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j. Cosmet. 58, 245-254 (May/June2007) sci.


Vehicleand enhancer effectson humanskin penetration of aminophylline from cream formulations: Evaluation vivo in

SU-CHING KUO, Department Applied of Chemistry, Chia Nan University Pharmacy Science, of and Tainan 71710, Taiwan R.O.C. Accepted p•/blication for March6, 2007.


The effects fouressential (rosemary, of oils ylang,lilacin,andpeppermint oils),andthreeplantoils(jojoba oil, corngerm oil, andoliveoil) on the permeation aminophylline of werestudied usinghumanskin.The permeation effects these werecompared those threechemical of oils with of penetration enhancers. Although all oilsenhanced permeation aminophylline, the of their effects werelessthan that of ethanol. Jojoba oil wasfoundto be the mostactive, causing abouta 32% peakheightdecrease N-H bending of absorbances in comparison with the control,whilepeppermint, lilacin,rosemary, ylangoilscaused and 28%, 24%, 18%, and 12% peakheightdecreases, respectively. Microemulsions containing 10% jojoba and30% corngerm oil oil werefoundto be superior vehicles thepercutaneous for absorption aminophylline. of Comparision with results obtained fromhigh-performance liquid chromatography shows goodagreement.


Theophylline a xanthene is derivative. When theophylline combined is with ethylene diaminein anhydrous alcohol, formsthe compound it aminophylline Aminophylline (1). hasbeenreported breakdownfat andcellulitefrom cellsby triggering to enzymes that helpthe bodyto release from stores certain fat in areas the body.It workswell on the of thighsand buttocks womenand hasa smoothing of effecton the skin. However,some individuals sufferfrom allergicreactions ethylenediamine to hydrochloride The (2). aminophylline creamhasbeenstudiedby Fouriertransform infraredFTIR (3), electrochemical detection reversed-phase (4), high-performance chromatography liquid (HPLC) (5), andspectral analysis Therewasonly onestudyappliedto aminophylline (6). cream, in which permeation throughrat skin in vitrowas performed a two-compartment in diffusion cell to a select enhancer Human skin is usuallythe preferred (7). skin membrance usein an absorption to study.No animalmodelgives absorption values identical to thoseobtained humanskin (8). The stratumcorneum long beenconsidered in has a major barrier to the penetration of topically applied chemicals. Studies have

Address correspondence Lai-Hao Wang. all to


) system. Samples of plant oils (jojobaoil. of PROCEDURES ATR-FTIR spectroscopy. Oil/water microemulsions four essentials of oils. no attenuatedtotal reflectionFouriertransform infrared(ATR-FTIR) spectroscopy stratumcorneum and strippingstudydealing with the interactionof aminophyllinecreamwith human skin in the presence and absence essential and chemical of oils penetration enhancers appeared the literahas in ture. The preparations were appliedto the humanskin i. some transdermal drug delivery systems have utilized enhancers accelerate to drug permeability(9). toiletries(and alsoin experimental and dermopharmacy) penetraas tion enhancers have been reported(10.Spectra obtained rep- resented average tenscans. in respectively. vivo. Although many studieson the essential oils in topical dermalpreparations.The effectsof microemulsions of containingIB-cyclodextrin hydrateand 1-methyl-2-pyrrolidone the percutaneous on absorption aminophylline of were compared with a controlformulationconsisting 50% ethanolsolution.These systems were usedas vehiclesfor the percutaneous absorption aminophylline. ATR-crystal (ZnSe. olive oil). Consequently. the amountof aminophylline the stratumcorneumlayerswasdeterminedusingATRin FTIR and HPLC. essential (rosemary ylang oil.). lilacin oil. threeplant oils. oils oil.11). cm x 1 cm. HPLC wasperv formedwith a Hitachi modelL-7100 pump and a model 7125 injectorequipped with a 20-pl sample loopanda modelL-7455 photodiode arraydetector (DAD). Chromatograms wereacquired peakareas and calculated means D-7000 chromatogram by ofa data integrator. namelyessential and oil soft(lipophilic skin)penetration enhancers. spectral analysis was performedusing a Spectrum 2.The chemicals investigated this study were 1-methyl-2-pyrrolidone.45 ø) An 7 enabled horizontal positioning the subject's of forearm(ventral). incorporated w/o emulsions. in IB-cyclodextrin hydrate.and after 30 to 60 rain. A recentreportby the European Centerfor Ecotoxicology and Toxicologyof Chemicals(ECETOC) statesthat general-purpose cosmeticcreamsare . peppermintoil). incident angle.0 (Perkin Elmer Ltd. All other chemicals were analytical reagentgrade. REAGENTS AND MATERIALS Aminophyllinewas purchased from Sigma (USA). EXPERIMENTAL APPARATUS Infraredspectra wererecorded with a Perkin Elmer System 2000/IRDM FTIR spectrophotometer with an attenuated total reflection (ATR) (accessory Graseby Specac from Inc.cosmetics. and cosmetics were bought from a numberof retail outlets in the southof Taiwan. an of collected a resolution4 cm Subsequent at of -•.246 JOURNAL OF COSMETIC SCIENCE shownthat most compounds have low permeabilities throughthe skin. purchased (Cs) fromTediaandAldich. The aim of this studywasto investigate skin absorption. corngerm oil. and three chemicalskin penetration enhancers containingSpanand Tween as emulsifying agent were prepared.dimensions. andtricaprylin .

