Shampoo with bacteriostatic, dandruff-removing and itch-relieving effects and preparation method thereof

Abstract

The present application belongs to the field of biotechnology. More particularly, it relates to a shampoo with the effects of inhibiting bacteria, removing dandruff and relieving itching and a preparation method thereof. The shampoo comprises the following components in mass percentage: 5-25% of surfactant, 0.1-0.3% of guar hydroxypropyltrimonium chloride, 0.3-0.5% of polyquaternium-100, 0.1-0.3% of sodium chloride, 0.1-0.5% of PPG-3 octyl ether, 1-3% of piper cubeba extract, 0.1-1.0% of piroctone olamine, 2-4% of humectant, 0.1-0.5% of chelating agent, 0.01-0.05% of pH regulator, 0.1-0.5% of essence and the balance of water. The human use evaluation test results show that the shampoo has more significant effects on conditions such as scalp swelling, dandruff and itching than market dandruff shampoo, and after 28 days of use by the test patients, the dandruff and itching phenomena are significantly improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology. More particularly, it relates to a shampoo with antibacterial, anti-dandruff and anti-itching effects and a preparation method thereof. BACKGROUND

[0002] Seborrheic Dermatitis (SD) is a chronic, papular and scaly skin disease with excessive sebum secretion and Malassezia infection. SD often occurs on the head, face, chest and back, and occurs on the scalp as Scalp seborrheic dermatitis / Dandruff (SSD). Seborrheic dermatitis is a chronic and recurrent inflammatory scalp disease characterized by the formation of erythema or mild to moderate scales, causing oily scalp and flaky dandruff with itching.

[0003] The main pathogenic bacteria of SSD is the abnormal colonization of Malassezia, which changes the position and leads to the disorder of scalp microbial ecosystem. The enzymes and corresponding metabolites produced by Malassezia can directly damage the cuticle of the skin, activate the immune mechanism of the scalp and produce chronic inflammation [1] . Its mechanism may be related to the activation of pattern recognition receptors, inflammasomes and NF-κB signaling pathway; at the same time, Malassezia can also form biofilm to block the effect of drugs, thereby producing drug resistance [2] . Therefore, effectively inhibiting the reproduction of Malassezia is one of the key points in the treatment of this disease. However, in the process of clinical treatment, chemical drugs have obvious side effects and drug resistance, especially the repeated and long-term use of antifungal drugs, which makes Malassezia lack sensitivity to them, often leading to treatment failure.

[0004] Alpinia katsumadai is the dried nearly mature seeds of the perennial herb Alpinia katsumadai of the Zingiberaceae family, which has the effects of drying dampness, promoting qi, warming the middle and stopping vomiting. Studies have shown that the volatile oil in Alpinia katsumadai has the effects of protecting gastric mucosa, anti-gastric ulcer, anti-inflammatory and antibacterial. It was found that the volatile oil of Alpinia katsumadai seed cluster had certain inhibitory effect on Escherichia coli, Staphylococcus aureus and Bacillus subtilis, but there was no report on the inhibitory effect of Alpinia katsumadai extract on Malassezia.

[0005] [1]Tao R,Li R,Wang R.Skin microbiome alterations in seborrheic dermatitis and dandruff:Asystematic review[J].Exp Dermatol,2021,30(10):1546-1553.

[0006] [2] Adalsteinsson JA, Kaushik S, Muzumdar S, et al. An update on the microbiology, immunology and genetics of seborrheic dermatitis [J]. Experimental dermatology, 2020, 29(5): 481-489. SUMMARY

[0007] The present application is first discovered that the extract of P. amalago has the effect of inhibiting Malassezia, especially Malassezia globosa, and further tests find that the extract of P. amalago has a satisfactory synergistic effect with traditional bacteriostatic agent piroctone olamine salt, thereby obtaining the present application.

[0008] The present application aims to provide a shampoo with the effects of inhibiting bacteria, removing dandruff and relieving itching, comprising the following components in mass percentage: surfactant 5-25%, guar hydroxypropyltrimonium chloride 0.1-0.3%, polyquaternium-10 0.3-0.5%, sodium chloride 0.1-0.3%, PPG-3 octyl ether 0.1-0.5%, P. amalago extract 1-3%, piroctone olamine salt 0.1-1.0%, humectant 2-4%, chelating agent 0.1-0.5%, pH regulator 0.01-0.05%, essence 0.1-0.5%, and water in residual amount.

[0009] Further, the P. amalago extract is the n-hexane extraction part of the alcohol extract of P. amalago.

