Preparation process of ganoderma lucidum compound hair-growing liniment

Abstract

The invention discloses a preparation process and quality evaluation of a ganoderma lucidum compound hair-growing liniment, which is characterized in that ganoderma lucidum, polygonum multiflorum, cacumen biotae and angelica sinensis are used as raw materials, 50% of ABTS free radical scavenging rate and 50% of sensory evaluation are used as assessment indexes, and the optimal formula of the ganoderma lucidum compound hair-growing liniment is optimized through a single factor test and a response surface test. The result is that the addition amount of the plant water extract is 54mL, the addition amount of minoxidil is 0.8 g, the addition amount of 95% ethanol is 18mL, the addition amount of 1, 2-propylene glycol is 27mL, the addition amount of edetate disodium is 0.05 g, and pure water is supplemented to 100mL. The hair-growing liniment has the effects of resisting oxidation, inhibiting bacteria and growing hair of mice, and is safe and non-irritant.

Description

Technical Field

[0001] The invention relates to the technical field of natural cosmetics, and in particular to the preparation and quality evaluation of a natural plant ganoderma lucidum compound hair growth liniment. Background Art

[0002] With the rapid development of modern life, the pressure of study and work increases, the spirit becomes tense, sleep is seriously lacking, the diet tends to be greasy and spicy, the hair becomes increasingly sparse, the hairline gradually moves back, the scalp is itchy and painful, and moisture is lost, causing great distress to people.

[0003] Minoxidil can improve local blood circulation, increase blood supply around hair follicles, provide more nutrients to hair follicles, and promote hair growth. It can be used for many types of hair loss, including androgenic alopecia, alopecia areata, postpartum hair loss, etc. However, in order to maintain the hair growth effect, it needs to be used continuously. If the drug is discontinued, the newly grown hair may gradually fall off. Although it has a good hair growth effect, long-term use can cause some side effects. Reflex sympathetic excitement can cause accelerated heart rate, arrhythmia, and skin flushing; water and sodium retention can cause weight gain and lower limb edema; hair hyperplasia, which is more obvious on the face, arms and back, often appears within 3-6 weeks after medication and disappears 1-6 months after discontinuation of medication.

[0004] Chinese herbal medicine can extend the length and cycle of hair growth, effectively slow down the rate of hair loss, and promote the growth and development of hair follicles. Chinese herbal hair growth and hair darkening ointments can promote hair growth and darkening, laying a foundation for the development of Chinese herbal compound in hair care products. [1] Among them, Ganoderma lucidum, Polygonum multiflorum, Angelica sinensis, and Platycladus orientalis leaves have attracted widespread attention in terms of hair growth.

[0005] Ganoderma Lucidum Karst is the fruiting body of the Polyporaceae fungus Ganoderma, which has the dual functions of immune activation and immune regulation. Ganoderma lucidum is combined with Chinese angelica, salvia miltiorrhiza, etc. to make a compound Ganoderma lucidum hair growth formula. This formula can stimulate local vasodilation, promote blood circulation, and regulate the immune function of cells near the hair bulb. It has a good effect in promoting hair regeneration. [2] .

[0006] He Shouwu can promote hair growth, prevent hair loss, and treat premature graying of hair. [3] .

[0007] The flavonoids and phthalides in Angelica have strong antioxidant activity. When used in shampoo, they can also protect hair and prevent hair loss. [4] .

[0008] Platycladus orientalis leaves are rich in active ingredients such as terpenes, volatile oils, and flavonoids, which can effectively improve scalp blood circulation and strengthen the metabolic function of hair follicles. Its total flavonoids can divide, proliferate, and differentiate, increase the growth rate of hair, restore the vitality of hair follicles, and further promote blood circulation. It is particularly suitable for treating permanent hair loss and premature graying of hair caused by blood heat. [5] .

[0009] The present invention introduces the theory of traditional Chinese medicine into the development of new products such as cosmetics and shampoo products. By rationally selecting Chinese medicinal materials, the hair growth can be promoted, the hair quality can be effectively improved, and the effect of hair growth and growth can be achieved. Modern Chinese medicine extraction and effective component detection and analysis technologies are applied to product research and development.

[0010] [1] Liu Xue. Screening of Chinese herbal compound for hair growth and hair darkening and its pharmacodynamics study[D]. China Pharmaceutical Industry Research Institute, 2020.

