Preparation process of white jade ointment

Abstract

The invention discloses a preparation process of a white jade ointment, and relates to the technical field of medicines. The white jade ointment is prepared from rhizoma bletillae, radix polygonati officinalis, ligusticum, red-knees herb, geranium wilfordii, salvia miltiorrhiza, lithospermum erythrorhizon, angelica sinensis and rhizoma zingiberis. The preparation process comprises the following steps: taking medicinal materials; performing supercritical fluid extraction; performing ultrasonic extraction; and forming the preparation. By adopting a modern preparation process and combining a large number of investigation experiments, an original preparation process of the white jade ointment is improved and optimized, so that the content of effective components in the white jade ointment is increased, meanwhile, the variety of auxiliary materials is reduced, and the dosage of the auxiliary materials is optimized; the method is of great significance in improving the medicine quality and improving the medicine safety, effectiveness and quality controllability.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to a preparation process of Baiyu Ointment. Background Art

[0002] Baiyu Ointment (Drug Approval Number: National Drug Approval Letter) has the effects of promoting blood circulation to dispel wind and nourishing blood and moistening the skin, and is used for skin chaps caused by wind-dryness of blood deficiency, such as cracks on hands and feet, frostbite, etc., and is one of the representative products of Guizhou Miao medicine. The prescription of Baiyu Ointment is: 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum sinense, 40g of Polygonum hydropiper, 40g of Geranium wilfordii, 20g of Salvia miltiorrhiza, 20g of Lithospermum erythrorhizon, 40g of Angelica sinensis, 40g of Dry Ginger. Its original preparation process is as follows: For the above nine flavors, take Salvia miltiorrhiza, Angelica sinensis, and Dry Ginger, use 80% ethanol as a solvent, soak for 12h, and then slowly percolate at a speed of 1 - 3 mL / min, collect 1200 mL of percolate, recover ethanol under reduced pressure and concentrate to a clear paste with a relative density of 1.15 (50°C) for standby; the remaining six flavors including Bletilla striata are decocted with water twice, 2h for the first time and 1.5h for the second time, combine the decoction liquids, filter, concentrate the filtrate to a clear paste with a relative density of 1.05 (60°C), and combine it with the above clear paste; add 60g of sorbitol, 60g of beta-cyclodextrin, 60g of glycerol, 20g of titanium dioxide (titanium white powder), 3g of sodium hyaluronate, heat (80°C) to dissolve for standby; separately take 10g of mink oil, 30g of lanolin, 60g of white petrolatum (petrolatum), 12g of mono- and diglycerides of fatty acids (monoglyceride), 10g of sucrose stearate (sucrose ester), 100g of liquid paraffin (paraffin oil), 50g of polysorbate 80, 50g of sorbitan oleate (span 80), 20g of emulsifying silicone oil, heat (80°C) to melt, mix with the above clear paste, emulsify for 15min, keep warm at 80°C for 30min, cool down to 40°C, then add 2g of elastase (elastase), 6g of benzoic acid, 1.5g of ethylparaben, mix well, and make 1000g to obtain the product.

[0003] The prescription of Baiyu Ointment consists of nine medicinal herbs in total. Modern research shows that the active ingredients and pharmacological effects of each medicinal herb in the prescription for treating skin chaps are as follows: The polysaccharides, bibenzyl compounds, and phenolic acids in Bletilla striata have the effects of promoting wound healing, enhancing the water-holding capacity of the stratum corneum, and repairing the skin barrier; The polysaccharides and steroidal saponins in Polygonatum odoratum have the effects of nourishing yin and moistening dryness, improving skin moisture retention, and relieving chaps; The volatile oils such as ligustilide and tetramethylpyrazine in Ligusticum sinense, as well as components such as ferulic acid, have the effects of expelling wind and dispelling cold, improving local microcirculation, and relieving frostbite pain; The tannins such as gallic acid and flavonoids such as quercetin in Polygonum hydropiper have the effects of astringing and promoting granulation and promoting the healing of cracks; The active ingredients such as tanshinone IIA and salvianolic acid B in Salvia miltiorrhiza have the effects of promoting blood circulation to remove blood stasis and improving local blood circulation; The active ingredients such as shikonin and acetylshikonin in Lithospermum erythrorhizon have the effects of cooling blood and detoxifying, anti-inflammatory and promoting repair, and preventing infection; The ferulic acid, angelica polysaccharides, and volatile oils such as ligustilide in Angelica sinensis have the effects of enriching blood and promoting blood circulation, moisturizing the skin, and regulating immune function; The gingerol (6-gingerol) and volatile oil components in Zingiber officinale have the effects of warming meridians and dispelling cold, promoting local blood circulation, and relieving frostbite. It can be seen that the main active substances in the prescription of Baiyu Ointment that play a role in treating skin chaps are mainly three categories, namely polysaccharides (Bletilla striata, Polygonatum odoratum, Angelica sinensis), volatile oils (Ligusticum sinense, Angelica sinensis, Zingiber officinale), and flavonoids (Polygonum hydropiper, Geranium wilfordii, Salvia miltiorrhiza). Among these three categories of substances, the polysaccharide substances have good water solubility, and water extraction method can be used for extraction in industrial production; The volatile oil is a fat-soluble component, and usually steam distillation method or organic solvent method is used for extraction; And the flavonoids are also fat-soluble components, and alcohol extraction method is suitable for extraction.

[0004] The original preparation process of Baiyu Ointment was established 30 years ago. With the continuous development of the research level of traditional Chinese medicine, modern research has proved that its original preparation process is not very reasonable and has the following drawbacks: (1) The retention rate of volatile oil in the finished product of the prescription is relatively low. Although the volatile oil of Angelica sinensis can be extracted after percolation with 80% ethanol, it is easily volatilized and lost due to heat during the subsequent ethanol recovery and concentration processes; Ligusticum sinense and Zingiber officinale did not collect volatile oil during the decoction process, and there is also a vacuum concentration process later, resulting in very little retention of their volatile oil. (2) The extraction rates of the active ingredients of flavonoids and polysaccharides in the prescription have not reached the maximum. Flavonoid components are contained in Polygonum hydropiper, Geranium wilfordii, and Salvia miltiorrhiza in the prescription, but only Salvia miltiorrhiza is extracted by alcohol extraction; Polysaccharide components are contained in Bletilla striata, Polygonatum odoratum, and Angelica sinensis in the prescription, but Angelica sinensis is missing in the extraction by water extraction method. (3) A total of 17 excipients are used in the original preparation process of Baiyu Ointment, with high production costs and it is not conducive to the quality control of Baiyu Ointment.

[0005] To solve the problems existing in the original preparation process of Baiyu Ointment, the invention team conducted a large number of investigations and experiments, combined with modern new production technologies and new materials, and improved and optimized the original preparation process of Baiyu Ointment, aiming to improve the quality of the drug, and enhance the safety, effectiveness and quality controllability of the drug. Summary of the Invention

[0006] The purpose of the present invention is to provide a preparation process of Baiyu Ointment to improve the quality of the drug, and enhance the safety, effectiveness and quality controllability of the drug.

