RESEARCH ARTICLE Kor. J. Aesthet. Cosmetol., Vol. 3 No., 3-, April 5 페녹시에탄올과알칸디올계화합물의혼합사용이항균활성과화장품방부력에미치는영향 최은영건국대학교일반대학원생물공학과 Effect of Phenoyethanol and Alkane Diol Miture on the Antimicrobial Activity and Antiseptic Ability in Cosmetics Eun-Young Choi Department of Bioengineering, Graduate School of Konkuk University Abstract The preservatives traditionally used in the cosmetics such as parabens are regarded as a component to be eliminated because of their harmful properties. Nonetheless, the addition of anti-microbial agents is unavoidable to prevent the microbial contamination. Alkane diols, one of the widely used moisturizing components of cosmetics, are now used as the alternative of anti-microbial agents. In this work, alkane diols such as,-pentanediol,,-heanediol,,-octanediol and phenoyethanol were tested to determine their anti-microbial activity and the possibility to be used as the preservatives in the cosmetics. The inhibition of microbial growth with single alkane diol was not strong enough ecept the,-octanediol, which could weakly inhibit the four tested microorganisms including E. coli, P. aeruginosa, S. aureus, C. albicans. For this reason, each alkane diol and phenoyethanol combination is not suitable for the cosmetic preservatives. On the other hand, the combined three chemicals,,-heanediol (.3%),,-octanediol (.3%) and phenoyethanol (.3%) could successfully inhibit the growth of the four microorganisms and this was confirmed by applying them to the cosmetic emulsion and showing that a 3 log reduction of microorganism was possible. On the basis of these results, the combination of alkane diols with phenoyethanol could be used as a good alternative of cosmetic preservatives. Keywords: Phenoyethanol, Alkane diols, Microorganisms, Anti-microbial activity, Antiseptic ability in cosmetics 화장품의품질은제품의성분, 조성및상태를위생상의성능및안전성에의하여평가되며화장품중많은성분은유화제품으로서동 식물유및광물유를비롯하여지방, 탄수화물등각종세균의영양원을함유하고있다 ( 류미숙, 99). 미생물의성장은적절한온도, 수분, ph 등의조건에영향을받으며, 살아있거나죽어있는유기체내에서 colossal rate로번식하므로대부분의화장품은미생물오염의우려가많다 ( 조춘 구등, ). 화장품의부패를막기위한방법으로방부제성분은매우중요하며화장품도식품, 의약품등과마찬가지로미생물에의한침투를막기위하여방부제를많이이용하고있다 (Shigeru et al., 3). 화장품에사용되는방부제의종류는매우다양하며화장품산업의증가로인해그사용량또한증가하는추세이다 ( 이후섭등, 4). 화장품은미생물이이용가능한성분들이풍부하여장기간보관및사용중에미생물에오염되기쉬우므로이를방지하기위해방부제를사용하는것은필수적이기때문이다. 일반적으로화장품의방부제로사용되는 Corresponding author: Eun-Young Choi, Department of Bioengineering, Graduate School of Konkuk University, Neungdong-ro, Gwangjin-gu, Seoul 43-7, Republic of Korea E-mail : eyoung4@daum.net Received February 5, 5; Revised April 7, 5; Accepted April 3, 5; Published April 3, 5 www.kosac.or.kr 3