[0011] [2] Chen Tianzhang. Clinical observation on the treatment of 40 cases of alopecia areata with compound Ganoderma lucidum hair growth tincture[J]. Journal of Nanjing Medical University, 1995, (01): 51.

[0012] [3] Li Tingting. Research on the formula of hair growth liquid and shampoo with hair growth effect[D]. Guangxi University of Science and Technology, 2021.

[0013] [4] Yang Dingquan, Liu Yunzhen, Ju Hai, et al. Clinical observation of hair growth tincture combined with Danggui Kushen pills in the treatment of androgenic alopecia[J]. World Journal of Integrated Traditional and Western Medicine, 2010, 5(04): 327-329+332.

[0014] [5] Ma Shihong, Shan Chengying, Nie Wei, et al. Study on the anti-hair loss efficacy of Platycladus orientalis leaves and Polygonum multiflorum composite extracts[J]. Chinese Wild Plant Resources, 2021, 40(05): 43-47. Summary of the invention

[0015] Long-term use of minoxidil may cause some side effects, such as dry scalp, itching, irritation and other problems. The purpose of this study is to solve the side effects of minoxidil and use the advantages of Chinese herbal medicine to compound the two. A preparation process of Ganoderma lucidum compound hair growth liniment is provided. First, the ABTS clearance rate is used as an indicator to optimize the extraction process of plant extracts; secondly, the antioxidant and properties are used as evaluation criteria to screen the addition amount of ethanol, 1,2-propylene glycol, and plant extracts to optimize the formula of Ganoderma lucidum compound liniment; finally, the functional evaluation of antioxidant, antibacterial, and hair growth effects and safety evaluation are investigated to prepare a hair growth liniment that has the effect of promoting hair growth and is safe and non-irritating.

[0016] To achieve the above purpose, the present invention adopts the following experimental scheme:

[0017] The invention provides a ganoderma lucidum compound hair growth liniment on one hand, which comprises plant extract, 95% ethanol, 1,2-propylene glycol, minoxidil tincture and disodium edetate; after compounding, the mixture is allowed to stand and then packaged to obtain the ganoderma lucidum compound hair growth liniment.

[0018] Preferably, the preparation of the plant extract: take dried Ganoderma lucidum, Polygonum multiflorum, Platycladus orientalis leaves, and Angelica sinensis, grind them, pass them through a 20-mesh sieve, add them in the mass ratio of 10.5:10.5:5.1:3.9 in sequence, and the solid-liquid ratio is 1:20 (g / mL). After soaking for 0.5 h, the temperature is 90°C, and the reflux extraction is heated for 3 h. The extract is filtered with 3 layers of gauze, and then placed in a centrifuge and centrifuged at a speed of 4800 r / min for 10 min, and the top clear liquid is taken.

[0019] Preferably, the Ganoderma lucidum compound hair growth liniment uses a variety of excipients, 95% ethanol and 1,2-propylene glycol dissolution agents, minoxidil hair growth agent, and edetate disodium stabilizer.

[0020] Preferably, the total amount of Ganoderma lucidum compound hair growth ointment is 100 mL, the amount of plant water extract added is 54 mL, the amount of minoxidil tincture added is 0.8 g (0.8%), the amount of 95% ethanol added is 18 mL, the amount of 1,2-propylene glycol added is 27 mL, and the amount of edetate disodium added is 0.05 g, and it is supplemented to 100 mL with 0.5% edetate disodium solution.

[0021] According to a second aspect of the present invention, the present invention provides a method for preparing a Ganoderma lucidum compound hair growth liniment, comprising the following steps:

[0022] a) mixing ethanol and 1,2-propylene glycol uniformly;

[0023] b) mixing minoxidil tincture with a), placing in an ultrasonic cleaner for ultrasonication for 10 min at a temperature of 40° C., after the mixture is dissolved, adding the plant extract, and shaking to mix well;

[0024] c) Dissolve edetate disodium in pure water using ultrasonic method, then add to b) and mix evenly, and ultrasonicate for 30 minutes until all the edetate is dissolved.

[0025] d) After cooling, the mixture is packaged to obtain the Ganoderma Lucidum Compound Hair Growth Liniment.

[0026] The ganoderma lucidum compound hair-growing liniment of the present invention has the effects of anti-oxidation, antibacterial and hair-growing.

