Preparation process of white jade ointment

Abstract

The application discloses a preparation process of Baiyu ointment and relates to the technical field of medicines. The Baiyu ointment prescription is composed of Radix Bupleuri, Rhizoma Polygonati, Ligusticum sinense, Polygonum hydropiper, Geranium wilfordii, Salvia miltiorrhiza, Arnebia euchroma, Angelica sinensis and Zingiber officinale. The preparation process comprises the following steps: taking medicinal materials; supercritical fluid extraction; ultrasonic extraction; and preparation forming. The original preparation process of the Baiyu ointment is improved and optimized by adopting the modern preparation process and combining with a large number of investigation experiments. The Baiyu ointment has the advantages of high content of effective components, reduced types of auxiliary materials and optimized dosage of the auxiliary materials. The Baiyu ointment has important significance for improving the medicine quality and enhancing the medicine safety, effectiveness and quality controllability.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a preparation process for a white jade ointment. Background Technology

[0002] Baiyu Ointment (Drug Approval Number: National Medicine Approval Number) has the effects of promoting blood circulation, dispelling wind, nourishing blood, and moisturizing the skin. It is used for skin cracking caused by blood deficiency and wind dryness, such as dry and cracked hands and feet, frostbite, etc., and is one of the representative products of Guizhou Miao medicine. The prescription for Baiyu Ointment is: Bletilla striata 80g, Polygonatum odoratum 40g, Ligusticum striatum 40g, Polygonum hydropiper 40g, Geranium wilfordii 40g, Salvia miltiorrhiza 20g, Lithospermum erythrorhizon 20g, Angelica sinensis 40g, and Zingiber officinale 40g. The original preparation process was as follows: For the above nine ingredients, take Salvia miltiorrhiza, Angelica sinensis, and dried ginger, and use 80% ethanol as a solvent. After soaking for 12 hours, slowly percolate at a rate of 1-3 mL / min, collect 1200 mL of percolate, recover the ethanol under reduced pressure, and concentrate to a clear extract with a relative density of 1.15 (50℃), for later use; for the remaining six ingredients, including Bletilla striata, decoct twice with water, the first time for 2 hours and the second time for 1.5 hours, combine the decoctions, filter, and concentrate the filtrate to a clear extract with a relative density of 1.05 (60℃), combine with the above clear extract; add 60 g of sorbitol, 60 g of beta-cyclodextrin, 60 g of glycerin, 20 g of titanium dioxide (titanium white powder), and 3 g of sodium hyaluronate. g, heat (80℃) to dissolve, set aside; separately take 10g mink oil, 30g lanolin, 60g white petrolatum, 12g mono- and di-glyceryl stearate, 10g sucrose stearate, 100g liquid paraffin, 50g polysorbate 80, 50g sorbitan oleate (Span 80), and 20g emulsified silicone oil, heat (80℃) to melt, mix with the above clear paste, emulsify for 15min, keep warm at 80℃ for 30min, cool to 40℃, add 2g elastase, 6g benzoic acid, and 1.5g ethylparaben, mix well, and make 1000g, which is the final product.

[0003] The Baiyu Ointment formula contains nine medicinal herbs. Modern research shows that the effective components and pharmacological effects of each herb in treating chapped skin are as follows: Bletilla striata polysaccharides, bibenzyl compounds, and phenolic acids promote wound healing, enhance the hydration capacity of the stratum corneum, and repair the skin barrier; Polygonatum odoratum polysaccharides and steroidal saponins nourish yin and moisten dryness, improve skin hydration, and relieve dryness and cracking; Ligusticum striatum contains ligustilide, ligustrazine, and other volatile oils, as well as ferulic acid, which dispel wind and cold, improve local microcirculation, and relieve frostbite pain; Polygonum hydropiper contains gallic acid and other tannins... Flavonoids such as quercetin have astringent and tissue-regenerating effects, promoting wound healing; Tanshinone IIA and Tanshinone B in Salvia miltiorrhiza have the effects of promoting blood circulation and removing blood stasis, improving local blood circulation; Shikonin and Acetylshikonin in Lithospermum erythrorhizon have the effects of cooling blood and detoxifying, anti-inflammatory and repair-promoting, and preventing infection; Ferulic acid, Angelica polysaccharide, and ligustilide in Angelica sinensis have the effects of nourishing blood and promoting blood circulation, moisturizing the skin, and regulating immune function; Gingerol (6-gingerol) and volatile oils in dried ginger have the effects of warming the meridians and dispelling cold, promoting local blood circulation, and relieving frostbite. Therefore, the effective substances in the Baiyu Ointment prescription that treat chapped skin mainly fall into three categories: polysaccharides (Bletilla striata, Polygonatum odoratum, Angelica sinensis), volatile oils (Ligusticum striatum, Angelica sinensis, dried ginger), and flavonoids (Polygonum hydropiper, Geranium wilfordii, Salvia miltiorrhiza). Of these three categories of substances, polysaccharides are relatively water-soluble and can be extracted using water extraction in industrial production; volatile oils are fat-soluble components and are usually extracted using steam distillation or organic solvent methods; while flavonoids are also fat-soluble components and are best extracted using alcohol extraction.

[0004] The original preparation process of Baiyu Ointment was established 30 years ago. With the continuous development of research in traditional Chinese medicine, modern research has shown that the original preparation process is not entirely reasonable and has the following drawbacks: (1) The retention rate of volatile oil in the finished product is low. Although the volatile oil of Angelica sinensis can be extracted after percolation with 80% ethanol, it is easy to lose due to heat during the subsequent ethanol recovery and concentration process; the volatile oil of Ligusticum striatum and dried ginger is not collected during the decoction process, and there is also a vacuum concentration process, which results in very little retention of volatile oil. (2) The extraction rate of flavonoids and polysaccharides in the prescription is not maximized. The prescription contains flavonoids in Polygonum hydropiper, Geranium wilfordii and Salvia miltiorrhiza, but only Salvia miltiorrhiza is extracted with alcohol; the prescription contains polysaccharides in Bletilla striata, Polygonatum odoratum and Angelica sinensis, but Angelica sinensis is not extracted by water extraction. (3) The original Baiyu Ointment preparation process used a total of 17 excipients, which resulted in high production costs and was also not conducive to the quality control of Baiyu Ointment.