Kor. J. Aesthet. Cosmetol., Vol. 3 No., 3-, April 5 대부분의화학방부제로는파라벤류, 페녹시에탄올, 클로로페네신등이사용되지만, 이러한화학방부제들은미생물의성장을억제하여제품의안정성을부여함과동시에민감성피부및접촉성피부염등피부알러지와환경호른몬및자극유발의원인이되기도한다 (Marouchoc, 98: Soni et al., 5). 또한안전지향의소비자들요구로인해화장품의표시성분으로제품에표기되는화학방부제에대한소비자의거부감이크게증가하고있다. 따라서화장품에사용되는모든화학방부제들은사용상의제한이필요한원료 ( 배합한도 ) 로지정하여관리되고있다. 이러한많은소비자들의안전성에관한요구로인해화장품업계에서는방부제를사용하지않은 preservative-free 제품, 자체방부기능을가능하게할수있는원료를사용하는 selfpreserving 제품등에관심이많았다 (Shigeru et al., 3). 최근에는자체항균작용이있는,-알칸디올계화합물의원료를사용하여화장품에서방부효과를나타낼수있는방법이지속적으로검토되고있다. 항균작용의,-알칸디올계화합물중대표적으로,-octanediol과,-heanediol 성분등이화장품내에서항균작용기능의원료로적용되어사용하고있다 (Orth & Kabara, 998; Gaonkar et al., 6; Shintre et al., 6; Levy, 8). 이들화합물에대한사용허용치가매우높고, 이허용치의수십배농도에서도별다른피부자극이없음이보고된바있다 (Levy et al., 9: Lee et al., ). 그러나이들방부제에대한항균효과에대해서는매우제한적인정보를얻을수있으나, 실제화장품에사용했을때기대할수있는방부력에대한챌린지테스트보고는전혀없는실정이다. 따라서본연구에서는화장품방부제인페녹시에탄올의사용량을줄이기위하여,-pentanediol,,-heanediol,,-octanediol과같은알칸디올계화합물질과페녹시에탄올을혼합하여사용했을때이들혼합물들의항균효과와방부력을조사해보았다.. 시험균주및배양본실험에서사용한시험균주 Candida albicans (C. albicans), ATCC 3, Escherichia coli (E. coli) ATCC 8739, Pseudomonas aeruginosa (P. aeruginosa) ATCC 97, Staphylococcus aureus (S. aureus) ATCC 65389는한국미생물보존센터에서분양받아사용하였다. C. albicans는 3, E. coli, P. aeruginosa, S. aureus 는 37 의 incubator에서 4 h 배양하였고, 3일간격으로 Tryptic Soy Broth (TSB), Potato Detrose Broth (PDB) 한천배지에계대하여균의활성을유지시켰다.. 사용배지및조성 본실험에이용한배지로는 DIFCO 사제품으로 Table (A), (B) 와같이 5 종의배지를사용하여시험균에대한항균력및방 부력테스트에사용하였다. Table. The composition of culture medium (A) (B) Medium E. coli, P. aeruginosa, S. aureus C. albicans Enrichment media TSB PDB Detection media TSA LT PDA LT Diluted solution TSB LT Ingredient TSB TSB LT TSA LT PDB PDA LT Trypticase soy broth 3 3 3 - - Potato starch Detrose Broth - - -.65.65 Agar - -.5 -.5 Lecithin -.7.7 -.7 tween 8 - - D.W.(mL) ph 7.3. 7.3. 7.3. 5.6. 5.6. T.S.B=Trypticase Soy Broth, T.S.A.=Trypticase Soy Agar, T.S.A. LT =Trypticase Soy Agar containing Lecithin and Tween 8, P.D.B=Potato starch Detrose Broth, P.D.A=Potato starch Detrose Agar, P.D.A. LT=Potato starch Detrose Agar Lecithin and Tween 8 Table. Composition of eperimental lotions including various preservatives A Ingredients Negative Eperimental sample,3 Butylene Glycol 3. Ethanol 3. Hydrogenated Phosphatidyl Choline. Cholesterol. B Carpryl / capryltriglyceride. C D E Tween 8.3 Positive control Propyl paraben - -. Methyl paraben - -. D.W Up to ml Phenoyethanol -.3 - -,-heanediol -.3 - -,-octanediol -.3 - - Sodium Polyacrylates & Ethyleneheyl stearate & Trideceth-6. 4 www.kosac.or.kr