[0027] Model group (shaving without medication), positive control group (2% minoxidil 0.26mL), negative control group (0.65mL of 0.8% compound liniment without Ganoderma lucidum), low-dose group (0.325mL of 0.8% compound liniment of Ganoderma lucidum), medium-dose group (0.65mL of 0.8% compound liniment of Ganoderma lucidum), high-dose group (1.3mL of 0.8% compound liniment of Ganoderma lucidum). The experiment was administered for 27 days. The model group grew light hair, but the hair did not grow completely. Compared with the positive and low-dose groups, the hair growth was almost complete. The hair growth of mice in the medium-dose and high-dose groups did not grow completely at the junction of the shaved area and the unshaved area. The low-dose group was relatively better. The scores from high to low were low-dose>positive group>medium dose>high dose>negative group>model group. The low-dose hair growth score reached 4.96, and the unit linear density was 2.92mg / mm, proving that the hair growth liquid has a good effect.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] (1) The present invention uses Ganoderma lucidum as the main ingredient and mixes it with Polygonum multiflorum, Angelica sinensis and Platycladus orientalis leaves. The mixed plant extract has certain antioxidant activity.

[0030] (2) Ethanol and 1,2-propylene glycol in a ratio of 3:4.5 can dissolve minoxidil better, and ultrasonic dissolution can dissolve it faster. The combination of plant extract and minoxidil can reduce the dosage of the latter, reduce costs, and improve safety. The combination of plant extract and minoxidil can achieve the effect of 1+1>2.

[0031] (3) The compound Ganoderma lucidum hair growth ointment has a good antibacterial effect, and synergistically promotes hair growth with antioxidants to create a good environment for the scalp, achieving the effect of hair growth and is safe and non-irritating.

[0032] (4) After adding plant extract, the hair growth ointment has a refreshing and non-greasy texture, is easier to be absorbed by the scalp, and does not cause greasiness or stickiness in the hair, thereby improving the convenience of use and maintaining the fresh fragrance of the plant. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 ABTS free radical scavenging rate curves of four traditional Chinese medicines

[0034] Figure 2 Response surface plot of plant extract addition amount and minoxidil tincture concentration

[0035] Figure 3 Response surface plot of plant extract addition amount and 1,2-propylene glycol addition amount

[0036] Figure 4 Response surface plot of minoxidil tincture concentration and 1,2-propylene glycol addition amount

[0037] Figure 5Hair growth in mice

[0038] Figure 6 Pathological sections of liver tissues of mice in normal group (a), model group (b), positive group (c), negative group (d), low (e), medium (f), and high (g) dose groups DETAILED DESCRIPTION

[0039] Example 1 Determination of active ingredient content

[0040] 1. Polysaccharide content determination method

[0041] Prepare 1 mg / mL glucose standard solution, dilute the standard solution with different concentrations of 0.015 mg / mL-0.075 mg / mL, add 1 mL of 5% phenol and 5 mL of 98% concentrated sulfuric acid, let stand for 30 minutes, and measure the absorbance at 490 nm.

[0042] Standard curve: y = 6.935x + 0.0959, R 2 =0.9992

[0043] 2. Flavonoid content determination method

[0044] Prepare 0.2 mg / mL rutin standard solution, accurately measure 1.5 mL, 2 mL, 3 mL, 4 mL, and 5 mL of the standard solution respectively and place them in a 25 mL volumetric flask, add 30% ethanol to 6 mL, add 5% sodium nitrite solution 1 mL and let it stand for 6 min, add 10% aluminum nitrate solution 1 mL and let it stand for 6 min, add 4% sodium hydroxide solution 10 mL, finally make up to volume with 30% ethanol, shake well and let it stand for 15 min, and measure the absorbance at a wavelength of 500 nm.

[0045] Standard curve y = 1.347x + 0.0517, R 2 =0.9991

[0046] Example 2 ABTS free radical scavenging experiment

[0047] Take 1.8mL ABTS solution and mix it with 0.2mL sample solution, shake it for 10s as the experimental sample solution, and let it stand in the dark for 6min. At the same time, take another 1.8mL ABTS solution and mix it with 0.2mL distilled water, shake it for 10s as the control solution, and let it stand in the dark for 6min. After that, adjust the wavelength of the UV spectrophotometer to 734nm, preheat it for 30min, measure the absorbance, and calculate the clearance rate. The higher the clearance rate, the stronger the antioxidant activity. Formula:

[0048]

[0049] Wherein, A0, A1 and A2 are the absorbance values ​​of the blank group, the control group and the sample group respectively. Example 3: Proportion screening experiment of plant extracts

