Preparation process of a sunscreen drug

Abstract

The present invention discloses a preparation process of a sunscreen drug, which relates to the field of pharmaceutical technology. The sunscreen drug is composed of honeysuckle, polygonum perfoliatum, sedum sarmentosum, commelina communis, polygonatum odoratum, lithospermum erythrorhizon, aloe vera, asparagus cochinchinensis, ganoderma lucidum, coix seed, and bee pollen, and is one of the representative products of Guizhou Miao medicine brands. The preparation process of the present invention comprises the following steps: taking medicinal materials, supercritical fluid extraction, water extraction, impurity removal and purification, concentration, and preparation forming. Through a large number of condition investigation tests, the present invention improves the preparation process of the sunscreen drug, increases the extraction rate of active ingredients, and optimizes the types and dosages of excipients for preparation forming, greatly reducing the process difficulty and production cost.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to a preparation process of a sunscreen medicine. Background Art

[0002] Sunburn prevention cream (Drug approval number: National Medicine Standard Z20025534) is one of the representative products of Guizhou Miao medicine brand. It has the effects of clearing away heat and detoxifying, cooling blood and removing spots. It is used to prevent and treat sunburn caused by heat and toxicity. The product has significant clinical efficacy and is favored by patients and doctors.

[0003] The sunscreen prevention and treatment ointment prescription is composed of honeysuckle, angelica sinensis, creeping vine, communis herb, polygonatum, lithospermum officinale, aloe, asparagus cochinchinensis, ganoderma lucidum, coix seed, and bee pollen. The original preparation process is: among the eleven medicinal materials in the prescription, ganoderma lucidum is used as 80% ethanol as a solvent, and after being immersed for 12 hours, it is slowly percolated at a rate of 1 to 3 mL per minute, the percolated liquid is collected, the ethanol is recovered under reduced pressure, and concentrated to a clear paste with a relative density of 1.15 (50°C); except for aloe, the other nine herbs 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 paste with a relative density of 1.05 (60°C). Take sorbitol, sodium alginate, glycerin, beta-cyclodextrin, titanium dioxide and aloe vera, add to the above-mentioned Ganoderma lucidum fluid extract and traditional Chinese medicine concentrate, and heat (80°C) to dissolve; separately take stearic acid, monoglyceride, vaseline, octadecyl alcohol, polysorbate-80, Span-80, lanolin, sucrose stearate, and emulsified silicone oil, heat (80°C) to melt, mix the above two solutions and emulsify for 15 minutes, keep at 80°C for 30 minutes, add benzoic acid and ethylparaben, mix well, and obtain the product.

[0004] As a drug, according to Article 5 of the "Administrative Measures for 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 the following problems existing in the original preparation process should be improved:

[0005] (1) Traditional Chinese medicines are not single components, but a collection of numerous active ingredients. Modern pharmacological studies have shown that the ingredients in the prescription that can prevent and treat sunburn include chlorogenic acid, the active ingredient in honeysuckle; quercetin and kaempferol, the active ingredients in sedum sarmentosum Bunge; β,β'-dimethylacryloylakannin, the active ingredient in lithospermum erythrorhizon Sieb. et Zucc.; quercetin, the active ingredient in polygonum perfoliatum L.; and ganoderma polysaccharide, the active ingredient in ganoderma lucidum. In the original preparation process, honeysuckle, sedum sarmentosum Bunge, polygonum perfoliatum L., and lithospermum erythrorhizon Sieb. et Zucc. were all extracted with water. However, studies have shown that the extraction rates of the active ingredients in the above four herbs are not high through simple water extraction processes. Ganoderma lucidum was extracted by percolation with an alcohol solution, but ganoderma polysaccharide has low solubility in alcohol and the extraction rate is not high, which will inevitably lead to waste of medicinal materials. Therefore, it is necessary to optimize the extraction process.

[0006] (2) A total of 16 excipients were used in the original preparation process of the ointment matrix, which was too complex and led to high costs. With the emergence of new materials and new processes, it is necessary to optimize the types and dosages of excipients.

[0007] Based on this, through a large number of conditional investigation experiments, the present invention has improved and optimized the preparation process of this sunscreen drug, aiming to simplify the production process, reduce production costs, and improve product quality and production efficiency. Summary of the Invention

[0008] The purpose of the present invention is to provide a preparation process for a sunscreen drug.

[0009] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0010] The preparation process of the sunscreen drug described in the present invention includes the following steps:

[0011] S1 Take medicinal materials:

[0012] 40 parts of honeysuckle, 40 parts of polygonum perfoliatum L., 30 parts of sedum sarmentosum Bunge, 30 parts of commelina communis L., 40 parts of polygonatum odoratum (Mill.) Druce, 20 parts of lithospermum erythrorhizon Sieb. et Zucc., 5 parts of aloe vera, 40 parts of asparagus cochinchinensis (Lour.) Merr., 40 parts of ganoderma lucidum, 20 parts of coix lacryma-jobi L., 10 parts of bee pollen.

[0013] S2 Supercritical fluid extraction:

[0014] Perform supercritical CO2-ethanol extraction on 5 medicinal materials including honeysuckle, polygonum perfoliatum L., sedum sarmentosum Bunge, lithospermum erythrorhizon Sieb. et Zucc., and ganoderma lucidum. After the extraction is completed, reduce the pressure and recover the precipitated extract for standby;

[0015] S3 Water extraction:

[0016] Decoct 5 medicinal materials including commelina communis L., polygonatum odoratum (Mill.) Druce, asparagus cochinchinensis (Lour.) Merr., coix lacryma-jobi L., and bee pollen with water twice, combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of 1.05;

[0017] S4 Impurity removal and purification:

[0018] Add the ZTC clarifying agent B solution to the clear extract obtained by the above water extraction method, stir evenly, and keep warm; then add the ZTC clarifying agent A solution, stir evenly, keep warm, stand for 24 h, filter, and reserve the filtrate.

[0019] S5 Concentration:

[0020] Combine the extract obtained by the supercritical fluid extraction method and the filtrate obtained after impurity removal and purification, and concentrate to a clear extract.

[0021] S6 Preparation and forming:

[0022] Add 2 - 4 parts of triethanolamine, 15 parts of glycerol, 1 part of ethyl paraben, 1 part of sodium benzoate, aloe vera, and an appropriate amount of distilled water to the clear extract obtained by the above concentration, heat to 80 °C as the aqueous phase; mix 25 parts of glyceryl monostearate, 40 - 80 parts of stearic acid, 20 - 40 parts of liquid paraffin, and 45 parts of stearyl alcohol, and heat to 80 °C to melt as the oil phase; slowly pour the aqueous phase into the oil phase at 80 °C, stir while adding, stir and emulsify for 20 min, keep warm, and make 1000 parts.

[0023] In the preparation process of the sunscreen drug described in the present invention, in step S2, the pressure of the supercritical CO2 - ethanol extraction is 10 - 40 MPa, the temperature is 40 - 60 °C, the CO2 - ethanol flow rate is 5 - 9 L / h, and the extraction time is 2 - 6 h.

[0024] Preferably, in the preparation process of the sunscreen drug described in the present invention, in step S2, the pressure of the supercritical CO2 - ethanol extraction is 30 MPa, the temperature is 50 °C, the CO2 - ethanol flow rate is 7 L / h, and the extraction time is 4 h.