페녹시에탄올과알칸디올계화합물의혼합사용이항균활성과화장품방부력에미치는영향 3. 합성방부제및알칸디올계화합물본연구에사용된합성방부제는비교적사용빈도가높은프로필파라벤, 메칠파라벤, 페녹시에탄올 ( 한빛화학, 한국 ) 을사용하였고알칸디올계화합물의표준시약,-pentanediol,,-heanediol,,-octanediol (Sigma, USA) 를사용하였다. 4. Broth culture method 배양된각균주의단일 colony를취해 ml 멸균된액체배지에접종하였다. C. albicans 배양은 3, E. coli, S. aureus, P. aeruginosa의배양은 37, 에서 4 h incubator 에서배양하였다. TSB, PDB 액체배지 9.5 ml에천연물질을 DMSO에각농도별로용해한.5 ml을넣어시험배지를준비한다. 배양한균주를 6-7 CFU/ml이되도록각시험배지에접종후 4 h incubator에서배양하였다. 그후 h 간격으로샘플을 ml씩취하여 spectrophotometer를이용하여 6 nm에서흡광도를측정하여미생물증식곡선을결정하였다. 5. Challenge test 합성방부제페녹시에탄올및알칸디올계화합물을함유한화장품의 challenge test는 The Cosmetic, Toiletry, and Fragrance Association (CTFA, 993) 에서제안한기준을따라수행하였다 (Levy et al., 9). Table 에나타낸처방설계대로방부제를첨가하지않은 g의로션제형과만들어대조군으로하고, 메칠파라벤.% 와프로필파라벤.% 가첨가된양성대조군과페녹시에탄올.3%,,-heanediol.3%,,-octanediol.3% 를혼합한시험군을제조하여사용하였다. 준비된각시험물질이첨가된화장품에 C. albicans, E. coli, S. aureus, P. aeruginosa 배양액을 6-7 CFU/g이되도록접종한후상온보관하였다. 접종직후,,, 3, 7, 4,, 8 일에각각 sample을취하여적당한비율로준비된희석액에희석한후멸균된검출용 agar 배지에도말하여 4 h incubator 에서배양한후 colony 수를계수하여경과일에따른접종균의사멸율을계산하여방부력을측정하였다. 6. 화장품의제조실험에사용된화장품의제조는 Table 의처방대로제조를하였다. 화장품제조의간략한과정을살펴보면, (A) 상을 6~8 에서완전용해한후, (B) 상을 (A) 상에투입한다. 그다음 (C) 상을 6 까지가온하고 (A)+(B) 상을 (C) 상에투입하며호모믹서 3, rpm에서 5 min간유화시킨다. 유화시킨후 (D) 상을 (A)+(B)+(C) 상에투입한다. 4 이하로냉각후 (E) 상을투입하여 5 min 동안 3, rpm으로교반하여점도가있는로션제형을얻어실험에사용하였다. 제조된화장품 제형은 5 항온조에서일주일간정치하여안정성테스트를시행하였다.. 페녹시에탄올과알칸디올계화합물의단독사용에따른항균효과페녹시에탄올과,-pentanediol,,-heanediol,,-octanediol을네가지균주 E. coli, P. aeruginosa, S. aureus, C. albicans에각각단독으로사용하여항균효과를조사하였고 E. coli의결과를나타내었다 (Figure ). E. coli 에서는페녹시에탄올.5% 이상,,-heanediol.3% 이상,,-octanediol.% 이상처리한경우균성장이효과적으로억제되었으나,,-pentanediol의경우에는 % 까지처리해도효과적인항균력을보이지못했다. 페녹시에탄올과,-pentanediol,,-heanediol,,-octanediol을단독으로사용하여 P. aeruginosa의항균효과를나타내었다 (Figure ). P. aeruginosa에서는페녹시에탄올.7%,,-heanediol.5% 까지처리했으나균주를완전히억제하지못하였으며,,-pentanediol의경우에는 % 까지처리해도항균효과가나타나지않았다. 반면에,-octanediol.3% 에서는균성장을효과적으로억제하였다. 페녹시에탄올과,-pentanediol,,-heanediol,,-octanediol을단독으로사용하여 S. aureus의항균효과를나타내었다 (Figure 3). S. aureus에서도 P. aeruginosa 와유사한항균효과를보였는데, 페녹시에탄올.7%,,-heanediol.5% 까지처리했으나균의성장을완전히억제하지못하였으며,,-pentanediol의경우에는 % 까지처리해도항균효과가거의나타나지않았다. 또한,-octanediol.3% 에서는균의성장을다소억제하였으나완전히억제하지는못하였다. 마지막으로페녹시에탄올과,-pentanediol,,-heanediol,,-octanediol을단독으로사용하여 C. albicans의항균효과를나타내었다 (Figure 4). C. albicans 에서는페녹시에탄올과,-pentanediol,,-heanediol,,-octanediol 네가지물질모두다균성장에매우약한억제력을보였다. 이러한결과로보아본연구에서실험한네가지화합물의항균력은,-octanediol이가장우수하였고, 그다음페녹시에탄올,,-heanediol,,-pentanediol의순으로낮아지는것으로보인다. 알칸디올계화합물화합물들이미생물에대해항균력을보이는것은대체로이들화합물들이미생물의세포막형성을방해하기때문인것으로알려져있 www.kosac.or.kr 5
