Six-ingredient skin-soothing lotion as well as preparation method and component detection method thereof

Abstract

The invention provides a six-ingredient skin-soothing lotion, a preparation method and a component detection method thereof, and belongs to the technical field of traditional Chinese medicine, every 1000 milliliters of the skin-soothing lotion is prepared by decocting and concentrating the following raw material medicines in parts by weight: 100 g of scutellaria baicalensis, 100 g of rhizoma impatientis, 100 g of mesiomeria sibirica, 100 g of rubus corchorifolius, 100 g of coptis chinensis and 100 g of sanguisorba officinalis. According to clinical medication requirements, the six-ingredient skin-soothing lotion is changed into lotion from an original water decoction, a single-factor test and an orthogonal test are adopted, the dry extract rate serves as an inspection index, extraction, concentration, liquid preparation and filling process parameters of the six-ingredient skin-soothing lotion are determined, a screening forming process is adopted, and finally the six-ingredient skin-soothing lotion is obtained through three-batch large-scale pilot scale production. The preparation process of the six-ingredient skin-soothing lotion is verified, and the result shows that the determined preparation process is scientific, reasonable and feasible.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a Liuwei Shufu lotion, a preparation method and a component detection method thereof. Background Art

[0002] Skin disease is one of the common diseases that seriously troubles the physical and mental health of patients. Most of the causes and pathogenesis have not been fully clarified. Although there are many treatments, drugs and methods, there is a lack of special and effective drugs. The disease recurs repeatedly and is difficult to cure. The skin is the surface of the human body. Traditional Chinese medicine adheres to the holistic concept and syndrome differentiation and treatment to diagnose and treat skin diseases. Traditional Chinese medicine has a history of thousands of years in recognizing and treating this disease, and it is constantly developing. There are many methods, fewer side effects than Western medicine, and it is easily accepted by the general public.

[0003] At present, Western medicine treats skin diseases mainly for symptomatic treatment. Although the first-line drug glucocorticoids are effective quickly, many patients have adverse reactions such as pigmentation, and the condition is prone to recurrence or aggravation, which limits clinical use. Since Guangxi is located in the subtropics and has a hot and humid climate, dampness, wind, insect and heat toxins are prone to invade the human body first, leading to frequent skin diseases. Zhuang medicine has the characteristics of attaching importance to external treatment, focusing on detoxification, early treatment when sick, early prevention when not sick, and specialization when treating skin. In addition, Guangxi Zhuang and Yao medicine resources are rich, and cold medicines are produced in abundance. They have outstanding effects of clearing heat, detoxifying and removing dampness. In the treatment of skin diseases, they have their unique therapeutic effects and are favored by the majority of patients. Therefore, it is necessary to design a six-flavor skin-soothing lotion, a preparation method and a component detection method thereof. Summary of the invention

[0004] The purpose of the present invention is to provide a Liuwei Shufu lotion, a preparation method and a component detection method thereof, so as to solve the technical problem that the antipruritic and anti-inflammatory effects of traditional Chinese medicine are not good.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0006] Liuwei Soothing Skin Lotion, every 1000 ml of skin lotion solution is made by decocting and concentrating the following raw materials by weight: 100g of Scutellaria baicalensis, 100g of Pengbei, 100g of Mei Nei Mei, 100g of Gou Mei, 100g of Coptis chinensis and 100g of Sanguisorba officinalis.

[0007] Liuwei Shufu Lotion, each 60 ml of skin lotion solution is made by water extraction and concentration of the following raw materials by weight: 6.0 kg of Scutellaria baicalensis, 6.0 kg of Pengbei, 6.0 kg of Mei Nei Mei, 6.0 kg of Gou Mei, 6.0 kg of Coptis chinensis, 6.0 kg of Sanguisorba officinalis and 0.18 kg of sodium benzoate.

[0008] The preparation method of Liuwei Shufu lotion is as follows: soak all the medicinal raw materials for 30 minutes, decoct twice, add 9 times the amount of water for the first time and 7 times the amount of water for the second time, decoct for 1 hour each time, filter, combine the filtrate, concentrate the filtrate to a concentrate with a relative density of 1.03 to 1.10, the temperature is 60°C, filter, stand overnight, take the supernatant and boil for 30 minutes, add 3g of sodium benzoate, add pure water to 1000ml, stir evenly, and fill.

[0009] The preparation method of Liuwei Shufu lotion comprises the following steps: step 1: adding six Chinese medicinal pieces of scutellaria baicalensis, purslane, charcoal motherwort, scutellaria baicalensis, magnolia officinalis and radix sanguisorbae into an extraction tank, soaking them in water for 30 minutes, decocting them twice, adding 9 times the amount of water for the first time and 7 times the amount of water for the second time, decocting them for 1 hour each time, during the decocting process, controlling the steam pressure at 0.01-0.25MPa before the medicinal liquid boils, and controlling the steam pressure at 0.01-0.10MPa after the medicinal liquid boils, so that the medicinal liquid keeps boiling, filtering the medicinal liquid with a 100-mesh filter, and combining the filtrate for standby use;

[0010] Step 2: Pump the filtrate in step 1 into a single-effect concentrator unit, with a concentration temperature of 75-90°C and a vacuum degree of -0.06--0.09MPa, and concentrate to a concentrate with a relative density of 1.03-1.10 at a temperature of 60°C. Filter the concentrate with a 200-mesh filter cloth and put it into a clean barrel, seal it with a cover, weigh it, attach a material card, and place it in a cool warehouse to stand overnight. Take the supernatant into a clean barrel, seal it with a cover, weigh it, attach a material card, and set it aside;

[0011] Step 3: Weigh sodium benzoate and dissolve it with purified water for later use. Take the supernatant of Liuwei Shufu Lotion Concentrate from the cool storage in a sandwich pot, heat and boil for 30 minutes, stop heating, add the completely dissolved sodium benzoate solution while stirring, stir for 30 minutes to mix, dilute to the prescribed amount with purified water, continue stirring for 30 minutes to mix, and the preparation is completed. Filter the prepared liquid with a 200-mesh filter cloth, put it in a clean bucket and set aside;

[0012] Step 4: Transfer the prepared liquid medicine to the liquid filling room of the external preparation workshop, start the automatic bottle unscrambler and the liquid filling machine linkage production line respectively, install and debug according to the batch production requirements, and debug the filling volume: 150ml / bottle. After the filling volume debugging is completed, confirm that the filling volume of each injection is qualified; the bottle cap is screwed in place, there is no crooked cap, no looseness, and no leakage in the seal; after the outer wall of the medicine bottle is clean and free of pollution, start filling. When the equipment is running at normal speed, during the filling process, and before the end of filling, check the filling volume and sealing conditions. Each time, check 10 bottles of products produced continuously, and check the filling volume and sealing conditions and record them. If the filling volume and sealing are unqualified, stop the machine immediately for inspection and adjust to qualified. The filled products are conveyed to the outer packaging room via the conveyor belt.

[0013] The TLC identification and detection method of schizonepeta in Liuwei Shufu lotion comprises the following steps: Step 1: Preparation of reference solution, taking 0.5 g of schizonepeta reference medicinal material powder, adding 30 mL of methanol, ultrasonically treating for 30 minutes, evaporating to dryness, adding 30 mL of water-saturated n-butanol to dissolve the residue, washing with ammonia test solution twice, 20 mL each time, evaporating the n-butanol solution after washing to dryness, adding 2 mL of methanol to dissolve the residue to prepare a reference medicinal material solution;

[0014] Step 2: Preparation of test sample and negative solution: take 20 mL of Zhuangyao Xiaoyan Zhiyou Lotion, shake it thoroughly with 30 mL of water-saturated n-butanol, take the n-butanol layer, wash it twice with ammonia test solution, 20 mL each time, evaporate the n-butanol solution after washing to dryness, add 2 mL of methanol to dissolve the residue, take the supernatant as the test sample solution, and prepare the negative control solution of Qujiu Biying in the same way;

[0015] Step 3: Thin layer chromatography: follow the thin layer chromatography test, take 1 μL of the test solution, 1 μL of the negative test solution, and 1 μL of the control medicinal material solution, and spot them on the same silica gel G thin layer plate respectively. Develop with chloroform-methanol-formic acid as the developing solvent, then spray with 10% sulfuric acid ethanol solution (chloroform: methanol: formic acid is 16:3:1), heat at 105°C for 3-5 minutes, and inspect under a fluorescent light.

[0016] The TLC identification and detection method of Chinese magnolia tree in Liuwei Shufu lotion comprises the following steps: Step 1: Preparation of reference solution: 0.1 g of Chinese magnolia tree reference medicinal material powder is taken, 5 mL of methanol is added, and ultrasonic treatment is performed for 30 minutes, and the mixture is allowed to stand, and the supernatant is taken as the reference medicinal material solution;

[0017] Step 2: Preparation of test sample and negative solution: take 20 mL of Zhuangyao Xiaoyan Zhiyou Lotion, shake it thoroughly with 30 mL of water-saturated n-butanol, take the n-butanol solution, wash it twice with ammonia test solution, 20 mL each time, evaporate the n-butanol solution after washing to dryness, add 2 mL of methanol to dissolve the residue, take the supernatant as the test sample solution, and prepare the negative control solution of L. magnoliae in the same way;

[0018] Step 3: Thin layer chromatography: According to the thin layer chromatography test, 2 μL of the test solution, 2 μL of the negative test solution, and 2 μL of the control medicinal material solution are respectively spotted on the same silica gel G thin layer plate to form strips. The plate is developed with the upper layer solution of n-butanol-glacial acetic acid-water as the developing agent, and then the plate is inspected under ultraviolet light after drying. The ratio of n-butanol: glacial acetic acid: water is 8:1:4.

