| Malassezia is a yeast-like fungus that resides on the surface of warm-blooded animals and human skin.Its growth depends on the physiological characteristics of lipids(except for Malassezia pachydermatis).Malassezia is mainly distributed in sebum-rich parts of the human body,such as the face,scalp,chest and back.It can participate in the pathogenesis of various skin diseases,such as tinea versicolor,Malassezia folliculitis,seborrheic dermatitis,atopic dermatitis,acne vulgaris,and psoriasis.Malassezia-induced skin disease is a fungal skin disease characterized by itching,desquamation,erythema,papules,and pustules,which seriously affects people’s health and quality of life.In addition,the production of dandruff is also most related to the infection of Malassezia,common scalp abnormal peeling,scalp itching and other symptoms.At present,the treatment of Malassezia-related skin diseases includes oral antifungal drugs such as ketoconazole,fluconazole,and itraconazole,and topical antifungal ointments such as Kangwang and Zele.Immunosuppressive agents and glucocorticoids can also be used for the treatment of Malassezia infection.However,the above antifungal drugs have many side effects,strong drug resistance and high irritation,which limit their application.Therefore,finding safe and effective natural drugs has become a hot and difficult point in the research of anti-Malassezia drugs.At present,the research on natural drugs of Malassezia in domestic and foreign literatures mainly focuses on traditional Chinese medicine extracts,traditional Chinese medicine compounds,single active ingredients of traditional Chinese medicine and plant essential oils.There are few reports on the antibacterial activity of volatile oil of traditional Chinese medicine against Malassezia,and the excellent antibacterial properties of volatile oil of traditional Chinese medicine make it expected to become a natural drug against Malassezia.The specific research contents and conclusions of this paper are as follows :Ⅰ.Study on the antibacterial activity of volatile oil of traditional Chinese medicine on Malassezia(1)Eight kinds of volatile oils(Cinnamon,Angelica,Clove,Mosla,Benzoin,Chuanxiong,Acorus tatarinowii,Angelica)with inhibitory effects on Malassezia furfur and Malassezia sympodialis were preliminarily screened by medium punching method.Cinnamon oil had strong inhibitory effects on Malassezia furfur and Malassezia sympodialis,and the diameters of inhibition zones were 48.25±1.09 mm and 45.88±2.65 mm,respectively,showing extreme sensitivity.For Malassezia furfur,the inhibition zone diameters of Angelica dahurica oil(29.50±0.66 mm),clove oil(26.08±0.88 mm),Mosla chinensis oil(25.17±0.76 mm)and benzoin oil(20.61±0.54 mm)were all greater than 20 mm,showing extreme sensitivity.The inhibition zone diameters of Ligusticum chuanxiong and Acorus tatarinowii oil were 11.92±0.14 mm and 11.25±0.66 mm,respectively,showing moderate sensitivity,while Angelica sinensis oil(9.25±0.66 mm)showed low sensitivity.For Malassezia sympodialis,the inhibition zone diameters of Angelica dahurica(35.38±0.88 mm),Mosla chinensis(33.38±1.24 mm)and Clove(27.13±0.18 mm)oil were all greater than 20 mm,showing extreme sensitivity.The inhibition zone diameters of Chuanxiong and Acorus tatarinowii oil were 18.38±1.94 mm and 16.25±0.35 mm,respectively,showing high sensitivity,while Angelica sinensis(14.50±1.06 mm)and Benzoin(13.63±0.18 mm)oil showed moderate sensitivity.Cinnamon oil,clove oil,Elsholtzia oil,Angelica dahurica oil and Sabin oil have good antibacterial effect on two kinds of Malassezia.Therefore,these five Chinese medicinal volatile oils were selected for further study.(2)The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of cinnamon,clove,Mosla chinensis,Angelica dahurica and benzoin oil were determined by double dilution method.The results showed that the MIC values of the five volatile oils against Malassezia furfur were 0.39,0.78,1.56,1.56 and 0.78 μL/m L,respectively,and the MIC values of the five volatile oils against Malassezia sympodial were 0.1,0.39,0.78,1.56 and 0.78 μL/m L,respectively.The MBC values of the five volatile oils against Malassezia furfur were 0.39,0.78,1.56,3.13 and 0.78 μL/m L,respectively.The MBC values of the five volatile oils against Malassezia sympodial were 0.1,0.39,0.78,1.56 and 0.78 μL/m L,respectively.Combined with the results of inhibition zone diameter,cinnamon oil,clove oil,elsholtzia oil and angelica oil were selected for the next step of volatile oil combined antibacterial activity research.