[0025] In the preparation process of the sunscreen drug described in the present invention, in step S3, the specific process of decocting with water is as follows: the decocting time is 2 h for the first time and 1.5 h for the second time, and 6 times the mass of the medicinal materials of water is added each time.

[0026] In the preparation process of the sunscreen drug described in the present invention, in step S3, the relative density is the measured value at 60 °C.

[0027] In the preparation process of the sunscreen drug described in the present invention, in step S4, the specific process of impurity removal and purification is as follows: add 2% - 4% of the ZTC clarifying agent B solution to the clear extract obtained by water extraction, stir evenly, keep warm at 40 °C - 60 °C for 20 min - 40 min; add 1% - 3% of the ZTC clarifying agent A solution, stir evenly, keep warm at 40 °C - 60 °C for 20 min - 40 min, stand for 24 h, filter, and reserve the filtrate.

[0028] Preferably, in the preparation process of the sunscreen drug of the present invention, the specific process of impurity removal and purification in step S4 is as follows: add 3% of ZTC clarifying agent B solution to the clear extract obtained by water extraction, stir evenly, and keep it warm at 40 °C for 20 min; add 2% of ZTC clarifying agent A solution, stir evenly, keep it warm at 40 °C for 20 min, let it stand for 24 h, filter, and reserve the filtrate.

[0029] In the preparation process of the sunscreen drug of the present invention, the concentration to clear extract in step S5 is specifically concentrated to a clear extract with a mass ratio of 1:1 to the medicinal materials.

[0030] Preferably, in the preparation process of the sunscreen drug of the present invention, the addition amount of triethanolamine in step S6 is 4 parts, the addition amount of stearic acid is 80 parts, and the addition amount of liquid paraffin is 40 parts.

[0031] In the preparation process of the sunscreen drug of the present invention, the heat preservation condition in step S6 is 80 °C, and the heat preservation time is 30 min.

[0032] The beneficial effects of the present invention:

[0033] Through experimental research, the present invention groups and extracts the prescription of the sunscreen drug, improves the extraction rate of active ingredients, and optimizes the excipients for preparation molding. Compared with the original preparation method, it has the following beneficial effects:

[0034] (1) The content of chlorogenic acid in the original process extract was 0.96 mg / mL, the content of quercetin was 0.18 mg / mL, the content of kaempferol was 0.018 mg / mL, the content of β,β'-dimethylacryloylalkanin was 0.08 mg / mL, and the content of ganoderma polysaccharide was 0.43 mg / mL; after using supercritical fluid extraction for the five medicinal materials of honeysuckle, sedum sarmentosum, polygonum perfoliatum, lithospermum erythrorhizon and ganoderma lucidum in the sunscreen drug prescription of the present invention, the content of chlorogenic acid in the dilution was 1.76 mg / mL, the content of quercetin was 0.39 mg / mL, the content of kaempferol was 0.040 mg / mL, the content of β,β'-dimethylacryloylalkanin was 0.17 mg / mL, and the content of ganoderma polysaccharide was 1.03 mg / mL, significantly improving the content of active ingredients.

[0035] (2) A total of 16 excipients were used in the original preparation process of the ointment matrix. After being improved by the present invention, only 8 excipients are used in the ointment matrix, significantly reducing the process difficulty and production cost.

[0036] (3) The results of pharmacological experiments show that when the dosage of the new process group is only 80% of that of the original process group, the treatment effect in the mouse solar dermatitis model is equivalent to that of the original process group. It shows that the present invention can reduce the usage amount of medicinal materials and save medicinal resources. Description of the Drawings

[0037] Figure 1 It is the standard curve graph for the determination of polysaccharide content; Specific implementation manners

[0038] The technical solutions of the present invention will be described in detail below in conjunction with specific embodiments. The following embodiments are only for explanation and illustration, and do not constitute a limitation to the technical solutions of the present invention.

[0039] Embodiment 1

[0040] The preparation process of the sunscreen drug is as follows:

[0041] 1) Take medicinal materials

[0042] Weigh 40 g of Flos Lonicerae, 40 g of Polygonum perfoliatum L., 30 g of Sedum sarmentosum Bunge, 30 g of Commelina communis L., 40 g of Polygonatum odoratum (Mill.) Druce, 20 g of Lithospermum erythrorhizon Sieb. et Zucc., 5 g of Aloe vera (L.) Burm. f., 40 g of Asparagus cochinchinensis (Lour.) Merr., 40 g of Ganoderma lucidum (Leyss. ex Fr.) Karst., 20 g of Coix lacryma-jobi L., and 10 g of bee pollen according to the prescription ratio.

[0043] 2) Supercritical fluid extraction

[0044] Perform supercritical extraction on 5 medicinal materials including Flos Lonicerae, Polygonum perfoliatum L., Sedum sarmentosum Bunge, Lithospermum erythrorhizon Sieb. et Zucc., and Ganoderma lucidum (Leyss. ex Fr.) Karst. under the conditions of a pressure of 30 MPa, a temperature of 50 °C, and a CO2-ethanol flow rate of 7 L / h for 4 h. After the extraction is completed, reduce the pressure and recover the precipitated extract for standby.

[0045] 3) Water extraction

[0046] Decoct 5 medicinal materials including Commelina communis L., Polygonatum odoratum (Mill.) Druce, Asparagus cochinchinensis (Lour.) Merr., Coix lacryma-jobi L., and bee pollen with water twice. The first time is for 2 h, and the second time is for 1.5 h. Add 6 times the mass of water of the medicinal materials each time. Combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60 °C).

[0047] 4) Impurity removal and purification

[0048] Add 3% of ZTC clarifying agent B solution to the clear paste prepared by the above water extraction, stir evenly, and keep it warm at 40 °C for 20 min; then add 2% of ZTC clarifying agent A solution, stir evenly, and keep it warm at 40 °C for 20 min; let it stand for 24 h, filter, and keep the filtrate for standby.

[0049] 5) Concentration

[0050] Combine the extract obtained by supercritical fluid extraction and the filtrate after impurity removal and purification, and concentrate it to a clear paste with a mass ratio of 1:1 to the medicinal materials.

[0051] 6) Preparation forming

[0052] Add 4 g of triethanolamine, 15 g of glycerol, 1 g of ethyl paraben, 1 g of sodium benzoate, 5 g of aloe vera to the clear extract obtained by the above concentration, and add distilled water to a total amount of 810 g. Heat to 80 °C as the aqueous phase. Melt 25 g of glyceryl monostearate, 80 g of stearic acid, 40 g of liquid paraffin, and 45 g of stearyl alcohol at 80 °C as the oil phase. Slowly pour the aqueous phase into the oil phase at 80 °C, stirring while adding, and stir for emulsification for 20 min. Keep warm at 80 °C for 30 min to make 1000 g.

[0053] Example 2

[0054] The preparation process of the sunscreen drug is as follows:

[0055] 1) Take the medicinal materials

[0056] Weigh 40 g of honeysuckle, 40 g of polygonum perfoliatum, 30 g of sedum sarmentosum, 30 g of commelina communis, 40 g of polygonatum odoratum, 20 g of lithospermum erythrorhizon, 5 g of aloe vera, 40 g of asparagus cochinchinensis, 40 g of ganoderma lucidum, 20 g of coix lacryma-jobi, and 10 g of bee pollen according to the prescription ratio.

