0101] The mixed matrix is ​​prepared as follows: 500 g of polyethylene glycol 4000 and 100 g of polyethylene glycol 400 are heated to melt at 70° C., and stirred at 70° C. for 15 min.

[0102] Example 4 Preparation process of a Fuzhiqing ointment pharmaceutical composition

[0103] (1) Obtaining medicinal materials

[0104] According to the prescription ratio, take 50g of Olea europaea, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Chrysanthemum indica, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Ilex kudingcha, 16g of Borneolum, and 3g of Menthol.

[0105] (2) Ethanol extraction

[0106] Take three medicinal materials, namely, Olea europaea, Phellodendron chinense and Sophora flavescens, add 8 times of 60% ethanol, heat and extract 3 times, extract for 0.5 hour, filter, recover the filtrate to obtain ethanol extract, and set aside.

[0107] (3) Water extraction

[0108] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and kudingcha tea, and boil them twice for 1 hour each time. Add 8 times more water each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60℃).

[0109] (4) Removal of impurities

[0110] When the water-extracted paste is cooled to 50°C, add 1% chitosan in 1% acetic acid solution under stirring, stir at 40°C for 20 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0111] (5) Concentration

[0112] Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0113] (6) Preparation molding

[0114] Add 45g of glycerol, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the mixed matrix prepared in advance, stir quickly at 70°C until mixed evenly, stop heating, add borneol and menthol when the temperature drops to 35°C, then add 12g of Tween-80, continue stirring until condensed, and make 1000g.

[0115] The mixed matrix is ​​prepared as follows: 300 g of polyethylene glycol 4000 and 60 g of polyethylene glycol 400 are heated to melt at 70° C., and stirred at 70° C. for 15 min.

[0116] Example 5 Preparation process of a Fuzhiqing ointment pharmaceutical composition

[0117] (1) Obtaining medicinal materials

[0118] According to the prescription ratio, take 50g of Olea europaea, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Chrysanthemum indica, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Ilex kudingcha, 16g of Borneolum, and 3g of Menthol.

[0119] (2) Ethanol extraction

[0120] Take three medicinal materials, namely, Olea europaea, Phellodendron chinense and Sophora flavescens, add 10 times 60% ethanol, heat and extract twice, extract for 1 hour, filter, recover the filtrate to obtain ethanol extract, and set aside.

[0121] (3) Water extraction

[0122] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and kudingcha tea, and boil them twice for 1 hour each time. Add 8 times more water each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60℃).

[0123] (4) Removal of impurities

[0124] When the water-extracted paste is cooled to 50°C, add 2% chitosan in 1% acetic acid solution under stirring, stir at 50°C for 10 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0125] (5) Concentration: Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0126] (6) Preparation molding

[0127] Add 45g of glycerin, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the mixed matrix prepared in advance, stir quickly at 70°C until mixed evenly, stop heating, add borneol and menthol when the temperature drops to 45°C, then add 12g of Tween-80, continue stirring until condensed, and make 1000g.

[0128] The mixed matrix is ​​prepared as follows: 300 g of polyethylene glycol 4000 and 60 g of polyethylene glycol 400 are heated to melt at 70° C., and stirred at 70° C. for 15 min.

[0129] Example 6 Preparation process of a Fuzhiqing ointment pharmaceutical composition

[0130] (1) Obtaining medicinal materials

[0131] According to the prescription ratio, take 50g of Olea europaea, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Chrysanthemum indica, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Ilex kudingcha, 16g of Borneolum, and 3g of Menthol.

[0132] (2) Ethanol extraction

[0133] Take three medicinal materials, namely, Olea europaea, Phellodendron chinense and Sophora flavescens, add 6 times of 60% ethanol, heat and extract 3 times, extract for 1.5 hours, filter, recover the filtrate to obtain ethanol extract, and set aside.

[0134] (3) Water extraction

[0135] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and kudingcha tea, and boil them twice for 1.5 hours each time, adding 8 times more water each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60℃).

[0136] (4) Impurity removal: When the water-extracted paste is cooled to 50°C, add 2% chitosan in 1% acetic acid solution while stirring, stir at 30°C for 30 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0137] (5) Concentration: Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0138] (6) Preparation molding

[0139] Add 45g of glycerol, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the mixed matrix prepared in advance, stir quickly at 70°C until mixed evenly, stop heating, add borneol and menthol when the temperature drops to 40°C, then add 12g of Tween-80, continue stirring until condensed, and make 1000g.

[0140] The mixed matrix is ​​prepared as follows: 300 g of polyethylene glycol 4000 and 60 g of polyethylene glycol 400 are heated to melt at 70° C., and stirred at 70° C. for 15 min.

[0141] Example 7 Preparation process of a Fuzhiqing ointment pharmaceutical composition

[0142] (1) Obtaining medicinal materials

[0143] According to the prescription ratio, take 50g of Olea europaea, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Chrysanthemum indica, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Ilex kudingcha, 16g of Borneolum, and 3g of Menthol.

[0144] (2) Ethanol extraction

[0145] Take three medicinal materials, namely, Olea europaea, Phellodendron chinense and Sophora flavescens, add 8 times 80% ethanol, heat and extract twice, extract for 1 hour, filter, and recover the filtrate to obtain ethanol extract for later use.

[0146] (3) Water extraction

[0147] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and kudingcha tea, and boil them for 3 times, each time for 0.5 h. Add 6 times more water each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60℃).

