Development Of Tea Tree Oil Disinfectant And Study On Its Sterilization Mechanism

Melaleuca alternifolia is a small evergreen tree or shrub belonging to the genus Melaleuca alternifolia of Myrtle family.Tea Tree Oil is an oily liquid extracted from the branches and leaves of Alternaria alternata by steam distillation.As a natural antimicrobial agent,tea tree oil plays a very important role in the clinical research of drugs.It is expected to become a natural drug that can alleviate the global drug resistance crisis and has a broad prospect of development and utilization.In this study,a new type of disinfectant with tea tree oil as the main component was developed,based on the properties of tea tree oil.The extraction technology,chemical composition,activity,content of characteristic components and sterilization mechanism of tea tree oil were studied in this paper.1.The optimum extraction technology of tea tree oil from leaves and branches of Melaleuca alternifolia was screened by steam distillation and Orthogonal test method.The extraction rate reached2.293%when the ratio of solid to liquid was 1:20,soaked for 6 hours and extracted for 6 hours.2.The chemical constituents of extracted tea tree oil and Main Camp brand tea tree oil were studied by GC-MS.The results showed that 62 constituents were separated from the former,50compounds were identified,and the relative content of the identified compounds accounted for 98.97%of the total;44 constituents were separated from the latter,35 compounds were identified,and the relative content of the identified compounds accounted for 99.49%of the total.Both of them contain more Terpinene-4-ol,which is similar to Terpinene-4-ol tea tree oil.3.The antimicrobial activity of tea tree oil extracted by tube butterfly method was compared with Main Camp brand tea tree oil.The results showed that the two tea tree oil have certain inhibitory effects on G~+(S.aureus,S.albus),G~-(E.coli,P.aeruginosa)and fungi(C.albicans).The former had a weaker inhibitory effect on 8 pathogenic microorganisms than the Main Camp tea tree oil(except E.coli and S.albus).The latter had a stronger inhibitory effect on 8 microorganisms than the positive control group(except E.coli and C.albicans).The inhibitory effect on S.aureus was more obvious.4.A method for the determination of Terpinene-4-ol and 1,8-Cineole in tea tree oil disinfectant by gas chromatography was established and its stability was studied.The results showed that there was a good linear relationship between the concentration of 1,8-Cineole and peak area in the range of 0.013-0.41 mg/mL(R~2=0.9999).The concentration of Terpinene-4-ol and peak area also showed a good linear relationship in the range of 0.175-5.575 mg/mL(R~2=0.9999);the average recovery of 1,8-Cineole was99.190%,RSD was 1.890%,the average recovery of Terpinene-4-ol was 102.580%,RSD was 1.450%.The disinfectant of tea tree oil should be carried out under the conditions of low temperature(below25?),shade,dryness and light avoidance during transportation and storage.It is sealed in polyethylene bottle and its validity period is more than one year.5.The antimicrobial mechanism of tea tree oil was studied by using Staphylococcus aureus as experimental strain.The results showed that tea tree oil had the ability to inhibit and remove the formation of Staphylococcus aureus biofilm.Its antimicrobial action site may be located on the bacterial cell membrane.Because of the integrity of cell membrane was severely damaged,permeability of cell membrane increaseed,causing cell splitted and leakage of cell contents,leading to cell death.