[0050] Take dried Ganoderma lucidum, Polygonum multiflorum, Platycladus orientalis leaves, and Angelica sinensis, crush them, pass through a 20-mesh sieve, accurately weigh them according to different mass ratios, fix the solid-liquid ratio to 1:20 (g / mL), soak them for 0.5h, heat at 90℃, and reflux for 3h. After the extraction is completed, filter them with 3 layers of gauze, put the filtrate into a centrifuge and centrifuge it at 4800r / min for 10min, and take the top clear liquid as the plant extract. The concentration of the raw medicinal material is 50mg / mL, and the raw medicinal material concentration is 3.3mg / mL after dilution 15 times for ABTS free radical scavenging rate experiment. Formula:

[0051]

[0052] Among them, A1 and A2 are the amount of raw medicinal materials and pure water used respectively.

[0053] 1. Single Factor Experiment

[0054] In the single-factor experimental screening experiment on the dosage of raw materials for the extract, 9g of Polygonum multiflorum, 6g of Platycladus orientalis leaves, and 3g of Angelica sinensis were fixed factors. The masses of Ganoderma lucidum were 9g, 10.5g, 12g, 13.5g, and 15g, respectively. The dosage of Ganoderma lucidum was initially screened, and then the dosage of Ganoderma lucidum was fixed. Polygonum multiflorum, Platycladus orientalis leaves, and Angelica sinensis were screened, and the clearance rate was used as an indicator of the plant extract.

[0055] The fixed factors were Ganoderma lucidum 10.5g, Platycladus orientalis leaves 6g, Angelica sinensis 3g, and Polygonum multiflorum added in amounts of 6g, 7.5g, 9g, 10.5g, and 12g, respectively;

[0056] The fixed factors were Ganoderma lucidum 10.5 g, Polygonum multiflorum 10.5 g, Angelica sinensis 3 g, and Platycladus orientalis leaves added in amounts of 4.2 g, 5.1 g, 6 g, 6.9 g, and 7.8 g, respectively;

[0057] The fixed factors were Ganoderma lucidum 10.5 g, Polygonum multiflorum 10.5 g, Platycladus orientalis 6 g, and Angelica sinensis added in amounts of 1.2 g, 2.1 g, 3 g, 3.9 g, and 4.8 g, respectively;

[0058] The results showed that when the amount of Ganoderma lucidum was 10.5g, the scavenging rate of the extract was the highest, which was 73.42%; when the amount of Polygonum multiflorum was 10.5g, the scavenging rate of the extract was the highest, which was 75.31%; when the amount of Platycladus orientalis leaves was 6.0g, the scavenging rate of the extract was the highest, which was 78.34%; when the amount of Angelica sinensis was 3.0g, the scavenging rate of the extract was the highest, which was 80.96%. Figure 1 .

[0059] 2. Orthogonal experiment

[0060] The dosage of Ganoderma lucidum, Polygonum multiflorum, Platycladus orientalis and Angelica sinensis was based on single factor, and three levels were selected for each dosage (see Table 1) according to L3 (3 4 ) Experimental table (see Table 2) was used to conduct orthogonal test to screen dosage, and the clearance rate was used as the evaluation index to select the optimal ratio of plant extract.

[0061] Table 1 Orthogonal factors and levels

[0062]

[0063] Table 2 Orthogonal experimental results and intuitive analysis

[0064]

[0065] The results are shown in Table 2. The influence of the four factors on the ABTS clearance rate is in the order of Polygonum multiflorum > Ganoderma lucidum > Angelica sinensis > Platycladus orientalis leaves; the best process of optimized plant extract: Ganoderma lucidum 10.5g, Polygonum multiflorum 10.5g, Platycladus orientalis leaves 5.1g, Angelica sinensis 3.9g, the clearance rate is 80.52±1.21%, the polysaccharide content is 16.32±0.54%, and the flavonoids content is 3.41±1.08%.

[0066] 3. Verification experiment

[0067] According to the results of the orthogonal experiment raw material dosage screening: Ganoderma lucidum 10.5g, Polygonum multiflorum 10.5g, Platycladus orientalis leaves 5.1g, Angelica sinensis 3.9g, three parallel verification experiments. When the concentration of the medicinal materials was 3.3mg / mL, the scavenging rate was 80.76±1.36%, when the VC concentration was 8.3ug / mL, the ABTS free radical scavenging rate was 80%, the polysaccharide content of the extract was 16.27±0.21%, and the flavonoid content was 3.46±0.64%, indicating that the plant extract has a certain antioxidant effect.

