Дисертації з теми "Polysaccharides used in cosmetics"

The presented master thesis deals with the monitoring of the influence of polysaccharides which are used as an additive in the cosmetic products, on the monitored types of bacteria which are part of the skin microbiome. And it also deals with the study the effect of polysaccharides as prebiotics on selected species of bacteria that are part of the skin microbiome. Two polysaccharides were selected to determine the effects on the skin microbiome: Nanomoist and PoLevan S. The first part of the thesis focuses on the literature search which deals with skin anatomy, skin diseases and skin microbiome and its function. The experimental part is focused on monitoring changes in the quantity of selected microorganisms of the skin microbiome, before and after application of polysaccharides to the skin using qPCR. Staphylococcus epidermidis, Cutibacterium acnes, Escherichia coli and Micrococcus luteus were monitored. The PCR products were detected by agarose gel electrophoresis. The bacterium Staphylococcus epidermidis was detected on the skin to the greatest extent, especially after the application of the polysaccharides Nanomoist and PoLevan S. Thus, a positive effect of both polysaccharides on the growth of this bacterium was found.

Doutoramento em Química
The use of plants with medicinal purposes is an ancient practice still very common in developing regions, and is rapidly spreading in industrialized countries. This fact is evidenced by the large number of ethnobotanical studies found in the literature referring that these plants are often used as decoctions and infusions. In most studies the reported biological activities are attributed to the presence of phenolic compounds, due to their antioxidant properties, and to polysaccharides, with its anti-tumoral properties. In “Trás-os-Montes” region, some of the most popular infusions used by the popular medicine are prepared with the dried leaves of Fraxinus angustifolia, the dried shoots of Mentha suaveolens, and the dried inflorescences of Pterospartum tridentatum. However, there are no studies about the polysaccharides present in these infusions. Thus, through the structural characterization of the polysaccharides present in the infusions of F. angustifolia, M. suaveolens, and P. tridentatum, the present PhD thesis intends to evaluate the possible relation between polysaccharides and the immunostimulatory activity that these infusions might present. In a preliminary phase, infusions of F. angustifolia were prepared according to the popular tradition, and it was observed that the obtained water soluble material contained approximately 85% of material non-retained in C18 cartridges, with hydrophilic characteristics, with the remaining 15% comprising retained-material with hydrophobic characteristics. It was also shown that the infusions only contained between 2 and 4% of high molecular weight material (HMWM), which comprised approximately 30% of carbohydrate material. Sugar and methylation analysis of the HMWM suggested the presence of pectic polysaccharides, together with type II arabinogalactans, mannans, and xyloglucans. However, the amount of material obtained is to low for the fractionation, and structural analysis of the polysaccharides present. The 4 h decoction, divided in two periods of 2 h, with water renewal, allowed to increase the HMWM yield, relatively to the infusions traditional infusions. It was also observed that the decoction also allowed to increase the HMWM proportion of carbohydrate material, due to an increase in the proportion of uronic acid present, although the neutral sugar residues seemed to be detected in similar proportions. Therefore, in all the experiments subsequently performed, the HMWM used was obtained through the decoction of F. angustifolia dried leaves, M. suaveolens dried shoots, and P. tridentatum dried inflorescences. x After the fractionation, through ethanol precipitation, and anion exchange chromatography, of the polysaccharides from the HMWM obtained by the decoction of the vegetable material of the distinct studied plants, it was observed the presence of high proportions of pectic polysaccharides, containing type I arabinogalactans, together with minor proportions of type II arabinogalactans, mannans, and xyloglucans. The presence of pectic polysaccharides in the extracts from F. angustifolia was also evidenced through endo-polygalacturonase treatment, and ESI-MS and ESI-MS/MS experiments. The detection of linked pentose and uronic acid residues, also seemed to suggest the presence of xylogalacturonan domains in the pectic polysaccharides from F. angustifolia. The extracts from F. angustifolia dried leaves also contained type II arabinogalactans that exhibited a higher structural diversity than those detected in the M. suaveolens, and P. tridentatum extracts, particularly in the substitution degree of the galactan backbone, and in the extension of the (1→5)-Araf side chains. Moreover, for all the plants studied, it was also observed that the type II arabinogalactans, extracted during the 2nd 2h of the extraction process, exhibited a substitution degree of the galactan backbone higher than those extracted during the 1st 2h. The extracts from P. tridentatum dried inflorescences contained higher proportions of mannans, and also of xyloglucans, both presenting a substitution degree higher than those, which were detected in lower proportion in the extracts of F. angustifolia and M. suaveolens. Through ESI-MS and ESI-MS/MS it was possible to evidence that the mannans present in the extracts of P. tridentatum presented acetyl groups on the O-2 of the mannosyl residues. It was also evidenced that the P. tridentatum mannans were more extensively acetylated than the mannans detected in the coffee infusion, LBG, and other non-conventional mannan sources. Moreover, it was detected the presence of oligosaccharides comprising hexose residues linked to non acetylated pentose residues, suggesting the possible presence of arabinose residues in the mannans from P. tridentatum extracts. The immunostimulatory activity of three fractions isolated from the extracts of F. angustifolia, M. suaveolens, and P. tridentatum, was tested and an increase in the NO production by macrophages, without compromising their cellular viability, was observed. The type I, and type II arabinogalactans detected in the extracts from F. angustifolia, and M. suaveolens seem to have contributed for the observed immunostimulatory activity. For the fraction from P. tridentatum, the mannans acetylation, and the presence of type I, and type II arabinogalactans seemed to contribute for the macrophage immunostimulatory activity observed. The possible presence of storage xyloglucans from the inflorescences seeds, also seems to have contributed for the immunostimulatory activity registered when the macrophages were stimulated with higher extract concentrations. The results obtained allow to conclude that the extracts of F. angustifolia dried leaves, M. suaveolens dried shoots, and P. tridentatum dried inflorescences contained high proportions of pectic polysaccharides, exhibiting type I arabinogalactans, together with other polysaccharides, such as type II arabinogalactans, mannans, and xyloglucans. This polysaccharide mixture seems to have contributed to the immunostimulatory activity of fractions isolated from the extracts of the studied plants. Therefore, as the same type of polysaccharides seem to be present in the decoctions and in the infusions, it seems possible that the polysaccharides might contribute for the therapeutic properties frequently associated by the popular tradition to the infusions of these plants.