pH 5. Determination /iqzddchromatography.thevalues theabsorption integrations noted N-H (1557cm of band were for -• absorbance) both treated and non-treated arms.Thesemeasurements servedas pretreatmentcontrol values. normal healthysubjects Six (meanage. 3M undera constant was weighedaccurately and spread over 14 cm area forearm cmx 2 cm). 250 x 4.this can be demonstrated the lack of the characteristic oil. of value.By means the injection -•.Taking into accountthe aminophylline contentof the formulation(approx.Therefore.Working standard in solutions were prepared froma standard stock solution methanol the range10-80 mg 1 in in -•.Theformulation maintained contact a 2 of (7 was in with the skinfor 30 min and60 min without occlusion. essential and fatty acids. markedskin The areaof the forearmwaspressed the ZnSe crystal(the window of the Skin Analyzer) on by its own weight. each and At time.50) as the mobile phaseat 1 ml rain andDAD wassetat 270 rim. The tapewasthen torn off.25-5.the stratumcorneum layerwasremoved stripping.SKIN PENETRATION OF AMINOPHYLLINE 247 typically applied skin 1 mg/cm(12). On the otherone. window was cleaned the with alcohol. Figure 1 is the infraredabsorbance spectraof human skin before(a) and after (b and c) contactwith fat-burning cream containing aminophylline. strong The C--O bandin the 1700 -1 cm range is mainly due to the C=O bondspresentin the aminophylline.0. v/v.there also weak is a N-H bond approximately cm at 1557 -1 in the stratumcorneum.ATR-FTIR measurements were recorded the site of application at beforetreatment.All in vivospectra were obtainedunderambientlaboratory conditions and recorded t: 0 at (before emulsion application) at t: 30 min andt-60 min afterapplication. therefore. respectively.0 years)were investigated before(drug-free)and after 30-min and 60-min periodsof treatmentwith a formulation fat-burning of cream.0%). vehicles the wereallowedto evaporate they would during as conditions of normal use. each to at 2 for formulation (0.3 g cream). In addition to a strong N-H stretch (3400cm-t).0 years)were investigated.plant oil. One forearm received no treatment and was used as a control. The skin wasnot occluded.Quantitation was basedon the peak height of the sample. by C=O band of the applied fat-burning creamin the skin print. themiddle each In of forearm. It is therefore on concluded that the adsorbed aminophylline .The presence spectral of bands frequencies at characteristic N-H bonds of provides evidence the adsorption for of the aminophylline the skin. RESULTS AND DISCUSSION The ATR-FTIR observed aminophylline absorbances the spectra the fat-burning in of creamon the human skin and is shownin Table I. by A standard stocksolutionwaspreparedby dissolving10 mg of aminophylline 10 ml of methanol. This operation wasrepeated several times. bands 2915cm 2847cm and1701cm 5% The at -1 -1 -1 correspond C-H andC=O vibrations.6 mm) columneluted methanol-phosphate buffer (20:80. for Stripping method. 22 + 1. to respOctively. The intensityof the N-H bondin the skin spectra aftercontact with the fat-burningcreamis much lower in the N-H bonds. 22 + 1. After eachmeasurement. RP-HPLC wasperformed a Phenomenex on LunaC•s (5u. sites 14 cm two of 2 were delineated. l•l of the 20 preparedsamplesolutionand standardsolutionwas chromatographed under the operating conditionsdescribed above. six normal healthy subjects (mean age.performed applying by by a pieceof adhesive tape (Scotch TM)8 cm in length for 15 seconds.