[0005] To address the problems in the original preparation process of Baiyu Ointment, the invention team conducted extensive investigations and experiments, combining modern new production technologies and materials to improve and optimize the original preparation process of Baiyu Ointment. The aim is to improve the quality, safety, efficacy, and quality control of the drug. Summary of the Invention

[0006] The purpose of this invention is to provide a preparation process for Baiyu Ointment, which improves drug quality and enhances drug safety, efficacy, and quality controllability.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] The preparation process of the white jade ointment of the present invention includes the following steps:

[0009] S1 retrieves medicinal materials:

[0010] Weigh out 80 parts of Bletilla striata, 40 parts of Polygonatum odoratum, 40 parts of Ligusticum striatum, 40 parts of Polygonum hydropiper, 40 parts of Geranium wilfordii, 20 parts of Salvia miltiorrhiza, 20 parts of Lithospermum erythrorhizon, 40 parts of Angelica sinensis, and 40 parts of dried ginger. Mix them well, pulverize them, and pass them through a 50-80 mesh sieve for later use.

[0011] S2 supercritical fluid extraction:

[0012] The fine powder obtained in step S1 was subjected to CO2 supercritical fluid extraction using a segmented method, and the extract was collected for later use.

[0013] The CO2 flow rate of the supercritical fluid extraction is 30 L / h;

[0014] The extraction parameters for the segmented method are as follows: 0–30 min, extraction temperature 35–40 °C, extraction pressure 25–30 MPa; 31–60 min, extraction temperature 40–45 °C, extraction pressure 30–35 MPa; 61–90 min, extraction temperature 45–50 °C, extraction pressure 35–40 MPa.

[0015] S3 ultrasonic extraction:

[0016] Take the residue after supercritical fluid extraction in step S2, add 6 to 10 times the amount of water, and ultrasonically extract for 30 to 50 minutes at a frequency of 20 to 40 kHz and a power of 150 to 250 W. Filter and concentrate the filtrate to a clear extract with a relative density of 1.10 for later use.

[0017] S4 Formulation:

[0018] Add 8-12 parts of sodium hyaluronate, 60-100 parts of carbomer, and 70-90 parts of betacyclodextrin to the clear paste obtained in step S3, and heat at 60°C to dissolve. Separately, take 80-120 parts of hydrogenated vegetable oil, 90-110 parts of olive oil, and 60-100 parts of polysorbate 80, heat at 80°C to melt, and mix with the clear paste above. Emulsify for 15-30 minutes, keep warm at 80°C for 30 minutes, and when the temperature drops to 40°C, add the supercritical fluid extract obtained in step S2 and 1-3 parts of ethylparaben, mix well, and the product is ready.

[0019] Preferably, in the preparation process of the white jade ointment of the present invention, the powder is pulverized and then passed through a 50-70 mesh sieve in step S1.

[0020] In a further preferred embodiment, in the preparation process of the white jade ointment of the present invention, the powder is pulverized and then passed through a 60-mesh sieve in step S1.

[0021] Preferably, in the preparation process of the white jade ointment of the present invention, the extraction parameters of the segmented method in step S2 are as follows: 0-30 min, extraction temperature is 38-40℃, extraction pressure is 28-30 MPa; 31-60 min, extraction temperature is 42-45℃, extraction pressure is 32-35 MPa; 61-90 min, extraction temperature is 47-50℃, extraction pressure is 37-40 MPa.

[0022] In a further preferred embodiment, in the preparation process of the white jade ointment of the present invention, the extraction parameters of the segmented method in step S2 are as follows: 0-30 min, extraction temperature is 40℃, extraction pressure is 30 MPa; 31-60 min, extraction temperature is 45℃, extraction pressure is 35 MPa; 61-90 min, extraction temperature is 50℃, extraction pressure is 40 MPa.

[0023] Preferably, in the preparation process of the white jade ointment of the present invention, step S3 ultrasonic extraction specifically involves: taking the residue after supercritical fluid extraction in step S2, adding 6 to 8 times the amount of water, and ultrasonically extracting for 40 to 50 minutes at a frequency of 20 to 30 kHz and a power of 200 to 250 W, filtering, and concentrating the filtrate to a clear ointment with a relative density of 1.10 for later use.

[0024] In a further preferred embodiment, in the preparation process of the white jade ointment of the present invention, step S3 ultrasonic extraction specifically involves: taking the residue after supercritical fluid extraction in step S2, adding 6 times the amount of water, and ultrasonically extracting for 40 minutes at a frequency of 30kHz and a power of 200W, filtering, and concentrating the filtrate to a clear ointment with a relative density of 1.10 for later use.

[0025] In the preparation process of the white jade ointment of the present invention, the relative density in step S3 is the measured value at 60°C.

[0026] Preferably, in the preparation process of the white jade ointment of the present invention, step S4, formulation forming, specifically involves: adding 10-12 parts of sodium hyaluronate, 80-100 parts of carbomer, and 70-80 parts of betacyclodextrin to the clear ointment obtained in step S3, heating at 60°C to dissolve, and setting aside; separately taking 100-120 parts of hydrogenated vegetable oil, 100-110 parts of olive oil, and 90-100 parts of polysorbate 80, heating and melting at 80°C, and mixing with the above clear ointment; emulsifying for 20-30 minutes, keeping warm at 80°C for 30 minutes, and cooling to 40°C, adding the supercritical fluid extract obtained in step S2 and 1.5-2.5 parts of ethylparaben, mixing well, and obtaining the final product.

[0027] In a further preferred embodiment, in the preparation process of the white jade ointment of the present invention, step S4, formulation forming, specifically involves: adding 10 parts of sodium hyaluronate, 100 parts of carbomer, and 70 parts of betacyclodextrin to the clear ointment obtained in step S3, heating at 60°C to dissolve, and setting aside; separately taking 120 parts of hydrogenated vegetable oil, 110 parts of olive oil, and 90 parts of polysorbate 80, heating and melting at 80°C, and mixing with the above clear ointment; emulsifying for 20 minutes, keeping warm at 80°C for 30 minutes, and cooling to 40°C, adding the supercritical fluid extract obtained in step S2 and 2 parts of ethylparaben, mixing well, and obtaining the final product.

[0028] The beneficial effects of this invention are:

[0029] 1. This invention provides a preparation process for Baiyu ointment. Using the original extraction method for Baiyu ointment, the contents of volatile oil, total flavonoids, and total polysaccharides in the resulting Baiyu ointment are 0, 0.45 mg / g, and 0.74 mg / g, respectively. However, using the extraction method provided by this invention, the contents of volatile oil, total flavonoids, and total polysaccharides in the resulting Baiyu ointment reach 3.42 mg / g, 1.07 mg / g, and 1.36 mg / g, respectively. This indicates that the preparation process of Baiyu ointment provided by this invention significantly increases the content of its effective components, which is of great significance for improving drug quality and enhancing drug safety, efficacy, and quality controllability.