[0057] 2) Supercritical fluid extraction

[0058] Perform supercritical extraction on 5 medicinal materials including honeysuckle, polygonum perfoliatum, sedum sarmentosum, lithospermum erythrorhizon, and ganoderma lucidum under the conditions of a pressure of 30 MPa, a temperature of 50 °C, and a CO2-ethanol flow rate of 7 L / h for 4 h. After the extraction is completed, reduce the pressure and recover the precipitated extract for standby.

[0059] 3) Water extraction

[0060] Decoct 5 medicinal materials including commelina communis, polygonatum odoratum, asparagus cochinchinensis, coix lacryma-jobi, and bee pollen with water twice. The first time is for 2 h, and the second time is for 1.5 h. Add 8 times the mass of the medicinal materials of water each time. Combine the decoction liquids, filter, and concentrate the filtrate to a clear extract with a relative density of about 1.05 (60 °C).

[0061] 4) Impurity removal and purification

[0062] Add 3% of ZTC clarifying agent B solution to the clear extract obtained by the above water extraction, stir evenly, and keep warm at 40 °C for 20 min; then add 2% of ZTC clarifying agent A solution, stir evenly, and keep warm at 40 °C for 20 min; let stand for 24 h, filter, and reserve the filtrate.

[0063] 5) Concentration

[0064] Combine the extract obtained by supercritical fluid extraction and the filtrate after impurity removal and purification, and concentrate to a clear extract with a mass ratio of 1:1 to the medicinal materials.

[0065] 6) Preparation forming

[0066] Add 4 g of triethanolamine, 15 g of glycerol, 1 g of ethylparaben, 1 g of sodium benzoate, 5 g of aloe vera to the clear extract obtained by the above concentration, add distilled water to a total amount of 810 g, heat to 80 °C as the aqueous phase, 25 g of glyceryl monostearate, 80 g of stearic acid, 40 g of liquid paraffin, 45 g of stearyl alcohol, heat to 80 °C and melt as the oil phase. Slowly pour the aqueous phase into the oil phase at 80 °C, stirring while adding, stir and emulsify for 20 min, keep warm at 80 °C for 30 min to make 1000 g.

[0067] Example 3

[0068] The preparation process of the sunscreen drug is as follows:

[0069] 1) Take the medicinal materials

[0070] Weigh 40 g of honeysuckle, 40 g of polygonum perfoliatum, 30 g of sedum sarmentosum, 30 g of commelina communis, 40 g of polygonatum odoratum, 20 g of lithospermum erythrorhizon, 5 g of aloe vera, 40 g of asparagus cochinchinensis, 40 g of ganoderma lucidum, 20 g of coix lacryma-jobi, and 10 g of bee pollen according to the prescription ratio.

[0071] 2) Supercritical fluid extraction

[0072] Perform supercritical extraction on 5 medicinal materials including honeysuckle, polygonum perfoliatum, sedum sarmentosum, lithospermum erythrorhizon and ganoderma lucidum under the conditions of a pressure of 10 MPa, a temperature of 40 °C, and a CO2-ethanol flow rate of 5 L / h for 6 h. After the extraction is completed, reduce the pressure and recover the precipitated extract for standby.

[0073] 3) Water extraction

[0074] Decoct 5 medicinal materials including commelina communis, polygonatum odoratum, asparagus cochinchinensis, coix lacryma-jobi and bee pollen with water twice, 2 h for the first time and 1.5 h for the second time, and add 6 times the mass of the medicinal materials of water each time. Combine the decoction liquids, filter, and concentrate the filtrate to a clear extract with a relative density of about 1.05 (60 °C).

[0075] 4) Impurity removal and purification

[0076] Add 2% of ZTC clarifying agent B solution to the clear extract obtained by the above water extraction, stir evenly, keep warm at 40 °C for 40 min; then add 1% of ZTC clarifying agent A solution, stir evenly, keep warm at 40 °C for 40 min; let stand for 24 h, filter, and reserve the filtrate.

[0077] 5) Concentration

[0078] Combine the extract obtained by supercritical fluid extraction and the filtrate after impurity removal and purification, and concentrate to a clear extract with a mass ratio of 1:1 to the medicinal materials.

[0079] 6) Preparation forming

[0080] Add 4 g of triethanolamine, 15 g of glycerol, 1 g of ethylparaben, 1 g of sodium benzoate, 5 g of aloe vera to the clear extract obtained by the above concentration, and add distilled water to make the total amount 810 g. Heat to 80 °C as the aqueous phase. Melt 25 g of glyceryl monostearate, 80 g of stearic acid, 40 g of liquid paraffin, and 45 g of stearyl alcohol at 80 °C as the oil phase. Slowly pour the aqueous phase into the oil phase at 80 °C, stirring while adding, and stir for emulsification for 20 min. Keep warm at 80 °C for 30 min to make 1000 g.

[0081] Example 4

[0082] The preparation process of the sunscreen drug is as follows:

[0083] 1) Take the medicinal materials

[0084] Weigh 40 g of honeysuckle, 40 g of polygonum perfoliatum, 30 g of sedum sarmentosum, 30 g of commelina communis, 40 g of polygonatum odoratum, 20 g of lithospermum erythrorhizon, 5 g of aloe vera, 40 g of asparagus cochinchinensis, 40 g of ganoderma lucidum, 20 g of coix lacryma-jobi, and 10 g of bee pollen according to the prescription ratio.

[0085] 2) Supercritical fluid extraction

[0086] Perform supercritical extraction on 5 medicinal materials including honeysuckle, polygonum perfoliatum, sedum sarmentosum, lithospermum erythrorhizon, and ganoderma lucidum under the conditions of a pressure of 40 MPa, a temperature of 60 °C, and a CO2-ethanol flow rate of 9 L / h for 2 h. After the extraction is completed, reduce the pressure and recover the precipitated extract for standby.

[0087] 3) Water extraction

[0088] Decoct 5 medicinal materials including commelina communis, polygonatum odoratum, asparagus cochinchinensis, coix lacryma-jobi, and bee pollen with water twice. The first time is for 2 h, and the second time is for 1.5 h. Add 6 times the mass of water of the medicinal materials each time. Combine the decoction, filter, and concentrate the filtrate to a clear extract with a relative density of about 1.05 (60 °C).

[0089] 4) Impurity removal and purification

[0090] Add 4% of ZTC clarifying agent B solution to the clear extract obtained by the above water extraction, stir evenly, and keep warm at 60 °C for 20 min; then add 3% of ZTC clarifying agent A solution, stir evenly, and keep warm at 60 °C for 20 min; let it stand for 24 h, filter, and keep the filtrate for standby.

[0091] 5) Concentration

[0092] Combine the extract obtained by supercritical fluid extraction and the filtrate after impurity removal and purification, and concentrate to a clear extract with a mass ratio of 1:1 to the medicinal materials.

[0093] 6) Preparation forming

[0094] Add 2 g of triethanolamine, 15 g of glycerol, 1 g of ethylparaben, 1 g of sodium benzoate, and 5 g of aloe vera to the clear extract obtained by the above concentration. Add distilled water to a total of 870 g, heat to 80 °C as the aqueous phase. Melt 25 g of glyceryl monostearate, 40 g of stearic acid, 20 g of liquid paraffin, and 45 g of stearyl alcohol by heating to 80 °C as the oil phase. Slowly pour the aqueous phase into the oil phase at 80 °C while stirring, stir and emulsify for 20 min, and keep warm at 80 °C for 30 min to prepare 1000 g.

