Studies On The Compond Eugenol Nanoemulsion

In this research, the traditional compound eugenol was recasted by using the nanoemulsion as a new carrier.It was purposed to develop a stable, safe and effective new drug, which can be used to cure pruritics, pain and inflammation of skin bited by insects like mosquito.At first, the establishment of analytical method, screening of nanoemulsion prescription and the inspection of influence factor were studied.Compound eugenol nanoemulsion was prepared by pseudotertiary phase diagram.The stability, security and pharmacodynamics were systemically studied.Part 1: The establishment of analytical method of compound eugenol nanoemulsion. Determin the eugenol content of compound eugenol nanoemulsion. Establishing the Ultraviolet analytical methed of compound eugenol nanoemulsion to study the recovery rate and accuracy. Results shown that eugenol has the maximal absorption in 280nm. The good linear range was 10~80μg/mL. The mean recovery, RSD of the recovery and the repeatability was 97.87%, 3.05% and 1.98%, respectively. The accuracies inter-day was less than 5% and the accuracies intra-day was less than 7%. The analytical method in this study has high recovery, good accuracy and repeatability, which can satisfy quality control research.Part 2: The preparation of compound eugenol nanoemulsion. Prescriptions of nanoemulsion were filtrated by the nanoemulsion evaluation standard. The influence factors of nanoemulsion were also investigated. The results of transdermal accumulative amount of compound eugenol nanoemulsion invitro by TT-6(B) pervasion pool showed that the best prescriptions of the nanoemulsion were: the surfactant, cosurfactant and the oil phase was RH40, the absolute alcohol and compound eugenol, respectively; The ratio of surfactant to cosurfactant was 7:1 and the surfactant to oil was 8:2. The preparation method of compound eugenol nanoemulsion was simply and easy to manipulate.Part 3: The evaluation of stability and quality of compound eugenol nanoemulsion. The quality was detected by electron microscope, photon correlation spectroscope, the viscosity statement, refraction and electric conductivity; The stability was evaluated by centrifuging, acceleration, light, low temperature, long-term and classical constant temperature experiments. The results showed that the compound eugenol nanoemulsion was clarified and transparent liquid. Its average diameter was (75.1±3.7)nm; The quality of compound eugenol nanoemulsion was stable; Its valid period can be conjected to be 1.8 years. The compound eugenol nanoemulsion of this study was stable.Part 4: The safety evaluation of compound eugenol nanoemulsion. Evaluate the safety of compound eugenol nanoemulsion by acute toxicity, skin irritation, eye irritation, cell toxicity experiment. It showed that the accumulation LD50 of the compound eugenol nanoemulsion test on mice was 11.273g/kg. The stimulation to the skin and eye of domestic rabbits were all nontoxic, no skin allergy response. The cell toxicity was 1 level, no obvious toxicity. The compound eugenol nanoemulsion was safety.Part 5: The pharmacodynamics evaluation of compound eugenol nanoemulsion. The pharmacodynamics of compound eugenol nanoemulsion was evaluated by antibacterial activity invitro, anti-inflammatory, analgesic and anti-pruritic. The mechanism of anti-inflammatory was investigsted by detecting the contents of MDA and active of GSH-Px, CAT, SOD in rat skin tissue. It showed that the compound eugenol nanoemulsion has prominent antibacterial effect on S.aureas and monilia albican. Its antibacterial diameter was more than 20mm, which had extreme significant diference (P<0.01) compared with compound eugenol normal mixture. Restrain rate on mice ear edema induced by dimethyBenzene was 90.9%, good control action on rat foot edema induced by egg white, has extreme significant diference compared with compound eugenol normal mixture(P<0.01). Analgetic rate on mice writhing reaction induced by acetic acid is 60.5%, Increase rate of pain threshold on pain reaction of mice induced by hot-plate is 96.5%, has extreme significant diference compared with compound eugenol normal mixture(P<0.01). Times of licking reaction on mice skin pruritic reaction induced by 4-AP is reduced, Itching threshold value on itch reaction induced by phosphoric acid histamine is enhanced; Active of GSH-Px, CAT, SOD in rat skin tissue is enhanced, the contents of MDA is reduced, showed that compound eugenol nanoemulsion has antioxidation, this is one of anti-inflammatory mechanism. The compound eugenol nanoemulsion of this study was effective in pharmacodynamics.Above all, the compound eugenol nanoemulsion was stable, safe and effective, according to the standards of microemulsion and drug delivery.