[0030] 2. The inventors investigated the factors affecting the extraction of the effective components of Baiyu Ointment and determined that the optimal extraction method is a dual-supercritical-ultrasonic extraction method. Specifically, the optimal process parameters for supercritical fluid extraction are: CO2 supercritical fluid extraction using a segmented extraction method; CO2 flow rate: 30 L / h; and the segmented extraction parameters are: 0–30 min, extraction temperature: 40℃, extraction pressure: 30 MPa; 31–60 min, extraction temperature: 45℃, extraction pressure: 35 MPa; 61–90 min, extraction temperature: 50℃, extraction pressure: 40 MPa. The optimal process parameters for ultrasonic extraction are: adding 6 times the weight of the residue in water; ultrasonic frequency: 30 kHz; ultrasonic power: 200 W; and ultrasonic time: 40 min.

[0031] 3. This invention improves the types and amounts of excipients in Baiyu Ointment. The original preparation process used 17 excipients, while the improved preparation process of this invention uses only 7 excipients. The optimal amounts of carbomer, hydrogenated vegetable oil and olive oil were determined through orthogonal experiments, which greatly reduces the process difficulty and production cost of Baiyu Ointment.

[0032] 4. Pharmacological experiments show that the treatment effect of the process method of the present invention in the rat skin fissure model is comparable to that of the original process method, even with only 50% of the amount of medicine used. This indicates that the process method provided by the present invention can reduce the amount of medicinal materials used and save medicinal resources. Attached Figure Description

[0033] Figure 1 The standard curve for the determination of total flavonoids;

[0034] Figure 2 This is a standard curve for the determination of total polysaccharides. Detailed Implementation

[0035] The technical solution of the present invention will be described in detail below with reference to specific embodiments. These embodiments are for explanation and illustration only and do not constitute a limitation on the technical solution of the present invention.

[0036] Example 1

[0037] The preparation process of Baiyu Ointment is as follows:

[0038] (1) Obtaining medicinal materials:

[0039] Weigh out 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum striatum, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, and 40g of dried ginger. Mix them well, pulverize them, and pass them through a 60-mesh sieve. Set aside for later use.

[0040] (2) Supercritical fluid extraction:

[0041] The fine powder obtained in step (1) was subjected to CO2 supercritical fluid extraction using a segmented method, and the extract was collected for later use.

[0042] The CO2 flow rate for supercritical fluid extraction is 30 L / h.

[0043] The extraction parameters for the fractional extraction method are as follows: 0–30 min, extraction temperature 40℃, extraction pressure 30 MPa; 31–60 min, extraction temperature 45℃, extraction pressure 35 MPa; 61–90 min, extraction temperature 50℃, extraction pressure 40 MPa.

[0044] (3) Ultrasonic extraction:

[0045] Take the residue after supercritical fluid extraction in step (2), add 6 times the amount of water, and extract it by ultrasonic extraction for 40 minutes at a frequency of 30kHz and a power of 200W. Filter the mixture and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use.

[0046] (4) Formulation:

[0047] Add 10g sodium hyaluronate, 100g carbomer, and 70g betacyclodextrin to the clear paste obtained in step (3), and heat at 60°C to dissolve it for later use; separately take 120g hydrogenated vegetable oil, 110g olive oil, and 90g polysorbate 80, heat at 80°C to melt them, and mix them with the clear paste above; emulsify for 20min, keep warm at 80°C for 30min, and when the temperature drops to 40°C, add the supercritical fluid extract obtained in step (2) and 2g ethylparaben, mix well, and the product is ready.

[0048] Example 2

[0049] The preparation process of Baiyu Ointment is as follows:

[0050] (1) Obtaining medicinal materials:

[0051] Weigh out 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum striatum, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, and 40g of dried ginger. Mix them well, pulverize them, and pass them through a 60-mesh sieve. Set aside for later use.

[0052] (2) Supercritical fluid extraction:

[0053] The fine powder obtained in step (1) was subjected to CO2 supercritical fluid extraction using a segmented method, and the extract was collected for later use.

[0054] The CO2 flow rate for supercritical fluid extraction is 30 L / h.

[0055] The extraction parameters for the fractional extraction method are as follows: 0–30 min, extraction temperature 35℃, extraction pressure 25 MPa; 31–60 min, extraction temperature 40℃, extraction pressure 30 MPa; 61–90 min, extraction temperature 45℃, extraction pressure 35 MPa.

[0056] (3) Ultrasonic extraction:

[0057] Take the residue after supercritical fluid extraction in step (2), add 6 times the amount of water, and extract it by ultrasonic extraction for 40 minutes at a frequency of 30kHz and a power of 200W. Filter the mixture and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use.

[0058] (4) Formulation:

[0059] Add 10g sodium hyaluronate, 100g carbomer, and 70g betacyclodextrin to the clear paste obtained in step (3), and heat at 60°C to dissolve it for later use; separately take 120g hydrogenated vegetable oil, 110g olive oil, and 90g polysorbate 80, heat at 80°C to melt them, and mix them with the clear paste above; emulsify for 20min, keep warm at 80°C for 30min, and when the temperature drops to 40°C, add the supercritical fluid extract obtained in step (2) and 2g ethylparaben, mix well, and the product is ready.

[0060] Example 3

[0061] The preparation process of Baiyu Ointment is as follows:

[0062] (1) Obtaining medicinal materials:

[0063] Weigh out 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum striatum, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, and 40g of dried ginger. Mix them well, pulverize them, and pass them through a 60-mesh sieve. Set aside for later use.

[0064] (2) Supercritical fluid extraction:

[0065] The fine powder obtained in step (1) was subjected to CO2 supercritical fluid extraction using a segmented method, and the extract was collected for later use.

[0066] The CO2 flow rate for supercritical fluid extraction is 30 L / h.

[0067] The extraction parameters for the fractional extraction method are as follows: 0–30 min, extraction temperature 38℃, extraction pressure 28 MPa; 31–60 min, extraction temperature 42℃, extraction pressure 32 MPa; 61–90 min, extraction temperature 47℃, extraction pressure 37 MPa.

[0068] (3) Ultrasonic extraction:

[0069] Take the residue after supercritical fluid extraction in step (2), add 6 times the amount of water, and extract it by ultrasonic extraction for 40 minutes at a frequency of 30kHz and a power of 200W. Filter the mixture and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use.

[0070] (4) Formulation:

[0071] Add 10g sodium hyaluronate, 100g carbomer, and 70g betacyclodextrin to the clear paste obtained in step (3), and heat at 60°C to dissolve it for later use; separately take 120g hydrogenated vegetable oil, 110g olive oil, and 90g polysorbate 80, heat at 80°C to melt them, and mix them with the clear paste above; emulsify for 20min, keep warm at 80°C for 30min, and when the temperature drops to 40°C, add the supercritical fluid extract obtained in step (2) and 2g ethylparaben, mix well, and the product is ready.

[0072] Example 4

[0073] The preparation process of Baiyu Ointment is as follows:

[0074] (1) Obtaining medicinal materials:

[0075] Weigh out 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum striatum, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, and 40g of dried ginger. Mix them well, pulverize them, and pass them through a 60-mesh sieve. Set aside for later use.