[0095] In order to further verify the reliability of the present invention and screen out the best solution, the inventor conducted a series of experiments, and some excerpts are as follows:

[0096] 1. Instruments and Reagents

[0097] 1.1 Main Instruments

[0098] LC-2010AHT fully automatic high-performance liquid chromatograph (Shimadzu, Japan); TU-1001 ultraviolet-visible spectrophotometer (Beijing Puxi); HH-4 digital display constant temperature water bath (Changzhou Putian Instrument Manufacturing Co., Ltd.); ultrasonic cleaner (Shanghai Shangyi).

[0099] 1.2 Main Reagents

[0100] Chlorogenic acid reference substance (batch number: 110753-202201), quercetin reference substance (batch number: 100081-202107), kaempferol reference substance (batch number: 110861-202214), β,β'-dimethylacryloyl alkannin reference substance (batch number: 111689-202203), provided by the National Institutes for Food and Drug Control; ZTC clarifying agent (Beijing Zheng Tian Cheng), methanol (chromatographic grade, Thermo Fisher Scientific), water (Yibao purified water); all medicinal materials used in the experiment were purchased from Guizhou Tongjitang Decoction Pieces Co., Ltd.

[0101] 2. Investigation of Extraction Methods

[0102] The prescription of the sunscreen drug consists of honeysuckle, polygonum perfoliatum, sedum sarmentosum, commelina communis, polygonatum odoratum, lithospermum erythrorhizon, aloe vera, asparagus cochinchinensis, ganoderma lucidum, coix lacryma-jobi, and bee pollen. The extraction rate of the active ingredients of honeysuckle, sedum sarmentosum, lithospermum erythrorhizon, polygonum perfoliatum, etc. in the prescription is relatively low by water extraction process, and the extraction rate of the active ingredients of ganoderma lucidum is relatively low by alcohol extraction process. In order to improve the extraction efficiency of the active ingredients, it is necessary to extract these 5 herbs by a suitable method in groups. The research team of the present invention took the contents of chlorogenic acid, quercetin, kaempferol, β,β'-dimethylacryloyl alkannin, and polysaccharides in the extract as the investigation indexes, and compared 4 different extraction processes, namely water extraction process, alcohol extraction process, enzyme extraction process, and supercritical fluid extraction process, through experiments, and finally optimized the best extraction process.

[0103] 2.1 Experimental Method

[0104] 2.1.1 Preparation of Test Solution

[0105] (1) Water extraction method

[0106] Weigh the medicinal materials according to the formula ratio (40 g of Flos Lonicerae, 40 g of Polygonum perfoliatum L., 30 g of Sedum sarmentosum Bunge, 20 g of Lithospermum erythrorhizon Sieb. et Zucc., and 40 g of Ganoderma lucidum), add water 6 times the mass of the medicinal materials, heat and extract for 1 h, filter, concentrate the filtrate and make up the volume to 500 mL.

[0107] (2) Ethanol extraction method

[0108] Weigh the medicinal materials according to the formula ratio (the same as the water extraction method), add 60% ethanol solution 6 times the mass of the medicinal materials, heat and extract for 1 h, filter, concentrate the filtrate and make up the volume to 500 mL.

[0109] (3) Supercritical fluid extraction method

[0110] Weigh the medicinal materials according to the formula ratio (the same as the water extraction method), carry out supercritical extraction under the conditions of a pressure of 30 MPa, a temperature of 50 °C, and a CO2-ethanol flow rate of 7 L / h, with an extraction time of 4 h. After extraction, reduce the pressure, recover the precipitated extract, dilute it with methanol and make up the volume to 500 mL.

[0111] (4) Enzymatic hydrolysis method

[0112] Weigh the medicinal materials according to the formula ratio (the same as the water extraction method), add water 6 times the mass of the medicinal materials and 2% cellulase, extract at 50 °C and a pH value of 5.0 for 2 h, boil and then filter, concentrate the filtrate and make up the volume to 500 mL.

[0113] 2.1.2 Preparation of Reference Solution

[0114] 2.1.2.1 Preparation of reference solutions for chlorogenic acid, quercetin, kaempferol, and β,β'-dimethylacrylalkannin

[0115] Take reference substances of chlorogenic acid, quercetin, kaempferol, and β,β'-dimethylacrylalkannin, and make solutions containing 40 μg, 80 μg, 60 μg, and 0.1 mg per 1 mL respectively with methanol, and you will get them.

[0116] 2.1.2.2 Preparation of reference solution for polysaccharide

[0117] Take 33 mg of anhydrous glucose reference substance dried to constant weight at 105 °C, accurately weigh it, place it in a 100 mL volumetric flask, dissolve it with water and dilute to the mark, shake well, and you will get it (containing 0.33 mg of anhydrous glucose per 1 mL).

[0118] 2.1.3 Content determination

[0119] The contents of chlorogenic acid, quercetin, kaempferol, and β,β'-dimethylacryloylalkanin were determined by high performance liquid chromatography, and the content of polysaccharide was determined by spectrophotometry.

[0120] 2.1.3.1 Determination of the contents of chlorogenic acid, quercetin, kaempferol, and β,β'-dimethylacryloylalkanin

[0121] Chromatographic operating conditions: a chromatographic column packed with octadecylsilane-bonded silica gel (250×4.6 mm, 5 μm); mobile phase: methanol as mobile phase A and 0.05% formic acid solution as mobile phase B, with the gradient elution program shown in Table 1; detection wavelength: 250 nm; flow rate: 1.0 mL / min; injection volume: 10 μL. The number of theoretical plates was not less than 5000 calculated based on the chlorogenic acid peak.

[0122] Table 1 Gradient elution program

[0123]

[0124] 2.1.3.2 Determination of the content of polysaccharide

[0125] (1) Preparation of the standard curve

[0126] Precisely measure 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, and 0.6 mL of the reference substance solution respectively, place them in 10-mL stoppered graduated test tubes, add water to 2.0 mL in each tube, shake well, slowly add 0.2% anthrone-sulfuric acid solution to the scale in an ice-water bath, mix well, cool in a water bath for 10 min after standing cold, take out, immediately cool in an ice-water bath for 10 min, take out, and use the corresponding reagent as the blank. According to the ultraviolet-visible spectrophotometry (General Principles 0401 of the Chinese Pharmacopoeia), measure the absorbance at a wavelength of 582 nm. Plot the standard curve with the absorbance as the ordinate and the concentration as the abscissa. The results are shown in Figure 1 .

[0127] (2) Determination method

[0128] Precisely measure 5 mL of the extract, place it in a 25-mL volumetric flask, and according to the method under the preparation of the standard curve, starting from "add water to 2.0 mL", measure the absorbance according to law, read the weight (mg) of anhydrous glucose contained in the test solution from the standard curve, and calculate to obtain the result.

[0129] 2.2 Results and discussion

[0130] It can be seen from the results that the extraction effects of the 4 extraction methods are ranked as: supercritical fluid extraction method > water extraction method > enzymatic hydrolysis method > alcohol extraction method. Therefore, the supercritical fluid extraction method was selected as the extraction method. The determination results of the contents of the active ingredients in the extracts by different extraction methods are shown in Table 2.

