Compound lithospermum ointment based on Yunnan lithospermum and preparation process of compound lithospermum ointment

Abstract

The invention discloses a compound lithospermum ointment based on Yunnan lithospermum erythrorhizon and a preparation process thereof, and relates to the technical field of traditional Chinese medicine external preparations, in terms of raw materials, the parameter range of Yunnan lithospermum erythrorhizon and the like is determined, and the quality stability of the raw materials is guaranteed; the extraction process adopts a 15-20 DEG C low-temperature light-shielding oxygen-isolation cold soaking combined pipeline type ultrasonic-assisted extraction process, core parameters are set, the extraction rate of alkannin is increased, and heat-sensitive active ingredients are reserved to the maximum extent; the compound compatibility strictly follows the principle of monarch, minister, assistant and guide, the weight part range of each functional component is defined, and multi-target synergistic interaction is realized; in the aspect of product safety, related parameters such as attributes of raw materials of the whole formula, pH value of the finished product and irritation are strictly limited; an exclusive HPLC fingerprint spectrum quality control system is established, and accurate regulation and control of functional components of a finished product and whole-process quality tracing are achieved; a complete process for preparing a traditional Chinese medicine dyeing product from radix arnebiae decoction dregs is developed, and high-valued cyclic utilization of solid wastes of traditional Chinese medicinal materials is realized.

Description

Technical Field

[0001] This invention relates to the field of external preparations of traditional Chinese medicine, specifically to a compound gromwell ointment based on Yunnan gromwell root and its preparation process. Background Technology

[0002] Purple gromwell ointment, a traditional Chinese medicine external preparation, has core effects of anti-inflammation and repairing skin wounds, and is widely used in daily skin care and emergency protection scenarios. However, existing technologies have many shortcomings: Poor raw material compatibility: Some products illegally use the plant Lithospermum erythrorhizon, violating resource protection policies; or they use ordinary Lithospermum erythrorhizon raw materials, which have low content of effective ingredients and poor efficacy.

[0003] The formulas are simple and the ingredients are mixed: most of the commercially available products are single-ingredient comfrey infusion preparations, lacking compound synergistic effects design, and 35% of them have exaggerated claims, while 28% have illegally added chemical ingredients such as hormones and preservatives, posing safety hazards.

[0004] Outdated extraction technology: Traditional extraction methods have low mass transfer efficiency, with an extraction rate of less than 70% for shikonin, and are prone to component degradation; some processes use organic solvents, and the residual toxicity conflicts with the safety requirements of maternal and infant care.

[0005] Limitations in efficacy and applicable scenarios: The core effects such as relieving itching and repairing are slow to respond and cannot meet immediate emergency needs; the product has poor compatibility and it is difficult to ensure safe use by sensitive groups such as children and pregnant women.

[0006] Low resource utilization: The extracted Lithospermum erythrorhizon residue has not been utilized as a resource, which is inconsistent with the concept of green circular economy. Moreover, most of the authentic medicinal materials are circulated as primary raw materials, and there is a lack of added value from deep processing. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a compound gromwell ointment based on Yunnan gromwell root and its preparation process. Regarding raw materials, the parameters of Yunnan gromwell root are clearly defined to ensure stable raw material quality. The extraction process employs a combination of low-temperature, light-proof, oxygen-free cold soaking at 15-20℃ and ultrasonic-assisted extraction in a pipeline, setting core parameters to improve the extraction rate of gromwell extract and maximize the retention of heat-sensitive active ingredients. The compound formulation strictly follows the principles of principal, assistant, adjuvant, and guide herbs, clearly defining the weight range of each active ingredient to achieve synergistic effects across multiple targets. Regarding product safety, strict limits are placed on the properties of all raw materials in the formula, the pH value of the finished product, and irritation-related parameters. A dedicated HPLC fingerprint quality control system is established to achieve precise control of the active ingredients in the finished product and full-process quality traceability. A complete process for preparing traditional Chinese medicine dyes from gromwell root residue is developed, realizing the high-value recycling of traditional Chinese medicine solid waste.

[0008] To solve the above-mentioned technical problems, the present invention provides the following technical solution: On the one hand, a compound gromwell ointment based on Yunnan gromwell root, comprising, by weight: 10-15 parts of Yunnan gromwell root extract, 3-5 parts of angelica root extract, 2-3 parts of rehmannia root extract, 2-4 parts of saposhnikovia root extract, 1-2 parts of myrrh extract, 1-2 parts of frankincense extract, 80-100 parts of cold-pressed virgin coconut oil, 15-20 parts of organic white beeswax, 10-15 parts of organic unrefined shea butter, 3-5 parts of organic vitamin E, and 1.1-2 parts of natural essential oil.

[0009] Furthermore, the Yunnan Lithospermum extract is derived from Lithospermum yunnanense, a plant belonging to the genus Lithospermum of the Boraginaceae family. It is dried at a low temperature of 30-40℃ until the moisture content is ≤8.0%, and the total shikonin content in the extract is ≥83%. The total shikonin is the sum of L-shikonin, β,β'-dimethylacryloyl shikonin, acetylshikonin, and isobutyrylshikonin.

[0010] Furthermore, the natural essential oil is composed of 0.5-1 parts organic real lavender essential oil, 0.3-0.5 parts organic peppermint essential oil, and 0.3-0.5 parts organic Roman chamomile essential oil; wherein, the real lavender essential oil contains ≥30% linalyl acetate, the peppermint essential oil contains ≥50% menthol, and the Roman chamomile essential oil contains ≥1.0% chamomile azulene.

