Dissertations / Theses on the topic 'Antioxidant and antimutagenic activity'

Presented diploma work is focused on study of antioxidants in different kinds of dried fruits. Analyses of ascorbate, tocopherols, carotenoids and flavonoids were performed using RP-HPLC with spectrophotometric detection. Analysis of dried fruits showed high level of vitamins and phenolocs mainly in berries. High level of carotenoids was observed in dried apricots and plums. Further, antioxidant activity of dried fruit extracts was tested by ABTS method. High antioxidant activity was found mainly in dried apples, cranberries and blueberries. The biological test with yeast Sascharomyces cerevisiae D7 was used for the analysis of antimutagenic efects of dried fruits. High antimutagenic activity exhibited dried cranberries and blueberries. Most of tested dried fruits with high antimutagenic effect exhibited also high antioxidant activity as well as high content of some antioxidants. No direct correlation was found among these parameters. Last part of this work was focused on study of influence of drying on antioxidant content in two types of apples. Drying at mild controlled conditions exhibited no significant negative effect on active substance content; in some samples their concentration was observed.

This diploma project deals with antioxidant and antimutagenic properties of selected herbal and fruit teas commonly used in Czech population. Influence of different tea packaging (bag teas and loose leaf teas) on bioactive compound content was compared. Further, effect of long-term storage in common household conditions was studied. Antioxidant properties of teas were characterized using some group parameters - total antioxidant activity ("Randox Total Antioxidant Status Kit"), total phenolics and total flavonoids - as well as some individual representatives of low molecular weight antioxidants. Higher antixidant content was found in herbal teas than in fruit teas. Comparing bag teas with loose leaf teas higher antioxidant activity was shown in loose leaf teas. Individual antioxidants were analyzed using HPLC method with spectrophotometric detection and verified by on-line LC/MS. IN all tea samples catechins - catechin, epicatechin, epicatechin gallate and other flavonoids - rutin, morin, quercetin, kaempferol, myricetin and luteolin were determined. In most of teas high level of catechin and rutin was detected. The highest level of flavonoids was determined in herbal poured teas. Ascorbic acid content was also determined by HPLC method. Higher vitamine C level was found in most of fruit teas and in rose hip tea. Antimutagenicity of tea extracts was tested by in vitro test using Saccharomyces cerevisiae D7 yeast. High antimutagenic activity showed mainly nettle tea, tutsan tea and most of fruit teas. During long-term storage (1 year, 20°C, darkness) a significant decrease of all analyzed antioxidant parameters was followed. Higher lost of antioxidants was found in fruit teas when compared with the herbal ones.

Qualitative and quantitative analysis of anthocyanins and other phenolic compounds from a purple corn extract was performed. The purple corn extract had cyanidin-3-glucoside, pelargonidin-3-glucoside, peonidin-3-glucoside and its respective acylated anthocyanin-glucosides. Cyadinin-3glucoside was the main constituent (44.4 ?? 4.7%) followed by the acylated cyanidin-3-glucoside (26.9 ?? 8.0%). Other phenolic compounds present in the purple corn corresponded to protocatechuic acid, vanillic acid, and p-coumaric acid. In addition, quercetin derivatives, a hesperitin derivative and pcoumaric and ferulic acid derivatives were found. Fractionation of phenolic compounds yielded two main fractions, an anthocyanin-rich water fraction (WF) and an ethyl acetate fraction (EAF). Evaluation of antimutagenic activity in both fractions revealed higher antimutagenic activity in the ethyl acetate fraction compared to the anthocyanin-rich fraction. On the other hand, antioxidant activity of the anthocyanin-rich fraction was higher compared to the ethyl acetate fraction. Further fractionation of the anthocyanin-rich fraction in a Toyopearl HW40 gel permeation column yielded five sub-fractions which showed no difference in antimutagenic activity except for the water sub-fraction WF-V. All the sub-fractions were active as antimutagens and antioxidants. Further fractionation of the ethyl acetate fraction yielded four sub-fractions that showed to be active as antimutagens and antioxidants. Ethyl acetate sub-fraction EAF-IV was the most active as an antimutagen. HPLC-DAD characterization of that sub-fraction revealed mainly the presence of a quercetin derivative with UV-visible spectral characteristics similar to rutin but with a little longer retention time. The mechanism of antimutagenic action by the phenolic compounds present either in the anthocyanin-rich fraction or the ethyl acetate fraction and sub-fraction EAFIV seems to be a contribution of a direct action on the enzymes involved in the activation of the mutagen and to the scavenging activity of the mutagen nucleophiles, as demonstrated by our assays.