[0131] Table 2 Determination Results of the Contents of Active Ingredients in Extracts by Different Extraction Methods

[0132]

[0133] 3. Investigation on Process Parameters of Supercritical Fluid Extraction Method

[0134] 3.1 Experimental Method

[0135] 3.1.1 Investigation on Extraction Pressure

[0136] Weigh 4 portions of medicinal materials according to the formula ratio (40 g of Flos Lonicerae, 40 g of Polygonum perfoliatum L., 30 g of Sedum sarmentosum Bunge, 20 g of Lithospermum erythrorhizon Sieb. et Zucc., and 40 g of Ganoderma lucidum). Conduct supercritical extraction at pressures of 10, 20, 30, and 40 MPa respectively, at a temperature of 50 °C and a CO2-ethanol flow rate of 7 L / h for 4 h. After extraction, reduce the pressure, recover the precipitated extract, dilute it with methanol and make up the volume to 500 mL. Use the solution under "2.1.2 Preparation of Reference Solution" as the reference solution, and conduct content analysis using the method under "2.1.3 Content Determination", and calculate the contents of each active ingredient.

[0137] 3.1.2 Investigation on Extraction Temperature

[0138] Weigh 3 portions of medicinal materials according to the formula ratio (40 g of Flos Lonicerae, 40 g of Polygonum perfoliatum L., 30 g of Sedum sarmentosum Bunge, 20 g of Lithospermum erythrorhizon Sieb. et Zucc., and 40 g of Ganoderma lucidum). Conduct supercritical extraction at temperatures of 40 °C, 50 °C, and 60 °C respectively, at a pressure of 30 MPa and a CO2-ethanol flow rate of 7 L / h for 4 h. After extraction, reduce the pressure, recover the precipitated extract, dilute it with methanol and make up the volume to 500 mL. Use the solution under "2.1.2 Preparation of Reference Solution" as the reference solution, and conduct content analysis using the method under "2.1.3 Content Determination", and calculate the contents of each active ingredient.

[0139] 3.1.3 Investigation on Extraction Time

[0140] Weigh 5 portions of medicinal materials according to the formula ratio (40 g of Flos Lonicerae, 40 g of Polygonum perfoliatum L., 30 g of Sedum sarmentosum Bunge, 20 g of Lithospermum erythrorhizon Sieb. et Zucc., and 40 g of Ganoderma lucidum). Conduct supercritical extraction at a temperature of 50 °C, a pressure of 30 MPa, and a CO2-ethanol flow rate of 7 L / h, with extraction times of 2, 3, 4, 5, and 6 h respectively. After extraction, reduce the pressure, recover the precipitated extract, dilute it with methanol and make up the volume to 500 mL. Use the solution under "2.1.2 Preparation of Reference Solution" as the reference solution, and conduct content analysis using the method under "2.1.3 Content Determination", and calculate the contents of each active ingredient.

[0141] 3.1.4 Investigation on CO2-Ethanol Flow Rate

[0142] Weigh 5 portions of medicinal materials according to the formula ratio (40 g of honeysuckle, 40 g of polygonum perfoliatum, 30 g of sedum sarmentosum, 20 g of lithospermum erythrorhizon, and 40 g of ganoderma lucidum), and perform supercritical extraction at a temperature of 50 °C, a pressure of 30 MPa, and CO2-ethanol flow rates of 5, 6, 7, 8, and 9 L / h respectively for 4 h. After the extraction is completed, reduce the pressure, recover the precipitated extract, dilute it with methanol and make the volume up to 500 mL. Using the solution under "2.1.2 Preparation of Reference Solution" as the reference solution, perform content analysis using the method under "2.1.3 Content Determination", and calculate the content of each active ingredient.

[0143] 3.2 Results and Discussion

[0144] 3.2.1 Investigation of Extraction Pressure

[0145] As can be seen from the results, the contents of chlorogenic acid, quercetin, kaempferol, β,β'-dimethylacryloyl alkannin, and polysaccharide continuously increase with the increase of extraction pressure. However, after the pressure exceeds 30 MPa, the increase in content slows down, and the requirements for instrument equipment under high pressure also gradually become higher. Therefore, 30 MPa is selected as the optimal process parameter for extraction pressure. The determination results of the contents of active ingredients under different extraction pressures are shown in Table 3.

[0146] Table 3 Determination Results of the Contents of Active Ingredients under Different Extraction Pressures

[0147]

[0148] 3.2.2 Investigation of Extraction Temperature

[0149] As can be seen from the results, when the extraction temperature is 50 °C, the contents of chlorogenic acid, quercetin, kaempferol, β,β'-dimethylacryloyl alkannin, and ganoderma polysaccharide are the highest. Therefore, 50 °C is selected as the optimal process parameter for extraction temperature. The determination results of the contents of active ingredients under different extraction temperatures are shown in Table 4.

[0150] Table 4 Determination Results of the Contents of Active Ingredients under Different Extraction Temperatures

[0151]

[0152] 3.2.3 Investigation of Extraction Time

[0153] As can be seen from the results, with the increase of extraction time, the determination results of the contents of chlorogenic acid, quercetin, kaempferol, β,β'-dimethylacryloyl alkannin, and polysaccharide show an upward trend. After 4 h of extraction, the increase in content slows down. Therefore, 4 h is selected as the optimal process parameter for extraction time. The determination results of the contents of active ingredients under different extraction times are shown in Table 5.

[0154] Table 5 Determination Results of the Contents of Active Ingredients under Different Extraction Times

[0155]

[0156] 3.2.4 Investigation of CO2 - ethanol flow rate

[0157] As can be seen from the results, when the flow rate is between 5 L / h and 7 L / h, the contents of chlorogenic acid, quercetin, kaempferol, β,β’ - dimethylacryloylakannin and polysaccharide gradually increase. When the flow rates are 7 L / h and 8 L / h, the contents are similar. When the flow rate is 9 L / h, the contact time between the extract and the supercritical fluid is reduced, and the mass transfer process is insufficient, resulting in a decrease in content. Therefore, 7 L / h is selected as the optimal process parameter for the CO2 - ethanol flow rate. The determination results of the contents of active ingredients under different CO2 - ethanol flow rates are shown in Table 6.

[0158] Table 6 Determination results of the contents of active ingredients under different CO2 - ethanol flow rates

[0159]

[0160] 4. Investigation of impurity removal and purification methods

[0161] In the prescription, Commelina communis, Polygonatum odoratum, Asparagus cochinchinensis, Coix lacryma - jobi and bee pollen are treated by water extraction method. Since there are many impurities in the water extract, it is necessary to carry out purification treatment. In order to determine the optimal purification process, the present invention compares the alcohol precipitation method, ZTC clarifying agent method and chitosan flocculation precipitation method with the retention rate and impurity removal rate of total polysaccharide in the extract as indexes.

[0162] Weigh the medicinal materials according to the formula ratio (90 g of Commelina communis, 120 g of Polygonatum odoratum, 120 g of Asparagus cochinchinensis, 60 g of Coix lacryma - jobi, 30 g of bee pollen), decoct twice with water, 2 hours for the first time and 1.5 hours for the second time. Combine the decoction, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60 °C). Determine the polysaccharide content and solid content, divide it into 3 equal parts on average, and prepare test solutions according to the following methods respectively to determine the polysaccharide retention rate and impurity removal rate.