Ⅱ.Study on the antibacterial activity of volatile oil against Malassezia(1)The four volatile oils of cinnamon,clove,Mosla chinensis and Angelica dahurica were compounded by chessboard dilution method,and the combined fractional inhibitory concentration(FIC)index of Malassezia was determined.The results showed that for Malassezia furfur,the FIC values of cinnamon oil-syringe oil,cinnamon oilsalsa oil,cinnamon oil-A.dahurica oil and clove oil-salsa oil were 2,which showed no correlation,while the FIC values of clove oil-A.dahurica oil and Elsholtzia oilA.dahurica oil were 3,which showed antagonistic effect.For Malassezia sympodialis,the FIC value of cinnamon oil-clove oil,cinnamon oil-Elsholtzia oil,cinnamon oilAngelica oil and clove oil-Elsholtzia oil was 1.5,and the FIC value of clove oil-Angelica oil and Elsholtzia oil-Angelica oil was 2,which showed independent effect.The results showed that the antibacterial activity of the above four volatile oils was not as strong as that of the single volatile oil,and there was no synergistic or synergistic effect.(2)The four volatile oils of cinnamon,clove,Mosla chinensis and Angelica dahurica were combined in different volume ratios(10:0,9:1,7:3,5:5,3:7,1:9,0:10)by medium punching method.The diameter of the inhibition zone was measured,and the volatile oil compound prescription with good antibacterial activity against Malassezia was screened.The results showed that the antibacterial activity of cinnamon oil-clove oil,cinnamon oil-Elsholtzia oil,cinnamon oil-Angelica oil and clove oilElsholtzia oil on the two kinds of Malassezia was not strong,and there was no synergistic or synergistic effect.The antibacterial activity of clove oil-Angelica oil and Elsholtzia oil-Angelica oil on the two kinds of Malassezia was worse than that of single volatile oil,and similar antagonistic effect appeared.The results showed that the above four kinds of volatile oils should not be compounded against Malassezia.Ⅲ.Preliminary study on cinnamon volatile oil micelles and related products(1)The system suitability test was carried out for the determination of the active ingredient cinnamaldehyde in cinnamon oil by high performance liquid chromatography.The linear relationship,precision,stability,repeatability and sample recovery rate were used to verify the applicability of the method.The results showed that the linear relationship between the concentration of cinnamaldehyde and the peak area was good in the range of 1 μg/m L-60 μg/m L.The precision,stability and repeatability met the requirements,and the recovery rate was 96.59 %.(2)Cinnamon oil micelles(CO-P407/Labrasol-MMs)were prepared by film dispersion method.Firstly,the carrier materials of P407 and caprylic acid polyethylene glycol glyceride(Labrasol)and their dosage were investigated.Secondly,the effects of drug loading ratio,rotary evaporation temperature,organic solvent volume and hydration volume on encapsulation efficiency,drug loading,particle size and polydispersity index(PDI)were investigated.Three factors that had great influence on the preparation of CO-P407/Labrasol-MMs were screened.Finally,the drug loading ratio(A),rotary evaporation temperature(B)and hydration volume(C)were used as independent variables,and the encapsulation efficiency(Y1)and drug loading(Y2)were used as dependent variables.The Box-Behnken design response surface method was used to optimize the preparation of CO-P407/Labrasol-MMs.The optimal prescription ratio was that the ratio of P407 to Labrasol was 9 : 1(w : w),the ratio of P407/Labarasol to cinnamon oil(CO)was 7 : 1(w : w),the rotary evaporation temperature was 25 ℃,and the hydration volume was 13 m L.The encapsulation efficiency was(85.15±0.81)%,the drug loading was(11.13±1.85)%,the particle size was(97.39±0.01)nm,the PDI was(0.59±0.01),and the Zeta potential was(-3.50±0.37)m V.(3)The types and dosages of main surfactant,cosurfactant and main-cosurfactant were investigated,and then CO-P407/Labrasol-MMs were added to shampoo products.The products were evaluated according to the current industry standards,and the test results met the corresponding standard requirements. |
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