The presented doctoral thesis is focused on the study of the biological effects of active compounds found in cereals, on the development of methods of analysis these effects, on the determination of the content of active substances and characterization of the relationship between composition and biological effects of cereals and cereal products. For the analysis several kinds of raw cereal samples (flakes, flour, germ, bran), flavored extruded cereal products and also samples of paddy and husked rice were chosen. To major types of analysed active compounds belong mainly phenolic compounds in the form of glycosides and aglycones, and also saccharides. Group parameters such as total polyphenols, flavonoids and total and reducing saccharides were determined spectrophotometrically, individual phenolics and saccharides were determined by high performance liquid chromatography (HPLC). TEAC, DPPH and CLAMS methods were used to determine the antioxidant activity. Indirect methods of determination of substances with antioxidant effect were used as well. Results of the total antioxidant activity were compared with values of antimutagenic/genotoxic activity obtained by several microbial test systems. Antimutagenic effect was expressed as a percentage of inhibition of effect of standard mutagen and could be considered as a potential preventive effect of cereals to DNA arising primarily by free radicals effect. The highest values of group and individual phenolics, antioxidant and antimutagenic activity were found in germs, bran, in buckwheat products and in coloured and raw rice. In flavoured cereal products addition of chocolate or fruit positively influences content of active phenolic substances as well as sugars, antixidant and antimutagenic activity. In a representative sample of Czech population, questionnairy study was performed to monitor interest in cereals and consumer preferences. The most of consumers consider cereals with chocolate flavour as less healthy than confirmed results of laboratory analyses. In this study some new food products were developed. Several types of model cereal products containing plant (fruit, vegetables) extracts were proposed. Extracts were added to cereals in freeze-dried and encapsulated form. The highest positive effect exhibited addition of local forrest fruit extract. Within preparation of encapsulated extracts several methods of preparing lipid or saccharides particles were also tested. The encapsulation efficiency of the methods and stability and size of particles were analysed. Optimal type of fortified cereal foods could be suggested based on the acquired results. Selected cereals were used as alternative carbon substrates (processed or raw) for the cultivation of microorganisms to produce enriched biomass usable in the feed industry. We can conclude that cereals in raw as well as processed form belong to universal foods and rich sources of biologically active substances. They can be processed by many ways. They can be used for direct consumption, as a part of new products and also undirectly as a substrate for feedstock.

Espécies de Psychotria encontradas no sul do Brasil produzem alcalóides do tipo monoterpeno indólicos, alguns deles com interessantes atividades biológicas e oriundos de novas rotas biossintéticas. P. leiocarpa Cham. & Schlecht. acumula N, b-D-glicopiranosilvincosamida (GPV), o primeiro alcalóide N-glicosilado desta classe a ser descrito. O extrato contendo GPV apresenta atividade analgésica inespecífica e, na planta, sua biossíntese é regulada pelo desenvolvimento e por luz. P. umbellata Vell., por sua vez, produz psicolatina, que apresenta alto potencial farmacológico, pois apresenta atividade analgésica do tipo opióide, ansiolítica e antipsicótica, interagindo com receptores de diversos sistemas de neurotransmissores no sistema nervoso central. Além disso, psicolatina é um eficiente agente redutor de peróxidos e quencher de oxigênio singlet in vitro. Os objetivos do presente trabalho foram estudar a fotorregulação de GPV em plântulas de P. leiocarpa, assim como avaliar os efeitos antioxidantes e antimutagênicos in vivo do extrato foliar bruto de P. umbellata e de psicolatina purificada, utilizando a levedura Saccharomyces cerevisiae. Essas duas últimas substâncias também foram avaliadas quanto à capacidade antioxidante contra o radical hidroxila in vitro. Em ensaios de transição luz-escuro realizados com plântulas assépticas de P. leiocarpa, o acúmulo de GPV mostrou ser responsivo a alterações na condição luminosa de cultivo. O papel negativo do escuro contínuo na biossíntese de GPV foi