Influence of Sesame Oil on Arbutin Release

Influence of benne Oil on Arbutin ReleaseThe Influence of Sesame Oil Addition on The Arbutin Release and Penetration in Carbomer Gel Base(Observation on Inhibition of Enzym Tyrosinase Activity)Tristiana Erawati, Widji Soeratri, Noorma Rosita, Wida Rukmanajati, Hanifa RahmaAbstractHydrophilic arbutin as lightening performer with log P regard as -1.35, make it gruelling to permeate through the skin and reach its site of action. Sesame oil appendage (3, 5, and 7% w/w) was expected to increase the arbutin release and penetrations. The aim of this admit was to investigate the diverge of benni oil addition on the arbutin release and penetrations in the Carbomer-940s jelly base. The release (flux) of arbutin, as initial process before penetration, expressiontion were studied using cellophane membrane and buffer phosphate pH 7.0 as media at 370.5C for six hours long. The penetration of arbutin was observation on inhibition of enzyme tyrosinase body process. Inhibition percent of t yrosinase by arbutin was determined in vitro by observing the absorbance value of dopachrome (an mediate product of melanin formation) as a response product between enzyme tyrosinase and L-tyrosine as a substrate using spectrophotometer. Conclusion of this study was benne oil addition 3 and 5% w/w decreased arbutin release, sesame oil addition 3, 5 and 7% w/w increased arbutin penetrations. Increasing of arbutin effectiveness more is caused by enhancer effect of sesame oil.Keyword Arbutin, Carbomer-940, Penetration, Release, Sesame oil, Tyrosinase-inhibitionIntroductionArbutin widely used in cosmetic as lightening agent to contain enzyme tyrosinase activity in basal membrane of the skin. Tyrosinase was known as enzyme that involved in melanin formation.1, 2 Because of the hydrophilic of arbutin with log P value 1.35 make it difficult to penetrate through the skin. To increase the penetration enhancer can be add in the formula. Sesame oil as an oily enhancer has f all in all pr otein (25%) and globulin (67.3%) its can increased penetration trough polar pathway by enlarge aqueous channel. Sesame oil also can use as healing effect from sunburn.3 It was known Sesame oil effective concentration as enhancer up to 10%.4 The aim of this study was to investigate the influence of sesame oil (3, 5, and 7% w/w) addition on the arbutin (3% w/w) penetrations in the Carbomer-940 change base through the modified lipid membrane. It was observation on inhibition of enzyme tyrosinase activity. However sesame oil is a viscous fluid can increase the viscosity of base so that it feared inhibits the release of arbutin and decrease penetration. In this study determined arbutin release from the base using cellophane membrane and buffer phosphate pH 7.0 as media at 370.5C for six hours long.Materials and Methods zeal of the arbutin mousse as Lightening productThe arbutin in Carbomer-940 jelly base formulas as lightening product was shown in table 1. In this research Carbomer-94 0 gelatin base contained tri ethanol ammine (TEA) as alkalizing agent, propylene-glycol as humectants, methyl-parabene and propyl-paraben as preservative, Na-EDTA as chelating agent, butylated hydroxyl toluene (BHT) as anti-oxidant and Tween-80 as surfactant. Arbutin 3% w/w in Carbomer-940 gel base was used as control. Arbutin 3% w/w with sesame oil 3% w/w in Carbomer-940 gel base named as F1, Arbutin 3% w/w with sesame oil 5% w/w in Carbomer-940 gel base named as F2 and Arbutin 3% w/w with sesame oil 7% w/w in Carbomer-940 gel base named as F3.The Characteristics determination of the arbutin gelThe Characteristics determination of arbutin gel included role of gel pHDetermination of the spreading-abilityDetermination of gel spreading-ability was performed using a pair of glass scale (20 X 20 cm). The gel preparation (1 gm) was pose in the middle of the first glass plate that given the scale. Then put the second glass plate on the first glass plate and measured the diameter of ge l spreading. After that put stabilize on the second glass plate therefore measured the diameter spreading-ability of the gel. The weight of ballast that put on the second plate was increased until spreading-ability of the gel was constant.Determination of arbutin releaseDetermination of arbutin release from the bases was done by the dissolution raveler Hanson inquiry SR-6 with paddle stirrer. Each cell dissemination fill with arbutin gel ( 2 grams), in 500mL buffer phosphate pH 7.0, temperature 37C, agitation 100 rpm. Samples (5mL) were taken at 5, 10, 15, 30, 45, 60, 90, 120, 180, 210, 240, 270, 300, 330, and 360 minutes, replace with 5mL buffer phosphate pH 7.0 to keep volume constant. The absorbance of arbutin in the sample measured by spectrophotometer. The arbutin release (flux) from the base obtained from the slope of the analog regression of the correlation curve between arbutin releases accumulations versus square root of time.The penetration evaluation arbutin gel (U nited Stated Pharmacopoeia, 2002)In vitro study for the penetration of the arbutin in Carbomer gel base was measured by the modification method of the penetration test USP XXV and British Pharmacopoeia, 2002 with dispersion apparatus ERWEKA DT 700.The in vitro study was evaluated as fol pitiables The arbutin gel (around 3 grams) was put in the diffusion cell then covers with the Millipore membrane which was impregnated with isopropyl-myristate as modified lipid membrane. Then the preparation of arbutin gel in diffusion cell was put into the penetration chamber contain 500 ml of phosphate buffer pH 6.5 0.05 at 