[0076] (2) Supercritical fluid extraction:

[0077] The fine powder obtained in step (1) was subjected to CO2 supercritical fluid extraction using a segmented method, and the extract was collected for later use.

[0078] The CO2 flow rate for supercritical fluid extraction is 30 L / h.

[0079] The extraction parameters for the fractional extraction method are as follows: 0–30 min, extraction temperature 40℃, extraction pressure 30 MPa; 31–60 min, extraction temperature 45℃, extraction pressure 35 MPa; 61–90 min, extraction temperature 50℃, extraction pressure 40 MPa.

[0080] (3) Ultrasonic extraction:

[0081] Take the residue after supercritical fluid extraction in step (2), add 10 times the amount of water, and extract it by ultrasonic extraction for 30 minutes at a frequency of 20kHz and a power of 150W. Filter the mixture and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use.

[0082] (4) Formulation:

[0083] Add 10g sodium hyaluronate, 100g carbomer, and 70g betacyclodextrin to the clear paste obtained in step (3), and heat at 60°C to dissolve it for later use; separately take 120g hydrogenated vegetable oil, 110g olive oil, and 90g polysorbate 80, heat at 80°C to melt them, and mix them with the clear paste above; emulsify for 20min, keep warm at 80°C for 30min, and when the temperature drops to 40°C, add the supercritical fluid extract obtained in step (2) and 2g ethylparaben, mix well, and the product is ready.

[0084] Example 5

[0085] The preparation process of Baiyu Ointment is as follows:

[0086] (1) Obtaining medicinal materials:

[0087] Weigh out 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum striatum, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, and 40g of dried ginger. Mix them well, pulverize them, and pass them through a 60-mesh sieve. Set aside for later use.

[0088] (2) Supercritical fluid extraction:

[0089] The fine powder obtained in step (1) was subjected to CO2 supercritical fluid extraction using a segmented method, and the extract was collected for later use.

[0090] The CO2 flow rate for supercritical fluid extraction is 30 L / h.

[0091] The extraction parameters for the fractional extraction method are as follows: 0–30 min, extraction temperature 40℃, extraction pressure 30 MPa; 31–60 min, extraction temperature 45℃, extraction pressure 35 MPa; 61–90 min, extraction temperature 50℃, extraction pressure 40 MPa.

[0092] (3) Ultrasonic extraction:

[0093] Take the residue after supercritical fluid extraction in step (2), add 8 times the amount of water, and ultrasonically extract for 50 minutes at a frequency of 40kHz and a power of 250W. Filter and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use.

[0094] (4) Formulation:

[0095] Add 10g sodium hyaluronate, 100g carbomer, and 70g betacyclodextrin to the clear paste obtained in step (3), and heat at 60°C to dissolve it for later use; separately take 120g hydrogenated vegetable oil, 110g olive oil, and 90g polysorbate 80, heat at 80°C to melt them, and mix them with the clear paste above; emulsify for 20min, keep warm at 80°C for 30min, and when the temperature drops to 40°C, add the supercritical fluid extract obtained in step (2) and 2g ethylparaben, mix well, and the product is ready.

[0096] Example 6

[0097] The preparation process of Baiyu Ointment is as follows:

[0098] (1) Obtaining medicinal materials:

[0099] Weigh out 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum striatum, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, and 40g of dried ginger. Mix them well, pulverize them, and pass them through a 60-mesh sieve. Set aside for later use.

[0100] (2) Supercritical fluid extraction:

[0101] The fine powder obtained in step (1) was subjected to CO2 supercritical fluid extraction using a segmented method, and the extract was collected for later use.

[0102] The CO2 flow rate for supercritical fluid extraction is 30 L / h.

[0103] The extraction parameters for the fractional extraction method are as follows: 0–30 min, extraction temperature 40℃, extraction pressure 30 MPa; 31–60 min, extraction temperature 45℃, extraction pressure 35 MPa; 61–90 min, extraction temperature 50℃, extraction pressure 40 MPa.

[0104] (3) Ultrasonic extraction:

[0105] Take the residue after supercritical fluid extraction in step (2), add 6 times the amount of water, and extract it by ultrasonic extraction for 40 minutes at a frequency of 30kHz and a power of 200W. Filter the mixture and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use.

[0106] (4) Formulation:

[0107] Add 8g sodium hyaluronate, 80g carbomer, and 80g betacyclodextrin to the clear paste obtained in step (3), and heat at 60°C to dissolve it for later use; separately take 100g hydrogenated vegetable oil, 100g olive oil, and 100g polysorbate 80, heat at 80°C to melt them, and mix them with the clear paste above; emulsify for 20 minutes, keep warm at 80°C for 30 minutes, and when the temperature drops to 40°C, add the supercritical fluid extract obtained in step (2) and 1.5g ethylparaben, mix well, and the product is ready.

[0108] To further verify the reliability of the present invention and select the optimal solution, the inventors conducted a series of experiments, some of which are excerpted below:

[0109] 1. Methods for determining the content of active ingredients

[0110] 1.1 Determination of total volatile oil

[0111] Take the sample to be tested, extract it twice with an appropriate amount of petroleum ether (30-60℃), evaporate the petroleum ether, weigh it, and the sample is obtained.

[0112] 1.2 Determination of total flavonoids

[0113] (1) Preparation of reference solution

[0114] Accurately weigh 50 mg of rutin reference standard and place it in a 25 mL volumetric flask. Add an appropriate amount of methanol, heat in a water bath to dissolve, cool, add methanol to the mark, and shake well. Accurately measure 10 mL and place it in a 100 mL volumetric flask. Add water to the mark and shake well to obtain the solution (each mL contains 0.2 mg of rutin).

[0115] (2) Plotting the standard curve

[0116] Accurately measure 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, and 6 mL of the reference solution into separate 25 mL volumetric flasks. Add water to each flask to a final volume of 6.0 mL. Add 1 mL of 5% sodium nitrite solution, mix well, and let stand for 6 min. Add 1 mL of 10% aluminum nitrate solution, shake well, and let stand for 6 min. Add 10 mL of sodium hydroxide solution, then add water to the mark, shake well, and let stand for 15 min. Using the corresponding reagents as blanks, measure the absorbance at 500 nm using UV-Vis spectrophotometry. Plot a standard curve with absorbance on the ordinate and concentration on the abscissa. Figure 1 As shown.

[0117] (3) Determination method

[0118] Accurately measure 5 mL of the drug solution for each test and place it in a 25 mL volumetric flask. Following the method under the preparation of the standard curve, starting from "add 1 mL of 5% sodium nitrite solution", determine the absorbance according to the method. Read the weight (μg) of rutin in the test solution from the standard curve and calculate the result.