1495 ----- C=C 95.0538 (correlation coefficient.0 2000.0 . = 0. ATR-FTIR spectra human skin treatedwith and without fat-burning creamcontaining5% of aminophylline/30%corn germ oil. In orderto quantifythe amountof adsorbed aminophylline.'oo . percentagedecreasesignal the of in strength wasplotted the mg/cm of aminophylline vs 2 cream applied the skin. 10. Treatedcreamat 60 min for N-H bendingcharacteristic. y = 0. (c) is largelyresponsible the N-H bondin the spectra skin treatedwith fat-burning for of cream. areaof 7 cm x 2 cm was markedon the an forearm.A 0.248 JOURNAL OF COSMETIC SCIENCE Table I Aminophylline Absorbances Observed Spectra in (ATR-FTIR) of Fat-Burning Sample HumanSkin on Absorption frequencies Absorbance band chosen for Functionality N-H O-H (cm-•) Primaryaminestretch Primaryaminebend Stretch (broad) quantitativeanalysis 3493.9902) was R obtained.64mg/cm2).a'oo Figure 1. a For calibratingabsorbance measurements.Effort on wasmadeto keepthe distribution the solution of uniform overthe 14 cm area 2 (0. 2849 1701 1649 1605. calibrationwasneeded. Afteronehour. IR scan the area obtained.(b) Treated creamat 0 min for N-H bending characteristic.3-g amountof aminophylline creamwasspread the markedarea. The approach determiningthe amountof drug remainingon the skin hasbeenused to for manyyears. 3380 1557 3398 -1557 -- C-H C=O Aliphaticcarbon stretch Ring Ring conjugated Ring benzene 2921. 70 60 50 %T 40 30 20. (a) Untreatedcream.8901 x + 0.1 • 90 • 80. an of was With the absorbance at 1557cm •(N-H fromskin). this experiment. 1525.05-0. C=( 4.A linear to calibration(Figure 2). evaluated amountof aminophylline In we the absorbed .

FTIR spectra The from2000 to 700 cm of aminophylline -• in stratumcorneum treatedwith 30% corngerm oil/10% essential are shownin Figure oil 3.0538 + o o 0.30% corngerm oil/10% jojobaoil/10% tricaprylin.200 "• 0. respectively. Figure 4 shows percentchange the peak heightsof the N-H bendingabsorthe in bances plant oils and essential of oils. cream (containing B [•-cyclodextrin enhancer). and candisturbintercellar lipid packing.9902 0. while peppermint.000 I I I a I 0. amithe .[•-cyclodextrin.200 0.SKIN PENETRATION OF AMINOPHYLLINE 249 R = 0.300 Aminophyiline / cm skin) ( mg • Figure 2.8901 •o = X0.100 0.100 o 0. 18% and 12% peakheightdecreases. 24%.causing abouta 32% peakheight decrease.300 Y0.000 0. and ylang oils caused 28%. hourafterapplication fat-burningcream (containing50% One of A ethanol). Treatment with plant oils and essentialoils in comparison with 5% aminophylline significantlyincreased permeabilityof the stratum corneumto aminophyllinein the comparison with treatmentwith 5% aminophyllinealone. and 1-methyl-2-pyrrolidone) tested enhanced absorption aminophyllinecompared a controlcreamcontainingoil the of to phase. Jojobaoil wasfoundto be the mostactive.and the control cream (without chemicalenhancers). Thesetreatments decreased peak the heightsof the N-H bendingabsorbances comparison in with the control(an aqueous vehicle). Aminophyllineamounts the stratumcorneum min after application. in 60 percutaneously recovering measuring amountof aminophyltine by and the that remained on the skinsurface.lilacin. aminophylline. cream (containing C 1-methyl2-pyrrolidoneenhancer). Plant oils suchasjojobaoil contain esters of unsaturated higheralcohols fatty acidsand terpenes terpenoids essential as and in oils. Three chemicalenhancers (ethanol. respectively.waterphase5%. and surfactants.The 30% corngerm oil/10% jojoba oil producedthe greatestenhancement the permeabilityof the epidermisto in aminophyllineamong the four essential oils studied.increasing flux of aminophylline the (13-15). rosemary.