[0068] Example 4: Screening of the formula of compound Ganoderma lucidum hair growth liquid

[0069] Single factor experiment: fixed factors: 3.5mL 95% ethanol, 5mL 1,2-propylene glycol, 9mL plant extract, 0.05% stabilizer EDTA-disodium, minoxidil concentration of 1.0%. Variable factors: 5% ethanol, 1,2-propylene glycol, plant extract and minoxidil addition, 5 levels for each factor. The method is to mix the 4 raw materials and shake them evenly, and place them in an ultrasonic cleaner for 40 minutes. The antioxidant activity and appearance of the mixed solution were comprehensively scored to investigate the effect of the amount of plant extract added on it.

[0070] Comprehensive score = (ABTS free radical scavenging rate × 0.5 / maximum ABTS free radical scavenging rate + appearance trait score × 0.5 / maximum appearance trait score) × 100.

[0071] Table 3 Scoring criteria for properties of Ganoderma lucidum compound hair growth liniment

[0072]

[0073] 1. Single Factor Experiment

[0074] (1) Screening of the amount of plant extract added

[0075] Taking the total score of antioxidant activity and properties as the evaluation index, the added amount of plant extract was investigated at 3mL, 6mL, 9mL, 12mL, and 15mL respectively.

[0076] Table 4 Results of investigation on the amount of liquid added

[0077]

[0078] As shown in Table 4, the third group had the highest comprehensive score, so the amount of plant extract added was selected to be 9 mL to prepare the Ganoderma lucidum compound hair growth liniment.

[0079] (2) Minoxidil concentration screening

[0080] Taking the total score of antioxidant activity and properties as the evaluation index, the concentrations of minoxidil tincture investigated were 0.6%, 0.8%, 1%, 1.2%, and 1.4%, respectively.

[0081] Table 5 Nordil tincture concentration investigation results

[0082]

[0083]

[0084] As shown in Table 5, the second group had the highest comprehensive score, so the minoxidil concentration of 0.8% (g / mL) was selected to prepare the Ganoderma lucidum compound hair growth liniment.

[0085] (3) Screening of ethanol addition amount

[0086] Taking the total score of antioxidant activity and properties as the evaluation index, the amount of ethanol added was investigated at 2.5mL, 3mL, 3.5mL, 4mL, and 4.5mL respectively.

[0087] Table 6 Results of investigation on ethanol addition

[0088]

[0089] As shown in Table 6, the second group had the highest comprehensive score, so the amount of ethanol added was selected to be 3 mL to prepare the Ganoderma lucidum compound hair growth liniment.

[0090] (4) Screening of the amount of propylene glycol added

[0091] Taking the total score of antioxidant activity and properties as the evaluation index, the addition amounts of 1,2-propylene glycol were investigated at 4mL, 4.5mL, 5mL, 5.5mL, and 6mL respectively.

[0092] Table 71,2 Results of investigation on the amount of propylene glycol added

[0093]

[0094]

[0095] From Table 7, the second group had the highest comprehensive score, so the amount of 1,2-propylene glycol added was selected to be 4.5 mL to prepare the Ganoderma lucidum compound hair growth liniment.

[0096] 2. Response surface experiment

[0097] Based on the single factor dosage of plant extract, minoxidil, ethanol, and 1,2-propylene glycol, the response surface experiment was designed using Design-Expert 8.0.6 software. The design table is shown in Table 8, and the appearance properties and ABTS free radical scavenging rate were used as evaluation indicators.

[0098] Table 8 Response surface design factor levels and coding

[0099]

[0100] Table 9 Response surface experiment results

[0101]

[0102]

[0103] Design-Expert8.0.6 software was used to perform variance analysis on the regression parameters of the response surface, and the quadratic fitting equation was:

[0104] M=94.82+0.71A+2.40B+3.16C-1.86D-5.60AB-3.66AC-2.68AD-6.67BC+1.61BD+0.16CD-11.63A 2 -7.89B 2 -0.48C 2 -5.75D 2 After performing variance analysis on the regression model, the key statistics of the model were obtained. Among them, the correlation coefficient R 2The RAdj value is 0.9585, which significantly proves the high consistency between the experimental prediction value and the observed value. At the same time, the RAdj value is 0.9170, which confirms the good fit of the model and shows that the experimental error is small, thus verifying the rationality of the model. In addition, the coefficient of variation CV of the model is 2.83%, which highlights the reliability of the experimental results. Figure 2-4 The optimal formula of process parameters of compound Ganoderma lucidum hair growth ointment is: 9 mL of plant extract, 0.8% concentration of minoxidil tincture, 3 mL of ethanol, and 4.5 mL of 1,2-propylene glycol.