[0148] (4) Removal of impurities

[0149] When the water-extracted paste is cooled to 50°C, add 1% chitosan in 1% acetic acid solution under stirring, stir at 40°C for 30 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0150] (5) Concentration: Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0151] (6) Preparation molding

[0152] Add 75g of glycerin, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the mixed matrix prepared in advance, stir quickly at 70°C until the mixture is evenly mixed, stop heating, add borneol and menthol when the temperature drops to 40°C, then add 20g of Tween-80, continue stirring until condensed, and make 1000g.

[0153] The mixed matrix is ​​prepared as follows: 500 g of polyethylene glycol 4000 and 100 g of polyethylene glycol 400 are heated to melt at 70° C., and stirred at 70° C. for 15 min.

[0154] Example 8 Preparation process of a Fuzhiqing ointment pharmaceutical composition

[0155] (1) Obtaining medicinal materials

[0156] According to the prescription ratio, take 50g of Olea europaea, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Chrysanthemum indica, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Ilex kudingcha, 16g of Borneolum, and 3g of Menthol.

[0157] (2) Ethanol extraction

[0158] Take three medicinal materials, namely, Olea europaea, Phellodendron amurense and Sophora flavescens, add 8 times of 40% ethanol, heat and extract 3 times, each time for 1.5 hours, filter, and recover the filtrate to obtain ethanol extract for later use.

[0159] (3) Water extraction

[0160] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and bitter melon tea, and boil them twice for 1.5 hours each time. Add 10 times the amount of water each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60℃).

[0161] (4) Removal of impurities

[0162] When the water-extracted paste is cooled to 50°C, add 2% chitosan in 1% acetic acid solution under stirring, stir at 40°C for 20 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0163] (5) Concentration

[0164] Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0165] (6) Preparation molding

[0166] Add 75g of glycerin, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the mixed matrix prepared in advance, stir quickly at 70°C until the mixture is evenly mixed, stop heating, add borneol and menthol when the temperature drops to 40°C, then add 20g of Tween-80, continue stirring until condensed, and make 1000g.

[0167] The mixed matrix is ​​prepared as follows: 500 g of polyethylene glycol 4000 and 100 g of polyethylene glycol 400 are heated to melt at 70° C., and stirred at 70° C. for 15 min.

[0168] In order to further verify the feasibility of the present invention, the inventor conducted a series of experiments, as follows:

[0169] 1. Investigation of extraction process

[0170] Fuzhiqing ointment is composed of the medicinal flavors of Jin Guolan, Rheum officinale, Phellodendron, Root of Cinnabar, Wild Chrysanthemum, Viola yedoensis, Snow Gallbladder, Sophora flavescens, Paris polyphylla, Dioscorea bulbifera, Curcuma longa, Sanguisorba officinalis, Kudingcha, etc. Modern pharmacological studies have shown that the effective ingredients of Jin Guolan, Phellodendron and Sophora flavescens in the prescription that play a bactericidal and anti-inflammatory role are all alkaloids, which are not very soluble in water. Therefore, the original process uses a water extraction process with a low extraction rate. In order to improve the efficiency of alkaloid extraction, it is necessary to extract these three medicines in groups using an appropriate method. The present invention team uses the content of total alkaloids in the extract, as well as the content of Gulunbin in Jin Guolan, berberine hydrochloride in Phellodendron, and oxymatrine in Sophora flavescens as investigation indicators. Through experimental comparison of four different extraction processes such as water extraction process, alcohol extraction process, enzyme extraction process and ultrasonic extraction process, the best extraction process is finally selected. The content of each indicator is determined as follows:

[0171] Determination of Gulumbin Content

[0172] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel is used as filler; acetonitrile-water (40:60) is used as mobile phase; detection wavelength is 210nm. The theoretical plate number is calculated based on the Gulomb peak and should not be less than 2500.

[0173] Preparation of test solution: Take about 0.5g of the product, accurately weigh, accurately add 10ml of 70% methanol, weigh the weight, ultrasonically treat (power 200W, frequency 59kHz) for 20 minutes, cool, weigh again, make up the lost weight with 70% methanol, shake well, filter, accurately measure 1ml of the filtrate, place it in a 10ml volumetric flask, add 70% methanol to the scale, shake well, and obtain.

[0174] Preparation of reference solution: Take an appropriate amount of columbin reference substance, accurately weigh it, and use 70% methanol to make a solution containing 0.25 mg per 1 ml.

[0175] Determination method: Accurately pipette 10 μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0176] Determination of berberine hydrochloride content

[0177] Chromatographic conditions and system suitability experiment: Octadecyl bonded silica gel was used as filler; acetonitrile-tetrahydrofuran-0.05 mol / L potassium dihydrogen phosphate solution (add 0.7% triethylamine, adjust pH to 3 with phosphoric acid) (28:12:164) was used as mobile phase; detection wavelength was 350 nm. The theoretical plate number calculated based on the berberine hydrochloride peak should be no less than 3500.

[0178] Preparation of reference solution: Take an appropriate amount of berberine hydrochloride reference substance, accurately weigh it, add methanol to make a 1 ml solution containing 0.015 mg, shake well, and obtain.

[0179] Preparation of test solution: Take 1g of the product, weigh accurately, place in a 50ml volumetric flask, add methanol to dilute to the scale, shake well, filter, take the filtrate, and pass it through a microporous filter membrane to obtain the solution.