comprovado pela redução dos níveis deste alcalóide em plântulas cultivadas na luz e transferidas para o escuro. Por outro lado, quando plântulas cultivadas no escuro foram expostas à luz, os níveis de GPVaumentaram, indicando o caráter promotor da luz na produção de GPV. Os efeitos das transições foram mais evidentes em plântulas cultivadas em meio sem sacarose do que em plântulas cultivadas com suprimento exógeno de carboidratos. A biossíntese de GPV é regulada por diferentes faixas de luz. As regiões do azul e do vermelho-extremo aumentaram os teores de GPV. A luz vermelha não afetou de forma significativa o teor de GPV. Os resultados revelam um padrão típico de VLFRs (Very Low Fluence Responses), possivelmente envolvendo ação de PhyA em conjunto com criptocromo.Tanto o extrato bruto foliar de P. umbellata quanto psicolatina apresentaram efeito antioxidante in vivo, reduzindo a inibição do crescimento de Saccharomyces cerevisiae sob estresse oxidativo induzido por peróxido de hidrogênio e paraquat. O extrato e o alcalóide purificado também apresentaram ótima atividade antioxidante in vitro, protegendo contra o ataque do radical hidroxila. Os índices de mutagênese induzida por peróxido de hidrogêncio foram significativamente reduzidos quando as células de S. cerevisiae foram co-cultivadas na presença tanto do extrato quanto de psicolatina.
Species of Psychotria founded in southern Brazil produce a set of novel monoterpene indole alkaloids (MIAs), several of which have interesting biological activities and originate from new metabolic pathways. P. leiocarpa Cham. & Schlecht. accumulates N, b-D-glucopyranosylvincosamide (GPV), the first N-glycosylated MIA described. Leaf extracts containing GPV display nonspecific analgesic activity and, in planta, its biosynthesis is regulated by development and light. P. umbellata Vell., in turn, produces psychollatine which has significant pharmacological potential, since it yields opioid-like analgesic, anxiolytic and antipsychotic activities, interacting with receptors of different neurotransmitter systems in the central nervous system. In addition, psychollatine is an efficient peroxide reducing agent and a singlet oxygen chemical quencher in vitro. This work aimed at studying the photoregulation of GPV in P. leiocarpa seedlings, as well as at investigating the antimutagenic and antioxidant in vivo effects of the crude foliar extract of P. umbellata and purified psychollatine using the yeast Saccharomyces cerevisiae. These last substances were also evaluated for their in vitro antioxidant properties against hydroxyl radicals.In light-dark transition assays with aseptic P. leiocarpa seedlings, GPV accumulation showed to be responsive to changes in light condition. The negative role of continuous dark on GPV biosynthesis was shown by reduction of the alkaloid contents when light growing seedlings were transferred to dark. On the other hand, dark growing seedlings increased GPV contents after light exposure, suggesting a positive light regulation of GPV production. Theseresults were more evident in seedlings cultivated in media without sucrose than in seedlings cultivated with carbohydrate supplementation. GPV biosynthesys is also regulated by different light qualities. Light in the blue and far-red regions increased GPV accumulation, whereas red ligh had no significant influence on GPV yield. These results are in agreement with the profile of VLFRs (Very Low Fluence Responses), mediated by PhyA with coaction of cryptochrome. Both the crude foliar extract of P. umbellata and psychollatine showed in vivo antioxidant effects by reducing the growth inhibition of Saccharomyces cerevisiae under hydrogen peroxide- and paraquat-induced oxidative stress. The extract and the purified alkaloid also showed strong in vitro antioxidant activity against hydroxyl radicals. The levels of hydrogen peroxide-induced mutagenicity were significantly reduced when S. cerevisiae cells were cocultivated with leaf crude extract or psychollatine.

Ispitivanja hemijskog sastava etarskih ulja iekstrakata izvedena su na vrsti Myrtus communisL. sa pet lokaliteta iz Crne Gore. Pored toga,ispitana je njihova antioksidantna aktivnost urazlicitim in vitro sistemima kako bi se utvrdiouticaj pomenutih ekstrakata i etarskih ulja naneutralizaciju DPPH, NO, OH i 2- radikala, kaoi njihov uticaj na lipidnu peroksidaciju ulipozomima i inhibiciju enzima ksantin-oksidaze.Takode, ispitana je i antibakterijska aktivnostetarskih ulja i ekstrakata ove vrste na 9bakterijskih sojeva, kao i njihov antimutagenipotencijal na bakterijskom soju Escherichia coliIC 202.