[0105] Example 5: Preparation method of Ganoderma lucidum compound hair growth liniment

[0106] Mix ethanol and 1,2-propylene glycol evenly; then add minoxidil and mix evenly, add plant extract and shake evenly, place in ultrasonic cleaner for 10 minutes, ultrasonic temperature 40°C; dissolve disodium edetate with pure water, add the solution to the solution containing minoxidil and mix evenly, ultrasonic again for 30 minutes until all dissolved. After cooling, seal and package to obtain Ganoderma lucidum compound hair growth liniment.

[0107] Example 6 Functional evaluation of Ganoderma lucidum compound hair growth liniment

[0108] 1. Antibacterial experiment

[0109] Place the agar medium in a microwave oven for high-temperature heating for about 3 minutes, then let it stand for a while, and then slowly pour it into the plate to ensure that there are no large bubbles and that it is evenly distributed. Subsequently, the culture media of Staphylococcus aureus, Bacillus subtilis, Bacillus pumilus, Saccharomyces cerevisiae, Candida albicans and Escherichia coli are subjected to corresponding high-temperature heating treatments, and left to stand for use. The prepared bacterial suspension is accurately aspirated with a pipette and evenly added to the culture medium, then gently shaken to ensure that it is evenly mixed, and then placed on top of the solidified agar plate. Among them, the amount of bacterial suspension of Candida albicans and Saccharomyces cerevisiae is 2.5mL, while other bacterial suspensions are taken in 1.5mL. After it is completely solidified, mark the name of the strain.

[0110] Use a hole puncher to make circular paper pieces (5 mm in diameter) and place 5 paper pieces of different strains in the corresponding area of ​​the plate. Use a pipette to take 15 μL of Ganoderma lucidum compound hair growth liniment and evenly drop it on the paper pieces. Keep the clean bench ventilated until the paper pieces are completely dry.

[0111] The culture plates of Candida albicans and Saccharomyces cerevisiae were placed in a lower incubator at 28°C for 24 hours; while the culture plates of Escherichia coli, Bacillus subtilis, Bacillus pumilus and Staphylococcus aureus were placed in an upper incubator at a constant temperature of 37°C for 16 hours, and the size of the inhibition zone and the antibacterial effect could be observed.

[0112] From the information in Table 10, it can be seen that the Ganoderma lucidum compound hair growth ointment has an antibacterial effect, and the Ganoderma lucidum compound hair growth ointment has the best antibacterial effect.

[0113] Table 10 Measurement results of the inhibition zone of Ganoderma lucidum compound hair growth liniment

[0114]

[0115] (2) Mouse hair growth experiment

[0116] Healthy male KM mice were taken and raised for one week. The mice were observed to be normal and randomly divided into 77 groups, with 10 mice in each group. They were normal group, model group, positive sample group, negative group, low-dose group, medium-dose group, and high-dose group. The normal group did not remove hair, and the other six groups were treated with hair removal in an area of ​​about 2cm×3cm on the back. The model group did not apply any sample liquid after hair removal, and it grew naturally, which served as a blank control. The high, medium, and low-dose groups were applied with 1.3mL / time, 0.65mL / time, and 0.325mL / time of Ganoderma lucidum compound hair growth liniment on the hair removal area of ​​the mice, respectively. The positive sample group was applied with 2% minoxidil hair growth liniment 0.26mL / time on the hair removal area, and the negative sample group was applied with 0.8% minoxidil hair growth liquid without Ganoderma lucidum 0.65mL / time on the hair removal area. All drug-treated groups were applied twice / day for 27 consecutive days to observe the hair growth of the mice. The skin color changes, skin allergic reactions and hair growth of the mice in each group were observed at each time period. The hair growth of the mice was scored and photographed during the experiment. Figure 5 .

[0117] Table 11 Scoring criteria

[0118]

[0119] Table 12 Mouse hair growth status score (n = 10, x ± SD)

[0120]

[0121] Compared with the model group, * P<0.05, ** P<0.01, **** P<0.0001

[0122] The experiment was administered for 27 days. The model group grew light hair, but the hair was not fully grown. Compared with the positive and low dose groups, the hair growth was almost complete. The hair growth of the mice in the shaved and unshaved areas in the medium and high dose groups was not fully grown. The low dose group was relatively better. The scores from high to low were low dose > positive group > medium dose > high dose > negative group > model group. This proves that the hair growth liquid has a good effect.