37 0.5C as diffusion medium, and then the paddle was stirred 100 rpm. The sample solution around 5 ml was collected at 360 minutes after it penetrated.Determination of enzyme tyrosinase activityL-tyrosine solution 0.5 ml added with 3.0 ml sample solution that collected from compartment receptor after 360 minutes penetrated through Millipore membrane which was impregnated wit h isopropyl-myristate. The mixture was oxygenized 5 minutes then added with 1.0 ml tyrosinase solution. After incubated for 10 minutes at 25C the mixture was inactivated with 0.5 ml TCA solution and then the absorption value measured at maximum wavelength of dophacrome.5The evaluation of inhibition of enzyme tyrosinase activityThe inhibition of enzyme tyrosinase activity was performed as inhibition percent, which free-base from calculation of absorption value per second enzymatic reaction with inhibitor, compared with absorption value per second enzymatic reaction without inhibitor, using the following equation6WhereasA = absorption value (A/second) at dophacrome maximum with inhibitorB = absorption value (A/second) at dophacrome maximum without inhibitorThe data (inhibition %) were analyzed with ANOVA one way method (p.Results and DiscussionThe result of this study, in table 2 shows that the pH of all formulas around 6 it mean appropriate with skin pH. The spreading profile of a rbutin gel preparation shows in Figure 1 and spreading-capasity of arbutin gels at 20 gram ballast shows in table 3. Spreading-capacity was formulas spreading-diameter at same ballast weight. The result of ANOVA one way test of spreading-capacity found the value of Fcalculation (13.741) Ftable (4.07). Its can argue there were significant deference minimal one pair of spreading-capacity formulas data. To know which spreading-capacity formulas was significant deference its tested by Honestly Significant respect (HSD) tests. The result of HSD test in table 4, that can concluded the spreading-capacity of formula 1 did not deference with control but higher than formula 2 and 3.Spreading-ability was the slope of linier-regression between spreading-diameter (cm) and ballast weight (gram), its shows in table 5. The slope value from its formulas was tested by ANOVA one way method, its found that the value of Fcalculation (0.274) table (4.07). So that can conclude its was no significant def erence between spreading-ability of all formulas. dining table1. Formulas of lightening productArbutin release (flux) was figure from the linier regression of the correlation curve between square root of time versus arbutin release accumulation. Slope (flux) of linier regression showed in table 6. To make sure if there is any difference of arbutin flux between formulas was done by statistical testing using ANOVA one way. It is showed that Fcalculation (50,918) Ftable (4.07). From HSD result know that flux formula 1 and 2 not statistically different, but if compare with control and formula 3 were decrease. This might be caused by interaction between arbutin, sesame oil, and Tween. Tween is anionic surfactant which is amphiphil, it have affinity towards polar or non polar substance, such as arbutin and sesame oil. This interaction makes arbutin more difficult to release from bases. some other factor that may influence was viscosity from formula 1 and 2 which is more viscous than c ontrol, it cause arbutin molecules difficult release from bases also. The increase of viscosity may inhibit the movement of molecules to release from bases.7Flux value of formula 3 not statistically different with control but higher than formula 1 and 2. It might be caused by addition of sesame oil reduce amount of water from formula. Decrease amount of water caused increase of arbutin concentration on water phase. Substance release from bases is known as diffusion. Based on Ficks law, diffusion is the process by which molecules moved from compartment with high concentration to low concentration.The arbutin effectiveness as lightening agent calculated as inhibition percent (%) of enzyme tyrosinase activity. The result of arbutin inhibition percent (%) with enhancer sesame oil in Carbomer gels shows in table 7. circuit card 2 The arbutin gel pH values Figure 1 The spreading profile of arbutin gel with various concentration of sesame oil. Each value represents the mean of 3 determinat ions.Tabel 3 Spreading-capasity of arbutin gels at 20 gram ballast* The result were obtained from an average of 3 times replication Table 4 HSD test result of spreading- capacity value of arbutin gelsTabel 5 Arbutin gels spreading-ability* The result were obtained from an average of 3 times replicationTabel 6. Flux of arbutin release from gel basesTable 7 The arbutin effectivity (inhibition %) in carbomer gel formulasThe result of ANOVA one way test of the arbutin effectiveness in carbomer gel formulas found the value of Fcalculation (23,582) Ftable (4.07), and from the HSD test result was found inhibition percent of control Table 8 The HSD test result of arbutin effectivity (inhibisi %)in carbomer gel formulasConclusionConclusion of this study was sesame oil addition 3 and 5% w/w decreased arbutin release, sesame oil addition 3, 5 and 7% w/w increased arbutin penetrations. Increasing of arbutin effectiveness more is caused by enhancer effect of sesame oil.AcknowledgementThis study was supported financially by Project divide of Faculty of Pharmacy, Airlangga University, Surabaya Indonesia.ReferencesTakada, K. and Tanaka, Y., 2000. Depigmentation Agents. In Elsner, P., Mailbach, H.I. (Eds.). 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