In this tessis the chemical analysis of the essential oils and methanolic extracts from five plant samples of Myrtus communis L., collected from different localities in Montenegro, have been investigate. Beside that, their antioxidant activity in differwnt in vitro systems has been study to establish their scavenging potential towards DPPH, NO, OH, and O2- free radicals, as wel as their effects on lipid peroxidation in liposoma and inhibition enzyme XOD. Also, the antibacterial activity of the essential oils and methanolic extract has been study on 9 bacterial strains, as wel as their antimutagenic effects on bacterial strain E. Colli IC202.

U ovoj doktorskoj disertaciji ispitan je hemijski sastav i biološke aktivnostiekstrakata deset  samoniklih  taksona roda  Allium  sect.  Codonoprasum:  A. carinatum subsp. pulchellum,  A. carinatum  subsp. carinatum,  A. fuscum  var. gracile,  A. fuscum  var. fuscum,  A. flavum subsp. flavum,  A. melanantherum,  A. paniculatum  subsp.  marginatum, A. pallens  subsp. tenuiflorum,  A. oleraceum  i  A. rhodopeum, sakupljenih na  27 lokaliteta u Srbiji. Cilj rada bio je da se dobiju podaci o sadržaju biološki aktivnih jedinjenja u ovim, do sada veoma malo ispitanim vrstama roda  Allium, i utvrdi njihova potencijalna lekovita vrednost.Analiza hemijskog sastava obuhvatila je: analizu volatilnih komponenti svežih lukovica primenom headspace GC-MS tehnike, kvalitativnu analizu metanolnih  ekstrakata primenom tečnohromatografskih metoda (LC-DAD-MS i LC-MS-MS),  kvantitativnu analizu odabranih fenolnih jedinjenja LC-MS-MS tehnikom,  određivanje sadržaja ukupnih  monomernih  antocijana  i određivanje aktivnosti aliinaze. Ispitivanja bioloških aktivnosti ekstrakata obuhvatila su: određivanje antioksidantne, antiinflamatorne, antimikrobne, antimutagene i genotoksične aktivnosti, kao i ispitivanje uticaja na rast zdravih i tumorskih ćelija i sposobnosti indukcije ćelijske smrti. Sumiranjem dobijenih rezultata može se zaključiti da ispitivani predstavnici roda Allium  sect.  Codonoprasum  predstavljaju bogate izvore biološki aktivnih jedinjenja sa širokim spektrom bioloških aktivnosti. Sa hemotaksonomskog aspekta značajno je da se dimetil-disulfid  izdvaja kao  najdominantnija  i često jedina  isparljiva komponenta, da ekstrakti većine vrsta  sadrže veliku količinu flavonoida (prvenstveno derivata kvercetina), da se vrste  A. pallens  i  A. oleraceum  izdvajaju od ostalih po tome što ne sadrže rutin  a sadrže  hiperozid,  da je vrsta  A. rhodopeum  siromašna fenolnim jedinjenjima i da su sve vrste, osim vrste  A.  flavum, bogate antocijanima. Aktivnost aliinaze je visoka u svim ispitivanim vrstama. Većina ispitivanih ekstrakata, izuzev ekstrakata vrsta  A. carinatum i A. melanantherum, pokazala je izraženu antioksidantnu aktivnost, dok su ekstrakti vrsta A. flavum, A. rhodopeum, A. oleraceum i A. paniculatum snažni antiinflamatorni agensi. Ekstrakti ispitivanih predstavnika sect.  Codonoprasum nisu pokazali antimikrobnu i antimutagenu aktivnost. Takođe, ovi ekstrakti nisu ispoljili genotoksični efekat na ćelije zdravog tkiva (izuzev slabog genotoksičnog efekta ekstrakta nadzemnih delova  A. flavum), što ukazuje na bezbednost upotrebe vrsta sect. Codonoprasum  kao hrane ili u obliku lekova. Ekstrakti celih biljaka A. paniculatum i A. rhodopeum, kao i ekstrakt nadzemnih delova  A. melanantherum  pokazali su snažnu antiproliferativnu aktivnost sa povoljnim ne-tumor/tumor koeficijentima i indukovali apoptozu u tumorskim ćelijama, iz čega se može zaključiti da imaju visok potencijal primene u antitumorskoj terapiji. 