[0011] A preparation process for a compound gromwell ointment based on Yunnan gromwell root includes the following steps: S1. Standardized pretreatment of raw materials: Yunnan gromwell is harvested, impurities are removed, dried at a low temperature of 30-40℃, fed at 20-25℃ and pulverized through a 40-60 mesh sieve to obtain Yunnan gromwell powder with a particle size D90 of 200-250μm; Angelica sinensis, Rehmannia glutinosa, Saposhnikovia divaricata, Myrrh, and Boswellia carterii are cleaned and dried respectively to prepare corresponding extracts for later use; S2. Low-temperature, light-protected cold maceration combined with pipeline ultrasonic-assisted extraction: Mix Yunnan purple gromwell powder with cold-pressed virgin coconut oil at a material-to-liquid ratio of 1:50-80, and purge with nitrogen 2-4 times under vacuum of -0.08~-0.1MPa until the oxygen content in the tank is 0.5-1.0%. Cold maceration is carried out at a constant temperature of 15-20℃ in the dark for 7-9 months, with stirring every 10-20 days. After cold maceration, ultrasonic-assisted extraction is carried out using a pipeline ultrasonic extraction unit. The prepared extracts of various excipients are added, and ultrasonication is continued for 20-30 minutes to obtain a mixture of active ingredients in the oil phase. S3. HPLC fingerprint chromatogram optimization: Establish exclusive fingerprint chromatograms for raw materials and finished products using high-performance liquid chromatography, adjust the proportions of each active ingredient, and ensure that the similarity between the fingerprint chromatogram of the test sample and the standard control fingerprint chromatogram is ≥0.95, and the batch-to-batch quality deviation is ≤5%. S4. Ointment Preparation and Aseptic Filling: The oil phase mixture is filtered through three stages to recover the purple gromwell residue, and the filtrate is transferred to an aseptic mixing tank. Organic white beeswax and organic unrefined shea butter are melted in a water bath at 50-60℃ and added to the mixing tank. After being mixed evenly, the mixture is cooled to 35-40℃, and organic vitamin E and natural essential oils are added. After being homogenized and emulsified under high pressure at 20-40MPa 1-3 times, the mixture is aseptically filled in a Class 100 clean environment to obtain the finished product.

[0012] Furthermore, in S1, the Angelica sinensis extract is prepared by supercritical CO2 extraction with the following parameters: extraction pressure 20-30 MPa, extraction temperature 30-40℃, extraction time 1.5-2.5 h, separation pressure 6-10 MPa, and separation temperature 25-35℃; the Rehmannia glutinosa extract is prepared by water extraction and alcohol precipitation with the following parameters: water volume 8-12 times, extraction temperature 30-40℃, extraction time 1-2 h, and alcohol precipitation concentration 55-65%; the Saposhnikovia divaricata extract is prepared by water extraction and alcohol precipitation with the following parameters: water volume 10-14 times, reflux extraction time 0.5-1.5 h, and alcohol precipitation concentration 65-75%; and the myrrh and frankincense extracts are prepared by low-temperature oil extraction with the following parameters: oil volume 6-10 times, extraction temperature 30-40℃, and extraction time 1.5-2.5 h.

[0013] Furthermore, in S2, the Yunnan gromwell powder and cold-pressed virgin coconut oil are mixed at a material-to-liquid ratio of 1:50-80.

[0014] Furthermore, in S2, the stirring parameters for the cold soaking process are: rotation speed 20-40 r / min, and stirring time 20-40 min per stirring session.

[0015] Furthermore, in S3, the HPLC detection parameters are as follows: C18 column; mobile phase: methanol-0.1% phosphoric acid aqueous solution, volume ratio 80:20-90:10; flow rate: 0.8-1.2 mL / min; detection wavelength: 270-280 nm; column temperature: 25-35℃; injection volume: 5-20 μL; running time: 30-50 min.

[0016] Furthermore, in S4, the three-stage filtration sequentially employs 180-220 mesh filter cloth, 280-320 mesh filter cloth, and 0.22-0.45μm polypropylene sterilization filter element; the aseptic filling environment parameters are: temperature 16-24℃, relative humidity 40-60%, filling volume deviation ≤±1%~±3%, and sealing temperature 110-140℃.

[0017] Furthermore, in step S4, the recovered Lithospermum erythrorhizon residue is used to prepare traditional Chinese medicine dyeing cultural and creative products. The specific process parameters are as follows: the recovered Lithospermum erythrorhizon residue is dried at a low temperature of 30-40℃ to a moisture content of 8.0-10.0%, pulverized through an 18-22 mesh sieve, and mixed with a 40-60% ethanol aqueous solution at a material-to-liquid ratio of 1:15-1:25. The mixture is stirred and extracted 1-3 times at 30-40℃, each time for 0.5-1.5 hours. After combining the extracts, the mixture is concentrated at a low temperature below 40℃ to a solid content of 8-12% to obtain Lithospermum erythrorhizon dyeing solution. The Lithospermum erythrorhizon dyeing solution is used to perform low-temperature traditional Chinese medicine dyeing on textile fabrics or special papers. The dyeing parameters are as follows: dyeing solution volume 15-25% (owf), salt 8-12 g / L, bath ratio 1:40-1:60, dyeing temperature 30-40℃, and dyeing time 40-80 min.

[0018] Compared with existing technologies, this compound gromwell ointment based on Yunnan gromwell root and its preparation process have the following beneficial effects: I. This invention ensures high activity and batch stability of the comfrey ointment from the source by precisely limiting the range of parameters throughout the entire process. The total shikonin content of the extract is high, and the content of active ingredients varies little between batches of finished products. It strictly follows the principle of monarch, minister, assistant, and guide in the formulation, scientifically sets the weight range of each functional ingredient, and achieves synergistic effects on multiple targets, significantly improving the repair efficiency of chronic eczema, etc. Through low-temperature, light-proof, oxygen-free cold soaking combined with pipeline ultrasonic-assisted extraction process, the parameters are precisely designed to achieve high extraction rate and activity retention rate of shikonin. It limits the safety parameter range of raw material properties and pH value of the finished product, uses organic and natural raw materials, and is suitable for all age groups without the risk of allergies. It establishes a dedicated HPLC fingerprint quality control system to achieve full-process quality traceability and ensure quality stability.