[0123] (3) Detection of hair length and weight of mice

[0124] After the experiment, use tweezers to pluck the hair in the longer area of ​​the hair removal area, select the longest 3-10 hairs and measure their length (first measure the longest 3 hairs, then measure the 10 hairs), and use scissors to cut off the new hair in an area of ​​1cmx2cm along the hair root. Use an analytical balance to weigh and calculate the average hair weight of each group of mice, and compare the hair length and weight among the groups.

[0125] On the 27th day of the experiment, the hair length and weight of each group of mice were measured, and the linear density was recorded in units of hair weight / hair length as shown in Table 13.

[0126] Table 13 Hair length and weight of mice in each group (n=10, x±SD)

[0127]

[0128] Compared with the model group, ** P<0.01, **** P<0.0001

[0129] The hair density was measured by dividing the gross weight by the hair linear density. The results showed that the low-dose group had the highest hair density, and the effect was close to that of the positive drug minoxidil. The medium-dose group had a greater hair density than the high-dose group. The negative-dose group had a greater hair density than the model group. The low-dose hair density was close to that of the normal group. Ganoderma lucidum compound hair growth ointment has the effect of growing hair.

[0130] Example 7 Safety Evaluation

[0131] (1) Comparison of organ index

[0132] The mice were weighed, killed, and the five organs of heart, liver, spleen, pancreas, glands, and kidneys were removed, weighed, and their relative weights were calculated. The weight and organ index of each group of mice at the end of the experiment were statistically analyzed, and the results are shown in Table 14.

[0133] Table 14 Comparison of mouse body weight and visceral index (n=10; x±SD)

[0134]

[0135] There was no significant difference in spleen index among the normal group, model group, positive group, negative group and drug administration group.

[0136] (2) Collection of blood biochemical indicators

[0137] After continuous administration for 27 days, blood was collected from the eye capillaries and the supernatant was collected after low-temperature centrifugation. The blood biochemical indicators such as ALT, AST and ALP in the serum were measured using an automatic biochemical analyzer. The results are shown in Table 15.

[0138] Table 15 Levels of AST, ALP and ALT in mice

[0139]

[0140]

[0141] Compared with the normal group, * P<0.05

[0142] There was no statistical difference in ALT and AST levels among the experimental groups; the ALP positive group had a higher level than the other groups ( * P<0.05), and there was no significant difference between the other groups and the normal group. This indicates that the Ganoderma lucidum compound hair-growth liquid has no damage to the liver function of mice.

[0143] (3) Pathological observation of liver tissue

[0144] The liver of mice in each experimental group was cut and fixed in a 4% paraformaldehyde solution, dehydrated, and embedded in paraffin. The tissue wax block was cut into slices with a thickness of 4um. After HE staining, the pathological changes of liver tissue were observed under an optical microscope (200 times). Microscopic observation showed clear structure. The hepatocyte cords were arranged radially around the central vein, and several mononuclear and binuclear hepatic cells were visible. No obvious cell degeneration and necrosis were observed in the liver of mice in each experimental group. Figure 6 .

[0145] Example 8 Stability Evaluation

[0146] pH value determination: According to the national standard for the detection of pH value of cosmetics (GB / T13531.1-2008), the pH value of the shampoo sample was measured at room temperature of 25°C. The pH meter was calibrated according to the instrument instructions, the electrode was inserted into the shampoo, the reading was taken, and the data was recorded. The experiment was repeated three times.

[0147] Centrifugal test: According to the experimental method described in the National Standard of the People's Republic of China for Shampoo and Shampoo Cream (GB / T29679-2013), take 10 mL of shampoo and centrifuge it at 2000r / min, 3000r / min, 4000r / min, and 5000r / min for 10 minutes respectively to observe whether the shampoo shows stratification, separation, etc.

[0148] Heat resistance test: According to the experimental method described in the National Standard of the People's Republic of China for Shampoo and Shampoo Cream (GB / T29679-2013), take two portions of shampoo, 10 mL each, place one portion of shampoo in a constant temperature incubator at 40±1℃ for 24 hours, then take it out, cool it to room temperature, and compare it with the other portion to observe whether the waterproofing becomes thinner, discolored, stratified, precipitated, etc.