In the present doctoral thesis the chemical composition and biological activities  of 10 wild growing taxa of genus  Allium  sect. Codonoprasum  (A. carinatum  subsp. pulchellum,  A. carinatum  subsp.  carinatum,  A. fuscum  var.  gracile,  A. fuscum  var. fuscum,  A. flavum  subsp.  flavum,  A. melanantherum,  A. oleraceum,  A. paniculatumsubsp. marginatum, A. pallens subsp. tenuiflorum and A. rhodopeum) were investigated. The samples were  collected from 27 locations in Serbia. The aim of the study was to obtain data on the content of biologically active compounds in extracts of  these unexplored species of the genus Allium and to determine their potential medicinal value.Phytochemical caracterisation included: headspace GC-MS analysis of fresh bulb volatiles, LC-DAD-MS and LC-MS-MS qualitative analysis of methanol extracts, LC-MS-MS quantitative analysis of 44 selected phenolic compounds in methanol extracts, determination of total monomeric anthocyanins content and alliinase activity. In order to assess the biological potential of methanol extracts, the antioxidant, anti-inflammatory, antimicrobial, antimutagenic, genotoxic and antiproliferative activities of  the extracts were studied.Summing up all the results obtained, it can be concluded that species of genus Allium  sect.  Codonoprasum  are rich sources of biologically active compounds with a broad spectrum of biological activities. Dimethyl disulfide is the most dominant and often the only volatile component of most species, which is significant from chemotaxonomic point of view. The methanol extracts of investigated species contain high levels of flavonoids (primarily quercetin derivatives). Specificity of  A. pallens  and A.  oleraceum  extracts is that they do not contain rutin, but contain hyperoside, while small quantity of phenolic compounds  is characteristic for  A. rhodopeum  extract. All investigated species, except of A. flavum, are rich in anthocyanins. Alliinase activity was high in  all examined species. Most of the extracts, except  A. carinatum  and  A. melanantherum  extracts, express considerable antioxidant activity,  while extracts of  A. flavum,  A. rhodopeum,  A. paniculatum  and  A. oleraceum  are potent anti-inflammatory agents. The investigated  Allium  extracts did not show antimicrobial and antimutagenic activity. Also, the extracts did not express genotoxic effect on healthy tissue cells (except the weak genotoxic effects of aerial parts extract of  A. flavum), indicating that the use  of these species as a food or as a drug is safe. Whole plant extracts of  A. paniculatum  and  A. rhodopeum, as well as aerial parts extract of  A. melanantherum showed strong antiproliferative activity (with a favorable  non-tumor/tumor ratios) and induced apoptosis in tumor cells, suggesting that these plants have a high potential for application in antitumor therapy.

Several seven-coordinate manganese complexes have been synthesised, characterised and tested for both superoxide dismutase and catalase activity. Macrocyclic ring contractions have led to a series of new seven coordinate mononuclear manganese(II) macrocycles that have potential for their use as working superoxide dismutase mimics. Numerous polynuclear seven coordinate manganese(II) macrocycles have been synthesised via Schiff base condensation. Subsequent reduction of the imine bonds has led to a variety of reduced amine analogues with varying axial ligands. The geometry has been compared about the manganese centres where possible. From the results of each complex tested for superoxide dismutase activity, the µ-chloro bridged tetranuclear complex [Mn2(C24H29N6O2)(Cl)2]2(ClO4)2 has proved to be the most efficient mimic with a calculated KMcCF value of 7.7 x 106 [M-1 s-1]. A method for measuring catalase activity has been developed, and the most efficient catalase active compound was found to be [Mn5(C24H29N6O2)2(OAc)2(ClO4)2](ClO4)2 with one molecule of complex breaking down approximately 59000 molecules of hydrogen peroxide after one minute. Catalase testing showed that a reduction of the imine bonds produced an increase in activity overall for the complexes of H2L1 (C24H29N6O2), but a decrease was observed for the reduced tripodal complexes. An increase in the number of manganese centres resulted in a rise in catalase activity. Many of the complexes tested for catalase activity showed an induction period prior to the activity being observed. This may suggest that the complexes undergo a change in structure, or that there is a rearrangement occurring before catalase activity may be observed. The results that are presented indicate that the axial ligands have an effect on the rate of catalase activity and the observed induction period. Of the molecules that were tested for both superoxide dismutase and catalase activity, the pentanuclear complex [Mn5(C24H29N6O2)2(OAc)2(ClO4)2](ClO4)2 showed high activity for both analyses. This may be due to the extra manganese centre within the complex and the axial ligands that are present when compared with other tetranuclear complexes. The complex [Mn5(HL1)(OAc)2(ClO4)2](ClO4)2 may prove to be a good candidate for a working superoxide dismutase mimic. Ring contracted complexes show high rates of superoxide dismutase activity but possess limited catalase activity. Attempts have been made to produce a direct method of measuring superoxide dismutase activity using a stop-flow technique to complement the results using the indirect NBT (Nitro blue Tetrazoleum) method. This was carried out by analysing low concentration solutions of both complex and superoxide on a millisecond timescale. Progress has been made for this method with preliminary results being obtained.