[0007] To achieve the above purpose, the technical solutions adopted by the present invention are specifically as follows:

[0008] The preparation process of the Baiyu Ointment described in the present invention includes the following steps:

[0009] S1 Take medicinal materials:

[0010] Weigh 80 parts of Bletilla striata, 40 parts of Polygonatum odoratum, 40 parts of Ligusticum sinense, 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 Dry Ginger, mix them evenly, pulverize them, and sieve them through a 50-80 mesh sieve for later use;

[0011] S2 Supercritical fluid extraction:

[0012] Use the segmented method to conduct CO2 supercritical fluid extraction on the fine powder obtained in step S1, collect the extract for later use;

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

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

[0015] S3 Ultrasonic extraction:

[0016] Take the medicinal residues after supercritical fluid extraction in step S2, add 6-10 times of water, and conduct ultrasonic extraction for 30-50 min under the conditions of a frequency of 20-40 kHz and a power of 150-250 W, filter, and concentrate the filtrate to a clear paste with a relative density of 1.10 for later use;

[0017] S4 Preparation forming:

[0018] Add 8 - 12 parts of sodium hyaluronate, 60 - 100 parts of carbomer, 70 - 90 parts of beta - cyclodextrin to the clear extract obtained in step S3, heat and dissolve at 60°C for standby; separately take 80 - 120 parts of hydrogenated vegetable oil, 90 - 110 parts of olive oil, 60 - 100 parts of polysorbate 80, heat and melt at 80°C, mix with the above - mentioned clear extract; emulsify for 15 - 30 min, keep warm at 80°C for 30 min, when the temperature is lowered to 40°C, add the supercritical fluid extraction extract obtained in step S2 and 1 - 3 parts of ethylparaben, mix evenly to obtain the product.

[0019] Preferably, in the preparation process of the white jade ointment of the present invention, in step S1, after crushing, it is sieved through a 50 - 70 mesh sieve.

[0020] More preferably, in the preparation process of the white jade ointment of the present invention, in step S1, after crushing, it is sieved through a 60 - mesh sieve.

[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 specifically: 0 - 30 min, the extraction temperature is 38 - 40°C, and the extraction pressure is 28 - 30 MPa; 31 - 60 min, the extraction temperature is 42 - 45°C, and the extraction pressure is 32 - 35 MPa; 61 - 90 min, the extraction temperature is 47 - 50°C, and the extraction pressure is 37 - 40 MPa.

[0022] More 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 specifically: 0 - 30 min, the extraction temperature is 40°C, and the extraction pressure is 30 MPa; 31 - 60 min, the extraction temperature is 45°C, and the extraction pressure is 35 MPa; 61 - 90 min, the extraction temperature is 50°C, and the extraction pressure is 40 MPa.

[0023] Preferably, in the preparation process of the white jade ointment of the present invention, the ultrasonic extraction in step S3 is specifically: take the medicinal residues after supercritical fluid extraction in step S2, add 6 - 8 times of water, and perform ultrasonic extraction for 40 - 50 min under the conditions of a frequency of 20 - 30 kHz and a power of 200 - 250 W, filter, and concentrate the filtrate to a clear extract with a relative density of 1.10 for standby.

[0024] More preferably, in the preparation process of the white jade ointment of the present invention, the ultrasonic extraction in step S3 is specifically: take the medicinal residues after supercritical fluid extraction in step S2, add 6 times of water, and perform ultrasonic extraction for 40 min under the conditions of a frequency of 30 kHz and a power of 200 W, filter, and concentrate the filtrate to a clear extract with a relative density of 1.10 for standby.

[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 for forming the preparation is specifically as follows: 10-12 parts of sodium hyaluronate, 80-100 parts of carbomer, and 70-80 parts of beta-cyclodextrin are added to the clear paste obtained in step S3, heated and dissolved at 60 °C, and reserved; another 100-120 parts of hydrogenated vegetable oil, 100-110 parts of olive oil, and 90-100 parts of polysorbate 80 are taken, heated and melted at 80 °C, and mixed with the above-mentioned clear paste; emulsified for 20-30 min, kept warm at 80 °C for 30 min, and when cooled to 40 °C, the supercritical fluid extraction extract obtained in step S2 and 1.5-2.5 parts of ethylparaben are added, and mixed evenly to obtain the product.

[0027] More preferably, in the preparation process of the white jade ointment of the present invention, step S4 for forming the preparation is specifically as follows: 10 parts of sodium hyaluronate, 100 parts of carbomer, and 70 parts of beta-cyclodextrin are added to the clear paste obtained in step S3, heated and dissolved at 60 °C, and reserved; another 120 parts of hydrogenated vegetable oil, 110 parts of olive oil, and 90 parts of polysorbate 80 are taken, heated and melted at 80 °C, and mixed with the above-mentioned clear paste; emulsified for 20 min, kept warm at 80 °C for 30 min, and when cooled to 40 °C, the supercritical fluid extraction extract obtained in step S2 and 2 parts of ethylparaben are added, and mixed evenly to obtain the product.

[0028] The beneficial effects of the present invention:

[0029] 1. The present invention provides a preparation process of white jade ointment. After extraction by the original process method of white jade ointment, the contents of volatile oil, total flavonoids, and total polysaccharides in the prepared white jade ointment are 0, 0.45 mg / g, and 0.74 mg / g respectively. However, after extraction by the process method provided by the present invention, the contents of volatile oil, total flavonoids, and total polysaccharides in the prepared white jade ointment reach 3.42 mg / g, 1.07 mg / g, and 1.36 mg / g respectively, indicating that the preparation process of the white jade ointment provided by the present invention greatly improves the content of active ingredients therein, which is of great significance for improving the quality of drugs and enhancing the safety, effectiveness, and quality controllability of drugs.

[0030] 2. The inventors investigated the factors affecting the extraction of the active ingredients of Baiyu Ointment and obtained that the optimal extraction method for the active ingredients of Baiyu Ointment is to use the double-super extraction method (i.e., supercritical-ultrasonic extraction method) for extraction. Among them, the optimal process parameters for supercritical fluid extraction are to use the segmented extraction method for CO2 supercritical fluid extraction, the CO2 flow rate is 30 L / h, and the extraction parameters for the segmented method are: 0 - 30 min, the extraction temperature is 40 °C, and the extraction pressure is 30 MPa; 31 - 60 min, the extraction temperature is 45 °C, and the extraction pressure is 35 MPa; 61 - 90 min, the extraction temperature is 50 °C, and the extraction pressure is 40 MPa. The optimal process parameters for ultrasonic extraction are: adding water with a mass 6 times that of the medicinal residues; the ultrasonic frequency is 30 kHz; the ultrasonic power is 200 W; the ultrasonic time is 40 min.

[0031] 3. The present invention improves the types and dosages of excipients in Baiyu Ointment. In the original preparation process, 17 kinds of excipients were used, while in the improved preparation process of the present invention, only 7 kinds of excipients are used, and the optimal usage amounts of carbomer, hydrogenated vegetable oil, and olive oil are determined through orthogonal experiments, greatly reducing the process difficulty and production cost of Baiyu Ointment.