[0180] Determination method: Accurately pipette 10 μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0181] Oxymatrine

[0182] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel is used as filler; acetonitrile-[0.01mol / L ammonium acetate solution (concentrated ammonia solution adjusted to pH 8.1)] (3:2) is used as mobile phase A, 0.01mol / L ammonium acetate solution (concentrated ammonia solution adjusted to pH 8.1) is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; the detection wavelength is 225nm. The number of theoretical plates calculated based on oxymatrine is not less than 4000.

[0183]

[0184] Preparation of test solution: Take about 1g of the product, accurately weigh it, put it in a stoppered conical flask, add 0.4ml of concentrated ammonia test solution, accurately add 25ml of chloroform, stopper it, weigh it, ultrasonically treat it (power 250W, frequency 33kHz) for 40 minutes, let it cool, weigh it again, make up the lost weight with chloroform, shake it well, filter it, accurately measure 10ml of the filtrate, recover the solvent to dryness, add anhydrous ethanol to dissolve the residue, transfer it to a 10ml volumetric flask, add anhydrous ethanol to the scale, shake it well, and you have it.

[0185] Preparation of reference solution: Take an appropriate amount of oxymatrine reference substance, weigh accurately, and add ethanol to make a solution containing 0.15 mg of oxymatrine per 1 ml.

[0186] Determination method: Accurately pipette 10μl of the above-mentioned reference solution and 10μl of the test solution, inject into liquid chromatograph, and determine.

[0187] Determination of total alkaloid content

[0188] Preparation of reference solution: Take about 15 mg of berberine hydrochloride reference substance, accurately weigh it, place it in a 50 ml volumetric flask, add about 40 ml of water, heat it in a water bath to dissolve it, dilute it to the scale with water, shake it well, and use it as the reference solution.

[0189] Preparation of standard curve: Take the reference solution, accurately measure 1.0ml, 2.0ml, 3.0ml, 4.0ml, and 5.0ml respectively, put them in 50ml volumetric flasks, add 3ml of acetone, dilute to the scale with water, and make a blank control at the same time. Measure the absorbance at a wavelength of 263nm, and regress the absorbance against the concentration to obtain the regression equation.

[0190] Preparation of test solution: Take 1g of the test sample, weigh it accurately, put it in a 50ml volumetric flask, add about 35ml of water, heat it in a water bath to dissolve it, cool it, add 3ml of acetone and then add water to the scale, shake it well, and it is ready.

[0191] Determination method: Take the test solution, measure the absorbance at a wavelength of 263nm, and calculate the berberine hydrochloride content.

[0192] 1. Preliminary selection of extraction method

[0193] (1) Water extraction method

[0194] Weigh the medicinal materials (50g of Jin Guolan, 20g of Huangbai, and 50g of Sophora flavescens) according to the proportion of the formula, add 6 times of water, heat and extract for 1.0 hour, filter, concentrate the filtrate and make it to 500ml, and determine the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine respectively. The results are shown in Table 1. Figure 1-Figure 3 .

[0195] (2) Alcohol extraction method

[0196] Weigh the medicinal materials according to the formula ratio (same as water extraction method), add 6 times 60% ethanol, heat and extract for 1.0 hour, filter, concentrate the filtrate and make it to 500ml, and determine the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine respectively. The results are shown in Table 1. Figure 4-Figure 6 .

[0197] (3) Enzymatic hydrolysis

[0198] Weigh the medicinal materials according to the formula ratio (same as water extraction method), add 6 times water and 2.0% cellulase, extract for 12 hours at 50℃ and pH 5.0, filter after boiling, concentrate the filtrate and make it to 500ml, and determine the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine respectively. The results are shown in Table 1. Figure 7-Figure 9 .

[0199] (4) Ultrasonic extraction method

[0200] Weigh the medicinal materials according to the formula ratio (same as water extraction method), add 6 times of water, ultrasonically extract for 0.5 hours, filter, concentrate the filtrate and make it to 500ml, and determine the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine respectively. The results are shown in Table 1. Figure 10-12 .

[0201] Table 1 Determination results of active ingredient content in extracts using different extraction methods

[0202]

[0203] From the test results, we can see that among the four extraction methods, the extraction effects are alcohol extraction > ultrasonic extraction > water extraction > enzyme extraction, so alcohol extraction can be selected as the best extraction method.

[0204] 2. Optimization of process parameters for alcohol extraction

[0205] The factors that affect the alcohol extraction effect are mainly ethanol concentration, alcohol addition amount, extraction times, extraction time, etc. This study used the single factor investigation method to investigate each index one by one.

[0206] (1) Ethanol concentration

[0207] Weigh 3 portions of medicinal materials (50 g of Rhizoma Chinensis, 20 g of Phellodendron chinense and 50 g of Sophora flavescens) according to the formula ratio, add 6 times of 40% ethanol, 60% ethanol and 80% ethanol respectively, extract for 1.0 hour, filter, concentrate the filtrate and make it to 500 ml, and determine the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine respectively. The results are shown in Table 2. Figure 13-Figure 21 .

[0208] Table 2 Investigation results of different ethanol concentrations

[0209]

[0210] The test results show that the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine in the prescriptions with ethanol concentrations of 60% and 80% do not change much, and the ethanol concentration of 60% can be selected as the optimal parameter.

[0211] (2) Amount of alcohol added

[0212] Weigh 3 portions of medicinal materials (50 g of Rhizoma Chinensis, 20 g of Phellodendron chinense and 50 g of Sophora flavescens) according to the formula ratio, add 6 times, 8 times and 10 times of 60% ethanol respectively, extract for 1.0 hour, filter, concentrate the filtrate and make it to 500 ml, and determine the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine respectively. The results are shown in Table 3. Figure 22-Figure 30 .