[0119] 1.3 Determination of total polysaccharides

[0120] (1) Preparation of reference solution

[0121] Accurately weigh 30 mg of anhydrous glucose reference standard dried to constant weight at 105℃, place it in a 100 mL volumetric flask, dissolve and dilute with water to the mark, and shake well to obtain (each 1 mL contains 0.30 mg of anhydrous glucose).

[0122] (2) Plotting the standard curve

[0123] Accurately measure 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, and 0.6 mL of the reference solution into separate 10 mL stoppered graduated test tubes. Add water to each tube to a final volume of 2.0 mL, shake well, and slowly add 0.2% anthrone-sulfuric acid solution to the mark in an ice-water bath. Mix well, cool, and then incubate in a water bath for 10 min. Remove from the water bath and immediately cool in an ice-water bath for 10 min. Use the corresponding reagents as blanks. Measure the absorbance at 582 nm using ultraviolet-visible spectrophotometry (General Chapter 0401 of the Chinese Pharmacopoeia). Plot a standard curve with absorbance as the ordinate and concentration as the abscissa, as shown below. Figure 2 As shown.

[0124] (3) Determination method

[0125] Accurately measure 5 mL of the extract and place it in a 25 mL volumetric flask. Following the method under the preparation of the standard curve, starting from "add water to 2.0 mL", determine the absorbance according to the method. Read the weight (mg) of anhydrous glucose in the test solution from the standard curve and calculate the result.

[0126] 2. Optimization of extraction process parameters

[0127] 2.1 Initial Selection of Extraction Methods

[0128] The Baiyu Ointment formula contains nine medicinal herbs. Modern research shows that the effective components and pharmacological effects of each herb in treating chapped skin are shown in Table 1.

[0129] Table 1. Effective components and pharmacological effects of each herb in Baiyu Ointment

[0130]

[0131]

[0132] As can be seen from the table above, the effective substances in the Baiyu Ointment formula that treat chapped skin mainly fall into three categories: polysaccharides (Bletilla striata, Polygonatum odoratum, Angelica sinensis), volatile oils (Ligusticum striatum, Angelica sinensis, dried ginger), and flavonoids (Polygonum hydropiper, Geranium wilfordii, Salvia miltiorrhiza). To maximize the extraction of these three categories of substances, the research team compared the following extraction methods:

[0133] (1) Fermentation extraction method

[0134] Weigh the medicinal materials according to the formula ratio (80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum striatum, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, and 40g of Zingiber officinale), grind them into coarse powder that passes through a 10-mesh sieve, add 8 times the total weight of the medicinal materials to water, boil and then cool, then add 12% of the total weight of the medicinal materials to glucose, inoculate with Carboxymycium chloride at an inoculum rate of 2.0%, ferment at 35℃ for 18 hours, filter, concentrate the filtrate and make up to 250mL, and determine the total volatile oil, total flavonoids and total polysaccharide content according to the method under "1. Determination of effective ingredient content".

[0135] (2) Supercritical fluid extraction method

[0136] Weigh the medicinal materials according to the formulation ratio (same as fermentation extraction method), and pulverize them into fine powder that passes through a 60-mesh sieve. Extract for 0.5 hours under the conditions of CO2 flow rate of 30 L / h, extraction temperature of 35℃, and extraction pressure of 30 MPa. Collect the extract and dilute it to 250 mL with 50% ethanol solution. Determine the content of total volatile oil, total flavonoids, and total polysaccharides according to the method under "1. Determination of Active Ingredient Content".

[0137] (3) Enzymatic hydrolysis

[0138] Weigh the medicinal materials according to the formula ratio (same as fermentation extraction method), crush them into coarse powder that passes through a 10-mesh sieve, add 8 times the amount of water and 30% cellulase, and extract for 10 hours at 55℃ and pH 5.0. After boiling, filter, concentrate the filtrate and make up to 250mL. Determine the content of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Determination of effective component content".

[0139] (4) Water extraction method

[0140] Weigh the medicinal materials according to the formula ratio (same as fermentation extraction method), crush them into coarse powder that passes through a 10-mesh sieve, add 8 times the amount of water of the medicinal materials and decoct for 1 hour (collect volatile oil at the same time), filter, concentrate the filtrate and make up to 250 mL, and determine the total volatile oil, total flavonoids and total polysaccharide content according to the method under "1. Determination of effective component content".

[0141] (5) Ultrasonic method

[0142] Weigh the medicinal materials according to the formula ratio (same as fermentation extraction method), crush them into coarse powder that passes through a 10-mesh sieve, add 10 times the amount of water of the medicinal materials, and extract by ultrasonic extraction for 30 minutes at a frequency of 40kHz and a power of 250W. Filter, concentrate the filtrate and make up to 250mL, and determine the content of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Determination of effective component content".

[0143] (6) Alcohol extraction method

[0144] Weigh the medicinal materials according to the formula ratio (same as fermentation extraction method), crush them into coarse powder that passes through a 10-mesh sieve, add 8 times the amount of medicinal material in 60% ethanol solution and reflux for 1 hour, filter, concentrate the filtrate and make up to 250 mL, and determine the content of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Determination of effective component content".

[0145] The content of active ingredients in the extracts obtained by the different methods described above was determined, and the results are shown in Table 2.

[0146] Table 2. Results of determination of active ingredient content in extracts from different extraction methods

[0147]

[0148] The results show that none of the six extraction methods could simultaneously extract all three active ingredients to the maximum extent. Although supercritical fluid extraction yielded the highest total volatile oil and total flavonoid content, it had the lowest polysaccharide extraction rate. Based on the experimental data, supercritical fluid extraction combined with water extraction or ultrasonic extraction could be used. However, considering the simplicity of the ultrasonic extraction process, the combination of supercritical fluid extraction and ultrasonic extraction was the preferred method. This involves first subjecting the medicinal material to supercritical fluid extraction, followed by ultrasonic extraction of the residue. To verify the feasibility of this approach, the research team conducted the following experiments:

[0149] Weigh out the medicinal materials according to the formulation ratio (same as fermentation extraction method), and weigh out a total of 3 portions. Crush the medicinal materials into fine powder that passes through a 60-mesh sieve. Two portions are repeated by supercritical fluid extraction and ultrasonic extraction methods as described above. The other portion is first extracted for 0.5 h under the conditions of CO2 flow rate of 30 L / h, extraction temperature of 35℃ and extraction pressure of 30 MPa. Collect the extract, then take the residue and add 10 times the mass of the residue to water. Ultrasonic extraction is carried out for 30 min under the conditions of frequency of 40 kHz and power of 250 W. Filter, concentrate the filtrate and mix it with the supercritical fluid extraction extract, and make up to 250 mL. Determine the content of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Determination of effective component content". The results are shown in Table 3.