[0019] The method for determining the content of syringin in Liuwei Shufu lotion comprises the following steps: Step 1: Preparation of reference solution: accurately weigh 10 mg of syringin reference substance, place it in a 100 ml volumetric flask and make up to volume with 50% methanol to prepare a reference solution with a concentration of 0.0952 mg / ml;

[0020] Step 2: Preparation of the test solution: use a pipette to measure 1 ml of Zhuangyao Xiaoyan Zhiyou Lotion, dilute to 10 ml volumetric flask with water, shake well, centrifuge at 10000r / min for 10 min, take the supernatant and filter through a 0.22μm microporous membrane to obtain the test solution. Prepare the negative control solution of Qujiu Biying in the same way;

[0021] Step 3: Take 5 μL of the reference solution, test solution and negative control solution, respectively, and inject them into the HPLC chromatograph for determination according to the law. The retention time of the chromatographic peak of syringin in the test sample is consistent with that of the reference solution, and there is no interference with the negative control;

[0022] Step 4: Linear relationship investigation: The reference solution with a syringin content of 0.0952 mg / ml obtained in step 1 was respectively taken and filtered through a 0.22 μm microporous filter membrane and injected into an Agilent high performance liquid chromatograph for detection. The injection volumes were 2 μL, 5 μL, 10 μL, 15 μL, 20 μL, 25 μL, and 30 μL, respectively. The syringin peak area was recorded. The syringin peak area was used as the abscissa and the syringin concentration was used as the ordinate. The least squares method was used for linear regression, and the regression equation for syringin was y=0.0004x-0.0339, R 2 =0.9995, the linear range of syringin is between 0.190μg and 2.856μg;

[0023] Step 5: Precision experiment, take the reference solution with a syringin content of 0.0952 mg / ml prepared in step 1 and inject it six times continuously according to the method under the chromatographic conditions, measure and record the syringin peak area, and calculate the RSD of the syringin peak area to be 2.57%, indicating that the instrument has good precision.

[0024] Step 6: Repeatability experiment, take six portions of the same batch of Zhuangyao anti-inflammatory and antipruritic lotion, prepare the test solution according to the method in step 2, and inject and test according to the method of chromatographic conditions, and calculate the RSD of the syringin peak area to be 2.11%;

[0025] Step 7: Stability test, prepare the test solution according to the method in step 2, and determine the peak area of ​​syringin according to the method when standing at room temperature for 0h, 3h, 6h, 12h, and 24h, and calculate the RSD of the peak area of ​​syringin to be 1.53%;

[0026] Step 8: Sample addition recovery experiment, take six portions of Zhuang medicine anti-inflammatory and antipruritic lotion with known content, 0.5 ml each, and place them in 10 ml volumetric flasks respectively, then weigh an appropriate amount of syringin reference substance, prepare a reference substance solution with a concentration of 0.952 mg / ml and accurately pipette 1 ml into the sample respectively, make up to volume with water and shake well, centrifuge at 10000 r / min for 10 min, take the supernatant and filter it through a 0.22 μm microporous filter membrane, and determine and record the syringin peak area according to the law, and calculate the average recovery rate to be 100.37%, and the RSD is 0.88%;

[0027] Step 9: Sample content determination, take 1 ml of each of the three batches of skin lotion, prepare the sample according to the sample preparation method in step 2, and test the sample according to the chromatographic conditions. The content of syringin in the Zhuang medicine anti-inflammatory and antipruritic lotion sample is calculated to be 0.225 mg / ml, 0.221 mg / ml, and 0.222 mg / ml, respectively.

[0028] The present invention has the following beneficial effects due to the adoption of the above technical solution:

[0029] (1) According to the clinical medication needs, the present invention changes the Liuwei Shufu lotion formula from the original water decoction to a lotion, adopts single factor experiment and orthogonal experiment, takes the dry cream rate as the investigation index, determines the extraction, concentration, liquid preparation, and filling process parameters of the Liuwei Shufu lotion, screens the molding process, and finally verifies the preparation process of the Liuwei Shufu lotion through three batches of scale-up pilot production. The results show that the determined preparation process is scientific, reasonable and feasible.

[0030] (2) The present invention establishes a TLC identification method for Herba Lycopodii and Herba Coptidis in Zhuang medicine anti-inflammatory and antipruritic lotion and a method for determining the content of syringin. The Herba Lycopodii and Herba Coptidis in Zhuang medicine anti-inflammatory and antipruritic lotion are qualitatively identified by thin layer chromatography (TLC); and the syringin in Zhuang medicine anti-inflammatory and antipruritic lotion is quantitatively analyzed by high performance liquid chromatography (HPLC). Chromatographic conditions: Luna 5μm C18 (2) (250mmx4.6mm, 5μm) column, acetonitrile (10%)-water (90%) as mobile phase, flow rate of 1.0ml / min; detection wavelength of 265nm; injection volume of 5μL; column temperature of 30℃. Syringin has a good linear relationship with its peak area in the range of 0.190μg to 2.856μg (R 2 =0.9995). The RSD values ​​of the precision, stability and repeatability experiments were all <2.5%, and the average recovery rate (n=6) was 100.37% (RSD=0.88%). This method is simple to operate, has strong specificity, good repeatability and stability, and can be used for the quality control of Zhuang medicine anti-inflammatory and antipruritic lotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a flow chart of the preparation process of the Liuwei Shufu lotion of the present invention;

[0032] Figure 2 It is the TLC chromatogram of Jiubiying of the present invention;

[0033] Figure 3 This is the TLC chromatogram of the Chinese magnolia tree of the present invention;

[0034] Figure 4 This is the HPLC chromatogram of the syringin content determination of the present invention, negative control (A), syringin reference (B), test sample (C). DETAILED DESCRIPTION

[0035] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and preferred embodiments. However, it should be noted that many details listed in the specification are only for the purpose of enabling the reader to have a thorough understanding of one or more aspects of the present invention, and these aspects of the present invention can be implemented even without these specific details.

[0036] Example 1: The prescription of this product is composed of six medicinal materials. According to the physical and chemical properties of each medicinal material, and referring to the relevant provisions of the "Implementation Rules for the Registration Management of Traditional Chinese Medicine and Ethnic Medicine Preparations Prepared by Traditional Technology in Guangxi Medical Institutions" and the "Technical Requirements for Research on Traditional Chinese Medicine and Ethnic Medicine Preparations in Guangxi Medical Institutions", the preparation process of Liuwei Shufu Lotion was studied and the detailed results are as follows.

[0037] Table 1 is the prescription parameter table of Liuwei Shufu lotion

[0038]

[0039] Preparation method

[0040] Soak the above six ingredients for 30 minutes, decoct twice, add 9 times the amount of water for the first time and 7 times the amount of water for the second time, decoct for 1 hour each time, filter, combine the filtrate, concentrate the filtrate to a concentrate with a relative density of 1.03-1.10 (60°C), filter, let stand overnight, take the supernatant and boil for 30 minutes, add 3g of sodium benzoate, add pure water to 1000ml, stir evenly, and fill.

[0041] Embodiment 2:

[0042] Table 2 is the prescription parameter table of Liuwei Shufu lotion

[0043]

[0044] Preparation process: receiving materials, pre-treatment, according to the batch preparation instructions and the material receiving list, receiving the qualified Chinese herbal medicine pieces such as Scutellaria baicalensis, Portulaca oleracea, Herba Cibotii, Herba Polygoni Multiflori, Herba Euphorbiae Lauraceae, and Radix Sanguisorbae Officinalis from the Chinese herbal medicine warehouse, weighing them and setting them aside.

[0045] Extraction and concentration: Extraction is carried out according to the batch preparation instruction. Six Chinese medicinal pieces of Radix Scutellariae, Herba Portulacae, Herba Eupatorii, Herba Salviae Miltiorrhizae, Herba Maluse and Herba Sanguisorbae are added to the extraction tank, soaked in water for 30 minutes, and decocted twice. The first time, 9 times the amount of water is added, and the second time, 7 times the amount of water is added. Each time, decocted for 1 hour. During the decoction process, the steam pressure is controlled at 0.01-0.25MPa before the liquid medicine boils; the steam pressure is controlled at 0.01-0.10MPa after the liquid medicine boils, so that the liquid medicine keeps boiling. The liquid medicine is filtered through a 100-mesh filter, and the filtrate is combined for standby use.

[0046] Concentrate, pump the filtrate into a single-effect concentrator, with a concentration temperature of 75-90°C and a vacuum degree of -0.06--0.09MPa, and concentrate to a concentrate with a relative density of 1.03-1.10 (60°C). Filter the concentrate with a 200-mesh filter cloth and put it into a clean barrel, seal it with a cover, weigh it, attach a material card, and place it in a cool warehouse to stand overnight. Take the supernatant into a clean barrel, seal it with a cover, weigh it, attach a material card, and set it aside.

[0047] Prepare the liquid. According to the batch preparation instruction, weigh the sodium benzoate and dissolve it with an appropriate amount of purified water for later use. Take the Liuwei Shufu lotion concentrate (supernatant) from the cool storage in a sandwich pot, heat and boil for 30 minutes, stop heating, add the completely dissolved sodium benzoate solution while stirring, stir for 30 minutes to mix, dilute to the prescribed amount with purified water, and continue stirring for 30 minutes to mix, and the liquid preparation is complete. Filter the prepared liquid with a 200-mesh filter cloth, put it in a clean bucket and set aside.

[0048] Filling: Transfer the prepared liquid medicine to the liquid filling room of the external preparation workshop, start the automatic bottle unscrambler and the liquid filling machine linkage production line respectively, install and debug according to the batch production requirements, and debug the filling volume: 150ml / bottle. After the filling volume debugging is completed, confirm that the filling volume of each injection is qualified; the bottle cap is screwed in place, there is no crooked cap, no looseness, and no leakage in the seal; after the outer wall of the medicine bottle is clean and free of pollution, start filling. When the equipment is running at normal speed, during the filling process, and before the end of filling, check the filling volume and sealing conditions. Each time, check 10 bottles of products produced continuously, and check the filling volume and sealing conditions and record them. If the filling volume and sealing are unqualified, stop the machine immediately for inspection and adjust to qualified. The filled products are conveyed to the outer packaging room via the conveyor belt.