[0019] Other advantages, objectives and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination or study, or may be learned from the practice of the invention. Attached Figure Description

[0020] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0021] Figure 1 A flowchart illustrating the preparation process of a compound gromwell ointment based on Yunnan gromwell root; Figure 2 This is a flowchart of a compound gromwell ointment based on Yunnan gromwell root and its preparation process. Detailed Implementation

[0022] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features, and effects of the present invention, in conjunction with the accompanying drawings and preferred embodiments, is provided below. Example

[0023] The compound gromwell ointment based on Yunnan gromwell root in this embodiment, by weight, uses the lower limit of the parameter range of this invention. The specific formula is as follows: 10 parts Yunnan gromwell root extract, 3 parts Angelica sinensis extract, 2 parts Rehmannia glutinosa extract, 2 parts Saposhnikovia divaricata extract, 1 part Myrrh extract, 1 part Frankincense extract, 80 parts cold-pressed virgin coconut oil, 15 parts organic white beeswax, 10 parts organic unrefined shea butter, 3 parts organic vitamin E, and 1.1 parts natural essential oils; wherein, the natural essential oils consist of 0.5 parts organic real lavender essential oil, 0.3 parts organic peppermint essential oil, and 0.3 parts organic Roman chamomile essential oil; the Yunnan gromwell root extract is obtained by low-temperature closed-loop hot air drying of Yunnan gromwell root at 35°C, and the total gromwell content in the extract is 83.1%.

[0024] like Figure 1 As shown, the preparation method in this embodiment is as follows: S1. Standardized pretreatment of raw materials: Microscopic identification and origin tracing of the raw material of Lithospermum yunnanense were performed to confirm that it was free of mold, insect infestation and pesticide residues. A closed-loop low-temperature hot air dryer is used, with an inlet air temperature of 30℃ and a drying time of 12 hours, drying the material to a moisture content of 7.9%. Throughout the drying process, the inlet and outlet of the equipment are wrapped with black light-proof cloth to prevent light exposure. A low-temperature ultrafine pulverizer was used, with the feed temperature set at 20℃ and the pulverizing speed at 3000 r / min. After pulverization, the powder was passed through a 40-mesh standard inspection sieve to obtain Yunnan Lithospermum powder. The particle size D90 of the powder was measured to be 248 μm using a laser particle size analyzer, and the moisture content was measured to be 6.9% using a Karl Fischer moisture analyzer. The powder was then sealed with food-grade nitrogen and stored in a light-proof cold storage at 4℃ for later use.

[0025] Angelica sinensis slices were cleaned and impurities removed, then cut into 2mm thick slices and dried at 30℃ with a forced air drying process until the moisture content reached 8.7%. A supercritical CO2 extraction device was used with an extraction pressure of 20MPa, an extraction temperature of 30℃, an extraction time of 2.5h, a separation pressure of 6MPa, and a separation temperature of 25℃. The pale yellow oily extract was collected, and the content of artemisinin was determined by HPLC to be 1.06%, which met the quality control requirements. The extract was then sealed in nitrogen and stored at 2℃ in the dark. Fresh Rehmannia glutinosa was washed, cut into 2mm thin slices, and extracted with 8 times the amount of deionized water. The mixture was stirred and extracted three times at 30℃ for 2 hours each time. The extracts were combined and filtered through a 200-mesh filter cloth. The filtrate was clarified by microfiltration through a 0.22μm ceramic membrane and then concentrated under reduced pressure at 35℃ to a relative density of 1.08 (25℃). Anhydrous ethanol was added until the alcohol concentration reached 55%. The mixture was allowed to stand at 2℃ for 14 hours. The supernatant was collected to recover the ethanol and dried under vacuum at 30℃ to a moisture content of 4.8%. The catalpol content was determined by HPLC to be 0.84%. Take the medicinal slices of Saposhnikovia divaricata, clean and select them, and cut them into 2mm thin slices. Add 10 times the amount of deionized water and reflux extract three times, 1.5h each time. Combine the extracts, filter through a 300-mesh filter, concentrate the filtrate to a relative density of 1.06 (25℃), add anhydrous ethanol to a precipitation concentration of 65%, let stand at 2℃ for 14h, take the supernatant to recover the ethanol, and dry it under vacuum at 35℃ to a moisture content of 4.7%. HPLC analysis showed that the content of cimicifugoside was 0.22%. Myrrh and frankincense resin were pulverized and passed through a 30-mesh sieve. Six times the amount of cold-pressed virgin coconut oil was added to each, and the mixture was stirred and extracted three times at 30°C in the dark for 2.5 hours each time. The mixture was filtered through a 200-mesh filter cloth, and the filtrates were combined to obtain myrrh and frankincense oil-soluble extracts. The volatile oil content of the myrrh extract was 5.3%, and the total boswellic acid content of the frankincense extract was 5.2%.

[0026] like Figure 2 As shown, S2, low-temperature, light-protected cold soaking combined with tubular ultrasonic-assisted extraction: A 500L food-grade 304 stainless steel light-proof and airtight cold soaking tank was used. 10kg of Yunnan purple gromwell powder and 500L of cold-pressed virgin coconut oil were added at a material-to-liquid ratio of 1:50. The tank opening was closed, and a vacuum pump was turned on to evacuate to a vacuum degree of -0.08MPa. The vacuum pump was then turned off, and food-grade nitrogen was introduced to atmospheric pressure. This process was repeated twice. Finally, the oxygen content inside the tank was measured to be 1.0%, and the tank opening was sealed. The jacket of the cold immersion tank is circulated with 15℃ constant temperature cooling water to precisely control the temperature of the material inside the tank to be constant at 15±0.5℃. The entire process is carried out in the dark for 7 months of cold immersion. Every 20 days, the pneumatic agitator at the bottom of the tank is turned on, with the stirring speed set at 20r / min and the stirring time at 40min to ensure that the powder and oil are in full contact and there is no sedimentation or agglomeration. After cold soaking, the cold soaking mixture is transferred to a food-grade pipeline ultrasonic extraction unit through a closed pipeline. The ultrasonic parameters are set as follows: power 300W, frequency 35kHz, jacket cooling water to control material temperature 15℃, material flow rate 30L / h, and continuous ultrasonic extraction for 60min. The ultrasonically extracted liquid was transferred to a 1000L sterile light-proof stirring tank. The pretreated Angelica sinensis, Rehmannia glutinosa, Saposhnikovia divaricata, Commiphora myrrha, and Boswellia carterii extracts were added according to the formula. The stirring was turned on at a low speed of 40r / min. The ultrasonic device in the tank was turned on at the same parameters as before. The ultrasonication was continued for 30min to obtain the oil phase mixture of the active ingredients. The total shikonin content in the oil phase mixture was determined by HPLC. The shikonin extraction rate was calculated to be 92.1%, and the total shikonin content was 1.22 mg / g.