[0150] Table 3 Validation results of different extraction methods

[0151]

[0152] As shown in Table 3, the combination of supercritical fluid extraction and ultrasonic extraction (dual-super-ultrasound extraction) can significantly improve the total polysaccharide extraction rate in the prescription, thereby maximizing the extraction of the three effective components in the Baiyu Ointment prescription.

[0153] 2.2 Optimization of Extraction Process

[0154] 2.2.1 Optimization of Supercritical Fluid Extraction Process

[0155] Research data indicates that the types of extracts obtained by supercritical CO2 fluid extraction vary under different pressures, temperatures, and flow rates. Generally, in the low-temperature, low-pressure stage, CO2 density is lower, preferentially extracting low-polarity, low-molecular-weight components (such as volatile oils and fat-soluble pigments). In the high-temperature, high-pressure stage, CO2 density increases, dissolving medium- to high-polarity components (such as flavonoids and alkaloids). Therefore, by adjusting pressure, temperature, and flow rate parameters, segmented extraction (i.e., stepwise extraction) of active ingredients can be achieved, thus selectively extracting the target components. Since the active ingredients in the Baiyu ointment formula include both low-polarity, low-molecular-weight volatile oils and high-polarity flavonoids, a segmented extraction method can be used to extract the active ingredients. Based on the commonly used process parameters of supercritical CO2 fluid extraction, the parameters in Table 4 are proposed for segmented extraction.

[0156] Table 4. Parameters for the CO2 Supercritical Fluid Extraction Method (Sectional Parameters)

[0157]

[0158] To verify the effectiveness of segmented extraction, the research team conducted a verification experiment according to the parameters in Table 5, and the results are shown in Table 6.

[0159] Table 5. Parameters of the segmented verification test for the CO2 supercritical fluid extraction method.

[0160]

[0161] Table 6 Results of CO2 Supercritical Fluid Fractional Extraction Experiment

[0162]

[0163] The results showed that fractional extraction using supercritical CO2 fluid extraction significantly improved the extraction rates of total volatile oils and total flavonoids in the formulation. Therefore, fractional extraction was selected as the method for supercritical fluid extraction.

[0164] 2.2.2 Optimization of Ultrasonic Extraction Process

[0165] The medicinal materials were weighed according to the formula ratio (960g of Bletilla striata, 480g of Polygonatum odoratum, 480g of Ligusticum striatum, 480g of Polygonum hydropiper, 480g of Geranium wilfordii, 240g of Salvia miltiorrhiza, 240g of Lithospermum erythrorhizon, 480g of Angelica sinensis, and 480g of dried ginger). The medicinal materials were pulverized into fine powder that passed through a 60-mesh sieve. Supercritical fluid extraction was carried out under the condition of CO2 flow rate of 30L / h according to the segmented extraction parameters in Table 7. The residue after extraction was divided into 12 equal portions for subsequent experiments.

[0166] Table 7. CO2 Supercritical Fluid Fractional Extraction Parameters

[0167]

[0168] (1) Investigation of water addition

[0169] Take three portions of the extracted residue, add 6, 8, and 10 times the amount of water respectively, and extract by ultrasonication for 30 min at a frequency of 40 kHz and a power of 250 W. Filter, concentrate the liquid and make up to 250 mL, and determine the total polysaccharide content. The results are shown in Table 8.

[0170] Table 8 Results of Water Addition Investigation

[0171]

[0172] The results showed that the amount of water added had little effect on the total polysaccharide content. Taking all factors into consideration, a water addition of 6 times the normal amount was chosen for subsequent studies.

[0173] (2) Ultrasonic frequency investigation

[0174] Take three portions of the extracted residue, add six times the amount of water to each, and ultrasonically extract for 30 min at frequencies of 20 kHz, 30 kHz and 40 kHz and power of 250 W respectively. Filter, concentrate the liquid and make up to 250 mL, and determine the total polysaccharide content. The results are shown in Table 9.

[0175] Table 9 Results of Ultrasonic Frequency Investigation

[0176]

[0177] The experimental results showed that the ultrasonic frequency had little effect on the total polysaccharide content. Taking all factors into consideration, an ultrasonic frequency of 30 kHz was chosen for subsequent research.

[0178] (3) Ultrasonic power assessment

[0179] Take 3 portions of the extracted residue, add 6 times the amount of water, and ultrasonically extract for 30 minutes at a frequency of 30kHz, with power of 150W, 200W and 250W respectively. Filter, concentrate the liquid and make up to 250ml, and determine the total polysaccharide content. The results are shown in Table 10.

[0180] Table 10 Results of Ultrasonic Power Evaluation

[0181]

[0182] The experimental results show that the ultrasonic power has a significant impact on the total polysaccharide content. The total polysaccharide content in the 150W sample is significantly lower than that in the 200W and 250W samples, but the total polysaccharide content in the 200W and 250W samples does not change much. Therefore, an ultrasonic power of 200W can be selected for subsequent research.

[0183] (4) Ultrasonic time study

[0184] Take three portions of the extracted residue, add six times the amount of water to each, and extract by ultrasonication for 30 min, 40 min and 50 min respectively at a frequency of 30 kHz and a power of 200 W. Filter, concentrate the liquid and make up to 250 mL, and determine the total polysaccharide content. The results are shown in Table 11.

[0185] Table 11 Results of Ultrasonic Time-Based Study

[0186]

[0187] The experimental results show that the ultrasonic time has a significant impact on the total polysaccharide content. The total polysaccharide content in the samples ultrasonicated for 40 min and 50 min is significantly higher than that in the samples ultrasonicated for 30 min, but the total polysaccharide content in the samples ultrasonicated for 40 min and 50 min does not change much. Therefore, an ultrasonic time of 40 min can be selected for subsequent studies.

[0188] 3. Optimal matrix selection

[0189] The original preparation process for Baiyu Ointment was as follows: Herbal extracts were prepared according to the prescription ratio, and 60g of sorbitol, 60g of beta-cyclodextrin, 60g of glycerin, 20g of titanium dioxide, and 3g of sodium hyaluronate were added. The mixture was heated (80℃) to dissolve the extract and set aside. Separately, 10g of mink oil, 30g of lanolin, 60g of petrolatum, 12g of monoglyceride, 10g of sucrose ester, 100g of paraffin oil, 50g of polysorbate 80, 50g of Span 80, and 20g of emulsified silicone oil were heated (80℃) to melt the mixture. This melted mixture was then combined with the above-mentioned ointment and emulsified for 15 minutes. The mixture was kept at 80℃ for 30 minutes, cooled to 40℃, and then 2g of elastase, 6g of benzoic acid, and 1.5g of ethylparaben were added and mixed thoroughly to produce 1000g of the ointment. This process used a large number of excipients, which was detrimental to the quality control of the formulation and resulted in excessively high production costs. With the development and application of new materials, it is necessary to optimize and improve the formulation process.