250 107. ATR-FTIR spectra aminophylline stratum of in corneum after60-min treatment with plantand essential oils. Comparative valuesof peak height decrease aminophylline the stratumcomeurn of in after 60-min treatmentwith plant and essential oils.0 2000. %T50. (b) 10% peppermintoil/30% corngerm oil. '"'•'""•. 40_ 30. . 60.8. 4O 35 30 20 '15 o Corn germ Olive Jojoba Peppermint Ylang Oil enhancers Lilacin Rosemary Figure 4. 20 10 4. 70. (a) 5% aminophylline/30% corngerm oil. (c) 10% jojobaoil/30% corngerm oil. JOURNAL OF COSMETIC SCIENCE 90.01900 1800 1700 li00 15'00 Figure 3.

creamB (containing[3-cyclodextrin enhancer). and [3-cyclodextrin consists six. nophyllinecomponents had penetratedthe stratum corneum(Figure 5). Comparative valuesof peak height decrease aminophylline the formulations: of in Cream A (containing50% ethanol). and creamC (containing 1-methyl-2-pyrrolidone enhancer). the high delivery rate of ethanol enhancers often produces skin irritation. andgreater thanwith [3-cyclodextrin a factorof 1. seven eight glucose of or unitslinkedtogether into a ring. A studyalsoreports linearrelationship a between flux andthermodythe namic activity of the drug in the vehicle with variousconcentrations ethanol (17).27%.penetration of enhancement be indirect by modification the thermodynamic can of activity of the vehicle.SKIN PENETRATION OF AMINOPHYLLINE 251 '"'4O e 20 Cream A Cream E• Forfnulation. Aminophyllineabsorption using 1-methyl-2-pyrrolidone lessthan with 50% ethawas nol by a factor1. C.suchasethanol.can leavethe permeantin a morethermodynamically activestatethan when the solventwas present (16). which hasa smallerring and polar molecule.sinceethanoldirectlyaffects the skin'smodification the formulation. and N-H absorbance at 1557cm • decreased 44.9%forcreams B.Rapid permeationof a goodsolventfrom the donorsolution. One-methylby 2-pyrrolidonehas a heterocyclic ring structure. The enhancement effectof 50% ethanoldemonstrated largestincrease permethe in ability whenthe threechemical enhancers wereused. of However. In addition.6.17. Cream C Figure 5. increasing lipid fluidity. One-methyl-2-pyrrolidone.6. respectively.which is a five-membered ring. 27. Basedon previousstudies . and and A. it canmodifythe intercellular lipid domainsto reducethe barrierresistance the thought to allow easierintercalationon the tightly packedlipids of the stratum corneumand a greaterdisruptionof theselipids. of Thus.43%.