[0213] Table 3 Results of investigation on different alcohol addition amounts

[0214]

[0215] The test results show that the amount of alcohol added has little effect on the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine, and 6 times can be selected as the optimal process parameter.

[0216] (3) Number of extractions

[0217] Weigh 3 portions of medicinal materials (50 g of Rhizoma Chinensis, 20 g of Phellodendron chinense and 50 g of Sophora flavescens) according to the formula ratio, add 6 times of 60% ethanol respectively, set the extraction times to 1, 2 and 3 times respectively, each extraction time is 1.0 hour, filter, concentrate the filtrate and make the volume to 500 ml, and determine the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine respectively. The results are shown in Table 4.

[0218] Table 4 Optimization results of extraction times

[0219]

[0220] The test results showed that the contents of total alkaloids, gulumbin, berberine hydrochloride and oxymatrine in the prescriptions extracted with ethanol twice and three times did not change much, but were significantly higher than those extracted once. Therefore, ethanol extraction twice was selected as the optimal parameter.

[0221] (4) Extraction time

[0222] Weigh 3 portions of medicinal materials (50 g of Rhizoma Chinensis, 20 g of Phellodendron chinense and 50 g of Sophora flavescens) according to the formula ratio, add 6 times of 60% ethanol respectively, extract twice, and set the extraction time for each time to 0.5 hour, 1.0 hour, and 1.5 hours respectively. Filter, concentrate the filtrate and make the volume to 500 ml, and determine the contents of phellodendron hydrochloride, berberine hydrochloride, matrine and oxymatrine. The results are shown in Table 5.

[0223] Table 5 Extraction time optimization results

[0224]

[0225] The test results show that the contents of phellodendronine hydrochloride, berberine hydrochloride, matrine and oxymatrine in the extraction prescriptions of 1.0 hour and 1.5 hours do not change much, and ethanol extraction of 1.0 hour can be selected as the optimal parameter.

[0226] 3. Purification process research

[0227] 1. Preliminary selection of purification method

[0228] After the alcohol extraction of the golden fruit olive, phellodendron and sophora flavescens in the prescription, the medicinal flavors except borneol and menthol are still extracted by water. Since the water extract has more impurities, it is necessary to perform purification treatment. In order to optimize the best purification process, the present invention compares the alcohol precipitation method, ZTC clarifier method and chitosan flocculation precipitation method with the retention rate and impurity removal rate of the total flavonoids in the extract as indicators. The method is:

[0229] Weigh the medicinal materials (150g of Rheum officinale, 90g of Cinnabar Root, 90g of Chrysanthemum indica, 120g of Viola yedoensis, 30g of Psoralea corylifolia, 30g of Paris polyphylla, 30g of Dioscorea bulbifera, 30g of Curcuma longa, 150g of Sanguisorba officinalis, 60g of Ilex kudingcha) according to the proportion of the formula, and decoct them twice with water according to the original process, the first time for 2 hours and the second time for 1.5 hours, adding 6 times of water each time, combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60°C), divide it into 3 parts on average, and purify it according to the following method.

[0230] (1) Alcohol precipitation method

[0231] Take 1 portion of the clear paste, add ethanol to make the alcohol content reach 60%, stir while adding, let stand for 24 hours, filter, concentrate the filtrate and make the volume to 500 ml, measure the total flavonoid retention rate and impurity removal rate, the results are shown in Table 6.

[0232] (2) ZTC clarifier method

[0233] Take 1 portion of the clear paste, add 2% ZTC clarifier B solution, stir, keep warm at 40°C for 10 minutes, then add 1% ZTC clarifier A solution, keep warm at 40°C for 10 minutes, let stand for 24 hours, filter, concentrate the filtrate and make up to 500 ml, determine the total flavonoid retention rate and impurity removal rate, the results are shown in Table 6.

[0234] (3) Chitosan flocculation precipitation method

[0235] Take 1 portion of the clear paste, add 1% chitosan in 1% acetic acid solution, flocculation temperature 40 ° C, stirring time 10 min, let stand for 24 hours, filter, concentrate the filtrate and make up to 500 ml, determine the total flavonoids retention rate and impurity removal rate, the results are shown in Table 6.

[0236] Table 6 Preliminary results of purification method

[0237]

[0238] The test results show that the impurity removal rate is ZTC clarifier method > chitosan flocculation precipitation method > alcohol precipitation method, and the total flavonoids retention rate after impurity removal is chitosan flocculation precipitation method > alcohol precipitation method > ZTC clarifier method, indicating that the ZTC clarifier method has the greatest loss of effective ingredients while removing impurities. Taking all factors into consideration, chitosan flocculation precipitation method can be selected as the best purification process.

[0239] 2. Optimization of process parameters for chitosan flocculation precipitation method

[0240] The factors that affect the purification effect of chitosan clarifier are mainly chitosan dosage, insulation temperature and insulation time. This experiment intends to investigate these three main factors separately to optimize the best impurity removal process parameters. The test method is as follows:

[0241] Weigh the medicinal materials (500g of Rheum officinale, 300g of Cinnabar Root, 300g of Chrysanthemum Wild, 400g of Viola yedoensis, 200g of Radix Codonopsis pilosulae, 200g of Paris polyphylla, 200g of Dioscorea bulbifera, 200g of Curcuma longa, 500g of Radix Sanguisorbae, 200g of Ilex kudingcha) according to the proportion of the formula, add water and boil twice, each time for 1 hour, adding 8 times of water each time, combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60°C), divide it into 10 equal portions, and set aside.