[0027] S3, HPLC fingerprint chromatogram ratio optimization: Weigh 10 mg each of levorotatory shikonin, β,β'-dimethylacryloyl arcinin, acetyl shikonin, isobutyryl shikonin, artemisinin lactone, and catalpol reference standards, and place them in separate 10 mL amber volumetric flasks. Dissolve them in chromatographic grade methanol and dilute to the mark to prepare 1 mg / mL single-standard stock solutions. Accurately measure 0.2 mL of each single-standard stock solution and place it in a 10 mL amber volumetric flask. Dilute to the mark with chromatographic grade methanol to prepare a mixed reference solution containing 20 μg of each reference standard per mL. Store at 4 °C protected from light. Measure 1.0 g of the oil phase mixture and place it in a 25 mL brown volumetric flask. Add chromatographic grade n-hexane to dissolve and dilute to the mark. Shake well and filter through a 0.22 μm organic phase filter membrane to obtain the test solution. The entire process should be carried out in the dark. An Agilent 1260 high-performance liquid chromatograph was used, with a C18 column (250 mm × 4.6 mm, 5 μm); the mobile phase was methanol-0.1% phosphoric acid aqueous solution; the flow rate was 0.8 mL / min; the detection wavelength was 270 nm; the column temperature was 25 ℃; the injection volume was 20 μL; and the run time was 50 min. Fingerprint chromatogram evaluation: The chromatogram of the test sample was recorded, relevant regulations were imported, and it was compared with the standard control fingerprint chromatogram established in this invention. The similarity was calculated to be 0.96, and the batch-to-batch quality deviation was 4.2%, both of which met the quality control requirements and no adjustment of the ratio was required.

[0028] S4. Paste preparation and aseptic filling: The qualified oil phase mixture was filtered in three stages using a plate and frame filter press. The first stage used 180-mesh food-grade polypropylene filter cloth, the second stage used 280-mesh food-grade polypropylene filter cloth, and the third stage used 0.45μm polypropylene sterilization filter cartridge. After filtration, the purple gromwell filter residue was sealed and recovered. The filtrate was transferred through a closed pipeline to a 500L sterile light-proof mixing tank in a Class 100 clean area. Weigh out 15 kg of organic white beeswax and 10 kg of organic unrefined shea butter according to the formula, add them to a water bath melting tank, turn on the 50℃ constant temperature water bath, set the stirring speed to 60 r / min, stir for 30 min until completely melted and no visible solid particles are left; after melting, filter through a 280 mesh filter cloth, transfer the filtrate to a sterile mixing tank, mix it with the oil phase filtrate, turn on the stirring, set the speed to 60 r / min, stir for 40 min until completely mixed and uniform, and control the temperature of the material in the tank to be stable at 50℃; Turn on the low-temperature cooling water in the jacket of the mixing tank to cool the material to 35°C at a uniform speed. Add 3kg of organic vitamin E and 1.1kg of natural essential oil according to the formula. Turn on the low-speed stirring at 40r / min and stir for 30min until completely mixed. Keep the process away from light and continuously fill with nitrogen for protection. High-pressure homogenization emulsification: The mixture is transferred into a sterile high-pressure homogenizer through a closed pipeline. The homogenization pressure is set to 20MPa, and the homogenization is carried out three times at room temperature. The material temperature is controlled to be ≤30℃ throughout the process to obtain a semi-finished paste with a fine texture and no particles. The semi-finished product underwent comprehensive testing. It was found to be a purplish-red, fine paste. The pH value of the 10% ethanol suspension at 25℃ was 5.6, and the total shikonin content was 1.07 mg / g. All indicators were qualified. The fully automatic aseptic tube filling and sealing machine is used for filling in a Class 100 clean environment. The filling environment is controlled as follows: temperature 16℃, relative humidity 60%, and settling bacteria ≤1 CFU / plate. The filling specification is set at 15g / tube, the filling volume deviation is ≤±3%, the sealing temperature is 110℃, and the sealing is tight without leakage or wrinkles. After filling, the tubes are inspected by light, and unqualified products are removed. After being sealed in aluminum foil bags and packaged, the finished compound comfrey ointment is obtained. Example

[0029] The compound gromwell ointment based on Yunnan gromwell root in this embodiment, by weight, uses the midpoint of the parameter range of this invention. The specific formula is as follows: 12.5 parts Yunnan gromwell root extract, 4 parts Angelica sinensis extract, 2.5 parts Rehmannia glutinosa extract, 3 parts Saposhnikovia divaricata extract, 1.5 parts Myrrh extract, 1.5 parts Frankincense extract, 90 parts cold-pressed virgin coconut oil, 17.5 parts organic white beeswax, 12.5 parts organic unrefined shea butter, 4 parts organic vitamin E, and 1.55 parts natural essential oils; wherein, the natural essential oils consist of 0.75 parts organic true lavender essential oil, 0.4 parts organic peppermint essential oil, and 0.4 parts organic Roman chamomile essential oil; the Yunnan gromwell root extract is dried by low-temperature closed-loop hot air drying of Yunnan gromwell root at 35℃, and the total gromwell content in the extract is 84.3%.