[0032] 4. The results of pharmacological experiments show that when the dosage of the process method group of the present invention is only 50% of that of the original process group, the therapeutic effect in the rat skin chapping model is equivalent to that of the original process group, indicating that the process method provided by the present invention can reduce the usage amount of medicinal materials and save medicinal material resources. Description of the Drawings

[0033] Figure 1 It is the standard curve graph for the determination of total flavonoids;

[0034] Figure 2 It is the standard curve graph for the determination of total polysaccharides. Detailed Description of the Invention

[0035] The technical solutions of the present invention will be described in detail below in combination 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.

[0036] Example 1

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

[0038] (1) Take medicinal materials:

[0039] Weigh 80 g of Bletilla striata, 40 g of Polygonatum odoratum, 40 g of Ligusticum sinense, 40 g of Polygonum hydropiper, 40 g of Geranium wilfordii, 20 g of Salvia miltiorrhiza, 20 g of Lithospermum erythrorhizon, 40 g of Angelica sinensis, and 40 g of Dryopteris crassirhizoma, mix them evenly, pulverize them, and pass through a 60-mesh sieve for standby.

[0040] (2) Supercritical fluid extraction:

[0041] The fine powder obtained in step (1) is subjected to CO2 supercritical fluid extraction by the segmented method, and the extract is collected for standby.

[0042] Among them, the CO2 flow rate of the CO2 supercritical fluid extraction is 30 L / h.

[0043] The extraction parameters of the segmented method are specifically as follows: 0 - 30 min, the extraction temperature is 40 °C, and the extraction pressure is 30 MPa; 31 - 60 min, the extraction temperature is 45 °C, and the extraction pressure is 35 MPa; 61 - 90 min, the extraction temperature is 50 °C, and the extraction pressure is 40 MPa.

[0044] (3) Ultrasonic extraction:

[0045] Take the medicinal residues after supercritical fluid extraction in step (2), add 6 times the amount of water, and perform ultrasonic extraction for 40 min under the conditions of a frequency of 30 kHz and a power of 200 W, filter, and concentrate the filtrate to a clear paste with a relative density of 1.10 at 60 °C for standby.

[0046] (4) Preparation of the preparation:

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

[0048] Example 2

[0049] The preparation process of the white jade ointment is as follows:

[0050] (1) Take the medicinal materials:

[0051] Weigh 80 g of bletilla striata, 40 g of polygonatum odoratum, 40 g of ligusticum sinense, 40 g of polygonum hydropiper, 40 g of geranium wilfordii, 20 g of salvia miltiorrhiza, 20 g of lithospermum erythrorhizon, 40 g of angelica sinensis, and 40 g of dried ginger, mix evenly, pulverize, and pass through a 60-mesh sieve for standby.

[0052] (2) Supercritical fluid extraction:

[0053] The fine powder obtained in step (1) is subjected to CO2 supercritical fluid extraction by the segmented method, and the extract is collected for standby.

[0054] Among them, the CO2 flow rate of the CO2 supercritical fluid extraction is 30 L / h.

[0055] The extraction parameters of the segmented method are specifically as follows: for 0 - 30 min, the extraction temperature is 35°C and the extraction pressure is 25 MPa; for 31 - 60 min, the extraction temperature is 40°C and the extraction pressure is 30 MPa; for 61 - 90 min, the extraction temperature is 45°C and the extraction pressure is 35 MPa.

[0056] (3) Ultrasonic extraction:

[0057] Take the medicinal residues after supercritical fluid extraction in step (2), add 6 times the amount of water, and perform ultrasonic extraction for 40 min under the conditions of a frequency of 30 kHz and a power of 200 W. Filter, and concentrate the filtrate to a clear paste with a relative density of 1.10 at 60°C for standby.

[0058] (4) Preparation of the preparation:

[0059] Add 10 g of sodium hyaluronate, 100 g of carbomer, and 70 g of beta-cyclodextrin to the clear paste prepared in step (3), heat and dissolve at 60°C for standby; separately take 120 g of hydrogenated vegetable oil, 110 g of olive oil, and 90 g of polysorbate 80, heat and melt at 80°C, and mix with the above-mentioned clear paste; emulsify for 20 min, keep warm at 80°C for 30 min, and when the temperature is lowered to 40°C, add the supercritical fluid extraction extract prepared in step (2) and 2 g of ethylparaben, and mix well to obtain the product.

[0060] Example 3

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

[0062] (1) Take the medicinal materials:

[0063] Weigh 80 g of Bletilla striata, 40 g of Polygonatum odoratum, 40 g of Ligusticum sinense, 40 g of Polygonum hydropiper, 40 g of Geranium wilfordii, 20 g of Salvia miltiorrhiza, 20 g of Lithospermum erythrorhizon, 40 g of Angelica sinensis, and 40 g of Dryopteris rhizome, mix well, pulverize, and pass through a 60-mesh sieve for standby.

[0064] (2) Supercritical fluid extraction:

[0065] Perform CO2 supercritical fluid extraction on the fine powder prepared in step (1) by the segmented method, collect the extract for standby.

[0066] Among them, the CO2 flow rate of CO2 supercritical fluid extraction is 30 L / h.

[0067] The extraction parameters of the segmented method are specifically as follows: for 0 - 30 min, the extraction temperature is 38°C and the extraction pressure is 28 MPa; for 31 - 60 min, the extraction temperature is 42°C and the extraction pressure is 32 MPa; for 61 - 90 min, the extraction temperature is 47°C and the extraction pressure is 37 MPa.

[0068] (3) Ultrasonic extraction:

[0069] Take the medicinal residues after supercritical fluid extraction in step (2), add 6 times the amount of water, and perform ultrasonic extraction for 40 min under the conditions of a frequency of 30 kHz and a power of 200 W. Filter, and concentrate the filtrate to a clear paste with a relative density of 1.10 at 60 °C for standby.

[0070] (4) Preparation and shaping:

[0071] Add 10 g of sodium hyaluronate, 100 g of carbomer, and 70 g of beta-cyclodextrin to the clear paste obtained in step (3), heat and dissolve at 60 °C for standby; separately take 120 g of hydrogenated vegetable oil, 110 g of olive oil, and 90 g of polysorbate 80, heat and melt at 80 °C, and mix with the above-mentioned clear paste; emulsify for 20 min, keep warm at 80 °C for 30 min, and when the temperature is lowered to 40 °C, add the supercritical fluid extraction extract obtained in step (2) and 2 g of ethylparaben, and mix evenly to obtain the product.

[0072] Example 4

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

[0074] (1) Take medicinal materials:

[0075] Weigh 80 g of Bletilla striata, 40 g of Polygonatum odoratum, 40 g of Ligusticum sinense, 40 g of Polygonum hydropiper, 40 g of Geranium wilfordii, 20 g of Salvia miltiorrhiza, 20 g of Lithospermum erythrorhizon, 40 g of Angelica sinensis, and 40 g of dried ginger, mix evenly, pulverize, and pass through a 60-mesh sieve for standby.

[0076] (2) Supercritical fluid extraction:

[0077] Perform CO2 supercritical fluid extraction on the fine powder obtained in step (1) by the segmented method, collect the extract for standby.

