Preparation And Anti-aging Effect Of Guarana Clove Oil Complex Nano-emulsion

Guarana（Paullinia cupana,Guarana）is a non-infested family,native to the Amazon Basin of Brazil.Its seed extract is rich in alkaloids,polyphenols,plant polysaccharides and other.It has long been widely used in daily diet and health care.Now,Guarana is also gradually developed by the cosmetics industry,and has a wide range of consumers.clove essential oil is a volatile aromatic substance extracted from the dried buds of myrtle plant clove（Syzygium aromaticum.）.modern studies have shown that clove essential oil is rich in active ingredients,such as eugenol,acetobutylphenol,β-strychnine,etc.it has anti-inflammatory and antibacterial effects,improves rough skin,promotes wound healing,and promotes percutaneous absorption of drugs.therefore,it is often used in medicine,food,cosmetics and other fields.The results showed that both guarana extract（GE）and clove essential oil（CO）had strong antioxidant effect,and external use could delay skin aging,but there were also some problems such as volatile,poor solubility and strong irritation.For the purpose of improving drug stability,enhancing drug,enhancing the therapeutic effect of the drug and reducing the irritation of the drug,a safe,effective and stable anti-skin aging transdermal drug delivery system was prepared by combining guarana and clove essential oils as model drugs,using nanoemulsification technology,The contents are as follows:1.Prescription screening and optimization of GE-CO-NEFirst,guarana extract prepared by ethanol reflux extraction,Among them,guarana g/m L;0.2 extraction of clove essential oil by steam distillation,The yield of essential oil is about 11.53.Using clove oil as the oil phase of nanoemulsion,RH 40,of nanoemulsion surfactant was selected by pseudo-triple phase diagram the surfactant is anhydrous ethanol.Adopt DOE full factor design,based on the mass ratio of surfactant,cosurfactant and oil phase,particle size and PDI as evaluation indicators,to optimize the formulation of nanoemulsion,then the optimal loading capacity of nanoemulsion is10%by pseudo ternary phase diagram,the final prescriptions for the GE-CO-NE were:RH 40（10%）,anhydrous ethanol（5%）,clove oil（1%）,guarana extract（10%）,water（74%）.2.Physical and chemical properties and stability evaluation of GE-CO-NEEvaluation of the physicochemical properties of the optimized GE-CO-NE,and compared with the physicochemical properties of blank clove essential oil nanoemulsion（BCO-NE）without drug loading.Turns out,GE-CO-NE brown-red,clear,transparent liquid,O/W nanoemulsion,pH 5.85±0.02,Viscosity 26.8±0.2 m Pa·s,Size 15.81±0.13 nm,PDI 0.109±0.012,Zeta potential-2.67±1.24 mV;BCO-NE colorless,clear,transparent liquid,pH 6.40±0.01,Viscosity 25.0±0.2 m Pa·s,The particle size is 14.09±0.04 nm,PDI 0.11±0.001,Zeta potential-2.44±1.22 mV.Compared with the BCO-NE,of unloaded drugs the addition of the drug changed the color of the nanoemulsion,while the effect on the pH,viscosity,particle size,PDI and Zeta potential of nanoemulsion was not significant（P>0.05）.The results showed that the GE-CO-NE was not stratified and demulsified in centrifugal test,freeze-thaw cycle test,heating-cooling cycle test,and the average particle size and PDI of nanoemulsion were stable in a certain range,which indicated that the GE-CO-NE had certain kinetic and thermodynamic stability;the storage stability test,GE-CO-NE were stored at 4,25 and 40℃for 90 d,.no delamination,demulsification and other unstable phenomena occurred in all the GE-CO-NE during the test period,and the average particle size and PDI were stable in a certain range,indicating that the GE-CO-NE a certain cold,heat resistance and storage capacity for a certain period of time.Comprehensive stability test results show that the GE-CO-NE stability is good and the formula of nanoemulsion is reasonable.3.In vitro release and skin irritation of GE-CO-NECatechin,caffeine and eugenol in GE-CO-NE were used as index components to establish a HPLC method for the determination of index components.The results showed that the method had a good linear relationship,and the specificity,precision,repeatability,stability and recovery test met the requirements.The permeation properties of GE-CO-NE were investigated by Franz diffusion tank.The transdermal properties of GECO-NE and GE-CO-M mixture were compared and analyzed.The results showed that the transdermal permeation of catechin,caffeine and eugenol in GE-CO-NE and GE-CO-M conformed to Higuchi equation,indicating that both drug carriers had sustained-release effect;however,the cumulative permeation volume per unit area,transdermal permeation rate and skin retention of catechin,caffeine and eugenol in GE-CO-NE were higher than those in GE-CO-M,indicating that the transdermal permeation performance of GE-CO-NE was better than that of GE-CO-M.The skin irritation of GE-CO-NE and blank nanoemulsion matrix（Blank-NE）was investigated and compared with GE-CO-M.The irritation of GE-CO-NE、Blank-NE and GE-CO-M was evaluated and compared by single and multiple administration skin irritation test.The results showed that both GE-CO-NE and Blank-NE had no irritation to the skin,but GE-CO-M showed mild irritation in single administration and multiple administration of skin irritation tests,indicating that after the preparation of GE and CO,the irritation of the drug to the skin was reduced.4.Pharmacodynamics of GE-CO-NE on anti-aging skinThe antioxidant capacity of GE-CO-NE was investigated by DPPH free radical scavenging test,and compared with GE-CO-M and unilateral GE and CO antioxidant capacity.The scavenging rate curves of GE-CO-NE、GE-CO-M、GE and CO on DPPH radicals were fitted,Calculate the half inhibitory concentration of each drug(IC₅₀),Turns out,IC₅₀:GE-CO-NE≈GE-CO-M<CO<GE,of four tested drugs indicating that there is no significant difference in IC50 between GE-CO-NE and GE-CO-M,They are lower than the GE and CO,of one side The results show that GE-CO-NE and GE-CO-M can achieve antioxidant effect at lower concentration,That is to say,the antioxidant capacity of GE-CO-NE and GE-CO-M is stronger than that of unilateral GE and CO,GE-CO-NE and GE-CO-M have the same antioxidant capacity,it shows that nano-emulsion excipients have no effect on the antioxidant ability of drugs.A mouse skin aging model was established by D-galactose combined with ultraviolet light irradiation,the effects of external coating GE-CO-NE、GE-CO-M、GE and CO intervention on aging skin were investigated.Turns out,the intervention of drugs can reduce the production of skin fold,increase the water content of skin,increase the activity of SOD enzyme in skin,increase the content of Hyp,reduce the production of MDA,thicken the dermis and increase the proportion of skin collagen fiber,Compared with the model group（P<0.05）.The results show that,the skin SOD enzyme activity,Hyp content and collagen fiber ratio in GE-CO-NE and GE-CO-M groups were higher than those in unilateral GE and CO groups,The thickening degree of epidermal dermis was more obvious than that of unilateral GE and CO group,The production of lipid peroxides MDA is less than that of unilateral GE and CO groups,The results showed that the effect of Guarana extract and clove essential oil on aging skin was better than that of unilateral GE and CO.Besides,the anti-aging effect of GE-CO-NE at the same concentration is also slightly stronger than that of GE-CO-M,it shows that nanoemulsion as a drug delivery carrier also has a positive effect on the efficacy of drugs.