[0030] The preparation method in this embodiment is as follows: S1. Standardized pretreatment of raw materials: Microscopic identification and origin tracing were performed on raw materials of Lithospermum erythrorhizon from Dali, Yunnan, confirming that they were free from mold, insect infestation, and pesticide residues. A closed-loop low-temperature hot air dryer is used, with the inlet air temperature set at 35℃ and the drying time at 10 hours, drying the material to a moisture content of 7.0%, and the entire drying process is conducted in the dark. A low-temperature ultrafine pulverizer was used, with the feed temperature set at 22℃ and the pulverizing speed at 3200 r / min. After pulverization, the powder was passed through a 50-mesh standard inspection sieve to obtain Yunnan Lithospermum powder. The particle size was measured to be D90=225μm by a laser particle size analyzer, and the moisture content was measured to be 6.5% by a Karl Fischer moisture analyzer. The powder was then sealed with nitrogen and stored in a light-proof cold storage at 4℃ for later use.

[0031] Angelica sinensis slices were cleaned and impurities removed, with a slice thickness of 2.5 mm, and dried at 35℃ to a moisture content of 8.2%. Supercritical CO2 extraction parameters were as follows: extraction pressure 25 MPa, extraction temperature 35℃, extraction time 2 h, separation pressure 8 MPa, and separation temperature 30℃. The extract was collected, and the content of artemisinin was determined by HPLC to be 1.18%. The extract was then stored under nitrogen and sealed at 4℃ in the dark. Take fresh Rehmannia glutinosa slices, add 10 times the amount of deionized water, and extract twice at 35℃ for 1.5 hours each time. Combine the extracts, filter through a 300-mesh filter, microfilter through a ceramic membrane, and concentrate under reduced pressure at 35℃ to a relative density of 1.10 (25℃). Add ethanol to a precipitation concentration of 60%, let stand at 4℃ for 12 hours, recover the ethanol from the supernatant, and dry under vacuum at 35℃ to a moisture content of 4.5%. HPLC analysis showed that the catalpol content was 0.91%. Store in a sealed container at 4℃. Take slices of Saposhnikovia divaricata, add 12 times the amount of deionized water, reflux and extract twice, 1 hour each time, combine the extracts, filter and concentrate to a relative density of 1.08 (25℃), add ethanol to a precipitation concentration of 70%, let stand at 4℃ for 12 hours, recover the ethanol from the supernatant, dry at 35℃ under vacuum to a moisture content of 4.6%, and detect the content of Cimicifuga glycosides by HPLC as 0.25%. Store in a sealed container. Myrrh and frankincense resin were pulverized and passed through a 40-mesh sieve. Eight times the amount of cold-pressed coconut oil was added to each, and the mixture was stirred and extracted twice at 35°C in the dark for 2 hours each time. The mixture was then filtered through a 300-mesh sieve. The filtrates were combined, and the volatile oil content of the myrrh extract was found to be 5.8%, and the total boswellic acid content of the frankincense extract was 5.6%. The mixtures were then stored in a nitrogen-filled, sealed container in the dark.

[0032] S2. Low-temperature, light-protected cold soaking combined with tubular ultrasound-assisted extraction: In a 1000L stainless steel light-proof cold soaking tank, add 12.5kg of Yunnan purple gromwell powder and 812.5L of cold-pressed coconut oil at a material-to-liquid ratio of 1:65. After vacuuming to -0.09MPa, purge with nitrogen three times to achieve an oxygen content of 0.75% in the tank and seal the tank opening. The jacket is circulated with constant temperature cooling water at 17.5℃, and the temperature inside the tank is controlled at 17.5±0.5℃. It is then cold-soaked in the dark for 8 months, and the agitator is turned on every 15 days at a speed of 30r / min for 30min. After cold soaking, the material is transferred to a pipeline ultrasonic extraction unit with the following parameters: 400W, 40kHz, material temperature 20℃, flow rate 50L / h, and continuous ultrasonication for 45min. The extract was transferred to a sterile mixing tank, and the extracts of each excipient were added. The mixture was stirred at 60 r / min and simultaneously sonicated for 25 min to obtain an oil phase mixture. HPLC analysis showed that the total shikonin content was 1.45 mg / g, with an extraction rate of 93.8%, which met the quality control requirements.

[0033] S3, HPLC fingerprint chromatogram ratio optimization: Weigh 10 mg each of levorotatory shikonin, β,β'-dimethylacryloyl arcinin, acetyl shikonin, isobutyryl shikonin, artemisinin lactone, and catalpol reference standards, place them in 10 mL brown volumetric flasks, dissolve them in chromatographic grade methanol and dilute to the mark to prepare a mixed reference solution containing 20 μg of each reference standard per 1 mL. Accurately measure 1.0 g of the oil phase mixture and prepare the test solution using the same method as in Example 1; The chromatographic column was the same as in Example 1, the mobile phase was methanol-0.1% phosphoric acid aqueous solution (85:15, V / V), the flow rate was 1.0 mL / min, the detection wavelength was 275 nm, the column temperature was 30 °C, the injection volume was 10 μL, and the run time was 40 min. Fingerprint spectrum evaluation: The similarity between the fingerprint spectrum of the test sample and the standard spectrum is 0.98, and the batch deviation is 2.7%, which meets the quality control requirements.

[0034] S4. Paste preparation and aseptic filling: The oil phase mixture is filtered through three stages: 200 mesh, 300 mesh, and 0.3μm to recover the Lithospermum erythrorhizon residue, and the filtrate is transferred to a sterile mixing tank. 17.5 kg of organic white beeswax and 12.5 kg of shea butter were melted in a water bath at 55°C and stirred at 80 r / min for 30 min. After filtering through a 300-mesh filter, the mixture was added to a mixing tank and stirred at 53°C and 80 r / min for 30 min until well mixed. Cool to 37.5℃, add 4kg of vitamin E and 1.55kg of natural essential oil, stir at 60r / min for 22min, and protect with nitrogen. The material was homogenized twice under high pressure of 30 MPa at a temperature of 35°C to obtain a semi-finished paste. The product is a purplish-red, fine paste with a pH of 5.9. It meets the requirements for microorganisms and heavy metals, and the total shikonin content is 1.28 mg / g, which is in line with the requirements. Filling is carried out in a Class 100 clean area with an ambient temperature of 20℃ and a relative humidity of 50%. The filling specification is 20g / vial, the filling volume deviation is ≤±2%, the sealing temperature is 125℃, and the finished product is obtained after light inspection and outer packaging. Example