[0078] Among them, the CO2 flow rate for CO2 supercritical fluid extraction is 30 L / h.

[0079] The specific extraction parameters of the segmented method are as follows: 0 - 30 min, the extraction temperature is 40 °C, and the extraction pressure is 30 MPa; 31 - 60 min, the extraction temperature is 45 °C, and the extraction pressure is 35 MPa; 61 - 90 min, the extraction temperature is 50 °C, and the extraction pressure is 40 MPa.

[0080] (3) Ultrasonic extraction:

[0081] Take the medicinal residues after supercritical fluid extraction in step (2), add 10 times the amount of water, and perform ultrasonic extraction for 30 min under the conditions of a frequency of 20 kHz and a power of 150 W. Filter, and concentrate the filtrate to a clear paste with a relative density of 1.10 at 60 °C for standby.

[0082] (4) Preparation and shaping:

[0083] Add 10g of sodium hyaluronate, 100g of carbomer, and 70g of beta-cyclodextrin to the clear paste prepared in step (3), heat at 60°C to dissolve, and set aside; take another 120g of hydrogenated vegetable oil, 110g of olive oil, and 90g of polysorbate 80, heat and melt at 80°C, and mix with the above-mentioned clear paste; emulsify for 20min, keep warm at 80°C for 30min, and when the temperature drops to 40°C, add the supercritical fluid extraction extract prepared in step (2) and 2g of ethyl hydroxybenzoate, mix well, and obtain the product.

[0084] Example 5

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

[0086] (1) Take medicinal materials:

[0087] Weigh 80g of Bletilla striata, 40g of Polygonatum odoratum, 40g of Ligusticum chuanxiong, 40g of Polygonum hydropiper, 40g of Geranium herb, 20g of Salvia miltiorrhiza, 20g of Lithospermum officinale, 40g of Angelica sinensis, and 40g of dried ginger, mix well, crush, and pass through a 60-mesh sieve for later use.

[0088] (2) Supercritical fluid extraction:

[0089] The fine powder obtained in step (1) is subjected to CO2 supercritical fluid extraction by a segmented method, and the extract is collected and set aside.

[0090] Among them, the CO2 flow rate of CO2 supercritical fluid extraction is 30L / h.

[0091] The specific extraction parameters of the segmented method are: 0-30 min, extraction temperature is 40°C, extraction pressure is 30 MPa; 31-60 min, extraction temperature is 45°C, extraction pressure is 35 MPa; 61-90 min, extraction temperature is 50°C, extraction pressure is 40 MPa.

[0092] (3) Ultrasonic extraction:

[0093] Take the medicinal residue after supercritical fluid extraction in step (2), add 8 times 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 paste with a relative density of 1.10 at 60°C for later use.

[0094] (4) Preparation molding:

[0095] Add 10 g of sodium hyaluronate, 100 g of carbomer, 70 g of beta-cyclodextrin to the clear extract obtained in step (3), heat and dissolve at 60 °C, and set aside; separately take 120 g of hydrogenated vegetable oil, 110 g of olive oil, 90 g of polysorbate 80, heat and melt at 80 °C, mix with the above clear extract; emulsify for 20 min, keep warm at 80 °C for 30 min, and add the supercritical fluid extraction extract obtained in step (2) and 2 g of ethylparaben when cooling to 40 °C, mix well to obtain the product.

[0096] Example 6

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

[0098] (1) Take medicinal materials:

[0099] Weigh 80 g of Bletilla striata, 40 g of Polygonatum odoratum, 40 g of Ligusticum sinense, 40 g of Polygonum hydropiper, 40 g of Geranium wilfordii, 20 g of Salvia miltiorrhiza, 20 g of Lithospermum erythrorhizon, 40 g of Angelica sinensis, 40 g of dried ginger, mix well, pulverize, and pass through a 60-mesh sieve, and set aside.

[0100] (2) Supercritical fluid extraction:

[0101] Perform CO2 supercritical fluid extraction on the fine powder obtained in step (1) by the segmented method, collect the extract, and set aside.

[0102] Among them, the CO2 flow rate of the CO2 supercritical fluid extraction is 30 L / h.

[0103] The extraction parameters of the segmented method are specifically: 0 - 30 min, the extraction temperature is 40 °C, and the extraction pressure is 30 MPa; 31 - 60 min, the extraction temperature is 45 °C, and the extraction pressure is 35 MPa; 61 - 90 min, the extraction temperature is 50 °C, and the extraction pressure is 40 MPa.

[0104] (3) Ultrasonic extraction:

[0105] Take the medicinal residues after supercritical fluid extraction in step (2), add 6 times the amount of water, perform ultrasonic extraction for 40 min under the conditions of a frequency of 30 kHz and a power of 200 W, filter, and concentrate the filtrate to a clear extract with a relative density of 1.10 at 60 °C, and set aside.

[0106] (4) Preparation for forming:

[0107] Add 8 g of sodium hyaluronate, 80 g of carbomer, and 80 g of beta-cyclodextrin to the clear extract obtained in step (3), heat and dissolve at 60 °C for standby; separately take 100 g of hydrogenated vegetable oil, 100 g of olive oil, and 100 g of polysorbate 80, heat and melt at 80 °C, mix with the above clear extract; emulsify for 20 min, keep warm at 80 °C for 30 min, and when the temperature is lowered to 40 °C, add the supercritical fluid extraction extract obtained in step (2) and 1.5 g of ethylparaben, mix well to obtain the product.

[0108] In order to further verify the reliability of the present invention and screen out the best solution, the inventors conducted a series of tests, and some of them are excerpted as follows:

[0109] 1. Determination method for the content of active ingredients

[0110] 1.1 Determination of total volatile oil

[0111] Take the test sample, extract it twice with an appropriate amount of petroleum ether (30 - 60 °C), evaporate the petroleum ether, weigh it, and obtain the result.

[0112] 1.2 Determination of total flavonoids

[0113] (1) Preparation of reference substance solution

[0114] Take 50 mg of rutin reference substance, accurately weigh it, place it in a 25 mL volumetric flask, add an appropriate amount of methanol, heat and dissolve it in a water bath, cool it, add methanol to the scale, and shake well. Accurately measure 10 mL and place it in a 100 mL volumetric flask, add water to the scale, and shake well to obtain (each 1 mL contains 0.2 mg of rutin).

[0115] (2) Plotting of standard curve

[0116] Accurately measure 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, and 6 mL of the reference substance solution respectively, place them in 25 mL volumetric flasks, add water to each to 6.0 mL, add 1 mL of 5% sodium nitrite solution, mix well, let stand for 6 min, add 1 mL of 10% aluminum nitrate solution, shake well, let stand for 6 min, add 10 mL of sodium hydroxide solution, and then add water to the scale, shake well, and let stand for 15 min. Using the corresponding reagent as the blank, according to the ultraviolet-visible spectrophotometry, measure the absorbance at a wavelength of 500 nm. Taking the absorbance value as the ordinate and the concentration as the abscissa, plot the standard curve, as Figure 1 shown.