Kor. J. Aesthet. Cosmetol., Vol. 3 No., 3-, April 5 (A) (B) O.D(6nm).8.6.4..%.5%.7% O.D(6nm).8.6.4..3% % % (C)( ( D).%.8.3%.5%.6.4 O.D(6nm) O.D(6nm).8.6.4.%.%.3%.. Figure. Effects of single preservatives on the microbial growth. Miroorganism: E. coli Preservatives; A) Phenoyethanol, B),-Pentanediol, C),-Heanediol, D),-Octanediol (A)( ( B) O.D(6nm).8.6.4..%.5%.7% O.D(6nm).8.6.4..3%.5% % (C)( ( D).%.8.3%.8.5%.6.6.4.4.. O.D(6nm) O.D(6nm).%.%.3% Figure. Effects of single preservatives on the microbial growth. Miroorganism: P. aeruginosa, Preservatives; A)Phenoyethanol, B),-Pentanediol, C),-Heanediol D),-Octanediol 다.(Kabara & Orth, 997). 따라서알칸디올계화합물의탄 소체인의길이가증가할수록세포막형성을방해하는효과가 증가하여항균효과또한증가한다고한다. 그러나항균력과 방부력은비례하지않는데탄소수가 8 개인,-octandediol 에비해 개인,-decanediol 은방부력에서오히려그효 과가떨어지는것으로알려져있는데, 이것은,-decanedio 6 www.kosac.or.kr
페녹시에탄올과알칸디올계화합물의혼합사용이항균활성과화장품방부력에미치는영향 (A)( ( B) O.D(6nm).8.6.4..%.5%.7% O.D(6nm).8.6.4..3%.5% % (C)( ( D).%.8.3%.5%.6.4 O.D(6nm) O.D(6nm).8.6.4.%.%.3%.. Figure 3. Effects of single preservatives on the microbial growth. Miroorganism: S. aureus, Preservatives; A)Phenoyethanol, B),-Pentanediol, C),-Heanediol, D),-Octanediol (A)( ( B) O.D(6nm).8.6.4..%.5%.7% O.D(6nm).8.6.4..3%.5% % (C)( ( D).%.8.3%.5% O.D(6nm).6.4. Figure 4. Effects of single preservatives on the microbial growth. Miroorganism: C. albicans, Preservatives; A)Phenoyethanol, B),-Pentanediol C),-Heanediol D),-Octanediol O.D(6nm).8.6.4..%.%.3% 이,-octandediol 에비해 water activity 가낮기때문이다 (Schmaus et al., 4). 페녹시에탄올과알칸디올계화합물의단독사용으로사용한 결과이네가지화합물들은주어진농도범위에서네가지균 주 E. coli, P. aeruginosa, S. aureus, C. albicans 의성장을 완전히억제하지못하는것으로나타났으며, 따라서이들단독 www.kosac.or.kr 7
Kor. J. Aesthet. Cosmetol., Vol. 3 No., 3-, April 5.7.9.6.8 O.D(6nm).5.4.3 O.D(6nm).7.6.5.4..3. 4 6, Heane diol.3, Caprylyl glycol.%, Heane diol.3, Caprylyl glycol.3%, Heane diol.5, Caprylyl glycol.%, Heane diol.5, Caprylyl glycol.3%, Heane diol.3, Caprylyl glycol.%, Phenoyethanol.% Figure 5. Effect of mied preservatives on the microbial growth. Miroorganism: E. coli 8.., Heane diol.3, Caprylyl glycol.%, Heane diol.3, Caprylyl glycol.3%, Heane diol.5, Caprylyl glycol.%, Heane diol.5, Caprylyl glycol.3%, Heane diol.3, Caprylyl glycol.%, Phenoyethanol.% Figure 7. Effect of mied preservatives on the microbial growth. Miroorganism: S. aureus Figure 6. Effect of mied preservatives on the microbial growth. Miroorganism: P. aeruginosa 물질로는화장품방부제로이용될수없음을확인하였다.. 페녹시에탄올과알칸디올계화합물혼합사용에따른항균 효과 O.D(6nm).9.8.7.6.5.4.3.., Heane diol.3, Caprylyl glycol.%, Heane diol.3, Caprylyl glycol.3%, Heane diol.5, Caprylyl glycol.%, Heane diol.5, Caprylyl glycol.3%, Heane diol.3, Caprylyl glycol.%, Phenoyethanol.% 방부제혼합사용실험에서는방부력이가장약한,-pentanediol 은제외하였다. 