O uso de plantas com fins medicinais é uma prática antiga, ainda bastante frequente em regiões de países em desenvolvimento, e que se tem vindo a disseminar em países mais industrializados. Este facto é suportado pelo elevado número de estudos etnobotânicos, onde se refere que estas plantas são frequentemente usadas na forma de decocções e infusões. Na maioria dos estudos as atividades biológicas são associadas à presença de compostos fenólicos e às suas propriedades antioxidantes, assim como aos polissacarídeos, nomeadamente devido às propriedades antitumorais. Na região de Trás-os-Montes, algumas das infusões mais utilizadas pela medicina popular têm por base as folhas secas do freixo (Fraxinus angustifolia), a parte aérea seca do mondrasto (Mentha suaveolens) e as inflorescências secas da carqueja (Pterospartum tridentatum). No entanto, não existe nenhum estudo sobre os polissacarídeos presentes nas infusões das folhas secas do freixo (Fraxinus angustifolia), da parte aérea do mondrasto (Mentha suaveolens) ou das inflorescências da carqueja (Pterospartum tridentatum). Deste modo, com a presente dissertação pretende-se, através da caracterização estrutural dos polissacarídeos presentes nas infusões de F. angustifolia, M. suaveolens e P. tridentatum, avaliar a possível relação entre os polissacarídeos e a atividade imunoestimuladora que estas infusões possam apresentar. Numa primeira fase, foram preparadas infusões tradicionais de F. angustifolia, tendo-se mostrado que o material solúvel das infusões continha cerca de 85% de material não retido em cartuchos C18 com características hidrofílicas, sendo os restantes 15% compostos por material retido de características hidrofóbicas. Mostrou-se ainda que as infusões obtidas apenas continham cerca de 2 a 4% de material de alto peso molecular (HMWM), que era composto por cerca de 30% de carboidratos. A análise de açúcares e a análise de metilação do HMWM sugeriu a presença de polissacarídeos pécticos, em conjunto com arabinogalactanas do tipo II, mananas e xiloglucanas. No entanto, a quantidade de material obtido é muito baixa para o fracionamento e a análise estrutural dos polissacarídeos presentes. A decocção de 4 h, divididas em 2 períodos de 2 h, com renovação da água, permitiu um aumento no rendimento em HMWM, relativamente às infusões tradicionais. Verificou-se ainda, que a decocção também permitiu aumentar a proporção de carboidratos no HMWM, particularmente devido ao aumento da proporção de ácido urónico presente, embora os resíduos de açúcares neutros se encontrassem em proporção semelhante. Deste modo, em todas as experiências realizadas posteriormente, o HMWM utilizado foi obtido através da decocção das folhas secas de F. angustifolia, da parte aérea seca de M. suaveolens e das inflorescências secas de P. tridentatum. Após fracionamento por precipitação em etanol e cromatografia de troca aniónica dos polissacarídeos presentes no HMWM obtido através da decocção dos materiais vegetais das diferentes plantas estudadas, verificou-se a presença de elevadas proporções de polissacarídeos pécticos, contendo arabinogalactanas do tipo I, em conjunto com menores proporções de arabinogalactanas do tipo II, mananas e xiloglucanas. A presença de polissacarídeos pécticos nos extratos de F. angustifolia foi também evidenciada através do tratamento enzimático com endopoligalacturonase e de experiências de ESI-MS e ESI-MS/MS. A deteção de resíduos de pentose ligados a resíduos de ácido urónico também pareceu indicar a presença de domínios de xilogalacturonana nos polissacarídeos pécticos de F. angustifolia. Os extratos das folhas secas de F. angustifolia mostraram conter arabinogalactanas do tipo II com uma maior diversidade estrutural do que as que foram detetadas nos extratos de M. suaveolens e P. tridentatum, particularmente no que diz respeito à substituição das cadeias de (1→3)-Galp e à extensão das cadeias laterais compostas por resíduos de (1→5)-Araf. Para além disso, verificou-se que, para todas as plantas estudadas, as arabinogalactanas do tipo II presentes nas frações obtidas durante a segunda parte do processo de extração apresentaram uma substituição das cadeias de (1→3)-Galp maior do que as extraídas durante a primeira parte da extração. Os extratos das inflorescências de P. tridentatum continham maiores proporções de mananas e de xiloglucanas, tendo-se detetado estruturas que apresentaram um maior grau de substituição do que as que foram detetadas em menores proporções nos extratos de F. angustifolia e M. suaveolens. Através das técnicas de ESI-MS e ESI-MS/MS, foi possível demonstrar que as mananas presentes nos extratos de P. tridentatum se encontravam acetiladas no O-2 dos resíduos de manose. Verificou-se que as mananas presentes nos extratos de P. tridentatum se encontravam acetiladas em maior extensão do que as mananas detetadas na infusão do café, na goma de alfarroba e em outras fontes não convencionais de mananas. Para além disso, também foram detetados oligossacarídeos contendo resíduos de pentose, ligados a resíduos de hexose, sugerindo a presença de resíduos de arabinose na estrutura das mananas de P. tridentatum. A atividade imunoestimuladora de três frações isoladas a partir dos extratos de F. angustifolia, M. suaveolens e P. tridentatum foi avaliada, tendo-se observado o aumento da produção de NO por parte de macrófagos, sem comprometer a sua viabilidade celular. As arabinogalactanas do tipo I e II parecem contribuir para a atividade imunoestimuladora observada nas frações dos extratos de F. angustifolia e de M. suaveolens No caso da fração obtida a partir dos extratos de P. tridentatum, a acetilação das mananas, bem como a presença de arabinogalactanas do tipo I e II parecem ter contribuído para a atividade imunoestimuladora dos macrófagos observada. A possível presença de xiloglucanas de armazenamento, provenientes das sementes das inflorescências de P. tridentatum, também poderá contribuir para a atividade imunoestimuladora observada quando os macrófagos foram estimulados com maiores concentrações de extrato. Os resultados obtidos permitem concluir que os extratos das folhas secas de F. angustifolia, da parte aérea de M. suaveolens e das inflorescências de P. tridentatum contêm elevadas proporções de polissacarídeos pécticos, contendo arabinogalactanas do tipo I, misturados com outros polissacarídeos, tais como arabinogalactanas do tipo II, mananas e xiloglucanas. Esta mistura de polissacarídeos mostrou contribuir para a atividade imunoestimuladora de frações isoladas a partir dos extratos das plantas estudadas. Deste modo, e como o mesmo tipo de polissacarídeos parece estar presente nas infusões e decoções, é possível que os polissacarídeos possam contribuir para as propriedades terapêuticas frequentemente atribuídas pela tradição popular às infusões destas plantas.