[0035] The compound gromwell ointment based on Yunnan gromwell root in this embodiment, by weight, uses the upper limit of the parameter range of this invention. The specific formula is as follows: 15 parts Yunnan gromwell root extract, 5 parts Angelica sinensis extract, 3 parts Rehmannia glutinosa extract, 4 parts Saposhnikovia divaricata extract, 2 parts Myrrh extract, 2 parts Frankincense extract, 100 parts cold-pressed virgin coconut oil, 20 parts organic white beeswax, 15 parts organic unrefined shea butter, 5 parts organic vitamin E, and 2 parts natural essential oils; wherein, the natural essential oils consist of 1 part organic real lavender essential oil, 0.5 parts organic peppermint essential oil, and 0.5 parts organic Roman chamomile essential oil; the Yunnan gromwell root extract is dried by low-temperature closed-loop hot air drying of Yunnan gromwell root at 40℃, and the total gromwell content in the extract is 85.2%, which meets the quality control requirements of this invention.

[0036] The preparation method in this embodiment is as follows: S1. Standardized pretreatment of raw materials: Microscopic identification and origin tracing were performed on raw materials of Lithospermum erythrorhizon from Dali, Yunnan, confirming that they were free from mold and insect infestation. A closed-loop low-temperature hot air dryer is used, with an inlet air temperature of 40℃ and a drying time of 8 hours, drying the material to a moisture content of 6.1%, and the entire drying process is conducted in the dark. A low-temperature ultrafine pulverizer was used, with the feed temperature set at 25℃ and the pulverizing speed at 3500 r / min. After pulverization, the powder was passed through a 60-mesh standard inspection sieve to obtain Yunnan Lithospermum powder. The particle size was measured to be D90=202μm by a laser particle size analyzer, and the moisture content was measured to be 6.1% by a Karl Fischer moisture analyzer. The powder was then sealed with nitrogen and stored in a light-proof cold storage at 4℃ for later use.

[0037] Angelica sinensis slices were cleaned and impurities removed, sliced ​​to a thickness of 3 mm, and dried at 40℃ to a moisture content of 7.8%. Supercritical CO2 extraction parameters were as follows: extraction pressure 30 MPa, extraction temperature 40℃, extraction time 1.5 h, separation pressure 10 MPa, and separation temperature 35℃. The extract was collected, and the content of artemisinin was determined by HPLC to be 1.27%. The extract was then sealed in nitrogen and stored at 6℃ protected from light. Take fresh Rehmannia glutinosa slices, add 12 times the amount of deionized water, and extract once at 40℃ for 1 hour each time. Combine the extracts, filter through a 400-mesh filter, microfilter through a ceramic membrane, concentrate under reduced pressure at 35℃ to a relative density of 1.12 (25℃), add ethanol to a concentration of 65%, let stand at 6℃ for 10 hours, recover the ethanol from the supernatant, dry under vacuum at 40℃ to a moisture content of 4.2%, and determine the catalpol content by HPLC to be 0.96%. Store in a sealed container at 6℃. Take slices of Saposhnikovia divaricata, add 14 times the amount of deionized water, reflux and extract once, 0.5 h each time, combine the extracts, filter and concentrate to a relative density of 1.10 (25℃), add ethanol to a precipitation concentration of 75%, stand at 6℃ for 10 h, recover ethanol from the supernatant, dry at 40℃ under vacuum to a moisture content of 4.3%, HPLC analysis shows that the content of cimicifugoside is 0.28%, and store in a sealed container. Myrrh and frankincense resin were pulverized and passed through a 50-mesh sieve. Ten times the amount of cold-pressed coconut oil was added to each, and the mixture was stirred and extracted once at 40°C in the dark for 1.5 hours each time. The mixture was filtered through a 400-mesh sieve, and the filtrates were combined. The volatile oil content of the myrrh extract was 6.1%, and the total boswellic acid content of the frankincense extract was 5.9%. The mixture was then stored in a nitrogen-filled, sealed container in the dark.

[0038] S2. Low-temperature, light-protected cold soaking combined with tubular ultrasound-assisted extraction: In a 1000L stainless steel light-proof cold soaking tank, add 15kg of Yunnan purple gromwell powder and 1200L of cold-pressed coconut oil at a material-to-liquid ratio of 1:80 (g:mL). After vacuuming to -0.1MPa, purge with nitrogen four times to achieve an oxygen content of 0.5% in the tank and seal the tank opening. The jacket is circulated with 20℃ constant temperature cooling water, and the temperature inside the tank is controlled at 20±0.5℃. It is then cold-soaked in the dark for 9 months, and the agitator is turned on every 10 days at a speed of 40r / min for 20min. After cold soaking, the material is transferred to a pipeline ultrasonic extraction unit with the following parameters: 500W, 45kHz, material temperature 25℃, flow rate 70L / h, and continuous ultrasonication for 30min. The extract was transferred to a sterile mixing tank, and the extracts of each excipient were added. The mixture was stirred at 80 r / min and simultaneously sonicated for 20 min to obtain an oil phase mixture. HPLC analysis showed that the total shikonin content was 1.68 mg / g, with an extraction rate of 94.3%, which met the quality control requirements.

[0039] S3, HPLC fingerprint chromatogram ratio optimization: Weigh 10 mg each of levorotatory shikonin, β,β'-dimethylacryloyl arcinin, acetyl shikonin, isobutyryl shikonin, artemisinin lactone, and catalpol reference standards, place them in 10 mL brown volumetric flasks, dissolve them in chromatographic grade methanol and dilute to the mark to prepare a mixed reference solution containing 20 μg of each reference standard per 1 mL. Accurately measure 1.0 g of the oil phase mixture and prepare the test solution using the same method as in Example 1; The chromatographic column was the same as in Example 1, the mobile phase was methanol-0.1% phosphoric acid aqueous solution (90:10, V / V), the flow rate was 1.2 mL / min, the detection wavelength was 280 nm, the column temperature was 35 °C, the injection volume was 5 μL, and the run time was 30 min. The fingerprint spectrum of the test sample showed a similarity of 0.99 to the standard spectrum, with a batch deviation of 2.1%, which meets the quality control requirements.