[0010] Further, the preparation method of the P. amalago extract is as follows:

[0011] S1, taking dried seeds of P. amalago, crushing, sieving, adding ethanol for heating reflux extraction, collecting the extract, filtering, concentrating the filtrate, recovering ethanol, drying to obtain dry extract;

[0012] S2, adding n-hexane to the above dry extract for extraction, concentrating to constant weight under reduced pressure, and drying to obtain the extract.

[0013] Further, in step S1, the heating reflux extraction conditions are as follows: solid-liquid ratio 1:10-30, reaction temperature 60-80℃; reaction time 1-3h; and ethanol volume fraction 60-85%.

[0014] Further, in step S2, the n-hexane extraction process parameters are as follows: temperature 40-70℃; extraction time 1.0-2.5h; and solid-liquid ratio 1:5-10.

[0015] Further, the surfactant is selected from one or a combination of two or more of sodium laureth sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine, cocamide MEA, sodium cocoyl glycinate, sodium lauroyl sarcosinate, sodium cocoyl glutamate and sodium myristoyl glutamate.

[0016] Further, the humectant is selected from one or a combination of two or more of allantoin, propylene glycol, sodium hyaluronate, panthenol, glycerin and keratin hydrolysate.

[0017] Further, the chelating agent is selected from one or a combination of two or more of EDTA, disodium EDTA and tetrasodium glutamate dicetate; and the pH adjusting agent is citric acid.

[0018] Another object of the present application is to provide a shampoo having antibacterial, dandruff-removing and itch-relieving effects, comprising the following components in the following mass percentages:

[0019] Sodium laureth sulfate 5-10%;

[0020] Sodium cocoyl methyl taurate 1-5%;

[0021] Cocamidopropyl betaine 1-5%;

[0022] Cocamide MEA 0.1-1%;

[0023] Guar hydroxypropyltrimonium chloride 0.1-0.3%;

[0024] Polyquaternium-100 0.3-0.5%;

[0025] Sodium chloride 0.1-0.3%;

[0026] PPG-3 octyl ether 0.1-0.5%;

[0027] Piper nigrum extract 1-3%;

[0028] Piroctone olamine 0.1-1.0%;

[0029] Panthenol 2-4%;

[0030] Disodium EDTA 0.1-0.5%;

[0031] Citric acid 0.01-0.05%;

[0032] Fragrance 0.1-0.5%; and

[0033] Water q.s.

[0034] Another object of the present application is to provide a method for preparing the shampoo, comprising the following steps:

[0035] Pre-disperse guar gum hydroxypropyltrimonium chloride in cold water, add disodium EDTA, stir evenly, heat to 50-70 DEG C, add sodium lauryl polyoxyethylene sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine and cocamide MEA in turn, after 20 min of insulation, add piroctone olamine, stir evenly, cool to 30-45 DEG C, add polyquaternium-10, PPG-3 octyl ether, stir evenly, then add pimenta racemosa extract, stir for 30 min, cool to room temperature, add panthenol and essence, stir for 30-40 min, add citric acid and sodium chloride respectively, product discharge, and the product is obtained.

[0036] The present application has the following beneficial effects:

[0037] (1) The inhibition zone test and MIC value test results show that the pimenta racemosa extract of the present application has an inhibition zone diameter of > 20 mm on Malassezia globosa, and the inhibition effect is most obvious, being extremely sensitive.

[0038] (2) The combined drug sensitivity test results show that the pimenta racemosa extract of the present application does not show synergistic effect when combined with traditional anti-dandruff agents, shows synergistic effect only when combined with OCT, FICI≤0.5, shows additive effect when combined with ketoconazole and climbazole, and shows unrelated effect when combined with ZPT and HD-100.

[0039] (3) The human use evaluation test results show that the shampoo of the present application has more significant effect on dandruff and itch than market anti-dandruff shampoo, and after 28 d of use by the test patients, dandruff and itch are significantly improved. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 The scalp dandruff change of the 10th patient before and after using the shampoo of example 1 for 28 d;

[0041] Figure 2 The average scalp oil change of the two groups of patients before and after using for 28 d;

[0042] Compared with the control group, * P < 0.05. DETAILED DESCRIPTION

[0043] The present application will be further described below in combination with specific examples, but the examples do not limit the present application in any form. Unless otherwise specified, the reagents, methods and devices used in the present application are conventional reagents, methods and devices in the technical field.

[0044] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0045] Malassezia solid medium: 1% proteose peptone, 2% agar, 1% glucose, 0.2% yeast extract, 0.05% glycerol monostearate, 0.5% Tween-60, 1% glycerol, 0.8% oxgall salt and 2% olive oil, constant volume to 1000 mL, pH 7.0-7.2; the liquid medium does not contain agar.