[0049] Table 3 Equipment and Instruments List

[0050] Serial number Main production equipment Specifications Main testing instruments Specifications 1 Multifunctional extraction tank TQNS-1500 Density Tester AR-300Y 2 Single effect concentrator unit TQNS-1500 Electric constant temperature blast drying oven PHG-9206A 3 Sandwich pot JCG-300 Mold incubator MJ-250-Ⅱ 4 Liquid filling machine ZCG-1 Waterproof constant temperature incubator GHP-9270 5 Bottle unscrambler ZC-GP High Pressure Steam Sterilizer SQ510C 6 Electronic scale TCS-300 Biological Safety Cabinets BSC-1604ⅡA2 7 Electronic balance ME155DU

[0051] Preparation process research data

[0052] 1. Dosage form selection: This prescription is an experience prescription of our hospital for many years. It is composed of six traditional Chinese medicines (ethnic medicines), including Scutellaria baicalensis, Portulaca oleracea, Herba Euphorbiae, Herba Euphorbiae, Herba Euphorbiae, and Herba Sanguisorbae. It has the effects of clearing away heat and dampness, cooling blood and detoxifying, promoting tissue regeneration and healing sores. It is used for external washing to treat acute dermatitis, eczema, contact dermatitis, pederus dermatitis, burns and scalds and other skin diseases. The main ingredients of Herba Euphorbiae in this prescription contain alkaloids, flavonoids and volatile oils. [1-5] The remaining herbs mainly contain flavonoids, volatile oils, polysaccharides, organic acids, saponins and tannins. [6-12] The main ingredients can be extracted by hot water, and decoctions are often used clinically. However, decoctions are temporarily prepared and applied according to doctors' prescriptions, which is not conducive to timely treatment of patients. The decoction is large in volume, inconvenient to use and carry, easy to mold and ferment after long-term storage, and difficult to store. Chinese medicine lotions have the characteristics of direct action, most of which do not pass through the circulatory system, can avoid the first-pass effect of the liver, and have little side effects on the body. In recent years, it has made significant progress in the treatment of skin diseases, postoperative rehabilitation and other auxiliary treatments for diseases. Taking into account the disease characteristics, route of administration and convenience of use of the clinical medication subjects, the recipe is processed and concentrated, and the effective ingredients are extracted to make an external lotion.

[0053] 2 Extraction process research: This study used an open reactor as a small-scale extraction tool and the dry paste rate as an evaluation index to investigate the relevant factors affecting the extraction of Liuwei Shufu Lotion. Through orthogonal experiments, the relevant influencing factors were combined and further optimized to obtain the optimal small-scale extraction process. Three batches of pilot process verification were carried out to finally determine the extraction process route and process parameters.

[0054] Liuwei Shufu Lotion (Zhuangyao Anti-inflammatory and Antipruritic Prescription) is a clinical prescription of Li Fangmei, chief physician of the Department of Dermatology. The prescription consists of 6 herbs, including Scutellaria baicalensis, Byaekbeiz (Purslane), Maexndeihmeij (Jiubiying), Gaeumei (Gaeumei), Maexvuengzlienz (Mahonlaomu), and Sanguisorba officinalis. It has been mainly used as a decoction in clinical practice, and has the effects of clearing away heat and detoxifying, cooling blood and dispelling wind, and drying dampness and relieving itching. It is used to treat superficial fungal diseases of the skin, acute eczema, atopic dermatitis, contact dermatitis, pederus dermatitis, psoriasis, urticaria, impetigo, I-II degree burns and scalds, and postoperative laser treatment of the skin. In order to maintain the basic consistency with the material basis of the original decoction, this product is decocted by water extraction.

[0055] Establishment of extraction process evaluation method: In this experiment, dry paste rate was selected as the extraction process evaluation index.

[0056] Determination of dry cream rate: Measure the volume of the extracted medicinal solution, then accurately draw 50 ml, place it in an evaporating dish that has been dried to constant weight, evaporate it in a water bath, dry it in a 105℃ electric constant temperature drying oven for 5 hours, move it to a desiccator, cool it for 30 minutes, accurately weigh it, dry it at the above temperature for 1 hour, cool it, and weigh it until the difference between two consecutive weighings does not exceed 5 mg. Calculate the dry cream rate according to the following formula.

[0057] Dry paste rate (%) = (dry paste mass × total volume of medicinal solution) / (total mass of medicinal materials × sampling volume) × 100%.

[0058] Water absorption test: weigh 90g of Scutellaria baicalensis, Sanguisorba officinalis, Portulaca oleracea, Mahonia lauraceae, Herba Glehniae, and Herba Glehniae according to the prescription ratio, add 8 times the amount of water to soak, observe the degree of penetration every 30 minutes, and measure the water absorption when all are soaked for 120 minutes, and repeat three times. The test results are shown in Table 4.

[0059] Table 4 Water absorption test results

[0060]

[0061] From the results, we can see that the water absorption rate of the medicinal materials after soaking is 163.35%, which is about 1.5 times the weight of the medicinal materials. Therefore, 1.5 times the weight of the medicinal materials should be added when adding water for the first time.

[0062] Single factor test investigation: Single factor test is to examine the influence of one experimental factor on the evaluation index by changing one experimental factor and fixing other factors. Through single factor test, the factors and levels of orthogonal test are determined, and then the extraction process is optimized. Common factors affecting the water extraction process of traditional Chinese medicine include soaking time, extraction time, amount of water added, and number of decoctions. The number of decoctions is generally selected as 1, 2, or 3 times. Therefore, the single factor test mainly examines the influence of soaking time, extraction time, and amount of water added on the evaluation index.

[0063] Soaking time investigation: according to the prescription ratio, weigh 6 portions of the daily prescription amount of medicinal materials, add 10 times the amount of water respectively, soak for 0, 0.5, 1.0, 1.5, 2.0, and 2.5 hours, heat and extract for 1.0 hour, filter, measure the volume of the filtrate, and then calculate the dry paste rate under each condition according to the method under "2.1". The results are shown in Table 5.

[0064] Table 5 Immersion time test results ( n=3)

[0065]

[0066]

[0067] The results show that the increase in soaking time has little effect on the dry paste rate, so soaking time is not considered as a factor for optimizing the extraction process. However, in actual production, the amount of medicinal materials is large, and in order to allow the medicinal materials to absorb water, the soaking time before extraction is determined to be 0.5h.

[0068] Investigation of the amount of water added: weigh 6 portions of the daily prescription amount of medicinal materials according to the prescription ratio, add 5, 7, 9, 11, 13, and 15 times the amount of water respectively, soak for 0.5 h, heat and extract for 1.0 h, filter, measure the volume of the filtrate, and calculate the dry paste rate under each condition. The results are shown in Table 6.

[0069] Table 6 Solvent multiple test results ( n=3)

[0070]

[0071] From the results, it can be seen that the change in the amount of water added has a great influence on the dry paste rate. When the amount of water added reaches 9 times, the increase in the dry paste rate tends to be gentle. Therefore, the amount of water added is selected as the investigation factor for the optimization of the extraction process, and the levels are 5, 7, and 9 times.

[0072] Extraction time investigation: weigh 6 portions of the daily prescription amount of medicinal materials according to the prescription ratio, add 9 times the amount of water respectively, soak for 0.5h, heat and extract for 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0h, filter, measure the volume of the filtrate, and then calculate the dry paste rate under each condition according to the method under "2.1". The results are shown in Table 7.

[0073] Table 7 Extraction time investigation test results ( n=3)

[0074]

[0075] From the results, it can be seen that the increase of extraction time has a greater impact on the dry paste rate. After the extraction time reaches 1.5h, the increase of the dry paste rate tends to be gentle. Therefore, the extraction time is selected as the investigation factor for the optimization of the extraction process, and the levels are selected as 0.5, 1.0, and 1.5h.

[0076] Orthogonal test: factor level design. According to the results of the influencing factor test, the factors affecting the extraction process include soaking time, water addition, extraction time and extraction times. Combined with the actual production, this study selected the water addition (A), extraction time (B), and extraction times (C) that have a greater impact on the extraction efficiency as the investigation factors of the orthogonal test, and used the dry paste rate as the evaluation index. 9 (3 4 ) Orthogonal design was used to optimize the extraction process of Liuwei Shufu Lotion. The experimental factor levels are shown in Table 8.

[0077] According to the results of single factor investigation, the amount of water added (A), extraction time (B), and number of extractions (C) were selected as the investigation factors of the orthogonal experiment. The levels of orthogonal design factors are shown in Table 8.

[0078] Table 8 Factor level table

[0079]

[0080] Orthogonal test results

[0081] The dry extract rate was used as the evaluation index, and the optimal extraction process was determined based on the evaluation results of the dry extract rate. The orthogonal test arrangement and results are shown in Table 9, and the variance analysis is shown in Table 10.

[0082] Table 9 Orthogonal experimental design and results of Liuwei Shufu lotion

[0083]

[0084] Results analysis: variance analysis was performed on the dry paste yield, and the analysis results are shown in Table 10.

[0085] Table 10 Analysis of variance of dry paste yield

[0086]

[0087]

[0088] The results of range and variance analysis of dry paste rate showed that influencing factors A, B, and C had significant effects on the extraction process (P<0.05), and the order of influencing factors was: C>B>A, that is, the number of extractions had the greatest impact, followed by extraction time and water addition amount. The best combination conditions were: A3B3C3.