02 - o 9 10 1! 12 13 ReteaQon time ( mia ) Figure 6. and •% •-mcthyl-2-pyrrolidoneenhancer). EVALUATION The aim of the different measurements. the stratumcorneum wasremoved succesby sivelytape strippingthe treatedareaand wasassayed aminophylline for content. and the drug was generallypresentin solutionform. 10% joio•a oil.aminophylline wasappliedto human skin. Rever•ed-phase HPLC and UV spectra a methanol-water of extract aminophylline.In Table II the data obtainedby ATR-FTIR and HPLC are compared.252 JOURNAL OF COSMETIC SCIENCE (16-18). •0% corn •erm oil. and afterperiodsof 30 or 60 min. was to evaluatethe amountsof aminophylline that havepenetrated through the skin by surface recovery._ •oo •-. pyrrolidones were utilized as solvents. In the presentstudy. at . Detection:HPLC-DAD.and the identificationof a peak wasmadeby reference retention to time at a maximum in the 270-nm region of the UV spectrum. With ATR-FTIR the average and .Figure 6 shows the chromatographic spectroscopic and behaviors aminophylline of from the creamformulation.. the enhancers were usedat a low concentration (<5%w/w) to obtain maximum therdynamicactivity. 250 \ \ ß ß \ {.76 min. :- 0.ATR-FTIR and HPLC-UV. Dottedline indicates aminophylline of (---) the peak is detected 270 nm and peak eluting at 11. With application a transdermal of deliverycream. :--.•0 . Aminophylline profile oœ creamC (containin••% aminophylline.

Chetoni. 95 (2006). J.. Saettone. REFERENCES (1) (2) (3) (4) (5) J. 19. Wang. maybe and employed with limited success whenformulated into a matrix-typetransdermal patchor in a gel.. G.5 Average 20. 19. L.canbe evaluated and considered preformulation in studies. R.J. The results of this studysuggest the choice the propercombination oil phase that of of lipidsmay allowachieving drug-controlled delivery from a topicalo/w microemulsion.Eds. F. J. N. 22. P. Pokol.J. D. standard deviations slightly higher than with HPLC because are FTIR detectionis not assensitive UV and the samples as werenot pretreated run on the ZnSecrystal. 1989). Yaoxue Xuebao.(MarcelDekker.However. 190 (1995). SolidStateIonics. E. Obesity. Gal. and H. Int. (8) R. and B.J. Haky. N. Marks. S Satoh.Int. and New York.P.01 -+0. Hori. and M. M. 23. Madarasz. Shore. s.Dermatol. Chromatogr.87. Foss. I. (6) B.69 22. 20. 19. Rel. M. Chinese Pharm. L. Haky.M. 22. Dinner. 587 (2004). Marks. 18. Liu. 197. M. 20. Najarro.4.52 % Decrease peakheightor peakarea= {(peak in heightor peakareadueto treatment--peak heightor peak areadue to control)/peak height or peakareadue to control}x 100. and J. J. M. Katz. (10) D.214 (1996). and S.90.93. M.92 -+1.. Pharm.J. Technol. 31. (11) D.20.J. Foss. Bronaugh andJ. Maibach. CreamC contains1-methyl-2-pyrrolidone enhancer. Abdullah. Maibach. 198 (1999). 172. Petrozziand R. . Microchimica Acta. 36(8).82. "Classification Percutaneous of Penetration Enhancers: ConA ceptualDiagram.Arch. Torok. p. Yourick. L. Liq.56.237.-E. and 209 (2002). 20.J. L. I. C&T.98. W.69 21. Artz. Ping. 153. 534 (2001).SKIN PENETRATION OF AMINOPHYLLINE 253 Table II Determination Aminophylline of after60-min Treatment with CreamC by Stripping Method % Decrease Analyticalmethods LC-UV ATR-FTIR Determinations 19. 47 (2000). W.70. Baan. and 23. 22. and The reproducibility the peakareaof aminophylline poor. I. 525 (1976).-C. Can. which the decrease the peakheightof the permeating in aminophylline determined the cream is in receiving phase. ThusLC-UV canbe used quantitation by for and FTIR for qualitativedetectionand spectroscopic identificationof aminophyllinein a mixture. Zhou.. O."in Percutaneous Absorption. datain a sample of is the areobtained bothLC-UV and FTIR. CONCLUSIONS In vivofindingson skin with an ATR crystalor a strippingmethod.3.48. Bronaugh H. 20.-H. Wang and C. L.S. Boldrini. E.47. G.J. ACKNOWLEDGMENTS Financialsupport this work by the Chia Nan Universityof Pharmacy of and Science is gratefullyacknowledged (CNAC-92-06). 2399 (1997).112. Burgalassi. Monti. 115. Liu and N. Plast. (7) J.L. W. (9) M. J.

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