[0046] Example 1-3, antibacterial and dandruff-removing shampoo (% by mass)

[0047]

[0048]

[0049] Preparation method:

[0050] After pre-dispersing guar gum hydroxypropyltrimonium chloride in cold water, add disodium EDTA, stir until uniform, heat to 70°C; add sodium laureth sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine and cocamide MEA in sequence, after 20 min of incubation, add piroctone olamine, stir until uniform; after cooling to 40°C, add polyquaternium-10 and PPG-3 octyl ether, stir until uniform, then add piper longum extract, stir for 30 min; after cooling to room temperature, add panthenol and fragrance, stir for 30 min, then add citric acid and sodium chloride, respectively, discharge the finished product, and the product is obtained.

[0051] Test Example 1, comparison of antibacterial activity of different extraction fractions of piper longum

[0052] 1.1. Preparation of different extraction fractions of piper longum: take dried seeds of piper longum, crush into coarse powder and sieve, soak in 65% ethanol solution according to the solid-liquid ratio of 1:10 for 30 min, then heat to reflux extraction at 70°C for 2 times, 1.5 h each time, collect the extraction liquid, filter and remove the residue; concentrate the filtered liquid, refrigerate to precipitate ethanol, recover the precipitated ethanol, dry the supernatant to obtain dry extract. Add 5 times the weight of n-hexane, petroleum ether, chloroform and ethyl acetate to the above dry extract, respectively, extract at 45°C for 2.0 h, concentrate to constant weight under reduced pressure, and dry to obtain n-hexane, petroleum ether, chloroform and ethyl acetate extraction fractions, respectively, for standby use.

[0053] 1.2 Test method

[0054] 1.2.1 Activation of strains and preparation of bacterial suspension: the test strains (Malassezia globosa CBS9757, purchased from the Medical Fungus Preservation Center of the Dermatology Institute of the Chinese Academy of Medical Sciences) were subcultured on Malassezia solid medium for 2 times, single colonies were picked and inoculated in Malassezia liquid medium, and incubated at 32°C in a 220 r / min shaker for 48 h. The bacterial suspension was prepared by diluting the bacterial liquid by the ratio of 1:10, 1:100 and 1:1000, respectively, to obtain the final concentration of 1×106 CFU / mL of bacteria solution as experimental bacteria suspension.

[0055] 1.2.2 Preparation of drug solution: The n-hexane, petroleum ether, chloroform and ethyl acetate extract fractions were dissolved in dimethyl sulfoxide to prepare drug solutions with a mass concentration of 100 mg / mL, which were stored at 4°C for later use. The positive drug piroctone olamine (OCT) was prepared at a concentration of 50 mg / mL.

[0056] 1.2.3 Bacteriostatic circle test: The Kirby-Bauer disk diffusion method was used to detect the antibacterial activity of each drug solution according to the procedure reported by the Clinical and Laboratory Standards Institute (CLSI). 200 μL of the bacteria suspension was taken with a pipette and evenly spread on the culture medium plate for later use. 20 μL of the drug solution was added to a sterile circular filter paper with dimensions of (6.0 mm x 1.0 mm). The filter paper was gently attached to the surface of the agar plate with tweezers. After incubation at 37°C for 48 h, the diameters of the bacteriostatic circles were observed and measured, and the average value was taken as the determination result. The edge of the bacteriostatic circle was limited to the point where no obvious bacterial growth could be observed with the naked eye. The results were determined according to the "Performance Standards for Antimicrobial Susceptibility Testing" issued by the National Committee for Clinical Laboratory Standards (NCCLS): a diameter > 20 mm was extremely sensitive, 15-20 mm was highly sensitive, 10-15 mm was moderately sensitive, and < 10 mm was low sensitive.

[0057] 1.3 MIC value determination: The reference standard was the M27 standard of the Clinical and Laboratory Standards Institute (CLSI) - Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts. Distilled water was used to dilute the drug solution in a two-fold dilution method with 10 gradients. A 96-well plate was taken, and 96 μL of liquid medium was added to columns 1-11. 4 μL of each drug solution was added to column 1, and the drug concentration was from high to low. 100 μL of bacteria solution was added to each well in columns 1-11, and three parallel samples were taken for each drug. Column 11 was used as a growth control, and an OCT positive control group and a negative control group were set up. After inoculation, the 96-well plate was incubated at 30°C for 48 h, and the results were observed. The positive wells showed a color change from yellow to red, and the lowest drug concentration at which the fungus was completely inhibited (no color change) was the MIC.