[0089] Taking into account economic cost, energy saving and actual production, combined with the range of dry paste rate and variance analysis results, the optimal extraction process of this product was determined to be A2B2C2, that is, weigh each medicinal material according to the prescription amount, soak it in water for 0.5h, decocted twice, adding 9 times the amount of water for the first time and 7 times the amount of water for the second time, and decocted for 1h each time.

[0090] Molding process research, preparation process:

[0091] The quality requirement of lotion is clarity, and a small amount of easily dispersed precipitation is allowed, so the molding process needs to consider the removal of impurities that are easy to precipitate. Since this product has many prescription flavors, complex chemical composition, and only uses traditional water extraction technology, precipitation will occur during the static process after the extract is concentrated, affecting the clarity of the product appearance. Combined with actual production conditions and cost-effective considerations, the "filtration method" is used to remove precipitation, and the change in the dry cream rate in the Liuwei Shufu lotion extract before and after filtration is examined to determine the molding process route of this product.

[0092] Weigh each medicinal material according to the prescription ratio (one-fifth), 18g each, weigh 3 portions, extract according to the process parameters optimized by the small test process, and repeat the test three times in a row. Filter the liquid with a 200-mesh filter cloth, and determine the dry paste content of the sample before and after filtration. The results are shown in Table 11.

[0093] Table 11 Investigation results before and after filtration

[0094]

[0095] The results showed that the change in the dry cream rate of the lotion was not much different, but the precipitation content of the concentrated solution after filtration was significantly reduced, and the clarification effect was better than that before filtration. This shows that filtering the liquid medicine with a 200-mesh filter cloth can not only retain the drug components of this product, but also obtain a liquid medicine with better clarity.

[0096] pH adjuster inspection shows that most lotions are prone to precipitation during storage and the clarity is unqualified. One of the reasons is the influence of pH value. Therefore, the pH value of the samples was inspected. The 2020 edition of the Chinese Pharmacopoeia stipulates under the general rules for preparations in Part IV, item 0127 of lotions, that lotions with water or dilute ethanol as solvents should generally be checked for pH value. Because changes in pH value may cause lotions to precipitate easily during storage and the clarity is unqualified. Therefore, the pH value of the samples was inspected, and the results are shown in Table 12.

[0097] Table 12 pH investigation of drug solution

[0098]

[0099] From the results, we can see that the pH value of the drug solution is between 5.0 and 5.1. Only a small amount of precipitation is produced during the sample placement process, which is dispersed by light shaking. Therefore, the addition of pH adjusters is not considered.

[0100] Selection and dosage of preservatives: Since this product needs to be opened repeatedly during use, it is easy to cause microbial contamination, so it is advisable to add an appropriate amount of preservatives. Sodium benzoate needs to be converted into benzoic acid under acidic conditions to work, so it has a better preservative effect on acidic lotions. [13-14] The pH value of this lotion is between 5.0 and 5.1. Considering the actual production and economic factors, sodium benzoate is selected as the preservative, and the dosage is 0.3%. The amount of sodium benzoate added meets the relevant requirements of the 2020 edition of the Chinese Pharmacopoeia, Part IV, General Rules for Preparations, Item 0127, Lotions.

[0101] In summary, after investigating the molding process, it was finally determined that the molding process of Liuwei Shufu Lotion is: extracting the concentrated liquid and filtering it, adding 0.3% sodium benzoate as a preservative.

[0102] Packaging materials, high-density polyethylene bottles (HDPE) for external liquid medicines are non-toxic, tasteless, odorless, not easy to break, and have good moisture-proof, oxygen-proof and display functions. They can protect drugs from the influence of surrounding environmental conditions and microorganisms, and have the advantages of light weight, low price, high safety in use, etc., which is conducive to the storage of liquid preparations. Therefore, this product intends to use high-density polyethylene bottles for external liquid medicines as the inner packaging material that directly contacts the drug.

[0103] Regarding the filling volume, according to the clinical use experience of the original prescription of the preparation in our hospital and comprehensive consideration of the actual production, the specification of Liuwei Shufu Lotion is set at 150ml per bottle, and each 1ml is equivalent to 0.6g of the medicinal piece.

[0104] Scale-up for pilot production:

[0105] Based on the process research, the process conditions were optimized and three batches of pilot production were carried out at 400 times the standard prescription amount. The trial production results are shown in Table 13.

[0106] Table 13 Results of three batches of pilot tests of Liuwei Shufu lotion

[0107]

[0108] The pilot results show that the yields of the three batches of pilot samples are all greater than 90%, and the identification and inspection items meet the requirements.

[0109] The production process is basically stable and basically meets the requirements of large-scale production, indicating that it is reasonable and feasible to use the above production process to prepare Liuwei Shufu Lotion.

[0110] According to the clinical medication needs, the Liuwei Shufu Lotion formula was changed from the original water decoction to a lotion. Single-factor experiment and orthogonal experiment were used, and the dry paste rate was used as the evaluation index to determine the extraction, concentration, liquid preparation, and filling process parameters of the Liuwei Shufu Lotion. The molding process was screened, and finally three batches of scale-up pilot production were carried out to verify the preparation process of the Liuwei Shufu Lotion. The results showed that the determined preparation process was scientific, reasonable and feasible.

[0111] Study on quality control of Zhuang medicine anti-inflammatory and antipruritic lotion:

[0112] 1 Experimental Materials

[0113] 1.1 Instrument

[0114] Agilent 1260 InfinityⅡ HPLC; Luna5μmC18(2) Chromatographic column (Agilent, USA, specifications 250×4.6mm, 5μm); ML204 Mettler analytical balance; Xiangyi H1650 desktop high-speed centrifuge; KQ-500DE ultrasonic cleaner; YOKO-ZS ultraviolet analysis camera (Wuhan Pharmaceutical New Technology Development Co., Ltd.); HWS-26 electric constant temperature water bath; Milli-Q pure water and ultrapure water integrated system.

[0115] 1.2 Reagents and Reference Materials

[0116] The reference medicinal materials of Jiubiying (batch number: 121076-201303), the reference medicinal materials of Mahonia (batch number: 121461-201503), and the reference substance of syringin (batch number: 111574-201605; purity: 95.2%, loading volume 20 mg / tube) were all provided by the China Food and Drug Inspection Institute; acetonitrile (chromatographic grade, Thermo Fisher Scientific Inc.); methanol (analytical grade, Merck KGaA); phosphoric acid (chromatographic grade, Shanghai Zhanyun Chemical Co., Ltd.); ether (AR. Chengdu Cologne Chemical Reagent Co., Ltd.); NaHCO3 (A R. Guangdong Guanghua Reagent Co., Ltd.); Na2CO3 (AR. Guangdong Guanghua Reagent Co., Ltd.); hydrochloric acid (AR. Guangdong Guanghua Reagent Co., Ltd.); ethyl acetate (AR. Chengdu Kelong Chemical Reagent Co., Ltd.); petroleum ether (AR. Chengdu Kelong Chemical Reagent Co., Ltd.); formic acid (AR. Tianjin Fuyu Fine Chemical Co., Ltd.); methanol (AR. Chengdu Kelong Chemical Reagent Co., Ltd.); aluminum chloride (AR. Tianjin Damao Chemical Reagent Co., Ltd.); sulfuric acid (AR. Guangdong Guanghua Reagent Co., Ltd.); vanillin (AR. Guangdong Guanghua Reagent Co., Ltd.).

[0117] 1.3 Medicinal materials and samples

[0118] All the medicinal materials used were purchased from Guangxi Xianzhu Chinese Medicine Technology Co., Ltd. and are qualified products tested according to the requirements of the Chinese Pharmacopoeia. Zhuang medicine anti-inflammatory and antipruritic lotion was made by the Zhuangyao Medicine Preparation Center of Guangxi International Zhuang Medicine Hospital. Batch number: 210601, 210602, 210603.

[0119] 2 Methods and Results

[0120] 2.1 TLC Identification of Jiubiying

[0121] 2.1.1 Preparation of reference solution

[0122] Take 0.5 g of Jiubing reference medicinal material powder, add 30 mL of methanol, ultrasonically treat for 30 minutes, evaporate to dryness, add 30 mL of water-saturated n-butanol to dissolve the residue, wash twice with ammonia test solution, 20 mL each time, evaporate the n-butanol solution after washing to dryness, add 2 mL of methanol to dissolve the residue to prepare a reference medicinal material solution.

[0123] 2.1.2 Preparation of test sample and negative solution

[0124] Take 20mL of Zhuangyao Xiaoyan Zhiyou Lotion, shake it thoroughly with 30mL of water-saturated n-butanol, take the n-butanol layer, wash it twice with ammonia test solution, 20mL each time, evaporate the n-butanol solution after washing to dryness, add 2mL of methanol to dissolve the residue, and take the supernatant as the test solution. Prepare the negative control solution of the lack of rescue in the same way.

[0125] 2.1.3 Thin layer chromatography

[0126] According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 Edition Part IV 0502), 1 μL of the test solution, 1 μL of the negative test solution, and 1 μL of the control medicinal material solution were respectively spotted on the same silica gel G thin layer plate, developed with chloroform-methanol-formic acid (16:3:1) as the developing agent, and then sprayed with 10% sulfuric acid ethanol solution after drying, heated at 105°C for 3-5 minutes, and inspected under a fluorescent lamp. The results are shown in Figure 2 , 1~3. Test solution (Batch No.: 210601, 210602, 210603) 4. Jiubiying negative solution 5. Jiubiying control drug.

[0127] 2.2 TLC Identification of Mahonia

[0128] 2.2.1 Preparation of reference solution

[0129] Take 0.1 g of the control medicinal material powder of Chinese magnolia vine, add 5 mL of methanol, and treat with ultrasound for 30 minutes. Let it stand, and take the supernatant as the control medicinal material solution.