[0190] The research team plans to use sodium hyaluronate, carbomer, beta-cyclodextrin, hydrogenated vegetable oil, olive oil, and polysorbate 80 as o / w matrix, and add ethylparaben as a preservative to carry out research on formulation molding process.

[0191] 3.1 Optimization Indicators and Scoring Criteria

[0192] (1) Appearance

[0193] The total score is 25 points. The evaluation is based on aspects such as color uniformity, fine texture, appropriate consistency, spreadability, and absence of graininess, with each aspect accounting for 5 points.

[0194] (2) Centrifugal stability

[0195] The maximum score is 25 points. Place each group of ointments in a centrifuge tube and centrifuge at 3000 rpm. -1 Centrifuge for 25 minutes and observe for any phenomena such as layering, clumping, bubbling, or water seepage.

[0196] (3) Cold resistance test

[0197] The maximum score is 5 points. Each group of ointments was placed in a plastic sample cup and stored in a 4℃ refrigerator for 1 month to observe for any phenomena such as layering, clumping, or water seepage.

[0198] (4) Heat resistance test

[0199] The maximum score is 5 points. Each group of ointments was placed in a plastic sample cup and stored in a 40℃ constant temperature oven for 1 month. The presence of any layering, discoloration, mold growth, or other phenomena was observed.

[0200] (5) Extensibility

[0201] The total score is 20 points. Take two clean glass plates, each 10cm x 10cm in size, and place a 125g weight on each plate (add the weight to make up the mass if necessary). Weigh 1g of each group of ointments that have been left at room temperature for 24 hours and place it between the two glass plates. After 1 minute, use a ruler to measure the diameter of the ointment after it has spread, and take the average of the ratio of the largest diameter to the smallest diameter.

[0202] (6) pH value

[0203] The total score is 20 points. Take 5g of each ointment and place it in a small beaker. Add an equal amount of distilled water to dissolve it completely and stir well. Use a pH meter to measure the pH value of each group. A pH value of 5-8 is full marks, and the rest is zero marks.

[0204] 3.2 Orthogonal Experiment

[0205] Three factors that significantly affect the physicochemical properties of the ointment—carbomer dosage (A), hydrogenated vegetable oil dosage (B), and olive oil dosage (C)—were investigated. The dosages of other excipients remained constant: sodium hyaluronate 10g, betacyclodextrin 70g, and polysorbate 80 90g. Based on the preliminary data, three levels were selected for each factor to conduct an orthogonal experiment. The factor levels are shown in Table 12, the experimental results are shown in Table 13, and the analysis of variance is shown in Table 14.

[0206] Table 12 Factors and Levels in the Orthogonal Experiment for Matrix Investigation

[0207]

[0208] Table 13 Orthogonal experimental design and results for matrix investigation

[0209]

[0210] Table 14 Results of matrix investigation analysis of variance

[0211]

[0212] Note: F 0.05 (2, 2) = 19.

[0213] As shown in Table 13, the influence of each factor on the ointment forming effect is A > B > C, and A3 > A2 > A1, B3 > B2 > B1, C3 > C1 > C2. The optimal matrix ratio is A3B3C3. Combined with the variance analysis results in Table 14, all three factors have a significant impact. Therefore, the optimal matrix ratio is carbomer dosage: hydrogenated vegetable oil dosage: olive oil = 10:12:11.

[0214] 4. Verification of optimal process parameters

[0215] (1) Weigh the medicinal materials

[0216] Weigh out 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum striatum, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, and 40g of Zingiber officinale according to the prescription, and mix them well.

[0217] (2) Crushing

[0218] The medicinal materials are pulverized into fine powder that passes through a 60-mesh sieve.

[0219] (3) Supercritical extraction

[0220] Under a CO2 flow rate of 30 L / h, the above fine powder was subjected to supercritical CO2 extraction using a segmented method. The extraction parameters are shown in Table 15. The extracted extract was collected for later use.

[0221] Table 15 Parameters for CO2 Supercritical Fluid Fractional Extraction

[0222]

[0223] (4) Ultrasonic extraction

[0224] Add 6 times the amount of water to the extracted residue and extract ultrasonically for 30 minutes at a frequency of 20kHz and a power of 150W. Filter and concentrate the liquid to a clear extract with a relative density of about 1.10 (60℃) for later use.

[0225] (5) Formulation

[0226] Add 10g sodium hyaluronate, 100g carbomer, and 70g betacyclodextrin to the clear extract and heat (60℃) to dissolve. Set aside. Separately, heat 120g hydrogenated vegetable oil, 110g olive oil, and 90g polysorbate 80 to melt (80℃), mix with the clear extract above, emulsify for 20 minutes, keep warm at 80℃ for 30 minutes, cool to about 40℃, add supercritical CO2 extract, add 2g ethylparaben, mix well, and the product is ready.

[0227] (6) Verification test

[0228] Three batches of samples were prepared according to the above optimal parameters, and the results are shown in Table 16.

[0229] Table 16 Validation Results of Optimal Process Parameters

[0230]

[0231] The results show that the white jade ointment prepared by the present invention has good performance in all aspects and the process is stable, indicating that the present invention is reasonable and feasible.

[0232] 5. Comparison test before and after process improvement

[0233] The same batch of medicinal materials were used to prepare ointments using the improved process and the original process, respectively. Each gram of ointment prepared by the two processes was equivalent to 0.36g of raw medicinal materials (i.e., 360g of medicinal materials per prescription, which was used to make 1000g of ointment). The contents of total volatile oil, total flavonoids and total polysaccharides in the two ointments were determined, and the results are shown in Table 17.

[0234] Table 17 Results of determination of effective component content by different preparation processes

[0235]

[0236] The experimental results show that, compared with the original preparation process of Baiyu Ointment, the improved process significantly increases the content of volatile oil, total flavonoids and total polysaccharides in the formula.

[0237] 6. Drug efficacy verification test

[0238] 6.1 Rat skin fissure model

[0239] (1) Experimental materials

[0240] Experimental animals: 56 male SD rats (SPF grade), weighing 200±20g.

[0241] Drugs and reagents: Baiyu ointment samples prepared according to the method of this invention; Baiyu ointment (Guizhou Lvtaiyang Pharmaceutical Co., Ltd.); physiological saline; urea ointment; enzyme-linked immunosorbent assay kit; electric thermostatic water bath; electric thermostatic drying oven; handheld shaver; high-speed refrigerated centrifuge; surgical scissors; enzyme-linked immunosorbent assay reader.