[0058] 2 Results and Discussion

[0059] 2.1 Bacteriostatic circle test results

[0060] The results of the bacteriostatic circle test are shown in Table 1 below. The n-hexane fraction of Monotropa rubida had a bacteriostatic circle diameter > 20 mm against Malassezia globosa, and the bacteriostatic effect was the most obvious, indicating extremely sensitive. The chloroform fraction of Monotropa rubida had a bacteriostatic circle diameter < 10 mm against Malassezia globosa at the same concentration. The petroleum ether extract fraction had a moderate sensitivity against Malassezia globosa, while the ethyl acetate fraction had no bacteriostatic activity.

[0061] Table 1 Comparison of the diameters of the inhibition zones of different extraction parts on Malassezia globosa

[0062] Drug solution Inhibition zone diameter (mm) E. cloacae 32.62±2.13 E. cloacae 13.52 ± 1.03 ** ]] E. cloacae 8.29 ± 0.75 *** ]] E. cloacae *** ]]> ​

[0063] Compared with the n-hexane part of Amomum xanthioides, ** P < 0.01, *** P < 0.001.

[0064] 2.2 MIC test results

[0065] Table 2 MIC test results of the n-hexane part of Amomum xanthioides on Malassezia globosa

[0066] Drug solution MIC (μg / mL) E. cloacae 125 OCT 62.5

[0067] Test Example Two, Combined Drug Sensitivity Test

[0068] The checkerboard dilution method was used to perform the combined drug sensitivity test, and the fractional inhibitory concentration index (FICI) was determined to evaluate the effect of the combined use of the n-hexane part of Amomum xanthioides (hereinafter referred to as Amomum xanthioides extract) and other drugs. The maximum stock solution concentration of the Amomum xanthioides extract was 100 mg / mL, the maximum stock solution concentration of piroctone olamine (OCT) and hexamidine diisethionate (HD-100) was 50 mg / mL, and the maximum stock solution concentration of ketoconazole, zinc pyrithione (ZPT), and climbazole was 25 mg / mL.

[0069] Distilled water was used to dilute the drug solution in 10 gradients by the two-fold dilution method. Liquid medium 96 μL was added to columns 1-10 of the 96-well plate, and the concentrations of the two drugs were added in two directions, each 2 μL. In the horizontal direction, the 10 concentration gradients of the Amomum xanthioides extract were arranged in 10 rows from high to low, and in the vertical direction, the 10 concentration gradients of the traditional antidandruff agent were arranged in 10 columns from high to low, so that the two drugs formed different combinations of drug concentrations in 80 wells. Then 100 μL of bacterial solution was added to each well in columns 1-11. The 11th well was used as a positive control, and the 12th well was used as a negative control. The 96-well plate was incubated at 30°C for 48 h, and the results were observed. The positive wells showed a color change from yellow to red. The fractional inhibitory concentration index (FICI) was used to evaluate the effect of the combined use of the Amomum xanthioides extract and other drugs, and the calculation is shown in the following formula.

[0070]

[0071] wherein A and B are the Amomum xanthioides extract and the traditional antidandruff agent, respectively, and MICA and MICB are the MIC values of A and B when used alone. MICA 联用 is the MIC value of A when A and B are used in combination, and MICB 联用The MIC value of B when A and B are used in combination. When FICI≤0.5, it is synergistic; 0.5

[0072] Table 3 MIC values of different drugs against Malassezia globosa

[0073] Drug solution MIC (μg / mL) E. cloacae 125 OCT 62.5 Ketoconazole 3.91 ZPT 15.63 Climbazole 7.81 Acetamiprid 31.25

[0074] Table 4 Test results of combined drug sensitivity test

[0075]

[0076] As can be seen from the above table, the combination of Volvariella volvacea extract and traditional antidandruff agents does not show synergistic effect in all cases, only the combination with OCT shows synergistic effect, FICI≤0.5, and the combination with ketoconazole and climbazole shows additive effect, and the combination with ZPT and HD-100 shows irrelevant effect.

[0077] Test three, human trial evaluation

[0078] 1.1 Inclusion criteria: 22 patients with seborrheic dermatitis of the scalp were selected, with an average age of 30±1 years old, and were randomly divided into 2 groups, 11 people in each group. Each patient has had moderate and above degree of scalp itching, dandruff, scalp redness and other symptoms for at least 3 months; the length of hair is ≥3 cm, there is no severe hair loss, and the hair root growth covers at least 70% of the scalp; the body is healthy and has no disease.