[0130] 2.2.2 Preparation of test sample and negative solution

[0131] Take 20mL of Zhuangyao Xiaoyan Zhiyou Lotion, shake it thoroughly with 30mL of water-saturated n-butanol, take the n-butanol solution, wash it twice with ammonia test solution, 20mL each time, evaporate the n-butanol solution after washing, add 2mL of methanol to dissolve the residue, and take the supernatant as the test solution. Prepare the negative control solution of the lack of magnolia in the same way.

[0132] 2.2.3 Thin layer chromatography

[0133] According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 Edition Part IV 0502), 2 μL of the test solution, 2 μL of the negative test solution, and 2 μL of the control medicinal material solution were respectively spotted on the same silica gel G thin layer plate to form strips, and the upper layer solution of n-butanol-glacial acetic acid-water (8:1:4) was used as the developing agent to develop it, and then dried and examined under ultraviolet light (365nm). The results are shown in Figure 3 , 1~3. Test solution (Batch No.: 210601, 210602, 210603) 4. Mahonia lactiflora negative solution 5. Mahonia lactiflora control medicinal materials.

[0134] 2.4 Determination of Syringin Content

[0135] 2.4.1 Chromatographic conditions Chromatographic column: Luna 5μm C18 (2) (4.6x250mm, 5μm); mobile phase: mobile phase A (acetonitrile) 10%, mobile phase B (water) 90%; flow rate is 1.0ml / min; detection wavelength is 265nm; injection volume is 5μL; column temperature is 30℃.

[0136] 2.4.2 Preparation of reference solution Accurately weigh 10 mg of syringin reference substance (batch number: 111574-201605; purity: 95.2%; purchased from China Food and Drug Inspection Institute), place it in a 100 ml volumetric flask and make up to volume with 50% methanol to prepare a reference solution with a concentration of 0.0952 mg / ml.

[0137] 2.4.3 Preparation of test solution Use a pipette to measure 1 ml of Zhuangyao Anti-inflammatory and Antipruritic Lotion, dilute to 10 ml volumetric flask with water, shake well, centrifuge at 10000r / min for 10 min, take the supernatant and filter it through a 0.22μm microporous filter membrane to obtain the test solution. Prepare the negative control solution of deficiency-relief by the same method.

[0138] 2.4.4 Specificity experiment Take 5 μL of the above reference solution, test solution and negative control solution, inject them into HPLC chromatograph, and measure according to the law. The retention time of the chromatographic peak of syringin in the test sample is consistent with that of the reference solution, and there is no interference with the negative control. The results are shown in Figure 4 .

[0139] 2.4.5 Investigation of linear relationship The reference solution with 0.0952 mg / ml syringin prepared under 2.4.2 was respectively taken and filtered through a 0.22 μm microporous membrane and injected into an Agilent high performance liquid chromatograph for detection. The injection volume was 2 μL, 5 μL, 10 μL, 15 μL, 20 μL, 25 μL, and 30 μL, respectively, and the syringin peak area was recorded. With the syringin peak area (X) as the abscissa and the syringin concentration (Y, μg) as the ordinate, a linear regression was performed using the least squares method, and the regression equation of syringin was y=0.0004x-0.0339, R 2 =0.9995. The experimental results showed that the linear range of syringin was good between 0.190 μg and 2.856 μg.

[0140] 2.4.6 Precision experiment: Take the reference solution with a syringin content of 0.0952 mg / ml prepared under 2.4.2 and inject it six times continuously according to the method under 2.4.1. Determine and record the syringin peak area. The RSD of the syringin peak area is calculated to be 2.57%, indicating that the instrument has good precision.

[0141] 2.4.7 Repeatability experiment Take six portions of Zhuang medicine anti-inflammatory and antipruritic lotion from the same batch (batch number 210602), prepare the test solution according to the method under "2.4.3", and test according to the method under 2.4.1. The RSD of the peak area of ​​syringin is calculated to be 2.11%, indicating that the method has good repeatability.

[0142] 2.4.8 Stability test: Prepare the test solution according to the method under "2.4.3". When standing at room temperature for 0h, 3h, 6h, 12h and 24h, measure the peak area of ​​syringin according to the method. The RSD of the peak area of ​​syringin was calculated to be 1.53%, indicating that the solution has good stability within 24h.

[0143] 2.4.9 Sample recovery experiment: take six portions of Zhuangyao anti-inflammatory and antipruritic lotion with known content, 0.5ml each, and place them in 10ml volumetric bottles, then weigh an appropriate amount of syringin reference substance, prepare a reference substance solution with a concentration of 0.952mg / ml and accurately pipette 1ml into the sample, dilute with water and shake well, centrifuge at 10000r / min for 10min, take the supernatant and pass it through a 0.22μm microporous filter membrane, and then determine and record the syringin peak area according to the law, and calculate the average recovery rate to be 100.37%, and the RSD is 0.88%. The experimental results are shown in Table 14.

[0144] Table 14 Syringin recovery rate (n=6)

[0145]

[0146] 2.4.10 Sample content determination Take 1 ml of each of three batches (batch numbers: 210601, 210602, 210603) of Zhuang medicine anti-inflammatory and antipruritic lotion, prepare the samples according to the test sample preparation method under 2.4.3, and perform sample injection and detection according to the chromatographic conditions under 2.4.1. The contents of syringin in the Zhuang medicine anti-inflammatory and antipruritic lotion samples are calculated to be 0.225 mg / ml, 0.221 mg / ml, and 0.222 mg / ml, respectively.

[0147] 3 Discussion and Conclusion

[0148] 3.1 Selection of index components and mobile phase Jiubiying is the main drug of Zhuangyao Xiaoyan Zhiyou Lotion, which contains multiple chemical components such as saponins, terpenes, organic acids, etc. Its characteristic components are syringin and long-pedunculated holly glycoside. Since the content of long-pedunculated holly glycoside is rarely determined, the detection limits of other components such as holly glycoside A and holly glycoside B2 are relatively low, so syringin is used as the quality control index in this study. [7-10] The author consulted relevant literature and found that the content of syringin in different harvesting periods and growing environments varies greatly. In addition, the large number of ingredients in this formula is likely to affect the separation of the index components. [11-14]Therefore, four different mobile phase systems, acetonitrile-water, acetonitrile-0.1% phosphoric acid aqueous solution, methanol-water, and methanol-0.1% phosphoric acid aqueous solution, were investigated for elution. The results showed that when acetonitrile-water solution was used as the mobile phase for elution, the chromatographic peak separation effect was optimal and the peak shape was better, so it was selected as the mobile phase.

[0149] 3.2 Selection of detection wavelength In this study, the DAD detector was used to perform full wavelength scanning of syringin at 190-800 nm. The results showed that the baseline was stable at 265 nm, the chromatographic peak shape of syringin was good, and there was no interference from impurities, so 265 nm was selected as the detection wavelength.

[0150] 3.3 Selection of mobile phase flow rate and column temperature This study investigated three different flow rates of 0.8 ml / min, 1.0 ml / min and 1.2 ml / min, and the effects of three different column temperatures of 25°C, 30°C and 35°C on the chromatographic behavior of syringin. The results showed that when the flow rate was 1.0 ml / min and the column temperature was 30°C, the baseline was stable, the chromatographic peak separation was optimal, and the peak shape was the best. Therefore, 1.0 ml / min was selected as the elution flow rate and 30°C was selected as the column temperature of this method.

[0151] In summary, this experiment established for the first time the TLC identification method of Herba Lycopodii and Herba Coptidis and the content determination method of syringin in Zhuang medicine anti-inflammatory and antipruritic lotion. Among them, Herba Lycopodii and Herba Coptidis had good TLC separation effect, neat and clear spots, moderate ratio shift, good reproducibility, no negative interference, and strong specificity; the HPLC content determination method of syringin was simple to operate, with good repeatability and stability, high precision, and strong specificity, which can be used for the quality control of Zhuang medicine anti-inflammatory and antipruritic lotion, and provide a scientific basis for its later market production.

[0152] Clinical observation:

[0153] 1. Clinical observation location: Guangxi International Zhuang Medicine Hospital

[0154] 2. Clinical observation doctors: Li Fangmei, Qi Xixi, Wei Fuqiao, Feng Qiao, Huang Minting, Yan Yan, Meng Fei, Chen Dongqiong, Wang Dongshan, Zhou Dongmei

[0155] 3. Clinical observation start and end dates: January 1, 2019 - August 31, 2023

[0156] 4. Medication plan: Zhuang medicine anti-inflammatory and antipruritic prescription (250ml / bag), diluted with warm water to 1000-2000ml, then wash the affected area, once a day, each time for 15-20 minutes.

[0157] 5. General information of patients: A retrospective analysis was conducted on 160 patients, all of whom were outpatients of the dermatology department of our hospital. The diagnosis of all cases met the diagnostic criteria for skin diseases with acute attacks and impaired skin barrier function, such as acute dermatitis eczema, contact dermatitis, pederer dermatitis, scabies, impetigo, skin infection, pityriasis rosea, urticaria, burns and scalds, and after laser surgery.

[0158] 6. Observation indicators: A clinical observation table is developed according to the type of disease, mainly observing the changes in the area, number, severity of skin lesions, and subjective symptoms.

[0159] 7. Clinical observation method: 160 patients were selected according to the diagnostic criteria to observe their safety and efficacy after medication.

[0160] 8. Evaluation criteria: Calculate the score according to PASI, and determine the efficacy based on the comparison of the scores before and after treatment.

[0161] 9. Statistical analysis method: Ridit analysis is used for grade data, and X 2 Test, measurement data were tested using t(u) test.

[0162] 10. Efficacy results: The total clinical effective rate is 81.25%, the cure rate is 63.13%, and the efficacy is stable.

[0163] 11. Safety results: No obvious adverse reactions were observed when using this prescription.