Plastics have a wide application field, where cosmetic products are one of them. Polymers which are building blocks to plastics exists in many variants, overall they can be categorized into two groups; microplastics and water soluble polymers. Both polymer types are important to study and understand since polymers in general are not covered by any legislation. To gain a more profound understanding of their impact on environment this study was conducted. In collaboration with SSNC (Naturskyddsföreningen), a database containing hundreds cosmetic products was processed. The most occurring polymers were quantified and prevalent ingredients having “poly” in their name were selected for further investigation namely Nylon 12-20 (microplast) and Acrylates C/10-30 Alkyl-crosspolymer (water soluble). A standardized analysis method OECD 301 F was performed to test the polymers biodegrading ability. Results from biodegradation method showed that, neither of the two polymers is readily biodegradable in aqueous environment, despite their different properties. In connection with the obtained results, a filtration analysis was performed, with the purpose to determine the possibility to capture the polymers using microfilters. Results mainly showed flowthrough of both polymers. Relating the results to reality implies that these polymers are not captured in waste water treatment plant due to inefficient filtration and thereby spread to the environment. In addition more research should be devoted to water-soluble polymers and their impact on nature. Based on all compiled results, it is proposed that legislation addressing microplastics should be edited and revised in such a way that water soluble polymers are included in future prohibitions (against microplastics).
Plast har många användningsområden varav kosmetiska produkter är ett av dem. I kosmetika används exempelvis naturliga ämnen, sådana som förekommer i naturen och således kan brytas ned. Sedan finns syntetiska polymerer, sådana som syntetiseras och tillverkas av människor. Det finns oerhört många varianter av syntetiska polymerer som används inom kosmetika, generellt kan de kategoriseras i två grupper; mikroplaster och vattenlösliga polymerer. Med hjälp av Naturskyddsföreningens databas som innehåller hundratals kosmetikaingredienser, söktes de komponenter som hade ”poly” i sitt namn eftersom de inte täcks av lagstiftning. Bland dessa, valdes två mest förekommande polymerer för att studeras vidare, nämligen Nylon 12-20 (mikroplast) och Acrylates C/10-30 alkyl crosspolymer (vattenlöslig). En standardiserad analysmetod OECD 301 F tillämpades för att testa deras biologiska nedbrytbarhet. Resultatet från nedbrytbarhetstestet visade att ingen av polymertyperna, trots deras åtskiljande egenskaper, är lättnedbrytbar i vattenmiljöer. I följd av resultatet från analysen genomfördes en filtreringsanalys på KTH, för att avgöra om respektive polymer går att fånga upp i olika storlekar av mikrofilter. Resultatet visade att ingen av dem fångades upp (med den utrustning som var tillgänglig på KTH). Baserat på resultaten föreslås det att lagstiftningar som innefattar mikroplaster bör redigeras och revideras på ett sådant sätt att vattenlösliga polymerer är inkluderade i framtida förbud (mot mikroplaster). Utöver detta bör mer forskning ägnas åt vattenlösliga polymerer och deras påverkan på naturen.

L’entreprise SEPROSYS est une société spécialisée dans le développement de solutions d'extraction et de séparation de molécules. Au cours de l’année 2011, l’entreprise a développé un procédé innovant de séparation et de purification fractionnée de biomolécules issues de macroalgues marines. Ce travail de thèse a pour objectif de trouver des applications aux fractions extraites selon le procédé d’extraction et de purification de la société SEPROSYS, et en particulier la fraction des polysaccharides sulfatés, dans deux domaines distincts : en pharmaceutique pour leur activité anti-coagulante et en cosmétique pour leur activité anti-âge. La première partie de nos travaux se penche sur le potentiel des polysaccharides sulfatés de macroalgues comme anti-coagulants. Pour cela, des extraits aqueux de différentes macroalgues brunes, rouges et vertes ont été préparés et leur activité anti-coagulante a été mesurée. Les algues présentant le meilleur potentiel ont ensuite été traitées au moyen du procédé SEPROSYS® afin de purifier leurs polysaccharides sulfatés et d’étudier leur activité anti-coagulante. De plus, l’objectif de ce travail consiste également à mieux comprendre la relation structure-fonction de l'activité anticoagulante d'ulvanes. A cette fin, des ulvanes issus d’Ulva sp. de pureté élevée ont été produits par le procédé SEPROSYS®, soumis à des modifications chimique (hypersulfatation) et physique (dépolymérisation) et leur activité anticoagulante a été étudiée. La deuxième partie de nos travaux concerne l’évaluation du potentiel de ces macroalgues comme agents actifs utilisables en dermo-cosmétique, en particulier pour leur activité anti-âge. Dans ce but, différentes fractions de macroalgues brunes, rouges et vertes ont été testées sur des lignées cellulaires de fibroblastes dermiques humains et leur effet sur la production de collagène par ces fibroblastes a été étudié. Enfin, le potentiel d’ulvanes extraits d’Uva sp. par le procédé de l’entreprise SEPROSYS pour stimuler la biosynthèse de collagène et d’acide hyaluronique par ces mêmes fibroblastes a été déterminé
SEPROSYS is a company specialized in the development of solutions for the extraction and separation of molecules. In 2011, the company has developed an innovative process of fractionated separation and purification of biomolecules from macroalgae. The purpose of this thesis work is to find applications for the purified fractions from the SEPROSYS® process and in particular the sulfated polysaccharides, for two distinct biological activities : as pharmaceutical for their anti-coagulant activity and as dermo-cosmetics for their anti-aging activity. The first part of our work focus on the potential of sulfated polysaccharides from macroalgae as anti-coagulant agents. To this end, aqueous extracts from brown, red and green macroalgae were prepared and their anti-coagulant activity was measured. The best macroalgae were then processed with the SEPROSYS procedure in order to purify the sulfated polysaccharides and study their activity. Furthermore, one of the objectives of this work was to acquire a better understanding of the structure/anti-coagulant function relationship of ulvans. Ulvans of high purity were extracted from Ulva sp. with the SEPROSYS® process, submitted to chemical (hypersulfatation) and physical (depolymerization) modifications and their anti-coagulant activity was measured. The second part of our work relates to the potential of the macroalgae as active agents to be used in dermo-cosmetics, and, in particular, for their anti-aging activity. Different brown, red and green macroalgae fractions were thus tested on normal human dermal fibroblasts cell lines and their effects on the collagen production were measured. Furthermore, the capacity of different molecular weight ulvans extracted from Ulva sp. to stimulate the collagen and hyaluronan biosynthesis was studied. To this end, we developed an efficient depolymerization procedure using ion exchange resins