[0117] (3) Determination method

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

[0119] 1.3 Determination of Total Polysaccharides

[0120] (1) Preparation of Reference Solution

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

[0122] (2) Preparation of Standard Curve

[0123] Accurately measure 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL of the reference solution respectively, place them in 10 mL stoppered graduated test tubes, add water to 2.0 mL for each, shake well, slowly add 0.2% anthrone - sulfuric acid solution to the mark in an ice - water bath, mix well, cool in a water bath for 10 min after cooling, 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 Chinese Pharmacopoeia), measure the absorbance at a wavelength of 582 nm. Take the absorbance value as the ordinate and the concentration as the abscissa to draw the standard curve, as Figure 2 shown.

[0124] (3) Determination Method

[0125] Accurately 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", determine 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.

[0126] 2. Optimization of Extraction Process Parameters

[0127] 2.1 Preliminary Selection of Extraction Methods

[0128] There are 9 herbs in the prescription of Baiyu Ointment. Modern research shows that the active ingredients and pharmacological effects of each herb in the prescription for treating skin chaps are shown in Table 1.

[0129] Table 1 Active Ingredients and Pharmacological Effects of Each Herb in Baiyu Ointment

[0130]

[0131]

[0132] It is not difficult to see from the above table that the effective substances in the prescription of Baiyu Ointment that play a role in treating skin chaps are mainly of 3 categories, namely polysaccharides (Bletilla striata, Polygonatum odoratum, Angelica sinensis), volatile oils (Ligusticum sinense, Angelica sinensis, Dry ginger), and flavonoids (Polygonum hydropiper, Geranium wilfordii, Salvia miltiorrhiza). In order to extract the above 3 categories of substances to the greatest extent, the research team compared the following extraction methods:

[0133] (1) Fermentation extraction method

[0134] Weigh the medicinal materials according to the formula ratio (80 g of Bletilla striata, 40 g of Polygonatum odoratum, 40 g of Ligusticum sinense, 40 g of Polygonum hydropiper, 40 g of Geranium wilfordii, 20 g of Salvia miltiorrhiza, 20 g of Lithospermum erythrorhizon, 40 g of Angelica sinensis, 40 g of Dry ginger), crush them into coarse powder passing through a 10-mesh sieve, add water 8 times the total mass of the medicinal materials, boil and then cool, add glucose accounting for 12% of the total mass of the medicinal materials, inoculate with Saccharomyces carlsbergensis, the inoculation amount is 2.0%, ferment at 35 °C for 18 h, filter, concentrate the filtrate and make the volume constant to 250 mL, and determine the contents of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Method for determining the content of active ingredients".

[0135] (2) Supercritical fluid extraction method

[0136] Weigh the medicinal materials according to the formula ratio (the same as the fermentation extraction method), crush the medicinal materials into fine powder passing through a 60-mesh sieve. Under the conditions of a CO2 flow rate of 30 L / h, an extraction temperature of 35 °C, and an extraction pressure of 30 MPa, extract for 0.5 h, collect the extract, and make the volume constant to 250 mL with 50% ethanol solution, and determine the contents of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Method for determining the content of active ingredients".

[0137] (3) Enzymolysis method

[0138] Weigh the medicinal materials according to the formula ratio (the same as the fermentation extraction method), crush them into coarse powder passing through a 10-mesh sieve, add water 8 times the mass of the medicinal materials and 30% cellulase, extract at 55 °C and a pH value of 5.0 for 10 h, boil and then filter, concentrate the filtrate and make the volume constant to 250 mL, and determine the contents of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Method for determining the content of active ingredients".[[ID=I9]]

[0139] (4) Water extraction method

[0140] Weigh the medicinal materials according to the formula ratio (the same as the fermentation extraction method), crush them into coarse powder passing through a 10-mesh sieve, add water 8 times the mass of the medicinal materials and decoct for 1 h (simultaneously collect the volatile oil), filter, concentrate the filtrate and make the volume constant to 250 mL, and determine the contents of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Method for determining the content of active ingredients".

[0141] (5) Ultrasonic method

[0142] Weigh the medicinal materials according to the formula ratio (same as the fermentation extraction method), crush them into coarse powder passing through a 10-mesh sieve, add water with a mass 10 times that of the medicinal materials, perform ultrasonic extraction for 30 min under the conditions of a frequency of 40 kHz and a power of 250 W, filter, concentrate the filtrate and make the volume up to 250 mL, and determine the contents of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Method for determining the content of active ingredients".

[0143] (6) Ethanol extraction method

[0144] Weigh the medicinal materials according to the formula ratio (same as the fermentation extraction method), crush them into coarse powder passing through a 10-mesh sieve, add a 60% ethanol solution with a mass 8 times that of the medicinal materials, perform reflux extraction for 1 h, filter, concentrate the filtrate and make the volume up to 250 mL, and determine the contents of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Method for determining the content of active ingredients".

[0145] Determine the contents of active ingredients in the extracts obtained by the above different methods respectively. The results are shown in Table 2.

[0146] Table 2 Determination results of the contents of active ingredients in the extracts by different extraction methods

[0147]

[0148] It can be seen from the results that among the 6 extraction methods, none of them can extract the three active ingredients to the maximum extent at the same time. Although the supercritical fluid extraction method has the highest contents of total volatile oil and total flavonoids, the polysaccharide extraction rate of this method is the lowest. Considering the comprehensive test data, the supercritical fluid extraction method can be combined with the water extraction method or the ultrasonic extraction method for co-extraction. However, considering that the ultrasonic extraction process is relatively simple, the supercritical fluid extraction method combined with the ultrasonic extraction method is preferred for co-extraction, that is, the medicinal materials are first subjected to supercritical fluid extraction, and then the medicinal residues are taken for ultrasonic extraction. In order to verify the feasibility of this scheme, the research team conducted the following tests:

[0149] Weigh the medicinal materials according to the formula ratio (same as the fermentation extraction method), and weigh a total of 3 portions. Crush the medicinal materials into fine powder passing through a 60-mesh sieve respectively. Two of them are subjected to repeated tests according to the above supercritical fluid extraction method and ultrasonic extraction method respectively. For the other portion, first perform extraction for 0.5 h under the conditions of a CO2 flow rate of 30 L / h, an extraction temperature of 35 °C and an extraction pressure of 30 MPa, collect the extract, then add water with a mass 10 times that of the medicinal residues to the medicinal residues, perform ultrasonic extraction for 30 min under the conditions of a frequency of 40 kHz and a power of 250 W, filter, concentrate the filtrate, mix it with the supercritical fluid extraction extract, and make the volume up to 250 mL. Determine the contents of total volatile oil, total flavonoids and total polysaccharides according to the method under "1. Method for determining the content of active ingredients". The results are shown in Table 3.