[0164] 12. Conclusion: The results of clinical observations show that the Zhuang medicine anti-inflammatory and antipruritic prescription has good improvements on acute and subacute skin diseases with heat syndrome as the main symptom: acute dermatitis eczema, contact dermatitis, pederer dermatitis, scabies, impetigo, skin infections, pityriasis rosea, urticaria, burns, scalds, laser surgery and other acute attacks and skin diseases with damaged skin barrier function. The total clinical effective rate is high, no toxic side effects were found, and the clinical use is safe and effective.

[0165] 2. Prescription Information

[0166] 1. Composition

[0167] 15g of Chinese magnolia officinalis, 15g of scutellaria baicalensis, 15g of purslane, 15g of sanguisorba officinalis, 15g

[0168] 2. Usage and Dosage

[0169] Zhuang medicine anti-inflammatory and antipruritic prescription (250ml / bag), diluted with warm water to 1000-2000ml, wash the affected area once a day, each time for 15-20 minutes.

[0170] 3. Function and indications

[0171] Traditional Chinese Medicine: Clears away heat and dampness, cools blood and detoxifies, promotes tissue regeneration and heals sores. It is used for acute and subacute skin diseases with heat syndrome as the main symptom, caused by wind-heat, damp-heat, heat toxins and insect toxins in the skin, with symptoms of skin erythema, papules, blisters, pustules, exudate, erosion, scratches, scabs, accompanied by itching or pain; it is suitable for acute dermatitis eczema, contact dermatitis, pederus dermatitis, scabies, impetigo, skin infection, pityriasis rosea, urticaria, burns and scalds, laser surgery and other acute attacks and skin barrier function damage skin diseases with the above symptoms, as well as post-photoelectric surgery wound care.

[0172] Zhuang Medicine: It can clear away heat and poison, remove dampness and poison, regulate the airway, cool blood and detoxify, astringe and promote tissue regeneration and heal sores. It is used for acute and subacute skin diseases caused by wind poison, heat poison, dampness poison and insect poison in the skin, with symptoms of skin erythema, papules, blisters, pustules, exudate, erosion, scratches, scabs, accompanied by itching or pain; it is suitable for acute dermatitis eczema, contact dermatitis, pederus dermatitis, scabies, impetigo, skin infection, pityriasis rosea, urticaria, burns and scalds, laser surgery and other acute attacks and skin barrier function damaged skin diseases with the above symptoms, as well as post-photoelectric surgery wound care.

[0173] 4. Formula theory

[0174] The applicants and research team of this project have been engaged in the clinical and scientific research work of Zhuang medicine for the treatment of skin diseases for a long time, in order to inherit and protect ethnic medicine, conduct theoretical research and organize literature. Under the guidance of Zhuang medicine theory, a large number of ethnic techniques and medicines for the treatment of skin diseases have been summarized in clinical work. Among them, the Zhuang medicine anti-inflammatory and antipruritic prescription has outstanding efficacy in the treatment of acute and subacute skin diseases with heat syndrome as the main symptom. The prescription is explained as follows:

[0175] 4.1 Principles of TCM prescription composition and explanations

[0176] The formula uses Chinese magnolia as the main drug, which is cold and bitter in nature, can clear away heat, dry dampness, and detoxify, and can treat carbuncle, eczema, and burns. The formula uses Chinese scutellaria, fire charcoal mother (gou berry), and purslane as the assistant drugs. Among them, Chinese scutellaria is cold and bitter, and has the effects of clearing away heat and detoxifying, promoting dampness and relieving pain. It can be decocted in water for external washing or ground into powder for external use, and can treat burns, eczema, carbuncle, furuncle, and furuncle; fire charcoal mother is cold, astringent, and sour in nature, and has the effects of clearing heat and promoting dampness, and invigorating qi and promoting blood circulation, and is used to treat carbuncle, eczema, and wind-heat influx of the skin; purslane is cold and sour in nature, and has the effects of clearing heat and detoxifying, dispersing blood and reducing swelling. It is used in dermatology to treat carbuncle, malignant ulcer, etc. The three drugs are used together to strengthen the main drug's effect of clearing heat and detoxifying and cooling blood. The prescription uses Sanguisorba officinalis and Scutellaria baicalensis as adjuvants. Sanguisorba officinalis is bitter, sour, astringent, and slightly cold. It enters the liver and large intestine meridians and has the effects of cooling blood and stopping bleeding, detoxifying and astringing sores. It is often used in dermatology for burns caused by water or fire, carbuncles and sores. Scutellaria baicalensis clears away heat and dampness, purges fire, detoxifies and stops bleeding. It is used for carbuncles and sores, and strengthens the effects of the main medicine in cooling blood and stopping bleeding, detoxifying and astringing sores.

[0177] In summary, the formula uses Chinese magnolia as the main drug, which can clear away heat, dry dampness, and detoxify, and treat carbuncle, eczema, and burns; uses scutellaria baicalensis, fire charcoal mother (gou berry), and purslane as the assistant drugs, which can strengthen the effects of clearing heat, detoxifying, and cooling blood, and treat the itching symptoms associated with damp-heat skin diseases; uses dandelions and scutellaria baicalensis as the assistant drugs, which can cool blood and stop bleeding, detoxify and heal sores. The combination of these drugs can play the effects of clearing away heat and dampness, cooling blood and detoxifying, promoting tissue regeneration and heal sores, and can be used to treat acute dermatitis eczema, contact dermatitis, pederus dermatitis, scabies, impetigo, skin infection, pityriasis rosea, urticaria, burns and scalds, and skin diseases with acute attacks and impaired skin barrier function after laser surgery, and has good efficacy.

[0178] 4.2 Zhuang Medicine Prescription Principles and Explanations

[0179] The main drug in the prescription is Herba Lycopodii, which is cold and bitter in nature, and has the effects of clearing away heat and toxins, removing dampness and toxins, regulating the qi channel, and treating carbuncle, eczema, and burns. The auxiliary drugs in the prescription are Herba Lycopodii, Herba Pyriferae, and Herba Portulacae. Herba Lycopodii is cold and bitter in nature, and has the effects of clearing away heat and toxins, removing dampness and toxins, and unblocking the gullet channel. It can be decocted in water for external washing or ground into powder for external use, and can treat burns, scalds, eczema, carbuncle, furuncle, and furuncle; Herba Pyriferae is cold, astringent, and sour in nature, and has the effects of clearing away heat and toxins, removing dampness and toxins, cooling blood and relieving pain, and is used to treat carbuncle, eczema, and wind-heat flow in the skin; Herba Portulacae is cold and sour in nature, and can clear away heat and toxins, regulate the qi channel, clear away heat and detoxify, disperse blood and reduce swelling, and is used in dermatology to treat carbuncle, malignant ulcer, etc. The three drugs are used together to enhance the effects of the main drug in clearing heat and detoxifying and cooling blood. The prescription uses Radix Sanguisorbae and Radix Scutellariae as the main medicine. Radix Sanguisorbae is bitter, sour, astringent, slightly cold, and enters the liver and large intestine meridians. It has the effects of cooling blood and stopping bleeding, detoxifying and astringing sores. It is used for blood in stool, hemorrhoids, bloody diarrhea, metrorrhagia, burns caused by water or fire, carbuncle, sore and poison; Radix Scutellariae is used to clear away heat and dampness, purge fire and detoxify, stop bleeding, stabilize the fetus, and lower blood pressure. It is used for damp-heat, summer-heat chest tightness and nausea, damp-heat fullness, diarrhea, jaundice, lung heat cough, high fever and thirst, blood heat vomiting and bleeding, carbuncle, sore and poison, and restless fetus. The two medicines are used together to enhance the effect of the main medicine in cooling blood and astringing sores.

[0180] The whole prescription is composed of the main medicine, auxiliary medicine and auxiliary medicine of Zhuang medicine. The main medicine has the effects of clearing away heat and toxins, removing dampness and toxins, regulating the airway, and treating carbuncle, eczema and burns; the three auxiliary medicines are used together to strengthen the effect of the main medicine in clearing heat, detoxifying and cooling blood, and can also treat other concurrent symptoms, detoxify and treat sores; the two auxiliary medicines can strengthen the effect of the main medicine in cooling blood and astringing sores. The combination of all the medicines has the effects of clearing away heat and toxins, removing dampness and toxins, regulating the airway, cooling blood and detoxifying, astringing, regenerating muscles and astringing sores. It is used to treat acute dermatitis eczema, contact dermatitis, pederus dermatitis, scabies, impetigo, skin infection, pityriasis rosea, urticaria, burns and scalds, laser surgery and other acute attacks and skin diseases with damaged skin barrier function, and has good efficacy.

[0181] 3. Clinical medication observation

[0182] This clinical observation was formulated and implemented in accordance with the "Implementation Rules for the Registration Management of Traditional Chinese Medicine and Ethnic Medicine Preparations Prepared by Traditional Crafts in Guangxi Medical Institutions", "Guidelines for Clinical Research of New Chinese Medicines", "Technical Guidelines for Clinical Research of Syndrome-based New Chinese Medicines" and other regulatory documents, as follows:

[0183] Patient information and clinical medication observation:

[0184] Table 15 Patient information and clinical medication observation

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191] (II) General Information

[0192] 160 patients were observed in the outpatient department of dermatology in our hospital. Among them, 64 were male and 96 were female. The oldest was 80 years old and the youngest was 0 years old. The age and gender distribution of patients are shown in Table 16, the year distribution of cases is shown in Table 17, and the distribution of diseases is shown in Table 18.