Doutoramento em Bioquímica
Os polissacarídeos são os componentes maioritários dos grãos de café verde e torrado e da bebida de café. Os mais abundantes são as galactomananas, seguindo-se as arabinogalactanas. Durante o processo de torra, as galactomananas e arabinogalactanas sofrem modificações estruturais, as quais estão longe de estar completamente elucidadas devido à sua diversidade e à complexidade estrutural dos compostos formados. Durante o processo de torra, as galactomananas e arabinogalactanas reagem com proteínas, ácidos clorogénicos e sacarose, originando compostos castanhos de alto peso molecular contendo nitrogénio, designados de melanoidinas. As melanoidinas do café apresentam diversas atividades biológicas e efeitos benéficos para a saúde. No entanto, a sua estrutura exata e os mecanismos envolvidos na sua formação permanecem desconhecidos, bem como a relação estrutura-atividade biológica. A utilização de sistemas modelo e a análise por espectrometria de massa permitem obter uma visão global e, simultaneamente, detalhada das modificações estruturais nos polissacarídeos do café promovidas pela torra, contribuindo para a elucidação das estruturas e mecanismos de formação das melanoidinas. Com base nesta tese, oligossacarídeos estruturalmente relacionados com a cadeia principal das galactomananas, (β1→4)-Dmanotriose (Man3), e as cadeias laterais das arabinogalactanas, (α1→5)-Larabinotriose (Ara3), isoladamente ou em misturas com ácido 5-Ocafeoilquínico (5-CQA), o ácido clorogénico mais abundante nos grãos de café verde, e péptidos compostos por tirosina e leucina, usados como modelos das proteínas, foram sujeitos a tratamento térmico a seco, mimetizando o processo de torra. A oxidação induzida por radicais hidroxilo (HO•) foi também estudada, uma vez que estes radicais parecem estar envolvidos na modificação dos polissacarídeos durante a torra. A identificação das modificações estruturais induzidas por tratamento térmico e oxidativo dos compostos modelo foi feita por estratégias analíticas baseadas principalmente em espectrometria de massa, mas também em cromatografia líquida. A cromatografia de gás foi usada na análise de açúcares neutros e ligações glicosídicas. Para validar as conclusões obtidas com os compostos modelo, foram também analisadas amostras de polissacarídeos do café obtidas a partir de resíduo de café e café instantâneo. Os resultados obtidos a partir dos oligossacarídeos modelo quando submetidos a tratamento térmico (seco), assim como à oxidação induzida por HO• (em solução), indicam a ocorrência de despolimerização, o que está de acordo com estudos anteriores que reportam a despolimerização das galactomananas e arabinogalactanas do café durante a torra. Foram ainda identificados outros compostos resultantes da quebra do anel de açúcares formados durante o tratamento térmico e oxidativo da Ara3. Por outro lado, o tratamento térmico a seco dos oligossacarídeos modelo (individualmente ou quando misturados) promoveu a formação de oligossacarídeos com um maior grau de polimerização, e também polissacarídeos com novos tipos de ligações glicosídicas, evidenciando a ocorrência de polimerização através reações de transglicosilação não enzimática induzidas por tratamento térmico a seco. As reações de transglicosilação induzidas por tratamento térmico a seco podem ocorrer entre resíduos de açúcares provenientes da mesma origem, mas também de origens diferentes com formação de estruturas híbridas, contendo arabinose e manose como observado nos casos dos compostos modelo usados. Os resultados obtidos a partir de amostras do resíduo de café e de café instantâneo sugerem a presença de polissacarídeos híbridos nestas amostras de café processado, corroborando a ocorrência de transglicosilação durante o processo de torra. Além disso, o estudo de misturas contendo diferentes proporções de cada oligossacarídeo modelo, mimetizando regiões do grão de café com composição distinta em polissacarídeos, sujeitos a diferentes períodos de tratamento térmico, permitiu inferir que diferentes estruturas híbridas e não híbridas podem ser formadas a partir das arabinogalactanas e galactomananas, dependendo da sua distribuição nas paredes celulares do grão e das condições de torra. Estes resultados podem explicar a heterogeneidade de estruturas de melanoidinas formadas durante a torra do café. Os resultados obtidos a partir de misturas modelo contendo um oligossacarídeo (Ara3 ou Man3) e 5-CQA sujeitas a tratamento térmico a seco, assim como de amostras provenientes do resíduo de café, mostraram a formação de compostos híbridos compostos por moléculas de CQA ligadas covalentemente a um número variável de resíduos de açúcar. Além disso, os resultados obtidos a partir da mistura contendo Man3 e 5-CQA mostraram que o CQA atua como catalisador das reações de transglicosilação. Por outro lado, nas misturas modelo contendo um péptido, mesmo contendo também 5-CQA e sujeitas ao mesmo tratamento, observou-se uma diminuição na extensão das reações transglicosilação. Este resultado pode explicar a baixa extensão das reações de transglicosilação não enzimáticas durante a torra nas regiões do grão de café mais ricas em proteínas, apesar dos polissacarídeos serem os componentes maioritários dos grãos de café. A diminuição das reações de transglicosilação na presença de péptidos/proteínas pode dever-se ao facto de os resíduos de açúcares redutores reagirem preferencialmente com os grupos amina de péptidos/proteínas por reação de Maillard, diminuindo o número de resíduos de açúcares redutores disponíveis para as reações de transglicosilação. Além dos compostos já descritos, uma diversidade de outros compostos foram formados a partir dos sistemas modelo, nomeadamente derivados de desidratação formados durante o tratamento térmico a seco. Em conclusão, a tipificação das modificações estruturais promovidas pela torra nos polissacarídeos do café abre o caminho para a compreensão dos mecanismos de formação das melanoidinas e da relação estrutura-atividade destes compostos.
Polysaccharides are the major components of green and roasted coffee beans, and coffee brew. The most abundant ones are galactomannans, followed by arabinogalactans. During the roasting process, galactomannans and arabinogalactans undergo structural modifications that are far to be completely elucidated due to their diversity and complexity of the compounds formed. During the roasting process, galactomannans and arabinogalactans react with proteins, chlorogenic acids, and sucrose, originating high molecular weight brown compounds containing nitrogen, known as melanoidins. Several biological activities and beneficial health effects have been attributed to coffee melanoidins. However, their exact structures and the mechanisms involved in their formation remain unknown, as well as the structure-biological activity relationship. The use of model systems and mass spectrometry analysis allow to obtain an overall view and, simultaneously, detailed, of the structural modifications in coffee polysaccharides promoted by roasting, contributing to the elucidation of the structures and formation mechanisms of melanoidins. Based on this thesis, oligosaccharides structurally related to the backbone of galactomannans, (β1→4)-D-mannotriose, and the side chains of arabinogalactans, (α1→5)-Larabinotriose, alone or in mixtures with 5-O-caffeoylquinic acid, the most abundant chlorogenic acid in green coffee beans, and dipeptides composed by tyrosine and leucine, used as models of proteins, were submitted to dry thermal treatments, mimicking the coffee roasting process. The oxidation induced by hydroxyl radicals (HO•) was also studied, since these radicals seem to be involved in the modification of the polysaccharides during roasting. The identification of the structural modifications induced by thermal and oxidative treatment of the model compounds was performed mostly by mass spectrometry-based analytical strategies, but also using liquid chromatography. Gas chromatography was used in the analysis of neutral sugars and glycosidic linkages. To validate the conclusions achieved with the model compounds, coffee polysaccharide samples obtained from spent coffee grounds and instant coffee were also analysed. The results obtained from the model oligosaccharides when submitted to thermal treatment (dry) or oxidation induced by HO• (in solution) indicate the occurrence of depolymerization, which is in line with previous studies reporting the depolymerization of coffee galactomannans and arabinogalactans during roasting. Compounds resulting from sugar ring cleavage were also formed during thermal treatment and oxidative treatment of Ara3. On the other hand, the dry thermal treatment of the model oligosaccharides (alone or when mixed) promoted the formation of oligosaccharides with a higher degree of polymerization, and also polysaccharides with new type of glycosidic linkages, evidencing the occurrence of polymerization via non-enzymatic transglycosylation reactions induced by dry thermal treatment. The transglycosylation reactions induced by dry thermal treatment can occur between sugar residues from the same origin, but also of different origins, with formation of hybrid structures, containing arabinose and mannose in the case of the model compounds used. The results obtained from spent coffee grounds and instant coffee samples suggest the presence of hybrid polysaccharides in these processed coffee samples, corroborating the occurrence of transglycosylation during the roasting process. Furthermore, the study of mixtures containing different proportions of each model oligosaccharide, mimicking coffee bean regions with distinct polysaccharide composition, subjected to different periods of thermal treatment, allowed to infer that different hybrid and non-hybrid structures may be formed from arabinogalactans and galactomannans, depending on their distribution in the bean cell walls and on roasting conditions. These results may explain the heterogeneity of melanoidins structures formed during coffee roasting. The results obtained from model mixtures containing an oligosaccharide (Ara3 or Man3) and 5-CQA and subjected to dry thermal treatment, as well as samples derived from spent coffee grounds, showed the formation of hybrid compounds composed by CQA molecules covalently linked to a variable number of sugar residues. Moreover, the results obtained from the mixture containing Man3 and 5-CQA showed that CQA acts as catalyst of transglycosylation reactions. On the other hand, in the model mixtures containing a peptide, even if containing 5-CQA and subjected to the same treatment, it was observed a decrease in the extent of transglycosylation reactions. This outcome can explain the low extent of non-enzymatic transglycosylation reactions during roasting in coffee bean regions enriched in proteins, although polysaccharides are the major components of the coffee beans. The decrease of transglycosylation reactions in the presence of peptides/proteins can be related with the preferential reactivity of reducing residues with the amino groups of peptides/proteins by Maillard reaction, decreasing the number of reducing residues available to be directly involved in the transglycosylation reactions. In addition to the compounds already described, a diversity of other compounds were formed from model systems, namely dehydrated derivatives formed during dry thermal treatment. In conclusion, the identification of the structural modifications in coffee polysaccharides promoted by roasting pave the way to the understanding of the mechanisms of formation of melanoidins and structure-activity relationship of these compounds.