페녹시에탄올과,-heanediol,,-octanediol 을혼합사용하였을때네 가지균주 E. coli, P. aeruginosa, S. aureus, C. albicans 에대한항균효과를조사한것이다. 먼저 E. coli 에서는,-heanediol.3% 이상,,-octanediol.% 이상을동 시처리할경우균성장이모두억제되었으며, 여기에페녹시에 탄올을첨가하면그효과가더욱더증대되었다 (Figure 5). 그다음 P. aeruginosa 에서는,-heanediol.5%, 이상 4 6 8 O.D(6nm).5.5.5, Heane diol.3, Caprylyl glycol.3%, Heane diol.5, Caprylyl glycol.%, Heane diol.5, Caprylyl glycol.3%, Heane diol.3, Caprylyl glycol.%, Phenoyethanol.%, Heane diol.3, Caprylyl glycol.3%, Phenoyethanol.% Figure 8. Effect of mied preservatives on the microbial growth. Miroorganism: C. albicans,-octanediol.3% 이상을동시처리한경우에만균성장이 억제되었으며, 그이하농도에서는효과적인항균력을나타내 지못하였다. 그러나,-heanediol.3%,,-octanediol.% 의농도에서도페녹시에탄올을.% 첨가한경우에는균 성장을효과적으로억제하는것으로나타났다 (Figure 6). S. aureus 에서는 P. aeruginosa 에서보다좀더우수한항 균효과를보였는데, 예를들어,-octanediol.3% 농도에 서,-heanediol.5% 까지처리하면균의성장을완전히억 제하였으며,,-heanediol.3%,,-octanediol.%, 페 녹시에탄올.% 를동시에처리하면균의성장이효과적으로 억제되었다 (Figure 7). 3 6 9 8 4 C. albicans 에서는위방부제들의혼합사용이다른균주 에비해덜효과적인것으로나타났으나, 페녹시에탄올.3%,,-heanediol.3%,,-octanediol.3% 를혼합하여사 용할경우균성장을확실히억제하였다 (Figure 8). 8 www.kosac.or.kr
페녹시에탄올과알칸디올계화합물의혼합사용이항균활성과화장품방부력에미치는영향 logcfu/ml logcfu/ml logcfu/ml (A) 8 6 4 (B) 7 6 5 4 3 (C) 8 6 4 3 7 5 8 Figure 9. Challenge test of mied preservatives in the cosmetics emulsions. ) (no preservatives), ) Positive control (Methyl paraben.%+propyl paraben.%), 3) Mied alkanediol preservatives (,-heanediol.3%+caprylyl glycol.3%+phenoyethanol.3%) 3. 화장품방부력테스트 위의실험결과를바탕으로, 페녹시에탄올과이들알칸디올 계화합물들의혼합물이과연화장품방부제로이용될수있을 지를확인하기위하여 oil in water 형태의화장품을제조하고, 여기에네가지균주들을접종하여화장품방부력테스트를실 험한후결과를나타내었다 (Figure 9). 화장품속에방부제를전혀첨가하지않은경우, 네가지 균모두화장품내에서 6 /ml 이상의콜로니를형성하며성 장한다는것을알수있었다. 화장품에사용되는전형적인방 부제인메칠파라벤.% 와프로필파라벤.% 를첨가한양 성대조군의경우, 보존기간 7 일이후네가지균주모두가 사멸하는것으로나타났다. 마지막으로페녹시에탄올.3%,,-heanediol.3%,,-octanediol.3% 를혼합하여첨 가한경우 E. coli, P. aeruginosa, S. aureus C. albicans 모 두에서 3 log 이상감소하는효과를보여이들혼합물의방부력 을확인할수있었다. Time(day) 3 7 5 8 Time(day) E. coli P.aeruginosa S.aureus C.albican 3 7 5 8 Time(day) E. coli P.aeruginosa S.aureus C.albican E. coli P.aeruginosa S.aureus C.albican 미생물오염을통해화장품이변질되거나분해되는것을방지하기위해사용되는살균, 보존제인화학방부제는소비자의안전을위해최근에는자체항균작용이있는알칸디올계화합물이대체방부제로다양하게이용되어질것으로기대되어지고있다. 본연구에서는화장품방부제인페녹시에탄올의사용량을줄이기위하여,-pentanediol,,-heanediol,,-octanediol과같은알칸디올계화합물물질과페녹시에탄올을단독으로사용하였을때와혼합하여사용했을때이들알칸디올계화합물의항균효과와화장품방부제로서의방부력을조사해보았다. 페녹시에탄올과알칸디올계화합물을단독으로사용한결과주어진농도범위에서 E. coli, P. aeruginosa, S. aureus, C. albicans 균들의성장을완전히억제하지못하는것으로나타나단독물질로는화장품방부제로이용될수없음을확인하였다. 혼합사용시 E. coli에서는,-heanediol.3% 와,-octanediol.% 이상을동시에처리한경우, P. aeruginosa에서는,-heanediol.5% 와,-octanediol.3% 이상을동시에처리한경우, S. aureus 에서는,-heanediol.5% 와,-octanediol.3% 이상을처리한경우, C. albicans에서는,-heanediol.3%,,-octanediol.3% 를혼합하여사용할경우처리균성장이효과적으로억제하는것으로나타났으며페녹시에탄올.