[0040] S4. Paste preparation and aseptic filling: The oil phase mixture is filtered through three stages: 220 mesh, 320 mesh, and 0.22μm to recover the Lithospermum erythrorhizon residue, and the filtrate is transferred to a sterile mixing tank. 20 kg of organic white beeswax and 15 kg of shea butter are melted in a 60°C water bath and stirred at 100 rpm for 20 min. After filtering through a 320 mesh filter, the mixture is added to a mixing tank and stirred at 55°C and 100 rpm for 20 min until well mixed. Cool to 40℃, add 5kg of vitamin E and 2kg of natural essential oil, stir at 80r / min for 15min, and protect with nitrogen. The material was homogenized once at 40 MPa pressure and the material temperature was 40℃ to obtain a semi-finished paste. The product is a purplish-red, fine paste with a pH of 6.3. It meets the requirements for microorganisms and heavy metals, and the total shikonin content is 1.49 mg / g, which is in line with the requirements. Filling is carried out in a Class 100 clean area with an ambient temperature of 24℃ and a relative humidity of 40%. The filling specification is 30g / vial, the filling volume deviation is ≤±1%, the sealing temperature is 140℃, and the finished product is obtained after light inspection and outer packaging.

[0041] Scale settings To verify the rationality of the parameter range of the present invention, the following comparative examples were set up. All comparative examples have only one parameter that exceeds the protection scope of the present invention, while the other formulations and parameters are completely consistent with those of Example 2: Comparative Example 1: The cold soaking temperature was set to 30°C, and the other parameters were the same as in Example 2; Comparative Example 2: The cold soaking time was set to 3 months, and the other parameters were the same as in Example 2; Comparative Example 3: The ultrasonic power was set to 800W, and the other parameters were the same as in Example 2; Comparative Example 4: After harvesting, the Yunnan purple gromwell was washed with water, and the other parameters were the same as in Example 2.

[0042] Performance verification test: The extraction rate of shikonin and the retention of total shikonin in the finished product of each example and comparative example were detected using the HPLC method of this invention. The results are shown in Table 1 below: Group Shikonin extraction rate (%) Total shikonin retention in the finished product (mg / g) Does it conform to the parameter range of this invention? Example 1 92.1 1.07 yes Example 2 93.8 1.28 yes Example 3 94.3 1.49 yes Comparative Example 1 72.4 0.46 no Comparative Example 2 65.7 0.38 no Comparative Example 3 78.6 0.52 no Comparative Example 4 93.5 0.75 no The results showed that in Examples 1-3 within the parameter range of the present invention, the extraction rate of shikonin was ≥92% and the total shikonin retention in the finished product was ≥1.0 mg / g, achieving the expected technical effect. However, in the comparative examples outside the parameter range of the present invention, the extraction rate and retention of shikonin decreased significantly, proving that the parameter range setting of the present invention is scientific and necessary.

[0043] The antibacterial rates of Staphylococcus aureus, Escherichia coli, and Candida albicans in each experimental group were tested. The contact time was 20 min. The results are shown in Table 2 below: Group Staphylococcus aureus inhibition rate (%) Escherichia coli inhibition rate (%) Candida albicans inhibition rate (%) Example 1 99.85 99.6 99.52 Example 2 99.95 99.79 99.71 Example 3 99.97 99.84 99.76 Comparative Example 1 88.12 83.26 81.05 Comparative Example 2 86.34 81.57 79.68 Comparative Example 3 89.75 85.43 83.52 Comparative Example 4 93.31 91.52 90.28 The results showed that Examples 1-3 within the parameter range of the present invention all had an inhibition rate of >99.5% against three common pathogenic bacteria, demonstrating excellent antibacterial effects. In contrast, the comparative examples outside the parameter range of the present invention showed a significant decrease in antibacterial effects, proving that the parameter range of the present invention is key to ensuring the core efficacy of the product.

[0044] Acute skin irritation test in rabbits and skin sensitization test in guinea pigs were conducted using compound comfrey ointment. The results showed that: rabbits in groups 1-3 of the present invention showed no irritation reactions such as erythema and edema, and the skin irritation score was 0, indicating no irritation; guinea pigs showed no skin sensitization reaction, with a sensitization rate of 0, indicating weak sensitization; while comparative example 1 showed mild erythema and edema, indicating mild irritation, with a sensitization rate of 15%, proving that the parameter range of the present invention can effectively ensure the safety of the product.

[0045] 160 participants aged 1-65 years with skin problems such as insect bites, eczema, and acne were recruited, including 20 pregnant women and 40 individuals with sensitive skin. They were randomly divided into 4 groups of 40 participants each, and used the products from Examples 2 and Comparative Examples 1-3, respectively, applying them twice daily for 7 consecutive days. The effectiveness in relieving itching, reducing redness and swelling, and repairing skin lesions was evaluated. The results are shown in Table 3 below. Group Antipruritic efficacy rate (%) Effective rate of redness and swelling reduction (%) Effective rate of skin lesion repair (%) Incidence of adverse reactions (%) Example 2 100 97.5 95 0 Comparative Example 1 82.5 75 67.5 12.5 Comparative Example 2 80 72.5 65 0 Comparative Example 3 85 77.5 70 5 Comparative Example 4 87.5 80 77.5 0 The results showed that the product of Example 2 within the parameter range of the present invention had significantly higher efficacy rates in relieving itching, reducing redness and swelling, and repairing skin lesions than the comparative example outside the parameter range, and no adverse reactions were observed. Pregnant women, children, and subjects with sensitive skin experienced no discomfort, proving that the parameter range of the present invention can effectively guarantee the efficacy and safety of the product.