[0242] (2) Test methods

[0243] ① Modeling, grouping, and drug administration:

[0244] Fifty-six rats were randomly divided into a blank control group, a model group, and a positive control group (urea ointment was applied evenly at a concentration of 0.1 g / cm³). 2 Original process group (uniformly applied 0.1g / cm) 2 ), this invention group 1 (0.1g / cm 2 Group 2 of the present invention (uniformly applied at 0.08 g / cm³) 2 Group 3 of the present invention (uniformly applied at 0.06 g / cm³) 2 Eight animals were placed in each group and were acclimatized for 7 days before the experiment.

[0245] Twenty-four hours prior to the experiment, a 4.0cm × 4.0cm area on the back of rats was shaved. After shaving, the control group was treated with physiological saline, while the other six groups were treated with 3mol / L sodium hydroxide solution for eight consecutive days to induce skin fissures and establish a wound model. Once the model was successfully established, the model groups were allowed to heal naturally without medication. The positive control group was treated with urea ointment, while the treatment groups were treated with the corresponding doses of the drug twice daily for seven consecutive days. Blood was collected from the abdominal aorta, centrifuged, and the supernatant serum was collected for later use. ELISA was used to detect relevant inflammatory factors, including interleukin-6 (IL-6), IL-10, nitric oxide (NO), nitric oxide synthase (NOS), and tumor necrosis factor-α (TNF-α), strictly following the kit instructions.

[0246] ② Visual observation: One hour after administering the medication on the last day, remove the medication, wash the skin on the back, and score the skin. The scoring criteria are as follows: 0 points: normal color, no redness or swelling; 1 point: slight redness and swelling; 2 points: redness, swelling, dryness, and wrinkles; 3 points: exudate, scratching of the skin on the back, and restlessness.

[0247] ③ Statistical methods: Data were processed using SPSS 17.0 statistical software. Data are expressed as mean ± standard deviation (x±s). The t-test was used for comparisons between groups, and the Ridit test was used for ordinal data. P < 0.01 was considered statistically significant.

[0248] (3) Test Results

[0249] ① Effects on a rat skin fissure model: The control group showed normal local skin condition. Ridit test showed significant redness, swelling, and exudation in the model group compared to the control group (P < 0.01). Compared to the model group, the original process group, group 1 of this invention, group 2 of this invention, and the urea ointment group all significantly inhibited skin redness, swelling, and exudation (P < 0.01). Group 2 of this invention showed comparable improvement to the original process group, while the amount of medicinal material used in group 2 was only 80% of that in the original process group, indicating that this invention can reduce the amount of medicinal material used. Results are shown in Table 18.

[0250] Table 18 Results of rat skin scoring (n=8)

[0251]

[0252] ② Effects on the levels of IL-6, IL-10, NO, NOS, and TNF-α in rat skin: Compared with the blank group, the levels of IL-6, IL-10, TNF-α, NOS, and NO in the skin of rats in the model group were significantly increased (P < 0.01); compared with the model group, the original process group, the urea ointment group, the present invention group 1, the present invention group 2, and the present invention group 3 significantly reduced the levels of IL-6, IL-10, TNF-α, NOS, and NO (P < 0.01); among them, the reduction in the present invention group 2 was comparable to that in the original process group, which further demonstrates that the present invention can reduce the amount of medicinal materials used. The results are shown in Table 19.

[0253] Table 19 Effects of each experimental group on the levels of IL-6, IL-10, TNF-α, NOS, and NO in a rat skin fissure model (x±s, n=8)

[0254]

[0255] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A preparation process for a white jade ointment, characterized in that, Includes the following steps: S1 retrieves medicinal materials: Weigh out 80 parts of Bletilla striata, 40 parts of Polygonatum odoratum, 40 parts of Ligusticum striatum, 40 parts of Polygonum hydropiper, 40 parts of Geranium wilfordii, 20 parts of Salvia miltiorrhiza, 20 parts of Lithospermum erythrorhizon, 40 parts of Angelica sinensis, and 40 parts of dried ginger. Mix them well, pulverize them, and pass them through a 60-mesh sieve. Set aside for later use. S2 supercritical fluid extraction: The fine powder obtained in step S1 was subjected to CO2 supercritical fluid extraction using a segmented method, and the extract was collected for later use. The CO2 flow rate of the supercritical fluid extraction is 30 L / h; The extraction parameters for the segmented method are as follows: 0~30 min, extraction temperature 35~40℃, extraction pressure 25~30 MPa; 31~60 min, extraction temperature 40~45℃, extraction pressure 30~35 MPa; 61~90 min, extraction temperature 45~50℃, extraction pressure 35~40 MPa. S3 ultrasonic extraction: Take the residue after supercritical fluid extraction in step S2, add 6 to 10 times the amount of water, and ultrasonically extract for 30 to 50 minutes at a frequency of 20 to 40 kHz and a power of 150 to 250 W. Filter and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use. S4 Formulation: Add 10 parts sodium hyaluronate, 100 parts carbomer, and 70 parts betacyclodextrin to the clear paste obtained in step S3, and heat at 60°C to dissolve. Set aside. Separately, take 120 parts hydrogenated vegetable oil, 110 parts olive oil, and 90 parts polysorbate 80, heat at 80°C to melt, and mix with the clear paste above. Emulsify for 20 min, keep warm at 80°C for 30 min, and when the temperature drops to 40°C, add the supercritical fluid extract obtained in step S2 and 2 parts ethylparaben, mix well, and the product is ready.

2. The preparation process according to claim 1, characterized in that, The extraction parameters for the segmented method described in step S2 are as follows: 0~30 min, extraction temperature 38~40℃, extraction pressure 28~30 MPa; 31~60 min, extraction temperature 42~45℃, extraction pressure 32~35 MPa; 61~90 min, extraction temperature 47~50℃, extraction pressure 37~40 MPa.

3. The preparation process according to claim 2, characterized in that, The extraction parameters for the segmented method described in step S2 are as follows: 0~30 min, extraction temperature is 40℃, extraction pressure is 30 MPa; 31~60 min, extraction temperature is 45℃, extraction pressure is 35 MPa; 61~90 min, extraction temperature is 50℃, extraction pressure is 40 MPa.

4. The preparation process according to claim 1, characterized in that, Step S3, ultrasonic extraction, is as follows: Take the residue after supercritical fluid extraction in step S2, add 6-8 times the amount of water, and ultrasonically extract for 40-50 min at a frequency of 20-30 kHz and a power of 200-250 W. Filter the mixture and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use.

5. The preparation process according to claim 4, characterized in that, Step S3, ultrasonic extraction, is as follows: Take the residue after supercritical fluid extraction in step S2, add 6 times the amount of water, and ultrasonically extract for 40 minutes at a frequency of 30 kHz and a power of 200 W. Filter the mixture and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60℃ for later use.

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