[0193] Table 16 Patient age and gender distribution

[0194]

[0195] Table 17 Annual distribution of cases

[0196]

[0197] Table 18 Distribution of diseases diagnosed by Western medicine

[0198]

[0199]

[0200] 3. Diagnostic criteria

[0201] 1. The Zhuang medicine diagnostic standards for facial eczema / dermatitis (can be stiff or tired) are formulated with reference to the relevant contents of "Chinese Zhuang Medicine" published by Guangxi Nationalities Publishing House, "Chinese Zhuang Medicine Internal Medicine" published by Guangxi Science and Technology Publishing House, "Chinese Zhuang Medicine Disease Diagnosis and Treatment Standards" published by Guangxi Nationalities Publishing House, "Concise Zhuang Medicine" published by Guangxi Nationalities Publishing House, and "Zhuang Medicine Trauma Science" published by China Traditional Chinese Medicine Press.

[0202] Clinical manifestations: Main symptoms: erythema, papules, scales, exudate, scabs, accompanied by sensitivity and itching, intolerance to environmental warmth and heat. Concomitant symptoms: poor appetite, loose stools, heavy limbs, sticky stools, yellow urine. Eye diagnosis signs: The veins on the white of the eye are dark red or light, and the veins are thin and curved. Nail diagnosis signs: The nail bed is dark or light, and the moon mark is exposed too much or too little.

[0203] Tongue and pulse: red tongue, yellow and greasy coating, stringy and slippery pulse. It is more common in those with congenital sensitivity, and is prone to recurrence in winter. This disease can occur in men, women, old and young, and is often caused by rheumatism, heat and toxicity blocking the skin, resulting in blockage of the "dragon path" and "fire path", and poor circulation of qi and blood. In addition, long-term illness leads to deficiency of vital energy, blood deficiency and wind dryness, obstruction of the "two paths", and lack of nourishment to the skin, which can also lead to the induction of this disease.

[0204] 2. The diagnostic standards of Zhuang medicine for acne (acne) are formulated with reference to the relevant contents of "Chinese Zhuang Medicine" published by Guangxi Nationalities Publishing House, "Chinese Zhuang Medicine Internal Medicine" published by Guangxi Science and Technology Publishing House, "Chinese Zhuang Medicine Disease Diagnosis and Treatment Standards" published by Guangxi Nationalities Publishing House, and "Concise Zhuang Medicine" published by Guangxi Nationalities Publishing House.

[0205] Clinical manifestations: Main symptoms: It is common in adolescents. The course of acne is often long, often one after another. Some patients can gradually recover after adolescence. Millet-like papules appear on the face, chest, back, etc., like thorns, or cysts and nodules. Some merge into pieces, red and swollen or with pus heads, which may be accompanied by slight itching or pain. White or yellow-white rice-like powder juice can be squeezed out. Some patients may develop cysts, nodules or form scars due to improper treatment or lack of hygiene. Concomitant symptoms: Pigmentation may appear in some affected areas. Eye diagnosis signs: "Le Da" The veins of the white eye are dark in color, and the veins of the white eye are less curved, small in curvature, and thick. Tongue and pulse: The tongue is red, the tongue is dark, or there are ecchymosis or petechiae, the tongue coating is thin and yellow, greasy, or yellow or yellow greasy, and the pulse is slippery or stringy.

[0206] Nail diagnosis signs: The nail bed is light red, bright red or pale white, the nail body is smooth and moist, the lunula is clear, the nail folds are symmetrical and complete, and the blood color returns to a uniform state when pressed. It is more common in young and middle-aged men and women, and is often caused by improper diet and invasion of external pathogens.

[0207] The efficacy evaluation criteria were formulated in accordance with the "Guidelines for Clinical Research of New Chinese Medicines" published by the State Administration of Traditional Chinese Medicine. The clinical efficacy evaluation is divided into 4 levels: clinical cure, marked effect, effective, and ineffective.

[0208] Score reduction index = (total score before treatment - total score after treatment) / total score before treatment × 100%.

[0209] ①Clinical cure: All skin lesions disappear and symptoms disappear; the score reduction index is ≥90%;

[0210] ② Significantly effective: Most of the skin lesions disappeared, the symptoms were significantly relieved, and the score index decreased by 60% to 89%;

[0211] ③ Improvement: The skin lesions are partially improved, the symptoms are improved, and the score index decreases by 30% to 59%;

[0212] ④ Ineffective: Facial skin lesions did not improve significantly, symptoms did not improve or worsened, and the score reduction index was <30%.

[0213] Effective rate = basic cure + percentage of marked effect.

[0214] 4. Statistical analysis: SPSS software was used to analyze the age, gender, course of disease and other general information of patients. Ridit analysis was used for graded data, and X-ray analysis was used for technical data. 2 Test, measurement data were tested using t(u) test.

[0215] 4. Results and Analysis

[0216] 1. Analysis of clinical efficacy

[0217] The overall clinical efficacy of Zhuangyao Xiaoyan Zhiyun prescription in 160 patients reached 81.25%, of which 101 were cured, 29 were markedly effective, 30 were improved, and 0 were ineffective. The specific analysis is shown in Table 19. The main diseases included dermatitis / eczema, pruritus, skin infection, scabies, urticaria, pityriasis rosea, and burns / laser surgery. The specific analysis of the total efficacy of each disease is shown in Table 20.

[0218] Table 19 The efficacy of Zhuang medicine anti-inflammatory and antipruritic prescriptions

[0219]

[0220] Table 20 The efficacy of Zhuang medicine anti-inflammatory and antipruritic prescriptions in treating various diseases

[0221]

[0222] (II) Safety

[0223] According to the statistics of adverse reaction observation, 160 patients were observed, and no serious adverse events, serious complications or adverse reactions occurred after taking the medicine.

[0224] (3) Typical Cases: Typical Case 1: Patient Lu Moumou, female, 1 month old, presented at the dermatology outpatient clinic of Guangxi International Zhuang Medicine Hospital on September 23, 2019 due to "facial rash for 10 days". The family member reported that the patient had had a facial rash for 10 days without treatment and no other discomfort. Special physical examination: Symmetrical erythema was seen on the face. The tongue was red, the coating was yellow, and the pulse was rapid. Diagnosis: Western medicine diagnosis: Eczema; Traditional Chinese medicine diagnosis: Damp sore - Damp-heat syndrome; Zhuang medicine diagnosis: Neng Han Neng Lei - Yang syndrome. Prescription: Zhuang medicine anti-inflammatory and antipruritic formula (250 ml / bag), 3 bags.

[0225] Dosage and usage: Each time, take 50 ml of the medicinal liquid, dilute it with 200 ml of warm water, then dip and wet compress the affected area, twice a day, 5 minutes each time. Instructions: Encourage breastfeeding, and the mother should have a light diet; pay attention to keeping the facial skin of the patient moist, and apply a moisturizing cream after each medicated bath. Telephone follow-up on September 30, 2019: After 1 week of treatment, the family member reported that the facial rash of the patient had completely subsided, with no other discomfort, and no adverse reactions were seen during the medication process. The curative effect of Lu Moumou was judged as clinical cure after treatment.

[0226] Typical Case 2: Qiu Moumou, female, 30 years old, presented at the dermatology outpatient clinic of Guangxi International Zhuang Medicine Hospital on February 11, 2023 due to "facial rash for 1 year". The patient reported that she had developed a facial rash without obvious cause 1 year ago, and was diagnosed with "acne" in an external hospital. After taking isotretinoin capsules, doxycycline capsules, etc., the condition improved; new rashes repeatedly appeared after stopping the medication, and the condition worsened when eating spicy and greasy foods, staying up late, and under great mental stress; new rashes repeatedly occurred, leaving scars. Symptoms: Erythema, follicular papules, pigmentation, and pitted scars were seen on the face. The tongue was light red, the coating was thin and white, and the pulse was slippery. Diagnosis: Western medicine diagnosis: Acne; Traditional Chinese medicine diagnosis: Acne - Damp-heat syndrome; Zhuang medicine diagnosis: Le Qiu - Yang syndrome.

[0227] Prescription: 1. Doxycycline hydrochloride enteric-coated capsules 0.1 g * 20, twice a day, 0.1 g each time, orally, for 7 consecutive days; 2. Compound lidocaine cream 10 g * 2, apply externally before laser surgery; 3. Human epidermal growth factor gel 100,000 IU (200 μg): 20 g, twice a day, 1 g each time, apply externally, for 7 consecutive days; 4. Zhuang medicine anti-inflammatory and antipruritic formula (250 ml / bag) * 7 bags, each time take half a bag (125 ml), dilute it with 500 ml of warm boiled water, then clean the postoperative wound surface, once in the morning and once in the evening, 3 - 5 minutes each time.

[0228] Disposal: 1. Zhuang medicine fire needle treatment; 2. CO2 fractional laser treatment. Instructions: Have a light diet, avoid scratching the affected area, and pay attention to moisturizing and sun protection. WeChat follow-up on February 18, 2023: The patient reported that the wound surface had completely scabbed and fallen off after the operation, the facial skin was not greasy or flushed, and no new rashes appeared. The curative effect of Qiu Moumou was judged as markedly effective after treatment.

[0229] 5. Conclusion: Zhuang medicine anti-inflammatory and antipruritic prescription has good improvement on acute and subacute skin diseases with heat syndrome as the main feature: acute dermatitis eczema, contact dermatitis, pederer dermatitis, scabies, impetigo, skin infection, pityriasis rosea, urticaria, burns, scalds, laser surgery and other acute attacks and skin diseases with damaged skin barrier function. The total clinical effective rate is high. So far, no adverse reaction events have been found for this prescription. The prescription is safe and effective for clinical use.

Claims

1. Six-flavor soothing lotion, characterized in that: Every 1000 ml of skin lotion solution is prepared by decocting and concentrating the following raw materials by weight: 100 g of Scutellaria baicalensis, 100 g of Psoralea corylifolia, 100 g of Meiniemei, 100 g of Rhizoma Coptidis, 100 g of Coptis chinensis and 100 g of Radix Sanguisorbae Officinalis.