碩士
嘉南藥理科技大學
化妝品科技研究所
96
The effects of polysaccharides (BSP) isolated from tubers of Bletilla striata on 3T3 mouse fibroblasts were examined in this study. We found that 0.5 mg/ml of BSP could up-regulate collagen III gene expression, leading to enhanced de novo synthesis of collagen III. The results also showed that MMP-2 gene expression increased in fibroblasts treated with BSP, while MMP-1 and MMP-9 gene expression remained a relatively low level. However, the activities of intracellular and extracellular MMP-2 maintained a constitutive status compared to control groups. In addition, we demonstrated that TIMP-1 gene expression was augmented in the presence of BSP, but a low expression level kept in TIMP-2. The data taken together suggested that increased TIMP-1 might inhibit MMP-2 activity, then bringing about the accumulation of neo-synthesized collagen III. Furthermore, we investigated that gene expression of SOD-1, SOD-2, and SOD-3 in fibroblasts treated with BSP remained unchanged. On the other hand, we proved that 10% of BSP dissolved in H2O possessed excellent water retention properties confirmed by the moisturizing test. In conclusion, the experimental data presented here intensely suggested that BSP could be used as a potential anti-aging ingredient in skincare cosmetics.

碩士
靜宜大學
應用化學研究所
89
Cosmetics are applied to moisturize skin and hair, stimulate smell,deodorize, and enhance normal appearance. Since cosmetics are applied on human skin, they should not contain any ingredients that may cause adverse health effects. Chinese herbs are common ingredients in cosmetics. Thus, the evaluation of safety for the usage of Chinese herbs as the ingredients in the cosmetics formulation is necessary and plays an important role. Both the aqueous and the ethanol extracts of fifteen Chinese herbs will investigate for the evaluation of safety. These fifteen Chinese herbs are:Angelicae dahuricae, Poria cocos, Bletilla striata, Armeniaca mandshurica,Angelicae sinensis, Trichosanthes kirilowii, Panax ginseng, Santalum album,Anemarrhena asphodeloides, Polygonum multiflorum, Asparagus cochinchinensis,Ophiopogon japonicus, Lithospermum erythrorhizon, Scutellaria baicalensis, and Hedysarum mongholicum. The Draize test and its modifications were used to identify irritants in this study. The test subjects were rabbits and white rats. The rabbit skin test results showed that only the 50% alcohol solutions containing Angelica sinensis and Lithospermum erythrorhizon had mild irritation, while no reactions were found with other solutions. The eyes of the rabbits had no reaction to all fifteen herbs while performing the low-volume eye irritation test. The acute toxicity test showed that within dose of 10000 mg/kg, all fifteen herbs had no significant toxicity reaction.

MENVSC
Department of Hydrology and Water Resources
Africa is endowed with various earthy materials, of which each community is uniquely identified. Inspite of the vast knowledge of western cosmetics, application of traditionally used cosmetics have been in existence and will be into practice from generation to generation, this is as a result of their various indigenous knowledge about traditional cosmetics. They are mostly applied during traditional rites and festive periods. Occasionally, earthy materials play a vital role in traditionally used cosmetics. Different reasons have been associated with the use of these earthy materials, they are: for skin cleansing, protection against ultra violet radiation, skin lightening, perfecting the distorted part of the skin and improving beauty. This study is focused on the physico-chemical, chemical and mineralogical characterisation of earthy materials indigenously used as cosmetics. This is achieved firstly by identifying the various earthy materials applied topically as well as their constituents. Samples for this study were obtained from six (6) different African countries namely: Botswana, Cameroon, Democratic Republic of the Congo, Nigeria, South Africa and Swaziland and were subjected to the following analyses: Scanning Electron Microscope and Electron Dispersive X-ray Spectroscopy for the morphology and elemental analysis, hydrometer method for the determination of particle sizes of each sample and a texture Auto Lookup Software Package (TAL Version 4.2) for the classification of their various textures (%clay, %sand and %silt). The X-ray diffractometry was applied for the identification of their mineral phases (secondary, major and minor). The Brunauer Emmett and Teller (BET) method was used for the physisorption analysis and classifying them according to their isotherm types (Type I, II, III, IV, V and VI). The chemical analysis was achieved by X-ray fluorescence (XRF). The result revealed that the morphology of the various representative samples varies, except CMR 1; 2, SCC 1; 2 and SLW 1; 2 which showed same structure and same elemental constituents, their resemblance indicated that they possess same geological formation. The XRF result indicated the presence of the following trace elements: Al, As, Au, Ba, Bi, Br, Ca, Cd, Ce, Cl, Co, Cr, Cs, Dy, Er, Eu, Fe, Ga, Gd, Ge, Hf, Hg, Ho, I, In, K, La, Lr, Lu, Mg, Mn, Mo, Na, Nd, Ni, Nb, Os, P, Pb, Pd, Pr, Pt, Re, Rh, Ru, S, Sc, Se, Sb, Si, Sm, Sn, Sr, Ta, Te, Tb, Th, Ti, U, V, W, Y, Yb, Zr and Zn, were measured in ppm. For example, Fe – 66.47 (Qwa-Qwa), Al – 4.64 (Swaziland), Si – 6.26 (Democratic Republic of Congo), Ti – 3.75 (Qwa-Qwa) and so on. From the particle size distribution analysis, some samples were identified to be clay (BTS 1, BTS 2, NG 1, SCC 4, SCC 5, SCC 9, SCC 11, SCC 12, SWL 1 and SWL 2) with their particle sizes < 2μm, whereas other samples showed other properties which are > 2 μm. Ten minerals were identified in the clay fraction samples: albite, goethite, halloysite, hematite, kaolinite (kaolinite-1Md, kaolinite 1) Mica (muscovite), sulphur, talc, chalcanthite and whewellite. The BET method vii for physical adsorption revealed that BTS 1, BTS 2, CMR 1, CMR 2, DRC A, DRC B, NG 1, NG 2, SWL 1, SWL 2, SCC 1, SCC 4, SCC 5, SCC 8, SCC 9, SCC 11 and SCC 12 countries possess Type II isotherm whereas samples from SCC 2, SCC 6, SCC 7 and SCC 10 had Type III isotherm. The physisorption analysis result also determined the various surface areas and porosity of each representative samples.