Mestrado em Biologia Aplicada - Biologia Molecular Celular
It is generally accepted that the principal roles of metallothioneins (MTs) lie in the detoxification of toxic metals and regulation of the metabolism of essential trace metals. However, there is increasing evidence that it can act as a free radical scavenger. Although the great number of studies on the antioxidant activity of MTs, the effective physiological role of this protein is still unclear. In order to understand the role of MTs in the protection against metal contamination and oxidative stress, the bivalve Cerastoderma edule was used to evaluate the response of MTs in both situations. Cadmium, a widely reported MT inducer, was used to simulate metal contamination whereas H2O2, an oxidizing compound, was used to simulate oxidative stress. In the first approach, cockles were exposed to a range of Cd and H2O2 concentrations and MTs and TBARS were quantified. Results showed that both treatments induced MT synthesis, confirming the involvement of MTs in metal contamination and oxidative stress. Indeed, the use of MTs as biomarkers for metal pollution was questioned due to the similar synthesis of MT in the two highest concentration used. At last, one concentration of Cd (10 μM) and of H2O2 (20 μM) were selected and cockles were exposed again. TBARS concentration and the intracellular amount of H2O2 were determined. Metal-MT complexes in the two conditions and control were isolated by size exclusion chromatography, and the binding of Zn and Cd to MTs and other cytosolic proteins was evaluated. Furthermore, MTs were quantified and their content in each treatment and control was compared to the amount of Zn associated to them. Results showed that the >H2O2 treatment induced high levels of oxidative stress, demonstrated by the high lipid peroxidation and intracellular concentration of H2O2. Data also indicated that Cd was mainly associated with MTs pool in the Cd treatment, confirming that the protective role of MTs in metal contamination in this bivalve species was due to the binding of MTs to Cd ions. Additionally, the percentage of Zn bound to MTs decreased in the H2O2 treatment, indicating Zn release in oxidative stress. Also, MTs molecules were not as metalated as in the control, confirming Zn release from MTs in oxidative stress and indicating that MTs were needed for demands other than Zn distribution. Further studies on the redox status of MTs are needed to determine the redox status of MTs in the oxidative stress, and understand if, in this bivalve, MTs are acting as ROS scavengers.