[0242] (1) Chitosan dosage

[0243] Take 3 portions of clear paste, add 1%, 2% and 3% chitosan 1% acetic acid solution respectively, stir, stir at 40℃ for 10min, let stand for 24h, filter, concentrate the filtrate and make the volume to 500ml, determine the total flavonoid retention rate and impurity removal rate, the results are shown in Table 7.

[0244] Table 7 Chitosan dosage optimization results

[0245]

[0246] The test results show that when the chitosan dosage reaches 3%, the impurity removal rate is improved, but the total flavonoids retention rate is greatly reduced. Taking all factors into consideration, the chitosan dosage can be selected to be 2%.

[0247] (2) Insulation temperature

[0248] Take 3 portions of clear paste, add 2% chitosan 1% acetic acid solution respectively, stir, stir at 30℃, 40℃ and 50℃ for 10min respectively, let stand for 24h, filter, concentrate the filtrate and make the volume to 500ml, determine the total flavonoid retention rate and impurity removal rate, the results are shown in Table 8.

[0249] Table 8 Optimization results of insulation temperature

[0250]

[0251] The test results show that different insulation temperatures have little effect on the flavonoid retention rate, but the impurity removal rate is lower at 30℃, and 40℃ can be selected as the optimal process parameter.

[0252] (3) Insulation time

[0253] Take 3 portions of clear paste, add 2% chitosan 1% acetic acid solution respectively, stir, keep warm at 40℃ for 10min, 20min and 30min respectively, let stand for 24h, filter, concentrate the filtrate and make up to 500ml, determine the total flavonoid retention rate and impurity removal rate, the results are shown in Table 9.

[0254] Table 9 Optimization results of holding time

[0255]

[0256] The test results show that different holding times have little effect on the impurity removal rate and total flavonoids retention rate, and 10 minutes can be selected as the optimal process parameter.

[0257] 4. Matrix Optimization

[0258] The preparation molding process of the original skin hemorrhoid clear ointment is as follows: extract the medicinal materials into a clear paste according to the prescription ratio, add sorbitol 60g, guar gum 4.2g, beta-cyclodextrin 30g, heat (80°C) to dissolve, and set aside; separately take stearic acid 10g, monoglyceride 80g, octadecanol 80g, 20g polysorbate 80, paraffin oil 20g, 40g Span 80, heat (80°C) to melt, add to the above solution, mix, emulsify for 15 minutes, keep warm at 80°C for 30 minutes, add borneol, menthol and laurocapram 10g, benzoic acid 6g, ethyl hydroxybenzoate 1.5g, mix, and make 1000g, that is. This process uses many types of auxiliary materials, which is not conducive to the quality control of the preparation, and the cost is too high. With the development of new materials and new technologies, it is necessary to optimize and improve the preparation molding process.

[0259] This study intends to use polyethylene glycol 4000, polyethylene glycol 400, Tween-80 and glycerin as the basic matrix, plus a conventional proportion of 0.3% ethylparaben as a preservative and 1% laurocapram as a transdermal absorption enhancer, and carried out research on the formulation molding process.

[0260] 1. Preferred indicators and scoring criteria

[0261] (1) Appearance

[0262] The full score is 20 points. The test is based on the uniform color, fine texture, appropriate consistency, spreadability and the presence of granularity, each of which accounts for 4 points.

[0263] (2) Centrifugal stability

[0264] The full score is 20 points. Each group of ointments was placed in a centrifuge tube and centrifuged at 2500 r / min. -1 Centrifuge for 30 minutes and observe whether there is stratification or foaming. Observe whether there is any abnormal phenomenon such as stratification, agglomeration, foaming or water discharge.

[0265] (3) Cold resistance test

[0266] The full score is 10. A certain amount of ointment from each group was placed in a plastic sample cup in a 4°C refrigerator for 1 month to observe whether there were any abnormal phenomena such as stratification, agglomeration, or water release.

[0267] (4) Heat resistance test

[0268] The full score is 10. A certain amount of ointment from each group was placed in a plastic sample cup in a constant temperature oven at 40°C for 1 month to observe whether there were any abnormal phenomena such as stratification, discoloration, mildew, etc.

[0269] (5) Ductility

[0270] The full score is 20 points. Take two clean glass plates of 10cm×10cm in size, with the upper one weighing 125g (add weights to make up for the insufficient mass). Weigh 1g of each group of ointments that have been placed at room temperature for 24 hours and place them between the two glass plates. After 1 minute, use a ruler to measure the diameter of the ointment after extension, and take the average value of the ratio of the maximum diameter to the minimum diameter.

[0271] (6) pH value

[0272] The full score is 20. Take 5g of each ointment in a small beaker, add an equal amount of distilled water to fully dissolve, stir evenly, and measure the pH of each group with a pH meter. pH 6-8 is the full score, and the rest is zero.

[0273] 2. Orthogonal test

[0274] Three factors that have a greater impact on the physical and chemical properties of the matrix, as well as the ratio of polyethylene glycol 4000 and polyethylene glycol 400 (A), the amount of Tween-80 (B) and the amount of glycerol (C) were determined as the factors to be investigated. According to the preliminary data, each factor was taken at 3 levels to carry out orthogonal experiments. The factor levels are shown in Table 10, the test results are shown in Table 11, and the variance analysis is shown in Table 12.