[0163] 4.1 Experimental method

[0164] 4.1.1 Sample preparation method

[0165] (1) Alcohol precipitation method

[0166] Take 1 part of the clear paste, add ethanol to make the alcohol content reach 60%, stir while adding, stand for 24 h, filter, and concentrate and make up the volume of the filtrate to 500 mL.

[0167] (2) ZTC clarifying agent method

[0168] Take 1 part of the clear extract, add 3% of the ZTC clarifying agent B solution, stir evenly, keep warm at 40°C for 20 min, then add 2% of the ZTC clarifying agent A solution, stir evenly, keep warm at 40°C for 20 min, let stand for 24 h, filter, and concentrate and make up the volume of the filtrate to 500 mL.

[0169] (3) Chitosan flocculation precipitation method

[0170] Take 1 part of the clear extract, add 1% acetic acid solution containing 1% chitosan, the flocculation temperature is 40°C, the stirring time is 10 min, let stand for 24 h, filter, and concentrate and make up the volume of the filtrate to 500 mL.

[0171] 4.1.2 Determination of polysaccharide retention rate

[0172] Determine the polysaccharide content in the concentrated solution before and after impurity removal according to the method under "2.1.3.2 Determination of polysaccharide content", and calculate the polysaccharide retention rate according to the following formula:

[0173]

[0174] 4.1.3 Determination of impurity removal rate

[0175] Precisely pipette 25 mL of the solution before and after impurity removal and the clear extract respectively, place them in an evaporating dish that has been dried to a constant weight, evaporate to dryness in a water bath, dry the residue at 105°C for 3 h, take it out, place it in a desiccator for 30 min, weigh it, calculate the dry extract weight, and calculate the impurity removal rate according to the following formula:

[0176]

[0177] 4.2 Results and discussion

[0178] The results show that the chitosan flocculation precipitation method has the highest impurity removal rate, but the ZTC clarifying agent method has the highest polysaccharide retention rate after treatment, indicating that the chitosan flocculation precipitation method has the greatest loss of active ingredients while removing impurities. Considering comprehensively, the ZTC clarifying agent method is selected as the best purification process. The results of the total polysaccharide retention rate and impurity removal rate after different purification methods are shown in Table 7.

[0179] Table 7 Total polysaccharide retention rate and impurity removal rate after different purification methods

[0180]

[0181] 5. Investigation on process parameters of the ZTC clarifying agent method

[0182] The main factors affecting the purification effect of the ZTC clarifying agent are the dosage of ZTC clarifying agent A, the dosage of ZTC clarifying agent B, the holding temperature and the holding time. This experiment intends to investigate these four main factors respectively to optimize the best impurity removal process parameters.

[0183] 5.1 Experimental method

[0184] Weigh the medicinal materials according to the formula ratio (Commelina communis 450 g, Polygonatum odoratum 600 g, Asparagus cochinchinensis 600 g, Coix lacryma-jobi 300 g, Bee pollen 150 g), decoct twice with water, 2 hours for the first time and 1.5 hours for the second time. Combine the decoction liquids, filter, and concentrate the filtrate to a clear paste with a relative density of about 1.05 (60 °C). Divide it into 12 equal parts on average, and prepare the test solutions according to the following methods respectively to determine the total polysaccharide retention rate and the impurity removal rate.

[0185] 5.1.1 Investigation on the dosage of ZTC clarifying agent B

[0186] Take 3 parts of the clear paste, add 2%, 3% and 4% of ZTC clarifying agent B solution respectively, stir, keep warm at 40 °C for 20 min, then add 2% of ZTC clarifying agent A solution, keep warm at 40 °C for 20 min, stand for 24 h, and concentrate and make up the volume of the filtrate to 500 mL.

[0187] 5.1.2 Investigation on the dosage of ZTC clarifying agent A

[0188] Take 3 parts of the clear paste, add 3% of ZTC clarifying agent B solution, stir, keep warm at 40 °C for 20 min, then add 1%, 2% and 3% of ZTC clarifying agent A solution respectively, keep warm at 40 °C for 20 min, stand for 24 h, and concentrate and make up the volume of the filtrate to 500 mL.

[0189] 5.1.3 Investigation on the heat preservation temperature

[0190] Take 3 parts of the clear paste, add 3% of ZTC clarifying agent B solution, stir, keep warm at 40 °C, 50 °C and 60 °C for 20 min respectively, then add 2% of ZTC clarifying agent A solution, keep warm at 40 °C, 50 °C and 60 °C for 20 min respectively, stand for 24 h, and concentrate and make up the volume of the filtrate to 500 mL.

[0191] 5.1.4 Investigation on the heat preservation time

[0192] Take 3 parts of the clear paste, add 3% of ZTC clarifying agent B solution, stir, keep warm at 40 °C for 20 min, 30 min and 40 min respectively, then add 2% of ZTC clarifying agent A solution, keep warm at 40 °C for 20 min, 30 min and 40 min respectively, stand for 24 h, and concentrate and make up the volume of the filtrate to 500 mL.

[0193] 5.1.5 Determination of the total polysaccharide content and the impurity removal rate

[0194] Determine the total polysaccharide content and the impurity removal rate according to the method under "4. Investigation on the impurity removal and purification method", and calculate the total polysaccharide retention rate and the impurity removal rate.

[0195] 5.2 Results and discussion

[0196] 5.2.1 Investigation on the dosage of ZTC clarifying agent B

[0197] As can be seen from the results, when the dosage of ZTC clarifying agent B is 4%, although the impurity removal rate increases, the reduction rate of the total polysaccharide retention rate is relatively large. Considering comprehensively, the dosage of ZTC clarifying agent B is selected as 3%. The results of the total polysaccharide retention rate and the impurity removal rate under different dosages of ZTC clarifying agent B are shown in Table 8.

[0198] Table 8 Total polysaccharide retention rate and impurity removal rate under different dosages of ZTC clarifying agent B

[0199]

[0200] 5.2.2 Investigation on the dosage of ZTC clarifying agent A

[0201] As can be seen from the results, when the dosage of ZTC clarifying agent A is 3%, although the impurity removal rate increases, the reduction rate of the total polysaccharide retention rate is relatively large. Considering comprehensively, the dosage of ZTC clarifying agent A is selected as 2%. The results of the total polysaccharide retention rate and the impurity removal rate under different dosages of ZTC clarifying agent A are shown in Table 9.

[0202] Table 9 Total polysaccharide retention rate and impurity removal rate under different dosages of ZTC clarifying agent A

[0203]

[0204] 5.2.3 Investigation on the heat preservation temperature

[0205] As can be seen from the results, the differences in the total polysaccharide retention rate and the impurity removal rate at each temperature are not significant. Considering comprehensively, 40 °C is selected as the process parameter of the heat preservation temperature. The results of the total polysaccharide retention rate and the impurity removal rate under different heat preservation temperature conditions are shown in Table 10.

[0206] Table 10 Total polysaccharide retention rate and impurity removal rate under different heat preservation temperatures

[0207]

[0208] 5.2.4 Investigation on the heat preservation time

[0209] As can be seen from the results, the differences in the total polysaccharide retention rate and the impurity removal rate under each heat preservation time condition are not significant. Considering comprehensively, 20 min is selected as the process parameter of the heat preservation time. The results of the total polysaccharide retention rate and the impurity removal rate under different heat preservation time conditions are shown in Table 11.