Os papéis geralmente associados às metalotioninas (MTs) resumem-se á desintoxicação de metais tóxicos e à regulação do metabolismo dos metais essenciais. No entanto, existem evidências cada vez mais acentuadas de que as MTs atuam na proteção contra o stresse oxidativo. Apesar do grande número de estudos que se focam na actividade antioxidante das MTs, o papel fisiológico efetivo destas proteínas não foi ainda clarificado. A fim de compreender o papel das MTs no stress oxidativo e na proteção contra o efeito dos metais, o bivalve Cerastoderma edule foi selecionado neste estudo para avaliar a resposta das MTs em ambas as situações. O cádmio, um forte indutor das MTs foi usado para causar contaminação metálica enquanto o H2O2, sendo um composto oxidante, foi usado para provocar stresse oxidativo. Numa primeira abordagem, os berbigões foram expostos a uma gama de concentrações de Cd e H2O2 e as MTs e os TBARS foram quantificados. Os resultados mostraram que apenas o H2O2 provocou peroxidação lipídica no berbigão e que ambos os tratamentos induziram a síntese de MTs, confirmando o envolvimento destas na contaminação metálica e no stresse oxidativo. De facto, a utilização das MTs como biomarcadores de poluição metálica foi neste estudo questionada devido á síntese de quantidades semelhantes de MTs nas duas concentrações mais elevadas de Cd e H2O2. Numa segunda abordagem, os berbigões foram novamente expostos a uma concentração selecionada para cada tratamento (10 μM de Cd e 20 μM de H2O2). A concentração dos TBARS e a quantidade intracelular de H2O2 foram determinados. Os complexos metal-MT em ambas as condições e no controlo foram isolados por cromatografia de exclusão molecular e a ligação entre os iões de Zn e Cd e as MTs e outras proteínas citosólicas foi avaliada. Para além disso, As MTs foram quantificadas em cada tratamento e no controlo, sendo o seu conteúdo comparado com a quantidade de Zn ligado. Os dados indicaram que o tratamento com H2O2 induziu elevados níveis de stresse oxidativo, demonstrado pela elevada peroxidação lipídica e pela grande concentração intracelular de H2O2. Relativamente aos resultados da cromatografia, os iões de Cd estavam principalmente ligados às MTs no tratamento com Cd, confirmando o efeito protector das MTs na contaminação metálica nesta espécie de bivalve. Adicionalmente, a percentagem de Zn ligado às MTs diminuiu no tratamento com H2O2, indicando que o stresse oxidativo impõe a libertação de Zn por parte das MTs. Em jeito de confirmação, as MTs estavam menos metaladas no tratamento com H2O2 do que no controlo. Seriam necessários estudos complementares para perceber se neste bivalve as MTs actuam como eliminadoras de ROS.

Reactive Oxygen Species (ROS) produced during normal aerobic metabolism, if not promptly removed by the detoxification mechanisms of the cells, can easily react with cell components, especially lipids, producing secondary cytotoxic molecules called reactive carbonyl species (RCSs). RCSs exhibit significant chemical reactivity and can cause protein modification and dysfunction. One of the most important ReSs is 4-hydroxinonenal (HNE), an unsaturated aldehyde that has been strongly linked to Alzheimer'S disease (AD). A new series of dipeptide histidyl hydrazide analogues of carnosine was prepared, with the aim of producing molecules with enhanced HNE scavenging activity. These compounds were demonstrated to scavenge HNE and to protect SH-SY5Y cells from HNE-induced tOxlcity and were superior in action to carnosine. The synthesis of an analogue of the best compound of the series containing caffeic acid, resulted in the generation of a new molecule exhibiting complete retention of HNE-scavenging activity and also possessing free Radical Scavenging Activity (RSA). Mitochondria are the major sites of high levels of oxidative stress and the targeting of a reactive carbonyl scavenger directly to these organelles would result in extinguishing the primary source of RCS, thus arresting any consequent cellular damage. In the present work, the possibility of specifying the cellular localization of histidyl hydrazide was investigated using previously described mitochondria penetrating peptides (MPPs) and new examples of such sequences I modified to be more resistant to protease degradation. The ligation of histidyl hydrazide to a previously reported MPP was successful in targeting the former to the mitochondria of HeLa cells. Finally. a particular chemical ligation approach was investigated for the development of a system of linking a common peptide vector and a variety of cargo molecules of choice through facile in situ coupling, without requiring the de novo synthesis of a new chemical conjugate in each instance.

Certain components of flaxseed are known to exhibit antioxidant activity. The objective of this research was to study the antioxidant activity of flaxseed proteins and protein hydrolysates. Proteins and their hydrolysates were prepared from defatted, non-defatted, and demucilaged flaxseed with and without dialysis, and the antioxidant activities were examined by DPPH scavenging assay, reducing power assay, and metal ion chelating assay. The degree of hydrolysis (DH) of the proteins using bacterial protease was higher than using trypsin. Using bacterial protease, higher DH was observed for demucilaged and defatted flaxseed protein/dialysis (DDFPD, 30% DH), defatted flaxseed protein/dialysis (DFPD, 28% DH), and non-defatted flaxseed proteins/dialysis with 14% DH, compared to flaxseed protein/non-dialysis which were DDFPND 28% DH, DFPND 25% DH, and NDFPND with 12% DH. Proteins from dialysis treatment as well as their hydrolysates showed positive effect on antioxidant activity. DDFPD hydrolysates using bacterial protease showed higher DPPH radical scavenging activity (73.23%) and reducing power activity (0.15) at the concentration of 2.5 mg/ml as well as Fe2+ chelating ability (75%) at of concentration 1 mg/ml. SDS-PAGE and native-PAGE results of non-hydrolyzed samples showed no change in electrophoretic behavior as a result of the treatments; a major band corresponding to MW 48 KDa and three minor bands with MW of 16, 23, and 34 KDa. SDS-PAGE of DDFPNDH and NDFPNDH hydrolysates obtained using trypsin showed one resistant band.