碩士
嘉南藥理科技大學
化妝品科技研究所
98
According to the previous dowments, many Chinese herbs are with skin thera peutic benefits. In recent years, the cosmetic products with Chinese herbs active ingredients have been a kind of tendency. Because Asiaw attach importance to skin whitening, whitening cosmetic products have a lot of requirement in ASIA market. This experiment used many kinds of Chinese medicinal herb, including Momordica charantia L., Nelumbo nucifera G., Nymphaea lotus L., Angelica sinensis D., Salvia miltiorrhiza B. etc.. We evaluated the extraction by antioxidation test, inhibition tyrosinase activity assay, MTT assay and so on. Next step, we chose the strong activty of extraction to add in whitening cosmetics. We evaluated the whitening cosmetic by in vivo. On the anti-oxidation tests, ABTS free radical scavenging ability of Nymphaea lotus L. showed SC50 was 0.0025 mg/mL, its better than Trolox. DPPH free radical scavenging ability showed the SC50 of Nymphaea lotus L. was 0.0067 mg/mL, and the SC50 of Trolox was 0.0045 mg/mL. Inhibition tyrosinase activity assay, the IC50 of Nymphaea lotus L. and Nelumbo nucifera G. mixture was 0.11 mg/mL. And the results show that add Vit C and Morus alba L. appropriately, can increase inhibition tyrosinase acitivity of Nelumbo nucifera G., Namely, whitening compound herbal extracts for inhibition tyrosinase activity, are better than the original single-whitening herbal extracts inhibit the effect more good. Apply the Chinese medicinal herb with whitening activity on the emulsion formula to perform skin nature appraisal and the skin chromatic aberration appraisal. Finally it demonstrated that the whitening can improve whitening effect. The emulsion B (active constituent increase Nymphaea lotus L, Nelumbo nucifera G.), not only had whitening effect but also color spot, the pore, the primary purple nature, the ultraviolet ray color spot can keep improvement after overall skin inspection . In the future, the Chinese medicine herb will have well develop in whitening and Multiple - effects cosmetic products.

碩士
中華醫事科技大學
生物醫學研究所
100
Purpose of this study was to investigate chitosan in cosmetics in the antimicrobial activity, respectively, at different concentrations and different molecular weight and pH were measured inhibitory effect, filter out the best inhibitory concentration and pH, and the cell survival rate test, cytotoxicity test, heavy metal testing and microbiological testing and other tests. Experimental use of sample to chitosan is mainly because of its better solubility than chitin. Chitosan non-toxic and biocompatible with Biodegradable, environmentally friendly polymer materials. In this study, we used three different molecular weight chitosan, the chitosan (MW:100,000~200,000 datons) were prepared as 0.5% (pH 3.2 and pH 5.6)、1% (pH3.3 and pH5.7) and 1.5% (pH5.7) chitosan (MW:100,000~300,000 datons) concentration of the preparation of 0.5% (pH 5.7), 1% (pH 5.7) and 2% (pH3.4) chitosan (MW:200,000~400,000 datons) concentration of the preparation into 1% (pH 5.7), and lactic acid as the solvent, the degradation of chitosan molecular weight, to improve the dissolution properties of chitosan the antibacterial agents antibacterial test analysis, experimental data and found that chitosan (MW:100,000~200,000 datons) at a concentration of 0.5% and the pH value of 5.6, that for Escherichia coli, Staphylococcus aureus all have antibacterial effects, the chitosan and chitosan (MW:200,000~400,000 datons) the inhibitory effect of poor; the same time, the concentration of 0.5%(pH5.6), 1%(pH5.7) chitosan (MW:100,000~200,000 datons) and a concentration of 1% (pH 5.7) of the chitin polymer sugar and lactic acid (control group) and a common antibacterial agent of cosmetics for cell survival and cell toxicity test, experimental results showed that lactic acid (control group) and common cosmetics and antibacterial agent with a cytotoxic cell-free survival, and different concentrations of chitosan on cell survival (MTT) and do not have good cytotoxicity (LDH). The concentration of 1.5%, pH 5.7 chitosan (MW:100,000~200,000 datons) for heavy metals testing, test data results for the ND, namely, that chitosan does not contain heavy metals. In addition, the preparation of a concentration of 1%, pH 5.6 and 0.5%, pH 5.6 and 1.5%, pH 5.7 chitosan for microbiological testing, results showed that chitosan on Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and total plate count detection is ND, namely, that the study sample dose not contain the above strains (that is not detected).

碩士
中華醫事科技大學
生物醫學研究所
100
This research investigates herb extracts as a potential sun screen ingredient in cosmetics. As cited in this experiment twenty herbs were tested for their sun protection and whitening factor as well as their anti-oxidizing properties. Pine bark, scutellaria, and turmeric extract showed viable results in ultraviolet ray absorption, in particular pine bark extract showed the most significant viability in spectrum absorption spectroscopy (spectrophotometer or spectrometer). By testing of MTT assay kit (MTT) and Cytotoxic Assay (LDH) in different concentrations of the pine bark extract, it was found that the cells in the epidermis (HS-68) had the highest survival rate and lowest toxicity levels in 3% concentration pine bark extract. Adding the same concentration of the extract into emulsion to test out its sun protection factor consequently yielded a SPF 8.81. The experimentation resulted in a practical application of the extract in the cosmetic formula. Assessment of in vitro tests revealed a differentiation in outcome, the results do show the pine bark extract can be used as cosmetic sunscreen.