[0275] Table 10. Orthogonal test factors and levels for matrix investigation

[0276]

[0277] Table 11 Orthogonal test design and results of matrix investigation

[0278]

[0279]

[0280] Table 12 Matrix investigation variance analysis results

[0281]

[0282] Note: F 0.05 (2, 2) = 19

[0283] From the orthogonal test results in Table 11, it can be seen that the influence of each factor on the matrix molding effect is A>B>C, and A3>A2>A1, B3>B2>B1, C3>C1>C2, and the optimal matrix ratio parameters are A3B3C3. Combined with the variance analysis in Table 12, it can be seen that the three factors have significant effects. Therefore, the optimal matrix ratio parameters are: polyethylene glycol 4000: polyethylene glycol 400: Tween-80: glycerol = 100:20:4:15.

[0284] 5. Verification of optimal process parameters

[0285] (1) Obtaining medicinal materials

[0286] Take 50g of Jin Guolan, 50g of Rheum officinale, 20g of Phellodendron chinense, 30g of Cinnabar Root, 30g of Wild Chrysanthemum, 40g of Viola yedoensis, 20g of Snow Gallbladder, 50g of Sophora flavescens, 20g of Paris polyphylla, 20g of Dioscorea bulbifera, 20g of Curcuma longa, 50g of Sanguisorba officinalis, 20g of Kudingcha, 16g of Borneolum, and 3g of Menthol according to the prescription ratio.

[0287] (2) Ethanol extraction

[0288] Add 6 times 60% ethanol to three medicinal materials, namely, Olea corylifolia, Phellodendron chinense and Sophora flavescens, and heat and extract twice for 1 hour. The extraction is filtered and the filtrate is recovered to obtain ethanol extract for later use.

[0289] (3) Water extraction

[0290] Add water to 10 ingredients including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow dandelion, parsley, dioscorea bulbifera, turmeric, dandelion, and bitter melon tea, and boil them twice for 1 hour each time, adding 8 times the amount of water each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60℃).

[0291] (4) Removal of impurities

[0292] When the water-extracted paste is cooled to 50°C, add 2% chitosan in 1% acetic acid solution under stirring, stir at 40°C for 10 minutes, let stand for 24 hours, filter, and use the filtrate for later use.

[0293] (5) Concentration

[0294] Combine the alcohol extract and the filtrate after impurity removal and concentrate to a clear paste with a ratio of 1:1 to medicinal materials.

[0295] (6) Preparation molding

[0296] Add 60g of glycerol, 3g of ethylparaben, and 10g of laurocapram to the clear paste, mix well, add to the matrix (matrix preparation: take 400g of polyethylene glycol 4000 and 80g of polyethylene glycol 400, heat to melt at 70°C, keep stirring at 70°C for 15min), stir quickly at 70°C until the mixture is uniform, stop heating, wait until the temperature drops to about 40°C, add 16g of Tween-80, continue stirring until condensed, and make 1000g.

[0297] (7) Verification test

[0298] Three batches of samples were prepared according to the above optimal parameters, and the results are shown in Table 13.

[0299] Table 13 Verification results of optimal water extraction process parameters

[0300]

[0301] The verification results show that the Fuzhiqing ointment prepared by the present invention has good indicators and a stable process, which shows that the present invention is reasonable and feasible.

[0302] VI. Drug efficacy verification test

[0303] 1. In vitro skin permeation test

[0304] (1) Experimental Materials

[0305] ①Instrument

[0306] TP-5 transdermal diffusion experiment instrument (Shanghai Kaikai Technology Trading Co., Ltd.); super constant temperature bath (Shanghai Hengping Instrument Factory); electric shaver (Oster Company, USA); electronic balance (Beijing Sartorius Instrument System Co., Ltd.); high performance liquid chromatography (HPLC) instrument (Shimadzu Corporation, Japan).

[0307] ②Reagents and drug testing

[0308] Acetonitrile (chromatographic grade, Tiandi Co., Ltd., USA), phosphoric acid, sodium dodecyl sulfate, methanol (analytical grade, Chongqing Wansheng Chuandong Chemical Co., Ltd.), water is ultrapure water. Berberine hydrochloride reference substance (batch number: 110713-202109, China Food and Drug Administration)

[0309] ③ Animals

[0310] 20 healthy Kunming mice, ♂, weight 18-20 g, SPF grade, provided by the Medical Experimental Animal Development Center of Guizhou Medical University, certificate number: Yidongzi No. 006.

[0311] (2) Experimental methods and results

[0312] ① Preparation of ex vivo mouse skin: After the mice were raised for 3 days, they were anesthetized by intraperitoneal injection of 30% chloral hydrate and killed by dislocation of the neck. The hair on the back skin of the mice was removed with a shaver. The skin of mice of appropriate size was taken and the subcutaneous adhesions were removed. The skin was rinsed with saline several times to remove excess tissue fragments, and the saline on the surface of the mouse skin was blotted with filter paper. The washed mouse skin was soaked in saline for use and used within 24 hours. Before the experiment, the mouse skin of appropriate size and undamaged was selected under a dissecting microscope for use.

[0313] ② In vitro transdermal absorption test: This test uses the modified Franz diffusion cell method. Place the mouse skin between the diffusion cell and the receiving cell, so that the stratum corneum of the mouse skin faces the supply chamber, and the dermis is in close contact with the receiving solution (30% ethanol-70% saline), and ensure that there are no bubbles. Accurately weigh 1g of Fuzhiqing ointment, place it in the supply cell, and apply it closely to the mouse skin so that it is evenly distributed on the surface of the mouse stratum corneum skin. Add 15ml of receiving solution (30% ethanol-70% saline) to the receiving chamber, keep a constant temperature of (37±0.5)℃, and stir at a speed of 350r / min. Take 1ml of receiving solution after 24h and set aside.