[0150] Table 3 Verification results of different extraction methods

[0151]

[0152] As can be seen from Table 3, the supercritical fluid extraction combined with ultrasonic extraction (double-super extraction method) can significantly improve the extraction rate of total polysaccharides in the prescription, maximizing the extraction of the three active ingredients 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 shows that the types of extracts are different when CO2 supercritical fluid extraction is carried out under different pressures, temperatures and flow rates. Generally speaking, at the low temperature and low pressure stage, the density of CO2 is small, and low-polarity and low-molecular-weight components (such as volatile oils, fat-soluble pigments, etc.) are preferentially extracted. At the high temperature and high pressure stage, the density of CO2 increases, and medium-high polarity components (such as flavonoids, alkaloids, etc.) can be dissolved. Therefore, the fractional extraction (i.e., stepwise extraction) of active ingredients can be achieved by adjusting the pressure, temperature and flow rate parameters, so as to selectively extract the target components. Since the active ingredients in the Baiyu Ointment prescription include both low-polarity and low-molecular-weight volatile oils and high-polarity component flavonoids, the fractional extraction method can be used to extract the active ingredients. According to the common process parameters of the carbon dioxide supercritical fluid extraction method, the fractional extraction is planned to be carried out according to the parameter settings in Table 4.

[0156] Table 4 Table of fractional parameters for CO2 supercritical fluid extraction method

[0157]

[0158] In order to verify the fractional extraction effect, the research team carried out a verification test according to the parameters in Table 5, and the results are shown in Table 6.

[0159] Table 5 Table of parameters for the fractional verification test of CO2 supercritical fluid extraction method

[0160]

[0161] Table 6 Results of CO2 supercritical fluid fractional extraction test

[0162]

[0163] The results show that the fractional extraction using the CO2 supercritical fluid extraction method can significantly improve the extraction rates of total volatile oils and total flavonoids in the prescription. Therefore, it is determined to use the fractional extraction method for supercritical fluid extraction.

[0164] 2.2.2 Optimization of ultrasonic extraction process

[0165] Weigh the medicinal materials according to the formula ratio (960 g of Bletilla striata, 480 g of Polygonatum odoratum, 480 g of Ligusticum sinense, 480 g of Polygonum hydropiper, 480 g of Geranium wilfordii, 240 g of Salvia miltiorrhiza, 240 g of Lithospermum erythrorhizon, 480 g of Angelica sinensis, 480 g of dried ginger), crush the medicinal materials into fine powder passing through a 60-mesh sieve, and carry out supercritical fluid extraction according to the segmented extraction parameters in Table 7 under the condition of a CO2 flow rate of 30 L / h. Then divide the medicinal residues after extraction into 12 equal parts for subsequent experiments.

[0166] Table 7 Segmented extraction parameters of CO2 supercritical fluid

[0167]

[0168] (1) Investigation of water addition

[0169] Take 3 portions of the medicinal residues after extraction, add 6 times, 8 times, and 10 times water respectively, carry out ultrasonic extraction for 30 min under the conditions of a frequency of 40 kHz and a power of 250 W, filter, concentrate the filtrate and make the volume constant to 250 mL, and determine the total polysaccharide content. The results are shown in Table 8.

[0170] Table 8 Results of the investigation of water addition

[0171]

[0172] The results show that the water addition has little effect on the total polysaccharide content. Considering comprehensively, 6 times the water addition is selected for subsequent research.

[0173] (2) Investigation of ultrasonic frequency

[0174] Take 3 portions of the medicinal residues after extraction, add 6 times water to each portion, carry out ultrasonic extraction for 30 min under the conditions of frequencies of 20 kHz, 30 kHz, and 40 kHz and a power of 250 W respectively, filter, concentrate the filtrate and make the volume constant to 250 mL, and determine the total polysaccharide content. The results are shown in Table 9.

[0175] Table 9 Results of the investigation of ultrasonic frequency

[0176]

[0177] The test results show that the ultrasonic frequency also has little effect on the total polysaccharide content. Considering comprehensively, an ultrasonic frequency of 30 kHz is selected for subsequent research.

[0178] (3) Investigation of ultrasonic power

[0179] Take 3 portions of the medicinal residues after extraction, add 6 times water, with a frequency of 30 kHz, carry out ultrasonic extraction for 30 minutes under the conditions of powers of 150 W, 200 W, and 250 W respectively, filter, concentrate the filtrate and make the volume constant to 250 ml, and determine the total polysaccharide content. The results are shown in Table 10.

[0180] Table 10 Results of the investigation of ultrasonic power

[0181]

[0182] The test results show that the ultrasonic power has a great influence on the total polysaccharide content. The total polysaccharide content in the sample with a power of 150W is significantly lower than that in the samples with 200W and 250W. However, the change in the total polysaccharide content between the samples with 200W and 250W is not significant. The ultrasonic power of 200W can be selected for subsequent research.

[0183] (4) Investigation of ultrasonic time

[0184] Take 3 portions of the extracted medicinal residues, add 6 times the amount of water to each portion, and under the conditions of a frequency of 30 kHz and a power of 200W, perform ultrasonic extraction for 30 min, 40 min, and 50 min respectively. Filter, concentrate the filtrate and make up the volume to 250 mL, and measure the total polysaccharide content. The results are shown in Table 11.

[0185] Table 11 Results of the investigation of ultrasonic time

[0186]

[0187] The test results show that the ultrasonic time has a great influence on the total polysaccharide content. The total polysaccharide content in the samples with ultrasonic times of 40 min and 50 min is significantly higher than that in the sample with an ultrasonic time of 30 min. However, the change in the total polysaccharide content between the samples with ultrasonic times of 40 min and 50 min is not significant. The ultrasonic time of 40 min can be selected for subsequent research.

[0188] 3. Optimization of the matrix

[0189] The preparation process of the original Baiyu Ointment is as follows: Take the medicinal materials according to the prescription ratio, extract them into a clear paste, add 60 g of sorbitol, 60 g of beta-cyclodextrin, 60 g of glycerol, 20 g of titanium dioxide, and 3 g of sodium hyaluronate, heat (80 °C) to dissolve, and set aside; separately take 10 g of mink oil, 30 g of lanolin, 60 g of vaseline, 12 g of monoglyceride, 10 g of sucrose ester, 100 g of paraffin oil, 50 g of polysorbate 80, 50 g of span 80, and 20 g of emulsifying silicone oil, heat (80 °C) to melt, mix with the above-mentioned clear paste, emulsify for 15 min, keep warm at 80 °C for 30 min, cool down to 40 °C, then add 2 g of elastase, 6 g of benzoic acid, and 1.5 g of ethylparaben, mix well, and make 1000 g, thus obtaining it. This process uses a relatively large variety of excipients, which is not conducive to the quality control of the preparation and has too high production costs. With the development and application of new materials, it is necessary to optimize and improve the preparation forming process.

[0190] The research team plans to select sodium hyaluronate, carbomer, beta-cyclodextrin, hydrogenated vegetable oil, olive oil, and polysorbate 80 as the o / w type matrix, and additionally use ethylparaben as the preservative to carry out research on the preparation forming process.

[0191] 3.1 Preferred indicators and scoring criteria

[0192] (1) Appearance

[0193] The maximum score is 25 points. The evaluation includes even color, fine texture, appropriate consistency, spreadability, and the presence of graininess, with each item accounting for 5 points.