% 를동시에처리하면 E. coli, P. aeruginosa, S. aureus, C. albicans 모두억제되는것을확인할수있었다. 혼합사용의결과를바탕으로화장품방부제로이용가능성을확인하기위하여제조한화장품에 E. coli, P. aeruginosa, S. aureus, C. albicans 를접종하여화장품방부력테스트를실시하였다. 페녹시에탄올.3% 와알칸디올계화합물인,-heanediol.3%,,-octanediol.3% 를혼합하여첨가한경우에는 E. coli, P. aeruginosa, S. aureus, C. albicans 모두에서 3 log이상감소하는효과를보여이들혼합물의방부력을확인할수있었다. 본실험에서는비교적안전한화합물로알려진알칸디올계화합물의물질들은화장품방부제로사용될수있으나이들의단독사용은불가능하며, 반드시 가지이상의알칸디올계화합물을페녹시에탄올과함께혼합하여사용하여야만그효과를극대화할수있을것으로사료된다. 류미숙. 화장품의미생물오염방지에대한방부력효과. 성균관 www.kosac.or.kr 9
Kor. J. Aesthet. Cosmetol., Vol. 3 No., 3-, April 5 대학교석사학위논문, 99. 이후섭, 이상훈, 김은죽, 정철희, 표형배. 가스크로마토그래피를이용한화장품중항균작용의 3종의직쇄상,- 알칸디올류의동시분석. 대한화장품학회지, 4: 37-44, 4. 조춘구, 김봉남, 홍세흠, 한창규. 합성항균제를대체하기위한천연물질의항균효과. 대한피부미용학회지, 8: 66-85,. Orth DS, Kabara JJ. Presevative-free and selfpreserving cosmetics and drugs. Cosmet. Toil., 3: 5-58, 998. Kabara J, Orth D. Preservative-free and self-preserving cosmetic and drug products: Principle and practices. Cosmetic Science and Technology Series, 6: 43-6, 997. Lee E, An S, Cho SA, Yun Y, Han J, Hwang YK, Kim HK, Lee TR. The influence of alkane chain length on the skin irritation potential of,-alkanediols. Int. J Cosmetic Sci., 33: 4-45,. Levy SB, Dulichan AM, Helman M. Safety of a preservative system containing, heanediol and capryly glycol. Cutaneous and occular toicology., 8: 3-4, 9. Soni MG, Carabin IG, Burdock GA. Safety assessment of esters of p-hydroybenzoic acid (parabens). Food Chem. Tocol., 43: 985-5, 5. Shintre MS, Gaonkar TA, Modak SM. Efficacy of an alcohol-based healthcare hand rub containing synergistic combination of farnesol and benzethonium chloride. Int. J. Hyg. Environ. Health., 9: 477-487, 6. Schmaus G et al..,-diol combinations: A safe way to reduce preservatives in cosmetics formulation. Cosmetic and Toiletries., 3: 53-64. 8. Shigeru Abe, Yuichi Sato, Shigeharu Inoue, Hiroko Ishibashi, Naruyama, Toshio Takizawa, Haruyuki Oshima, Hideyo Yamaguchi. Anti-Candida albicans Activity of essential Oils Including Lemongrass Oil and its Component. Citral. Jpn J. Med. Myeol., 44: 9-4. 3. Marouchoc SR. Cosmetic preservation. Cosmet. Technol., : 38, 98. Levy S. Safety of a new preservative system containing,-heanediol and,-octanediol for use in cosmetic products. J. Am. Acad. Dermatol., 58: 55, 8. Gaonkar TA, Geraldo I, Shintre M, Modak SM. In vivo efficacy of an alcohol-based surgical hand disinfectant containing a synergistic combination of ethylheylglycerin and preservatives. J. Hosp. Infect., 63: 4-47, 6. www.kosac.or.kr