[0314] ③ Determination of berberine hydrochloride:

[0315] Chromatographic conditions and system suitability test: octadecylsilane bonded silica gel is used as filler; acetonitrile-0.1% phosphoric acid solution (50:50) (0.1g sodium dodecyl sulfonate is added per 100ml) is used as mobile phase; detection wavelength is 265nm. The theoretical plate number calculated based on the berberine hydrochloride peak should be no less than 4000.

[0316] Preparation of reference solution: Take an appropriate amount of berberine hydrochloride reference substance, weigh it accurately, add mobile phase to make a solution containing 0.05 mg per 1 ml.

[0317] Preparation of test solution: Accurately pipette 0.5 ml of the above receiving solution into a 100 ml conical flask with a stopper, evaporate to dryness in a water bath, cool, accurately add 15 ml of methanol, stopper, weigh, ultrasonically treat (power 250 W, frequency 40 kHz) for 30 minutes, cool, weigh again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the solution.

[0318] Determination method: Accurately pipette 10μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0319] The measurement results are shown in Table 14.

[0320] Table 14 Comparison of transdermal absorption of Fuzhiqing ointment by two preparation methods

[0321]

[0322] The test results show that, compared with the Fuzhiqing ointment prepared by the preparation method of the present invention and the Fuzhiqing ointment prepared by the original preparation method, in an in vitro transdermal absorption test, the transdermal absorption rate is higher and the transdermal absorption effect is better.

[0323] 2. Mouse hemorrhoid model test

[0324] (1) Experimental Materials

[0325] ① Animals: SPF grade KM mice, half male and half female, weighing 22-25 g, 60 mice, half male and half female.

[0326] ② Drugs and reagents: Fuzhiqing ointment sample prepared according to the preparation method of the present invention; Fuzhiqing ointment (Guizhou Green Sun Pharmaceutical Co., Ltd., batch number: 20230316); rat interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), nitric oxide (NO) enzyme-linked immunosorbent assay (ELISA) kit (Shanghai Youxuan Biotechnology Co., Ltd., batch number 20230315SR).

[0327] (2) Test methods

[0328] ① Modeling grouping and drug administration: 60 mice were taken, and 10 mice were randomly selected as the blank group after adaptive feeding for 3 days. The perianal skin of other animals was first disinfected, and 0.05 ml of 20% acetic acid solution was pushed into the anus of each mouse for 1 minute. The blank group was pushed into 0.05 normal saline. After 24 hours of modeling, white ulcer surface, perianal swelling and inflammatory exudation were visible, indicating that the modeling was successful. The mice with successful modeling were randomly divided into 5 groups: model group, original process group, present invention group 1 (0.6 times the original process dosage), present invention group 2 (0.8 times the original process dosage) and present invention group 3 (the same dosage as the original process). After grouping, perianal external application of drug administration began, the blank group and the model group were externally applied with normal saline, and the other groups were externally applied with ointment. Each group applied the drug 3 times a day for 7 consecutive days.

[0329] ② Anal perianal scoring after mouse modeling: 1 hour after drug administration on the last day, the drug was removed, the anal perianal area was cleaned, and the anal perianal area was scored. The scoring criteria were: 0 point for complete healing, 1 point for basic healing, 2 points for a little ulcer exudate, and 3 points for ulcer exudate.

[0330] ③ Determination of IL-1β and TNF-α content in mouse serum: 6 hours after the last medication, remove the drug, clean the medication site, remove the eyeballs and collect blood, and detect the serum IL-1β and TNF-α content.

[0331] ④Statistical methods: SPSS17.0 statistical software was used for data processing. The data were expressed as x±s. The t-test was used for comparison between groups. The Ridit test was used for rank data. P < 0.05 was considered statistically significant.

[0332] (3) Test results

[0333] ① Effect on mouse hemorrhoid model: The blank group was normal locally. After Ridit test, compared with the blank group, the model group had significant ulcer exudate locally (P < 0.01); compared with the model group, the original process group, the present invention group 2 and the present invention group 3 all significantly inhibited perianal swelling and ulcers, promoted ulcer healing, and improved perianal pathological conditions (P < 0.01), among which the present invention group 2 and the original process group had similar improvement effects, and the amount of medicinal materials used in the present invention group 2 was only 80% of that in the original process group, indicating that the present invention can reduce the amount of medicinal materials used. See Table 15.

[0334] Table 15 Effect of Fuzhiqing Ointment on Perianal Score of Mice (n=10)

[0335]

[0336] ② Effect on the content of IL-1β and TNF-α in mouse serum: Compared with the blank group, the levels of TNF-α and IL-1β in the serum of the model group mice were significantly increased (P < 0.01); compared with the model group, the original process group, the present invention group 2 and the present invention group 3 can significantly reduce the levels of IL-1β and TNF-α in the serum of the mouse hemorrhoid model (P < 0.01); the reduction of the present invention group 2 is comparable to that of the original process group, which again shows that the present invention can reduce the amount of medicinal materials used. See Table 16.