2. Six-flavor soothing lotion, characterized in that: Each 60 ml skin lotion solution is prepared by water extraction and concentration of the following raw materials by weight: 6.0 kg of Scutellaria baicalensis, 6.0 kg of Psoralea corylifolia, 6.0 kg of Meiniemei, 6.0 kg of Rhizoma Coptidis, 6.0 kg of Coptis chinensis, 6.0 kg of Radix Sanguisorbae and 0.18 kg of sodium benzoate.

3. The method for preparing the Liuwei Shufu lotion according to claim 1, characterized in that: Soak all the medicinal raw materials for 30 minutes, decoct twice, add 9 times the amount of water for the first time and 7 times the amount of water for the second time, decoct for 1 hour each time, filter, combine the filtrate, concentrate the filtrate to a concentrate with a relative density of 1.03-1.10, the temperature is 60°C, filter, let stand overnight, take the supernatant and boil for 30 minutes, add 3g of sodium benzoate, add pure water to 1000ml, stir evenly, and fill.

4. The method for preparing the Liuwei Shufu lotion according to claim 2, characterized in that: The steps include: Step 1: add six Chinese medicinal pieces of Radix Scutellariae, Herba Portulacae, Herba Eupatorii, Herba Euphorbiae, Herba Mahoniae and Herba Sanguisorbae to an extraction tank, soak them in water for 30 minutes, decoct them twice, add 9 times the amount of water for the first time and 7 times the amount of water for the second time, decoct them for 1 hour each time, and during the decoction process, control the steam pressure at 0.01-0.25MPa before the boiling of the medicinal liquid, and control the steam pressure at 0.01-0.10MPa after the boiling of the medicinal liquid, so that the medicinal liquid is kept boiling, filter the medicinal liquid with a 100-mesh filter, combine the filtrate and set aside; Step 2: Pump the filtrate in step 1 into a single-effect concentrator unit, with a concentration temperature of 75-90°C and a vacuum degree of -0.06--0.09MPa, and concentrate to a concentrate with a relative density of 1.03-1.10 at a temperature of 60°C. Filter the concentrate with a 200-mesh filter cloth and put it into a clean barrel, seal it with a cover, weigh it, attach a material card, and place it in a cool warehouse to stand overnight. Take the supernatant into a clean barrel, seal it with a cover, weigh it, attach a material card, and set it aside; Step 3: Weigh sodium benzoate and dissolve it with purified water for later use. Take the supernatant of Liuwei Shufu Lotion Concentrate from the cool storage in a sandwich pot, heat and boil for 30 minutes, stop heating, add the completely dissolved sodium benzoate solution while stirring, stir for 30 minutes to mix, dilute to the prescribed amount with purified water, continue stirring for 30 minutes to mix, and the preparation is completed. Filter the prepared liquid with a 200-mesh filter cloth, put it in a clean bucket and set aside; Step 4: Transfer the prepared liquid medicine to the liquid filling room of the external preparation workshop, start the automatic bottle unscrambler and the liquid filling machine linkage production line respectively, install and debug according to the batch production requirements, and debug the filling volume: 150ml / bottle. After the filling volume debugging is completed, confirm that the filling volume of each injection is qualified; the bottle cap is screwed in place, there is no crooked cap, no looseness, and no leakage in the seal; after the outer wall of the medicine bottle is clean and free of pollution, start filling. When the equipment is running at normal speed, during the filling process, and before the end of filling, check the filling volume and sealing conditions. Each time, check 10 bottles of products produced continuously, and check the filling volume and sealing conditions and record them. If the filling volume and sealing are unqualified, stop the machine immediately for inspection and adjust to qualified. The filled products are conveyed to the outer packaging room via the conveyor belt.

5. The TLC identification and detection method of schizonepeta tenuifolia in Liuwei Shufu lotion according to claim 1 or 2, characterized in that: The steps include: Step 1: Preparation of reference solution: 0.5 g of Jiubing reference medicinal material powder was added with 30 mL of methanol, and ultrasonically treated for 30 minutes, evaporated to dryness, and the residue was dissolved in 30 mL of water-saturated n-butanol, and washed twice with ammonia test solution, 20 mL each time, and the n-butanol solution after washing was evaporated to dryness, and the residue was dissolved in 2 mL of methanol to prepare a reference medicinal material solution; Step 2: Preparation of test sample and negative solution: take 20 mL of Zhuangyao Xiaoyan Zhiyou Lotion, shake it thoroughly with 30 mL of water-saturated n-butanol, take the n-butanol layer, wash it twice with ammonia test solution, 20 mL each time, evaporate the n-butanol solution after washing to dryness, add 2 mL of methanol to dissolve the residue, take the supernatant as the test sample solution, and prepare the negative control solution of Qujiu Biying in the same way; Step 3: Thin layer chromatography: follow the thin layer chromatography test, take 1 μL of the test solution, 1 μL of the negative test solution, and 1 μL of the control medicinal material solution, and spot them on the same silica gel G thin layer plate respectively. Develop with chloroform-methanol-formic acid as the developing solvent, then spray with 10% sulfuric acid ethanol solution (chloroform: methanol: formic acid is 16:3:1), heat at 105°C for 3-5 minutes, and inspect under a fluorescent light.

6. The TLC identification and detection method of the Chinese magnolia officinalis in Liuwei Shufu lotion according to claim 1 or 2, characterized in that: The steps include: Step 1: Preparation of reference solution: Take 0.1 g of Chinese magnolia officinalis reference medicinal material powder, add 5 mL of methanol, ultrasonically treat for 30 minutes, let stand, and take the supernatant as the reference medicinal material solution; Step 2: Preparation of test sample and negative solution: take 20 mL of Zhuangyao Xiaoyan Zhiyou Lotion, shake it thoroughly with 30 mL of water-saturated n-butanol, take the n-butanol solution, wash it twice with ammonia test solution, 20 mL each time, evaporate the n-butanol solution after washing to dryness, add 2 mL of methanol to dissolve the residue, take the supernatant as the test sample solution, and prepare the negative control solution of L. magnoliae in the same way; Step 3: Thin layer chromatography: According to the thin layer chromatography test, 2 μL of the test solution, 2 μL of the negative test solution, and 2 μL of the control medicinal material solution are respectively spotted on the same silica gel G thin layer plate to form strips. The plate is developed with the upper layer solution of n-butanol-glacial acetic acid-water as the developing agent, and then the plate is inspected under ultraviolet light after drying. The ratio of n-butanol: glacial acetic acid: water is 8:1:

4.

7. The method for determining the content of syringin in Liuwei Shufu Lotion according to claim 1 or 2, characterized in that: The steps include: Step 1: Preparation of reference solution: accurately weigh 10 mg of syringin reference substance, place it in a 100 ml volumetric flask and dilute it with 50% methanol to make a reference solution with a concentration of 0.0952 mg / ml; Step 2: Preparation of the test solution: use a pipette to measure 1 ml of Zhuangyao Xiaoyan Zhiyou Lotion, dilute to 10 ml volumetric flask with water, shake well, centrifuge at 10000r / min for 10 min, take the supernatant and filter through a 0.22μm microporous membrane to obtain the test solution. Prepare the negative control solution of Qujiu Biying in the same way; Step 3: Take 5 μL of the reference solution, test solution and negative control solution, respectively, and inject them into the HPLC chromatograph for determination according to the law. The retention time of the chromatographic peak of syringin in the test sample is consistent with that of the reference solution, and there is no interference with the negative control; Step 4: Linear relationship investigation: The reference solution with a syringin content of 0.0952 mg / ml obtained in step 1 was respectively taken and filtered through a 0.22 μm microporous filter membrane and injected into an Agilent high performance liquid chromatograph for detection. The injection volumes were 2 μL, 5 μL, 10 μL, 15 μL, 20 μL, 25 μL, and 30 μL, respectively. The syringin peak area was recorded. The syringin peak area was used as the abscissa and the syringin concentration was used as the ordinate. The least squares method was used for linear regression, and the regression equation for syringin was y=0.0004x-0.0339, R 2 =0.9995, the linear range of syringin is between 0.190μg and 2.856μg; Step 5: Precision experiment, take the reference solution with a syringin content of 0.0952 mg / ml prepared in step 1 and inject it six times continuously according to the method under the chromatographic conditions, measure and record the syringin peak area, and calculate the RSD of the syringin peak area to be 2.57%, indicating that the instrument has good precision. Step 6: Repeatability experiment, take six portions of the same batch of Zhuangyao anti-inflammatory and antipruritic lotion, prepare the test solution according to the method in step 2, and inject and test according to the method of chromatographic conditions, and calculate the RSD of the syringin peak area to be 2.11%; Step 7: Stability test, prepare the test solution according to the method in step 2, and determine the peak area of ​​syringin according to the method when standing at room temperature for 0h, 3h, 6h, 12h, and 24h, and calculate the RSD of the peak area of ​​syringin to be 1.53%; Step 8: Sample addition recovery experiment, take six portions of Zhuang medicine anti-inflammatory and antipruritic lotion with known content, 0.5 ml each, and place them in 10 ml volumetric flasks respectively, then weigh an appropriate amount of syringin reference substance, prepare a reference substance solution with a concentration of 0.952 mg / ml and accurately pipette 1 ml into the sample respectively, make up to volume with water and shake well, centrifuge at 10000 r / min for 10 min, take the supernatant and filter it through a 0.22 μm microporous filter membrane, and determine and record the syringin peak area according to the law, and calculate the average recovery rate to be 100.37%, and the RSD is 0.88%; Step 9: Sample content determination, take 1 ml of each of the three batches of skin lotion, prepare the sample according to the sample preparation method in step 2, and test the sample according to the chromatographic conditions. The content of syringin in the Zhuang medicine anti-inflammatory and antipruritic lotion sample is calculated to be 0.225 mg / ml, 0.221 mg / ml, and 0.222 mg / ml, respectively.

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