[0210] Table 11 Total polysaccharide retention rate and impurity removal rate under different heat preservation times

[0211]

[0212] 6. Matrix optimization

[0213] The original forming process of the sunscreen pharmaceutical preparation was as follows: Herbs were taken according to the prescription ratio and extracted into clear paste. Sorbitol, sodium alginate, glycerol, beta-cyclodextrin, titanium dioxide and aloe were taken and added to the above-mentioned Ganoderma lucidum liquid extract and traditional Chinese medicine concentrated liquid, and heated (80°C) to dissolve; Separately, stearic acid, monoglyceride, vaseline, stearyl alcohol, polysorbate 80, span 80, lanolin, sucrose stearate, and emulsifying silicone oil were heated (80°C) to melt. The above two solutions were mixed and emulsified for 15 minutes, kept at 80°C for 30 minutes, and benzoic acid and ethylparaben were added and mixed evenly to obtain the product. This process uses a large variety of excipients, which is not conducive to the quality control of the preparation and has too high a cost.

[0214] In this study, triethanolamine, glycerol, glyceryl monostearate, stearic acid, liquid paraffin and stearyl alcohol were selected as the o / w type matrix, and ethylparaben and sodium benzoate were used as preservatives to carry out the research on the forming process of the preparation.

[0215] 6.1 Preferred indicators and scoring criteria

[0216] 6.1.1 Appearance

[0217] The full score is 20 points. It is inspected from aspects such as color, texture, consistency, spreading dispersibility and presence or absence of granularity, with each item accounting for 4 points.

[0218] 6.1.2 Centrifugal stability

[0219] The full score is 20 points. Each group of ointments was placed in a centrifuge tube and centrifuged at 2500 r·min －1 for 30 minutes, and abnormal phenomena such as layering, caking, foaming or water separation were observed.

[0220] 6.1.3 Cold tolerance test

[0221] The full score is 10 points. A certain amount of each group of ointments was placed in a plastic sample cup and placed in a 4°C refrigerator for 1 month, and abnormal phenomena such as layering, caking and water separation were observed.

[0222] 6.1.4 Heat resistance test

[0223] The full score is 10 points. A certain amount of each group of ointments was placed in a plastic sample cup and placed in a 40°C constant temperature oven for 1 month, and abnormal phenomena such as layering, color change and mildew were observed.

[0224] 6.1.5 Ductility

[0225] The full score is 20 points. Two clean glass plates of 10 cm × 10 cm were taken, and the upper one weighed 125 g (make up with weights if the mass is insufficient). 1 g of each group of ointments placed at room temperature for 24 h was weighed and placed between the two glass plates. After 1 minute, the diameter of the extended ointment was measured with a ruler, and the average value of the ratio of the maximum diameter to the minimum diameter was taken.

[0226] 6.1.6 pH value

[0227] The full score is 20 points. Take 5 g of each group of ointment and place it in a small beaker, add an equal amount of distilled water to dissolve it fully, stir evenly, and measure the pH of each group with a pH meter. The pH value between 6 and 8 gets full marks, and the rest gets zero points.

[0228] 6.2 Orthogonal test design

[0229] Three factors that have a greater impact on the physical and chemical properties of the matrix, namely the dosage of triethanolamine (A), the dosage of stearic acid (B), and the dosage of liquid paraffin (C), are defined as the investigation factors. According to the preliminary selected data, each factor takes 3 levels to carry out the orthogonal test. The factor level table is shown in Table 12.

[0230] Table 12 Factor and level table of orthogonal test for matrix investigation

[0231]

[0232] 6.3 Results and discussion

[0233] It can be seen from the results that the influence of each factor on the matrix forming effect is A > B > C, and A3 > A2 > A1, B3 > B2 > B1, C3 > C1 > C2. The best matrix ratio parameter is A3B3C3. Combining the variance analysis, it can be seen that all 3 factors have significant effects. Therefore, the best matrix ratio parameter is: triethanolamine: stearic acid: liquid paraffin = 4:80:40. The orthogonal test design and results of matrix investigation are shown in Table 13, and the variance analysis of matrix investigation is shown in Table 14.

[0234] Table 13 Orthogonal test design and results of matrix investigation

[0235]

[0236]

[0237] Table 14 Variance analysis of matrix investigation

[0238]

[0239] 7. Verification of the best process parameters

[0240] 7.1 Experimental method

[0241] (1) Take medicinal materials

[0242] Weigh 40 g of Flos Lonicerae, 40 g of Polygonum perfoliatum L., 30 g of Sedum sarmentosum Bunge, 30 g of Commelina communis L., 40 g of Polygonatum odoratum (Mill.) Druce, 20 g of Lithospermum erythrorhizon Sieb. et Zucc., 5 g of Aloe vera (L.) Burm.f., 40 g of Asparagus cochinchinensis (Lour.) Merr., 40 g of Ganoderma lucidum (Leyss. ex Fr.) Karst., 20 g of Coix lacryma-jobi L. var. ma-yuen (Roman.) Stapf, and 10 g of bee pollen according to the prescription ratio.

[0243] (2) Supercritical fluid extraction

[0244] Five kinds of medicinal materials, namely honeysuckle, polygonum perfoliatum, sedum sarmentosum, lithospermum erythrorhizon and ganoderma lucidum, are subjected to supercritical extraction under the conditions of a pressure of 30 MPa, a temperature of 50 °C and a CO2-ethanol flow rate of 7 L / h for 4 h. After the extraction is completed, the pressure is reduced, and the precipitated extract is recovered for standby.

[0245] (3) Water extraction

[0246] Commelina communis, polygonatum odoratum, asparagus cochinchinensis, coix lacryma-jobi and bee pollen are decocted with water twice. The first decoction is for 2 h and the second is for 1.5 h, and 6 times the mass of the medicinal materials of water is added each time. The decoction liquids are combined, filtered, and the filtrate is concentrated to a clear paste with a relative density of about 1.05 (60 °C).

[0247] (4) Impurity removal and purification

[0248] 3% of the ZTC clarifying agent B solution is added to the clear paste obtained by the above water extraction, stirred evenly, and kept warm at 40 °C for 20 min; then 2% of the ZTC clarifying agent A solution is added, stirred evenly, and kept warm at 40 °C for 20 min; left standing for 24 h, filtered, and the filtrate is for standby.

[0249] (5) Concentration

[0250] The extract obtained by supercritical fluid extraction and the filtrate after impurity removal and purification are combined and concentrated to a clear paste with a mass ratio of 1:1 to the medicinal materials.

[0251] (6) Preparation and shaping

[0252] 4 g of triethanolamine, 15 g of glycerol, 1 g of ethylparaben, 1 g of sodium benzoate, 5 g of aloe are added to the clear paste obtained by the above concentration, and distilled water is added to a total amount of 810 g, heated to 80 °C as the water phase, 25 g of glyceryl monostearate, 80 g of stearic acid, 40 g of liquid paraffin, 45 g of stearyl alcohol are heated to 80 °C and melted as the oil phase. At 80 °C, the water phase is slowly poured into the oil phase, stirring while adding, stirring and emulsifying for 20 minutes, and kept warm at 80 °C for 30 minutes to prepare 1000 g.

[0253] (7) Verification test

[0254] Three batches of samples are prepared according to the above optimal parameters, and scored according to the indicators and scoring criteria under "6. Matrix optimization".

[0255] 7.2 Results and discussion

[0256] It can be seen from the results that the sun protection drugs prepared according to the optimal process parameters provided by the present invention have good indicators and stable processes. The verification results of the optimal process parameters are shown in Table 15.