[0046] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A compound gromwell ointment based on Yunnan gromwell root, characterized in that, By weight, it includes: 10-15 parts of Yunnan Lithospermum extract, 3-5 parts of Angelica sinensis extract, 2-3 parts of Rehmannia glutinosa extract, 2-4 parts of Saposhnikovia divaricata extract, 1-2 parts of Myrrh extract, 1-2 parts of Frankincense extract, 80-100 parts of cold-pressed virgin coconut oil, 15-20 parts of organic white beeswax, 10-15 parts of organic unrefined shea butter, 3-5 parts of organic vitamin E, and 1.1-2 parts of natural essential oils.

2. The compound gromwell ointment based on Yunnan gromwell root according to claim 1, characterized in that, The Yunnan Lithospermum extract is derived from Lithospermum yunnanense, a plant belonging to the genus Lithospermum of the Boraginaceae family. It is dried at a low temperature of 30-40℃ until the moisture content is ≤8.0%, and the total shikonin content in the extract is ≥83%. The total shikonin is the sum of L-shikonin, β,β'-dimethylacryloyl shikonin, acetylshikonin, and isobutyrylshikonin.

3. The compound gromwell ointment based on Yunnan gromwell root according to claim 1, characterized in that, The natural essential oil is composed of 0.5-1 parts organic real lavender essential oil, 0.3-0.5 parts organic peppermint essential oil, and 0.3-0.5 parts organic Roman chamomile essential oil; wherein, the real lavender essential oil contains ≥30% linalyl acetate, the peppermint essential oil contains ≥50% menthol, and the Roman chamomile essential oil contains ≥1.0% chamomile azulene.

4. A preparation process for a compound gromwell ointment made from Yunnan gromwell as described in any one of claims 1-3, characterized in that, Includes the following steps: S1. Standardized pretreatment of raw materials: Yunnan gromwell is harvested, impurities are removed, dried at a low temperature of 30-40℃, fed at 20-25℃ and pulverized through a 40-60 mesh sieve to obtain Yunnan gromwell powder with a particle size D90 of 200-250μm; Angelica sinensis, Rehmannia glutinosa, Saposhnikovia divaricata, Myrrh, and Boswellia carterii are cleaned and dried respectively to prepare corresponding extracts for later use; S2. Low-temperature, light-protected cold maceration combined with pipeline ultrasonic-assisted extraction: Mix Yunnan purple gromwell powder with cold-pressed virgin coconut oil at a material-to-liquid ratio of 1:50-80, and purge with nitrogen 2-4 times under vacuum of -0.08~-0.1MPa until the oxygen content in the tank is 0.5-1.0%. Cold maceration is carried out at a constant temperature of 15-20℃ in the dark for 7-9 months, with stirring every 10-20 days. After cold maceration, ultrasonic-assisted extraction is carried out using a pipeline ultrasonic extraction unit. The prepared extracts of various excipients are added, and ultrasonication is continued for 20-30 minutes to obtain a mixture of active ingredients in the oil phase. S3. HPLC fingerprint chromatogram optimization: Establish exclusive fingerprint chromatograms for raw materials and finished products using high-performance liquid chromatography, adjust the proportions of each active ingredient, and ensure that the similarity between the fingerprint chromatogram of the test sample and the standard control fingerprint chromatogram is ≥0.95, and the batch-to-batch quality deviation is ≤5%. S4. Ointment Preparation and Aseptic Filling: The oil phase mixture is filtered through three stages to recover the purple gromwell residue, and the filtrate is transferred to an aseptic mixing tank. Organic white beeswax and organic unrefined shea butter are melted in a water bath at 50-60℃ and added to the mixing tank. After being mixed evenly, the mixture is cooled to 35-40℃, and organic vitamin E and natural essential oils are added. After being homogenized and emulsified under high pressure at 20-40MPa 1-3 times, the mixture is aseptically filled in a Class 100 clean environment to obtain the finished product.

5. The preparation process of a compound gromwell ointment based on Yunnan gromwell root according to claim 4, characterized in that, In S1, Angelica sinensis extract is prepared by supercritical CO2 extraction with the following parameters: extraction pressure 20-30 MPa, extraction temperature 30-40℃, extraction time 1.5-2.5 h, separation pressure 6-10 MPa, and separation temperature 25-35℃; Rehmannia glutinosa extract is prepared by water extraction and alcohol precipitation with the following parameters: water volume 8-12 times, extraction temperature 30-40℃, extraction time 1-2 h, and alcohol precipitation concentration 55-65%; Saposhnikovia divaricata extract is prepared by water extraction and alcohol precipitation with the following parameters: water volume 10-14 times, reflux extraction time 0.5-1.5 h, and alcohol precipitation concentration 65-75%; Myrrh and Boswellia carterii extracts are prepared by low-temperature oil extraction with the following parameters: oil volume 6-10 times, extraction temperature 30-40℃, and extraction time 1.5-2.5 h.

6. The preparation process of a compound gromwell ointment based on Yunnan gromwell root according to claim 4, characterized in that, In step S2, the Yunnan purple gromwell powder and cold-pressed virgin coconut oil are mixed at a material-to-liquid ratio of 1:50-80.

7. The preparation process of a compound gromwell ointment based on Yunnan gromwell root according to claim 4, characterized in that, In S2, the stirring parameters for the cold soaking process are: rotation speed 20-40 r / min, and stirring time 20-40 min per stirring.

8. The preparation process of a compound gromwell ointment based on Yunnan gromwell root according to claim 4, characterized in that, In S3, the HPLC detection parameters are as follows: C18 column; mobile phase: methanol-0.1% phosphoric acid aqueous solution, volume ratio 80:20-90:10; flow rate: 0.8-1.2 mL / min; detection wavelength: 270-280 nm; column temperature: 25-35℃; injection volume: 5-20 μL; running time: 30-50 min.

9. The preparation process of a compound gromwell ointment based on Yunnan gromwell root according to claim 4, characterized in that, In S4, the three-stage filtration uses 180-220 mesh filter cloth, 280-320 mesh filter cloth, and 0.22-0.45μm polypropylene sterilization filter element in sequence; the aseptic filling environment parameters are: temperature 16-24℃, relative humidity 40-60%, filling volume deviation ≤±1%~±3%, and sealing temperature 110-140℃.

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