[0079] 1.2 Test method: the shampoo of Example 1 and the control group (commercially available Head & Shoulders Cool & Calm Dandruff Shampoo, Guangdong Cosmetic Network Preparation No. 2022100003) were used, 3 times a week, each time massaging the scalp for at least 1 min before rinsing thoroughly, for 28 consecutive days. The degree of dandruff, the degree of scalp redness and the degree of scalp oiliness were evaluated before the test and on the 28th day, respectively, and were recorded as T0D and T28D.

[0080] Evaluation method: the changes of dandruff were observed using a scalp image analyzer; the oil content was detected using a skin sebum tester Sebum Scale; the patients evaluated the degree of scalp redness and scalp itching using a 0-5 point scoring standard, and the results are shown in Figures 1-2 and Table 5.

[0081] 2 Results

[0082] Table 5 Score of the degree of scalp redness and scalp itching

[0083]

[0084] Compared with the control group, * P<0.05;** P<0.01.

[0085] From Figures 1-2 As can be seen from Table 5, the shampoo of the present application and the commercially available Head & Shoulders Cool Menthol Dandruff Shampoo, although both containing piroctone olamine, the shampoo of the present application can better improve the itching, redness, dandruff and oiliness of the scalp of the patients with seborrheic dermatitis.

[0086] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited to the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application should be equivalent replacement methods, and are all included in the protection scope of the present application.

Claims

1. A shampoo having antibacterial, anti-dandruff, and anti-itching effects, characterized by comprising: Components comprising the following mass percentages: surfactant 5~25%, guar hydroxypropyltrimonium chloride 0.1~0.3%, polyquaternium-10 0.3~0.5%, sodium chloride 0.1~0.3%, PPG-3 octyl ether 0.1~0.5%, pimento extract 1~3%, piroctone olamine 0.1~1.0%, humectant 2~4%, chelating agent 0.1~0.5%, pH adjuster 0.01~0.05%, essence 0.1~0.5% and water balance; the preparation method of the pimento extract is: S1, take pimento dry seeds, crush, sieve, add ethanol to heat reflux extraction, collect the extract, filter, concentrate the filtrate, recover ethanol, dry to obtain dry extract; S2, add n-hexane to the above dry extract for extraction, reduce pressure concentration to constant weight, dry, to obtain; The surfactant is selected from one or more than two combinations of sodium laureth sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine, cocamide MEA, sodium cocoyl glycinate, sodium lauroyl methylsulfate, sodium cocoyl glutamate and sodium myristoyl glutamate; the humectant is selected from one or more than two combinations of allantoin, propylene glycol, sodium hyaluronate, panthenol, glycerol and keratin hydrolysate; the chelating agent is selected from one or more than two combinations of EDTA, disodium EDTA and tetrasodium glutamate diacetate; the pH adjuster is citric acid.

2. The shampoo according to claim 1, characterized in that, In the step S1, the heating reflux extraction conditions are: solid-liquid ratio 1:10~30, reaction temperature 60~80℃; reaction time 1~3h; ethanol volume fraction 60~85%.

3. The shampoo according to claim 1, characterized in that, In the step S2, the n-hexane extraction process parameters are: temperature 40~70℃; extraction time 1.0~2.5h; solid-liquid ratio 1:5~10.

4. The shampoo with bacteriostatic, dandruff-removing and itching-relieving effects according to claim 1, characterized in that, Components comprising the following mass percentages: Sodium laureth sulfate 5~10%; Sodium cocoyl methyl taurate 1~5%; Cocamidopropyl betaine 1~5%; Cocamide MEA 0.1~1%; Guar hydroxypropyltrimonium chloride 0.1~0.3%; Polyquaternium-10 0.3~0.5%; Sodium chloride 0.1~0.3%; PPG-3 octyl ether 0.1~0.5%; Pimento extract 1~3%; Piroctone olamine 0.1~1.0%; Panthenol 2~4%; Disodium EDTA 0.1~0.5%; Citric acid 0.01~0.05%; Essence 0.1~0.5%; and Balance water.

5. A method of preparing a shampoo according to claim 4, characterized in that, Including the following steps: Guar gum hydroxypropyltrimonium chloride is pre-dispersed in cold water, then disodium EDTA is added, stirred evenly, heated to 50~70℃, then sodium lauryl polyoxyethylene sulfate, sodium cocoyl methyl taurate, cocamidopropyl betaine and cocamide MEA are added in sequence, after 20min of incubation, piroctone olamine is added and stirred evenly, then the temperature is lowered to 30~45℃, polyquaternium-10 and PPG-3 octyl ether are added and stirred evenly, then pimenta racemosa extract is added, stirred for 30min, then the temperature is lowered to room temperature, panthenol and fragrance are added, stirred for 30~40min, then citric acid and sodium chloride are added, the finished product is discharged, and the product is obtained.

Images