[0257] Table 15 Verification results of the optimal process parameters

[0258]

[0259] 8. Drug efficacy verification test

[0260] 8.1 Mouse model of solar dermatitis

[0261] 8.1.1 Experimental Materials

[0262] ① Animals: 70 specific pathogen-free BALB / c female mice, weighing 18-20 g.

[0263] ② Drugs and reagents: sunscreen drug samples prepared according to the preparation method of the present invention; sunscreen drugs (Guizhou Green Sun Pharmaceutical Co., Ltd.); 5% sodium chloride solution; calamine lotion; enzyme-linked immunosorbent assay kit; electric constant temperature water bath; electric constant temperature blast drying oven; hand-held shaver; high-speed refrigerated centrifuge; surgical scissors; microplate reader.

[0264] 8.1.2 Experimental methods

[0265] ① Modeling and grouping and drug administration: 50 mice were divided into 7 groups, with 10 mice in each group. A flat area of ​​4 cm × 4 cm on the back was selected for hair removal using a handheld shaver. The mice in the blank control group were not treated after hair removal; the mice in the animal model group were given 0.1 mL / cm2 of normal saline within the hair removal area. 2 , once a day; the original process group evenly applied 0.1g / cm 2 , Group 1 of the present invention is evenly applied with 0.1g / cm 2 (same dosage as the original process), the present invention group 2 is evenly applied with 0.08g / cm 2 (0.8 times the dosage of the original process) and the present invention group 3 are evenly applied with 0.1g / cm 2 (0.6 times the original dosage), once a day; Calamine lotion group evenly applied 0.1mL / cm 2 , once a day, for 8 consecutive days according to the above scheme. On the 6th day, the 5 groups of mice were placed at a distance of 10 cm from the 308nm ultraviolet rays for 1.5 hours. After irradiation, the mice were given the corresponding drugs according to the previous doses. After the model was successfully established, the experimental results were observed. After the mice in the animal model group were continuously smeared with normal saline for 6 days, they were irradiated with ultraviolet rays under the same conditions and methods as other drug groups. Skin redness and swelling appeared after a 48-hour incubation period, indicating that the mouse model of solar dermatitis was successfully established.

[0266] ② Visual observation: One hour after the last day of drug administration, the drug was removed, the back skin was cleaned, and the skin was scored according to the following criteria: 0 point, normal color, no redness or swelling; 1 point, slight redness or swelling; 2 points, redness or swelling, dryness or wrinkles; 3 points, exudation of fluid, scratching of the back skin, and restlessness.

[0267] ③ Determination of the contents of NO, NOS, and TNF-α in the skin of mice: After observing the intuitive morphological changes in the skin of the mice in the depilated area, the skin in the corresponding irradiated area was taken with surgical scissors. The obtained skin tissue was homogenized at 3000 r / min and stopped after 10 min. The levels of nitric oxide (NO), nitric oxide synthase (NOS), and TNF-α were detected by enzyme-linked immunosorbent assay.

[0268] ④ Statistical method: The data were processed using SPSS 17.0 statistical software. The data were expressed as x±s. The t-test was used for comparison between groups, and the Ridit test was used for ranked data. A P value < 0.01 was considered statistically significant.

[0269] 8.2 Results and discussion

[0270] ① Effects on the mouse model of solar dermatitis: In the blank group, everything was normal locally. After Ridit test, compared with the blank group, significant local swelling and exudation were observed in the model group (P < 0.01); compared with the model group, the original process group, Invention Group 1, Invention Group 2, and calamine lotion group all significantly inhibited skin swelling, liquid exudation, etc. (P < 0.01). Among them, the improvement effects of Invention Group 2 and the original process group were comparable, and the amount of medicinal materials used in 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. The results are shown in Table 16.

[0271] Table 16 Effects of sunscreen drugs on the skin score of mice (n = 10)

[0272]

[0273]

[0274] ② Effects on the contents of NO, NOS, and TNF-α in the skin of mice: Compared with the blank group, the levels of TNF-α, NOS, and NO in the skin of mice in the model group were significantly increased (P < 0.01); compared with the model group, the original process group, calamine lotion group, Invention Group 1, Invention Group 2, and Invention Group 3 could significantly reduce the levels of TNF-α, NOS, and NO (P < 0.01); among them, the reduction amplitude of Invention Group 2 was comparable to that of the original process group, further indicating that the present invention can reduce the amount of medicinal materials used. See Table 17.

[0275] Table 17 Effects of each experimental group on the levels of TNF-α, NOS, and NO in the mouse model of solar dermatitis (x±s, n = 10)

[0276]

[0277] Although the present invention has been described in detail above with general descriptions, specific embodiments and experiments, some modifications or improvements can be made thereto based on the present invention, which are obvious to those skilled in the art. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection of the present invention.

Claims

1. A process for preparing a sunscreen drug, characterized in that: The steps include: S1 Take herbs: 40 parts of honeysuckle, 40 parts of Radix Angelicae Pubescentis, 30 parts of Herba Codonopsise, 30 parts of Commelinae, 40 parts of Polygonatum odoratum, 20 parts of Lithospermum officinale, 5 parts of Aloe, 40 parts of Asparagus cochinchinensis, 40 parts of Ganoderma lucidum, 20 parts of Coix seeds, and 10 parts of Bee Pollen; S2 Supercritical Fluid Extraction: The five medicinal materials of honeysuckle, Radix Angelicae Pubescentis, Herba Lycopodii, Radix Lithospermi and Ganoderma Lucidum were subjected to supercritical CO2-ethanol extraction, after the extraction, the pressure was reduced, and the precipitated extract was recovered for standby use; The supercritical CO2-ethanol extraction pressure is 30 MPa, the temperature is 50°C, the CO2-ethanol flow rate is 7 L / h, and the extraction time is 4 h. S3 Water Extract: Add water to decoct the five medicinal materials, namely, Commelina communis, Polygonatum odoratum, Asparagus cochinchinensis, Coix seeds, and Bee pollen, for 2 hours the first time and 1.5 hours the second time, adding 6 to 8 times the weight of the medicinal materials each time; combine the decoctions, filter, and concentrate the filtrate to a clear paste with a relative density of 1.05 at 60°C; S4 impurity removal and purification: Add 3% ZTC clarifier B solution to the clear paste obtained by water extraction, stir evenly, and keep warm at 40℃ for 20min; add 2% ZTC clarifier A solution, stir evenly, keep warm at 40℃ for 20min, let stand for 24h, filter, and use the filtrate for later use; S5 Concentrate: The extract obtained by supercritical fluid extraction and the filtrate obtained after impurity removal and purification are combined and concentrated to a clear paste having a mass ratio of 1:1 to the medicinal material; S6 preparation molding: Add 4 parts of triethanolamine, 15 parts of glycerol, 1 part of ethylparaben, 1 part of sodium benzoate, aloe vera and appropriate amount of distilled water to the concentrated clear paste, and heat to 80°C as the water phase; mix 25 parts of glyceryl monostearate, 80 parts of stearic acid, 40 parts of liquid paraffin and 45 parts of stearyl alcohol, and heat to 80°C to melt as the oil phase; slowly pour the water phase into the oil phase at 80°C, stirring while adding, stirring and emulsifying for 20 minutes, and keep warm at 80°C for 30 minutes to make 1000 portions.

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