[0337] Table 16 Effect of serum TNF-α and IL-1β levels in mouse hemorrhoid model in each experimental group (x±s, n=10)

[0338]

[0339] Although the present invention has been described in detail above by means of general description, specific implementation methods and experiments, it is obvious to those skilled in the art that some modifications or improvements may be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A preparation process of a Fuzhiqing ointment pharmaceutical composition, the pharmaceutical composition is composed of 50 parts of golden fruit olive, 50 parts of rhubarb, 20 parts of phellodendron, 30 parts of cinnabar roots, 30 parts of wild chrysanthemum, 40 parts of Viola yedoensis, 20 parts of snow gall, 50 parts of sophora flavescens, 20 parts of parsley, 20 parts of Dioscorea bulbifera, 20 parts of turmeric, 50 parts of sanguisorba officinalis, 20 parts of kudingcha, 16 parts of borneol, and 3 parts of menthol. The raw materials are characterized in that The preparation process comprises the following steps: S1, according to the prescription ratio, take three medicinal materials, namely, Olea europaea, Phellodendron chinense, and Sophora flavescens, add 6 to 10 times of 40% to 80% ethanol, heat and extract 1 to 3 times, extract for 0.5 to 1.5 hours, filter, and recover the filtrate to obtain an ethanol extract for standby use; S2, according to the prescription ratio, take 10 kinds of herbs including rhubarb, cinnabar root, wild chrysanthemum, Viola yedoensis, snow dandelion, Paris polyphylla, Dioscorea bulbifera, Curcuma longa, Sanguisorba officinalis, and Kudingcha, add water and boil for 1 to 3 times, each time for 0.5 to 1.5 hours, add 6 to 10 times of water each time, combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of 1.05 to 1.10 at 60°C; S3, when the water-extracted paste is cooled to 50°C, add 1% to 3% chitosan in 1% acetic acid solution under stirring, stir at 30 to 50°C for 10 to 30 minutes, let stand for 24 hours, filter, and use the filtrate for later use; S4, combining the alcohol extract and the filtrate after impurity removal, and concentrating them into a clear paste with a 1:1 ratio of medicinal materials; S5, add 40-80 parts of glycerin, 3 parts of ethylparaben, and 10 parts of laurocapram to the clear paste, mix well, add to a mixed matrix consisting of 200-600 parts of polyethylene glycol 4000 and 60-100 parts of polyethylene glycol 400, quickly stir at 70°C until the mixture is uniform, stop heating, add borneol and menthol when the temperature drops to 40°C, then add 12-20 parts of Tween-80, continue stirring until condensed, and make 1000 parts.

2. The preparation process of the Fuzhiqing ointment pharmaceutical composition according to claim 1, characterized in that, The step S1 is: taking three medicinal materials of Oleracea chinensis, Phellodendron chinense and Sophora flavescens according to the prescription ratio, adding 6 times of ethanol, heating and extracting twice, each time for 1 hour, filtering, and recovering the filtrate to obtain ethanol extract for standby use.

3. The preparation process of the Fuzhiqing ointment pharmaceutical composition according to claim 1, characterized in that, The concentration of ethanol in step S1 is: 60% ethanol.

4. The preparation process of the Fuzhiqing ointment pharmaceutical composition according to claim 1, characterized in that: The step S2 comprises: taking 10 kinds of medicinal materials including rhubarb, cinnabar root, wild chrysanthemum, viola yedoensis, snow twig, parsley, dioscorea bulbifera, turmeric, dandelion and kudingcha according to the prescription ratio, adding water and boiling for 2 times, each time for 1 hour, adding 8 times of water each time, combining the decoctions, filtering, and concentrating the filtrate to a clear paste with a relative density of 1.05-1.10 at 60°C.

5. The preparation process of the Fuzhiqing ointment pharmaceutical composition according to claim 1, characterized in that: The step S3 is: when the water-extracted paste is cooled to 50° C., 2% chitosan solution in 1% acetic acid is added under stirring, stirred at 40° C. for 10 minutes, allowed to stand for 24 hours, filtered, and the filtrate is used for later use.

6. The preparation process of the Fuzhiqing ointment pharmaceutical composition according to claim 1, characterized in that: The step S5 is as follows: adding 60 parts of glycerol, 3 parts of ethylparaben and 10 parts of laurocapram to the clear paste, mixing well, adding to a mixed matrix consisting of 400 parts of polyethylene glycol 4000 and 80 parts of polyethylene glycol 400, rapidly stirring at 70° C. until the mixture is uniformly mixed, stopping heating, adding borneol and menthol when the temperature drops to 40° C., then adding 16 parts of Tween-80, continuing stirring until condensation, and making 1000 parts.

7. The preparation process of the Fuzhiqing ointment pharmaceutical composition according to claim 1, characterized in that: The ratio of polyethylene glycol 4000, polyethylene glycol 400, Tween-80 and glycerol in step S5 is: polyethylene glycol 4000: polyethylene glycol 400: Tween-80: glycerol = 100: 10-20: 2-4: 5-15.

8. The preparation process of the Fuzhiqing ointment pharmaceutical composition according to claim 7, characterized in that: The ratio of polyethylene glycol 4000, polyethylene glycol 400, Tween-80 and glycerol in step S5 is: polyethylene glycol 4000: polyethylene glycol 400: Tween-80: glycerol = 100:20:4:

15.

9. The preparation process of the Fuzhiqing ointment pharmaceutical composition according to claim 1, characterized in that: The preparation method of the mixed matrix in step S5 is as follows: polyethylene glycol 4000 and polyethylene glycol 400 are taken according to the formula ratio, heated to melt at 70° C., and stirred at 70° C. for 15 minutes.

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