碩士
嘉南藥理科技大學
化妝品科技研究所
96
Taiwanofungus camphoratus is an medicinal mushroom in Taiwan. It is well known that the major effective components in this medical fungus are polysaccharides and triterpenoids. Recently we found that low molecular weight (less than 30,000) of exopolysaccharides from the mycelium culture of T. camphoratus could suppress the activities of matrix metalloproteinases (MMPs). Therefore, this study was designed to utilize endogenous enzymes to degrade high molecular weight of expolysaccharides from the submerged culture of T. camphoratus into low molecular weight of polysaccharides. The optimal condition of polysaccharide-degrading enzymes for the production of low molecular weight of polysaccharides with the anti-aging properties will be explored. 　　By using endo-β-1,3-glucanases (40 units/ml) at 40℃ for 6 h, high molecular weight of exopolysaccharides was transformed into low molecular weight, and less than 31,542 Da of polysaccharides was obtained. These low molecular weight of polysaccharides were applied to investigate the capacities for inhibiting MMPs secreted from 3T3 fibroblasts by gelatin-based zymography. The results showed that at the concentration of 500 ?慊/ml, low molecular weight of polysaccharides did inhibit the activities of pro-MMP-2, MMP-2, and MMP-9. 　　The experiment for the production of exopolysaccharides from T. camphoratus was further scale-up to 5-L fermentor. The biomass and exopolysaccharides content could be increased to 27.8 g/L and 6.9 g/L, respectively. The different submerged culture of exopolysaccharides were also applied to utilize endogenous enzymes to decompose into low molecular weight of polysaccharides, and the inhibitory ability on the activities of MMP-2 and MMP-9 was evaluated. The results presented here show that low molecular weight of polysaccharides exhibit the markedly repressive ability on the activities of pro-MMP-2, MMP-2, and MMP-9. 　　Consequently, we suggest that the low molecular weight of polysaccharides derived from exopolysaccharides of T. camphoratus mycelial culture degraded with glucanases exhibit the antiaging capacities and can be an effective ingredient used in skincare products.

碩士
嘉南藥理科技大學
化妝品科技研究所
98
Previous studies reported that the matrix metalloproteinases (matrix metalloproteinases, MMPs) play an important role both in addition to the maintenance of normal physiological function and the pathogenesis of many diseases. Because MMPs can be broken down collagen, elastin and other extracellular matrix, they may be involved in the process of skin aging. In this study, we selected the alcohol extracts from thirty-four kinds of Chinese herbal medicines, and analyzed MMPs activity secerted from mouse 3T3 fibroblasts by gelatin-based zymography. We found that the alcohol extracts of Panax ginseng、Angelica sinensis、Spilanthes acmella、Cucurbita moschata could inhibit the activity of MMP-2 significantly, and the alcohol extracts of Aquilaria malaccensis、Androraphis paniculata decreased the activity of MMP-9 significantly. Because MMP-2 is able to damage collagens Ⅰ and Ⅲ, and MMP-9 can destory collagen IV, we consider that the alcohol extract of P. ginseng, A. sinensis, S. acmella, C. moschata, A. malaccensis, A. paniculata have intensive anti-aging effects and they can be applied in anti-aging skin care products. On the other hand, we found that alcohol extracts of 14 kinds of Chinese herbal medicines (Arctium lappa, Aquilaria malaccensis, Nelumbo nucifera, Andrographis paniculata, Aenthum graveolens、Nelumbo nucifera, Lonicera japonica, Agrimonia pilosa, Tribulus ierrestris, Crataegus pinnatifida, Ginkgo biloba, Lilium lancifolium, Diospyros kali, Hedyotis diffusa) were tested by MTT assay and the results showed the phenomenon of cell proliferation. These Chinese herbal medicine extracts can be used in cosmetics for cell activation.

碩士
嘉南藥理科技大學
化妝品科技研究所
100
In this study, ultraviolet - visible absorption spectroscopy and high-performance liquid chromatography with ultraviolet detection were used to investigate the permeability of methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, phenoxyethanol, methyl isothiazolinone, dimethylol-dimethylhydantoinc, imidazolidinyl urea and two commerical composite preservatives through thin film of plastic packaging materials, such as polyethylene (PE), linear low density polyethylene (LLDPE) or non-oriented casting polypropylene (CPP) In ultraviolet absorption spectroscopy, the absorption behavior of preservatives in the wavelength range of UV radiation(200-400nm) were first measured and the detection limit and quantification limit were calculated from the concentration and absorbance relationship at maximum absorption wavelength of respected preservatives. In high performance liquid chromatography, the influence of components and pH of the mobile phase on the chromatogram were studied to determine the optimum chromatographic condition. The detection limit and quantification limit of high performance liquid chromatographic method were also calculated. The above methods with the use of Franz cell were applied to study the permeability of preservatives through thin plastic film of packaging materials for cosmetics, The results showed that, ultraviolet absorption spectrometry shows higher sensitivity than high performance liquid chromatographic method, but it is not possible to resolve the absorption of individual component in a mixture. However, ultraviolet absorption spectrometry is still a better method to study the interation of a single preservative and thin plastic film. In the results of preservatives permeability through PE film from ultraviolet absorption spectrometry, methyl isothiazolinone shows much higher permeation rate than other preservatives and Imidazolidinyl Urea shows the least. However, phenoxyethanol shows the highest permeation rate in the study of the prentration behavior of mixed preservatives and analyzed by high performance liquid chromatography and the permeration rate of p-hydroxybenzoate esters were inverse proporational to the molecular size of the preservatives. Therefore, whether the permeability of preservative is changed after mixed is worth furth study. Compared the permeability of a single preservative through other plastic films, methyl isobutyl thiazolinone shows highest permeability through the CPP film and most of all test preservatives shows lower permeability through LLDPE film. Therefore, it is suggested that LLDPE film has better barrier effect for preservatives. It is concluded that this study provides informations on the permeability of preservative through plastic film, These informations could be an important reference when choosing the appropriate packaging material for cosmetics. But the penetration behavior of mixed preservatived is worth furth study.

碩士
國立臺灣海洋大學
食品科學系
94
The purpose of this study is to explore the antioxidativity and rheological properties of crude polysaccharide aqueous solution of Isochrysis sp. CCMP 1324 and its application in vital moisture cream. First, using boiled water extracted crude polysaccharide of Isochrysis sp. CCMP 1324. And then the crude extracted solution were monitored its polysaccharide content, antioxidativity and rheological properties. The stability, safety and effectivity of cosmetics made from crude polysaccharide were also monitored. From the results, the reducing power, scavenging effect of DPPH radical, chelating Fe2+ effect and trolox equivalent antioxidant capacity (TEAC) of the crude polysaccharide solutions were found enhancing with the concentration increasing. After treating with pepsin or trypsin, the crude polysaccharide aqueous solution was declining tendency of antioxidativity ability. The rheological behavior of crude polysaccharide aqueous solution is pseudoplastic fluids. Increasing the solute concentration enhanced its shear thinning properties. The apparent viscosity of crude polysaccharide aqueous solution decreased and increased with increasing NaCl concentration and pH, respectively. The stability of vital moisture cream containing with or without Isochrysis sp. CCMP 1324 crude polysaccharides was stable for more a month. Based on the Daize score test, there were no erythema found in user. As increasing the Isochrysis sp. CCMP 1324 crude polysaccharides concentration enhanced the apparent viscosities and electrical capacitance decrease ratio increased.