[0194] (2) Centrifugal stability

[0195] The full score is 25 points. Each group of ointments was placed in a centrifuge tube and centrifuged at 3000 r·min. -1 Centrifuge for 25 minutes and observe for stratification, agglomeration, foaming or water discharge.

[0196] (3) Cold resistance test

[0197] The maximum score is 5. A certain amount of ointment from each group was placed in a plastic sample cup and placed in a 4°C refrigerator for 1 month to observe whether there was any delamination, caking, or water release.

[0198] (4) Heat resistance test

[0199] The full score is 5. A certain amount of ointment from each group was placed in a plastic sample cup and placed in a constant temperature oven at 40°C for 1 month to observe for delamination, discoloration, mildew, etc.

[0200] (5) Ductility

[0201] The maximum score is 20. Take two clean glass plates measuring 10 cm x 10 cm and place a 125g weight on each plate (add weight if the weight is insufficient). Weigh 1g of each ointment, which has 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 stretched ointment and take the average ratio of the maximum to minimum diameter.

[0202] (6) pH value

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

[0204] 3.2 Orthogonal experiment

[0205] Three factors that have a greater impact on the physical and chemical properties of the ointment, namely, carbomer dosage (A), hydrogenated vegetable oil dosage (B) and olive oil dosage (C), were selected as investigation factors. The dosages of other excipients remained unchanged, the dosage of sodium hyaluronate was 10g, the dosage of beta-cyclodextrin was 70g, and the dosage of polysorbate 80 was 90g. Based on the preliminary data, each factor was taken at 3 levels for orthogonal experiments. The factor levels are shown in Table 12, the test results are shown in Table 13, and the variance analysis is shown in Table 14.

[0206] Table 12 Orthogonal Test Factors and Levels of Matrix Investigation

[0207]

[0208] Table 13 Orthogonal Test Design and Results of Matrix Investigation

[0209]

[0210] Table 14 ANOVA Results of Matrix Investigation

[0211]

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

[0213] From the orthogonal test results in Table 13, it can be seen that 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 best matrix ratio parameter is A3B3C3. At the same time, combined with the ANOVA results in Table 14, all three factors have significant effects. Therefore, the best matrix ratio parameter is carbomer dosage: hydrogenated vegetable oil dosage: olive oil = 10:12:11.

[0214] 4. Verification of the Best Process Parameters

[0215] (1) Weigh the medicinal materials

[0216] Weigh 80 g of Bletilla striata, 40 g of Polygonatum odoratum, 40 g of Ligusticum sinense, 40 g of Polygonum hydropiper, 40 g of Geranium wilfordii, 20 g of Salvia miltiorrhiza, 20 g of Lithospermum erythrorhizon, 40 g of Angelica sinensis, and 40 g of Dryopteris crassirhizoma according to the formula, and mix them evenly.

[0217] (2) Pulverize

[0218] Pulverize the medicinal materials into fine powder passing through a 60-mesh sieve.

[0219] (3) Supercritical extraction

[0220] Under the condition of a CO2 flow rate of 30 L / h, perform CO2 supercritical extraction on the above-mentioned fine powder by the segmented method. The extraction parameters are shown in Table 15. Collect the extracted extracts and set aside.

[0221] Table 15 Segmented Extraction Parameters of CO2 Supercritical Fluid

[0222]

[0223] (4) Ultrasonic extraction

[0224] Add 6 times the amount of water to the residue after extraction, and perform ultrasonic extraction for 30 min under the conditions of a frequency of 20 kHz and a power of 150 W. Filter, and concentrate the filtrate to a clear paste with a relative density of about 1.10 (60 °C) and set aside.

[0225] (5) Preparation of the Formulation

[0226] Add 10 g of sodium hyaluronate, 100 g of carbomer, and 70 g of beta-cyclodextrin to the clear extract, heat (60 °C) to dissolve, and set aside. Separately, take 120 g of hydrogenated vegetable oil, 110 g of olive oil, and 90 g of polysorbate 80, heat (80 °C) to melt, mix with the above clear extract, emulsify for 20 min, keep warm at 80 °C for 30 min, add the CO2 supercritical extract when cooling to about 40 °C, add 2 g of ethylparaben, mix well, and obtain the product.

[0227] (6) Verification Test

[0228] Prepare 3 batches of samples according to the above optimal parameters, and the results are shown in Table 16.

[0229] Table 16 Verification Results of the Optimal Process Parameters

[0230]

[0231] The results show that all indicators of the white jade ointment prepared by the present invention are good and the process is stable, indicating that the present invention is reasonable and feasible.

[0232] 5. Comparative Test before and after Process Improvement

[0233] Take the same batch of medicinal materials and prepare ointments according to the improved process and the original process respectively. The amount of crude drug equivalent per gram of the ointment prepared by both processes is 0.36 g (i.e., 360 g of medicinal materials in each prescription amount are made into 1000 g of ointment). Measure the contents of total volatile oil, total flavonoids, and total polysaccharides in the two kinds of ointments, and the results are shown in Table 17.

[0234] Table 17 Determination Results of the Contents of Active Ingredients in Different Preparation Processes

[0235]

[0236] The test results show that compared with the original preparation process of the white jade ointment, the improved process significantly increases the contents of volatile oil, total flavonoids, and total polysaccharides in the prescription.

[0237] 6. Efficacy Verification Test

[0238] 6.1 Rat Skin Chapping Model

[0239] (1) Experimental Materials

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

[0241] Drugs and reagents: The white jade ointment sample prepared according to the preparation method of the present invention; white jade ointment (Guizhou Green Sun Pharmaceutical Co., Ltd.); physiological saline; urea ointment; enzyme-linked immunosorbent assay kit; electrothermal constant temperature water bath; electrothermal constant temperature forced air drying oven; hand-held hair clipper; high-speed refrigerated centrifuge; surgical scissors; microplate reader.

[0242] (2) Test method

[0243] ①Model establishment, grouping and drug administration:

[0244] Fifty-six rats were randomly divided into a blank group, a model group, a positive group (urea ointment, evenly applied 0.1 g / cm 2 ), the original process group (evenly applied 0.1 g / cm 2 ), the present invention group 1 (0.1 g / cm 2 ), the present invention group 2 (evenly applied 0.08 g / cm 2 ), the present invention group 3 (evenly applied 0.06 g / cm 2 ); there were 8 rats in each group, and they were adaptively fed for 7 days before the experiment.

[0245] Twenty-four hours before the experiment, the back of the rats with an area of 4.0 cm × 4.0 cm was depilated. After depilation, the blank group was smeared with physiological saline, and the remaining 6 groups were continuously smeared with 3 mol / L sodium hydroxide solution at the depilated area for 8 days to cause the exposed skin of the rats to crack and establish a wound model. After the model was successfully established, the model group was not given medicine and allowed to recover naturally. The positive group was smeared with urea ointment, and the administration groups were smeared with the corresponding doses of drugs, once in the morning and once in the evening. After 7 consecutive days, blood was taken from the abdominal aorta, and the upper serum was taken after centrifugation for standby. The relevant inflammatory factors interleukin IL-6, IL-10, nitric oxide (NO), nitric oxide synthase (NOS), and tumor necrosis factor TNF-α were detected by ELISA method, and the operation was carried out strictly according to the requirements of the kit instructions.