[0149] Cold resistance test: According to the test method described in the National Standard of the People's Republic of China for Shampoo and Shampoo Cream (GB / T29679-2013), take two portions of shampoo, 10 mL each, place one sample in a refrigerator at -15±1℃ for 24 hours, then take it out and cool it to room temperature. Then compare it with the other portion to observe whether the shampoo becomes thinner, discolored, stratified, precipitated, etc.

[0150] According to the evaluation results in Table 16, it is concluded that the Ganoderma lucidum compound hair growth liniment has a uniform appearance and a pH value of 4.0 to 9.0. It can maintain its original state after centrifugation tests and heat and cold resistance tests and has good stability.

[0151] Table 16 Evaluation of the stability of hair growth liquid

[0152]

Claims

1. A preparation method for a Ganoderma lucidum compound hair growth liniment, wherein the raw materials include plant extract and minoxidil, and the auxiliary materials include 95% ethanol, 1,2-propylene glycol, and disodium edetate.

2. The Ganoderma lucidum compound hair growth liniment according to claim 1, characterized in that: The volume percentage of the plant extract with a crude drug concentration of 3.3 mg / mL in the hair growth ointment is 36% to 72%, the mass concentration of minoxidil is 0.6% to 1.0%, the volume percentage of 95% ethanol is 15% to 21%, and the volume percentage of 1-2 propylene glycol is 24% to 30%.

3. The Ganoderma lucidum compound hair-growth liniment according to claim 1, wherein the compound plant extract is prepared by the following method: 10.5 g of Ganoderma lucidum, 10.5 g of Polygonum multiflorum, 6.9 g of Platycladus orientalis, and 2.1 g of Angelica sinensis are crushed, with a solid-liquid ratio of 1:20 (g / mL), soaked for 0.5 h, and then the temperature is 90° C. and the extraction time is 3 h; the plant extract is filtered through 3 layers of gauze, centrifuged at 4800 rpm for 10 min, and the extract with a crude drug concentration of 50 mg / mL is the plant extract in the formula. When the crude drug concentration is 3.3 mg / mL, the ABTS free radical scavenging rate is 80.76±1.36%.

4. The Ganoderma lucidum compound hair growth liniment according to claim 1, wherein the amount of plant water extract added is 54 mL, the amount of minoxidil added is 0.8 g, the amount of 95% ethanol added is 18 mL, the amount of 1,2-propylene glycol added is 27 mL, and the amount of edetate disodium added is 0.05 g.

5. The Ganoderma lucidum compound hair growth liniment according to claim 1 is a light yellow clear solution with a light Chinese herbal medicine fragrance, a pH value of 6.3±0.2, and good stability.

6. A method for preparing the Ganoderma lucidum compound hair growth liniment according to claim 1, comprising the following steps: a) mixing ethanol and 1,2-propylene glycol uniformly; b) mixing minoxidil with a), placing in an ultrasonic cleaner for 10 min at a temperature of 40° C., and after the mixture is dissolved, adding the plant extract and shaking to mix well; c) Dissolve edetate disodium in pure water using ultrasonic method, then add to b) and mix evenly. Ultrasonication time is 30 min until all edetate is dissolved. d) After cooling, seal the mixture to obtain the Ganoderma Lucidum Compound Hair Growth Liniment.

7. A pharmacodynamic study on the Ganoderma lucidum compound hair growth liniment according to claim 1. The experimental administration lasted for 27 days. The model group grew light hair, but the hair was not fully grown. Compared with the positive and low dose groups, the hair growth was almost complete. The connection between the shaved area and the unshaved area in the medium and high dose groups was not fully grown. The scores from high to low were low dose > positive group > medium dose > high dose > negative group > model group. The hair growth score of the low dose was 4.96, and the unit linear density was 2.92 mg / mm, which proved that the hair growth liquid had a good effect.

8. A safety evaluation of the Ganoderma lucidum compound hair-growing liniment according to claim 1, wherein there is no significant difference in spleen organ index, liver blood biochemical indexes, and liver tissue pathological observation between the drug-treated group and the normal group; microscopic observation of each drug-treated group and the normal group shows clear liver cell cords arranged radially around the central vein, several mono- and binucleate liver cells can be seen, and no obvious cell degeneration and necrosis are observed, indicating that the Ganoderma lucidum compound hair-growing liniment is relatively safe.

9. A use of the Ganoderma lucidum compound hair growth liniment according to claim 1, characterized in that The prepared ointment has antioxidant, antibacterial and hair growth effects.

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