碩士
嘉南藥理科技大學
化妝品科技研究所
96
The ultimate purpose of this research is to develop a on-line electrochemical analysis system used for in-situ transdermal absorption studied for cosmetics. The manufacturing and performance analysis of electrode which is the key component in electrochemical analysis are the main scope in this study. Compared to platinum electrode, the cost of materials used for fabricating carbon electrode is much lower. Therefore, this study is focused on the manufacturing process and performance analysis of carbon electrode. The effect of conductive carbon content on the activity of electrode was studied. Electrochemical Impedance Spectroscopy(EIS) was used to characterize the electrode. Cyclic Voltammetry(CV) was used to analysis the response of the electrode to K3[Fe(CN)6] and vitamin C and its derivatives which are the common active ingredient of in cosmetics. The effect of supporting electrolyte’s concentration on response current was also studied. The polarization resistance and capacity of the carbon electrode obtained from electrochemical impedance analysis show good reproducibility. The response of self-made carbon electrode to K3[Fe(CN)6] shows well redox behavior and the responses to ascorbic acid, magnesium ascorbyl phosphate and ascorbyl glucoside show competent linear relation to the concentration of the analyte in the range of 5.0x10-2~1.0x10-4 M with correlation coefficient higher than 0.9988. Furthermore, the response of carbon electrode to ascorbic acid has the highest sensitivity than it to ascorbyl glucoside and the response to magnesium ascorbyl phosphate got the lowest sensitivity among these three vitamin C related substances. In summary, the self-made carbon electrode in this study shows good stability and competent reproducible response to analyte. The applicable concentration range for active ingredient in cosmetics can be established by analyzing the linear relation and sensitivity of the response to the analyte. It is promising that there are great potential for the utilization of this electrode in the developing of on-line electrochemical in-situ transdermal absorption analysis system. The test and research result found the carbon electrode of manufacture, make use of the Electrochemical Impedance Spectroscopy method to get characteristic parameters, such as...etc, representability the carbon electrode of manufacture have fairly good and stability. And analysis result of Cyclic Voltammetry method have rather consistent and stability at different contained respondence of redox.As for analyze the often used active ingredient - vitamin C and its derivatives, we found concentrations of within the scope of, its respond of value of line relationship factor,all reaches 0.9988 above, simultaneously at vitamin C, vitamin C phosphoric acid magnesium and the vitamin C glucose between the vitamin C have the highest sensitivity,vitamin C glucose secondly, the vitamin C phosphoric acid magnesium lowest. Summary above result, this research of the carbon electrode have an equal, moreover analysis respondence of redox with fairly good reproducibility and stability,to utilize the electrode analyze sensitivity of samples and line relation of samples respond that we can build up a scope of detected with electrode application in the active ingredient,therefore applies this electrode establishment electrochemical the on-line and in-situ absorbed system through the skin that should have a good development potential.

碩士
中原大學
生物醫學工程研究所
104
Artificial antibacterial agents (such as Methyl Peraben(M.P.) and Propyl Paraben(P.P.)) were important factor in causing skin allergies and skin irritation. Therefore, searching for alternatives and decreases of the artificial antibacterial agents have become a major issue in cosmetics industry. The interactions between antibiotic medicines and plant components have already been investigated by domestic and foreign scholars. Nonetheless, there isn’t relevant reports that relative to interactions between artificial agents in cosmetics and the extracts of Chinese herbal. This study evaluates the in-vitro interactions (synergism, addition effect, antagonism ) between methanol extracts of Pogostemon cablin, Almond acid and two kinds of common ingredients used in cosmetics, against Escherichia coli and Propionibacterium acne. The results of this study were expected that as a reference of type and that dose for adding antibacterial agents in cosmetics and Chinese herbals in order to reduce damage to human skin. In this study, we extracted Pogostemon cablin with methanol, then practiced respectively the alone and one-on-one combined Antibacterial Susceptibility Testing (AST) of four kinds of reagents (Pogostemon cablin, almond acid, M.P. and P.P.) of vary concentrations (0.1 %, 1.0 %, 2.0 % and so on) with the Disk-diffusion method, and at last assessed the interactions between these four reagents with the Combination Index (CI). The results of this study showed that Pogostemon cablin possessed good synergism or addition effects when it was combined respectively with Almond acid, M.P. and P.P.. This means that the formula amounts of M.P. and P.P. can be reduced and substituted by the increased using of Pogostemon cablin extracts and Almond acid in cosmetics, and the overall antibacterial ability of cosmetics could still be able to maintain. In addition to assess for anti-pimples of cosmetics in dose study that Almond acid which is commonly used could be replaced by adding more ingredients of Pogostemon cablin, and the best combination formula is 1.0 % Pogostemon cablin to 0.1 % Almond acid. As for the components of combined artificial antibacterial agents, M.P. and P.P., this study indicated that there is obvious antibacterial synergism while combining these two artificial antibacterial agents. Thus, even without the addition of Chinese herbal extracts; the original amount of M.P. and P.P. can be moderately reduced.

碩士
美和技術學院
健康與生技產業研究所
98
Photoaging is known to be correlated with different kinds of oxidative stress. The extent of protein oxidation could be used as a biomarker for oxidative stress. An in vivo method for evaluating the anti-oxidative properties of compounds based on a simple immunoanalysis protocol to detect the extent of protein oxidation of stratum corneum by measuring the level of exposed carbonyl group on the proteins. The application of this method was used for evaluating the properties of various anti-oxidants in cosmetic formulations. Eight commonly used anti-oxidants in cosmetic formulation and one combination of anti-oxidants have been evaluated. The results indicated that pure vitamin E and vitamin C showed the best anti-oxidative capabilities. They could reduce the level of exposed carbonyl group by 90 %. Vitamin E-acetate and lipoic acid fall behind with 50~60% reduction, and ferulic acid with 22% reduction. Vitamin C-palmitate and AA2G showed poor performance. Glutathione increased the level of carbonyl group by 38 %. In the study of combination of anti-oxidants, vitamin E-acetate and lipoic acid showed synergistic effect. Our Study also indicated that vitamin C-palmitate and AA2G showed significant capability for scavenging DPPH; however, they did not perform well for reducing the level of exposed carbonyl group in vivo. Vitamin E-acetate and Lipoic acid showed no DPPH scavenging capabilities. On the other hand, they showed capability for reducing the level of exposed carbonyl group. It is very likely that vitamin E-acetate could be converted to vitamin E in stratum corneum based on a time course study of vitamin E-acetate.