[0246] ②Gross observation: One hour after drug administration on the last day, the drug was removed, the back skin was washed, and the skin was scored. The scoring criteria were: 0 points, normal color, no swelling; 1 point, slight swelling; 2 points, swelling, dry and wrinkled; 3 points, exudate, scratching the back skin and showing restlessness.

[0247] ③Statistical method: The data were processed using SPSS 17.0 statistical software. The data were expressed as x±s. The t-test was used for inter-group comparison, and the Ridit test was used for ranked data. P < 0.01 was considered statistically significant.

[0248] (3) Test results

[0249] ① Effects on the rat skin chapping model: In the blank group, everything was normal locally. After Ridit test, compared with the blank group, there were significant local redness and swelling, and exudation in the model group (P < 0.01); compared with the model group, the original process group, Invention Group 1, Invention Group 2, and urea ointment group all significantly inhibited skin redness and swelling, and 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 18.

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

[0251]

[0252] ② Effects on the contents 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, urea ointment group, Invention Group 1, Invention Group 2, and Invention Group 3 could significantly reduce the levels of IL-6, IL-10, 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, indicating again 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 test group on the levels of IL-6, IL-10, TNF-α, NOS, and NO in the rat skin chapping model (x ± s, n = 8)

[0254]

[0255] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, based on the present invention, some modifications or improvements can be made, 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 required by the present invention.

Claims

1. A preparation process of white jade ointment, characterized in that It includes the following steps: S1 Taking medicinal materials: Weigh 80 parts of Bletilla striata, 40 parts of Polygonatum odoratum, 40 parts of Ligusticum sinense, 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 evenly, pulverize them, and sieve them through a 50 - 80 mesh sieve for standby; S2 Supercritical fluid extraction: Use the segmented method to perform CO2 supercritical fluid extraction on the fine powder obtained in step S1, and collect the extract for standby; The CO2 flow rate of the CO2 supercritical fluid extraction is 30 L / h; The specific extraction parameters of the segmented method are as follows: 0 - 30 min, the extraction temperature is 35 - 40 °C, and the extraction pressure is 25 - 30 MPa; 31 - 60 min, the extraction temperature is 40 - 45 °C, and the extraction pressure is 30 - 35 MPa; 61 - 90 min, the extraction temperature is 45 - 50 °C, and the extraction pressure is 35 - 40 MPa; S3 Ultrasonic extraction: Take the medicinal residues after supercritical fluid extraction in step S2, add 6 - 10 times the amount of water, and perform ultrasonic extraction for 30 - 50 min under the conditions of a frequency of 20 - 40 kHz and a power of 150 - 250 W. Filter, and concentrate the filtrate to a clear paste with a relative density of 1.10 for standby; S4 Preparation forming: Add 8 - 12 parts of sodium hyaluronate, 60 - 100 parts of carbomer, and 70 - 90 parts of beta - cyclodextrin to the clear paste obtained in step S3, heat and dissolve it at 60 °C for standby; Separately, take 80 - 120 parts of hydrogenated vegetable oil, 90 - 110 parts of olive oil, and 60 - 100 parts of polysorbate 80, heat and melt them at 80 °C, and mix them with the above - mentioned clear paste; Emulsify for 15 - 30 min, keep warm at 80 °C for 30 min, and when the temperature drops to 40 °C, add the supercritical fluid extraction extract obtained in step S2 and 1 - 3 parts of ethylparaben, and mix evenly to obtain the product.

2. The preparation process according to claim 1, characterized in that, In step S1, after pulverization, sieve through a 50 - 70 mesh sieve.

3. The preparation process according to claim 2, wherein In step S1, after pulverization, sieve through a 60 mesh sieve.

4. The preparation process according to claim 1, wherein The specific extraction parameters of the segmented method in step S2 are as follows: 0 - 30 min, the extraction temperature is 38 - 40 °C, and the extraction pressure is 28 - 30 MPa; 31 - 60 min, the extraction temperature is 42 - 45 °C, and the extraction pressure is 32 - 35 MPa; 61 - 90 min, the extraction temperature is 47 - 50 °C, and the extraction pressure is 37 - 40 MPa.

5. The preparation process according to claim 4, characterized in that, The specific extraction parameters of the segmented method in step S2 are as follows: 0 - 30 min, the extraction temperature is 40 °C, and the extraction pressure is 30 MPa; 31 - 60 min, the extraction temperature is 45 °C, and the extraction pressure is 35 MPa; 61 - 90 min, the extraction temperature is 50 °C, and the extraction pressure is 40 MPa.

6. The preparation process according to claim 1, characterized in that, The ultrasonic extraction in step S3 is specifically as follows: Take the medicinal residues after supercritical fluid extraction in step S2, add 6 - 8 times the amount of water, and perform ultrasonic extraction for 40 - 50 min under the conditions of a frequency of 20 - 30 kHz and a power of 200 - 250 W. Filter, and concentrate the filtrate to a clear paste with a relative density of 1.10 for standby.

7. The preparation process according to claim 6, characterized in that, Step S3 ultrasonic extraction is specifically as follows: Take the medicinal residues after supercritical fluid extraction in step S2, add 6 times the amount of water, and perform ultrasonic extraction for 40 min under the conditions of a frequency of 30 kHz and a power of 200 W. Then filter, and concentrate the filtrate to a clear paste with a relative density of 1.10 for standby.

8. The preparation process according to any one of claims 1 or 6 or 7, characterized in that, The relative density mentioned in step S3 is the measured value at 60 °C.

9. The preparation process according to claim 1, wherein, Step S4 formulation molding is specifically as follows: Add 10 - 12 parts of sodium hyaluronate, 80 - 100 parts of carbomer, and 70 - 80 parts of beta-cyclodextrin to the clear paste obtained in step S3, heat and dissolve at 60 °C for standby; separately take 100 - 120 parts of hydrogenated vegetable oil, 100 - 110 parts of olive oil, and 90 - 100 parts of polysorbate 80, heat and melt at 80 °C, and mix with the above-mentioned clear paste; emulsify for 20 - 30 min, keep warm at 80 °C for 30 min, and when the temperature is lowered to 40 °C, add the supercritical fluid extraction extract obtained in step S2 and 1.5 - 2.5 parts of ethylparaben, and mix evenly to obtain the product.

10. The preparation process according to claim 9, characterized in that, Step S4 formulation molding is specifically as follows: Add 10 parts of sodium hyaluronate, 100 parts of carbomer, and 70 parts of beta-cyclodextrin to the clear paste obtained in step S3, heat and dissolve at 60 °C for standby; separately take 120 parts of hydrogenated vegetable oil, 110 parts of olive oil, and 90 parts of polysorbate 80, heat and melt at 80 °C, and mix with the above-mentioned clear paste; emulsify for 20 min, keep warm at 80 °C for 30 min, and when the temperature is lowered to 40 °C, add the supercritical fluid extraction extract obtained in step S2 and 2 parts of ethylparaben, and mix evenly to obtain the product.

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