Certaines composantes des graines de lin sont reconnues pour avoir des effets antioxydants. L'objectif du projet de recherche ci-présent est d'étudier les effets antioxydants des protéines des graines de lin et de leurs hydrolysâtes. Les protéines et leurs hydrolysâtes ont été préparé à partir de graines de lin dégraissées ou natures et démucilaginées, avec et sans dialyse. Leurs effets antioxydants ont été examinés avec le test de scannage DPPH, le test du pouvoir de réduction, et le test des ions métalliques chélates. Le degré d'hydrolyse (DH) des protéines était plus élevé avec l'utilisation d'une protéase bactériale qu'avec l'utilisation de trypsine. En utilisant la protéase bactériale, un DH plus élevé a été observé avec les graines démucilaginées et dégraissées avec dialyse/protéine (DDFPD, 30% DH), les graines dégraissées avec dialyse/protéine (DFPD, 28% DH), et les graines nature avec dialyse/protéines (14% DH), comparé aux graines protéine/sans dialyse qui ont eu comme résultat DDFPND 28%, DFPND 25% DH, et NDFPND avec 12% DH. Les protéines et leurs hydrolysâtes du traitement avec dialyse ont montré qu'il y avait bel et bien un effet antioxydant. Les hydrolysâtes, du test DDFPD et de l'utilisation du protéase bactériale, ont montré un niveau d'activité de radicaux libres plus élevé (DPPH 73.23%) ainsi qu'un pouvoir de réduction de (0.15) à une concentration de 2.5 mg/ml, de même qu'une habilité au Fe2+ de 75% à la concentration de 1mg/ml. Les résultats de SDS-PAGE et PAGE-original des échantillons non hydrolysés montrent qu'il n'y a aucun changement dans le comportement électrophorétique résultant du traitement: une bande majeure qui correspond au MW 48 KDa et trois bandes mineures avec un MW de 16, 23, et 34 KDa. Dans les échantillons d'hydrolysâtes SDS-PAGE du DDFPNDH et les hydrolysâtes du NDFPNDH obtenus en utilisant la trypsine ont montré une bande résistante.

Mn-salophen complex with superoxide-scavenging activity was prepared from manganese(III) acetate dihydrate and salophen in ethanol. Visible absorption spectrum of the red-brown solution exhibited a broad absorption band at 430 - 450 nm with two shoulders between 500 and 600 nm which were absent with either salophen or manganic acetate alone. Titration of salophen with manganese(III) was consistent with a 1:1 Mn to salophen stoichiometry of the complex based on changes in the absorbance at 500 nm or of superoxide scavenging activity. The SOD-like activity of the complex in the xanthine-xanthine oxidase/cytochrome c assay was 1450 units/mg salophen. The SOD activity of the complex was suppressed 50% in the presence of EDTA (1 mM), but was not altered in the presence of bovine serum albumin (1 mg/ml) or crude protein extract of E. coli QC779 sodA sodB (1 mg/ml). E. coli QC779 sodA sodB grew scantily after an 8 hour lag phase in aerobic M63 glucose minimal medium.
Ph. D.

The objective of this research was to study the antioxidant activity of soybean protein hydrolysates (SPH) and chickpea protein hydrolysates (CPH) at different concentrations, and to measure the antioxidant activity of fractions collected from the RP-HPLC analysis of SPH and CPH. Protein hydrolysates were prepared by the proteolytic enzyme trypsin. The hydrolysates obtained were subjected to DPPH (1, 1-diphenyl-2 picrylhydrazyl) radical scavenging assay. The SPH and CPH at concentration of 2.5-10 mg/ml showed antioxidant activity of 16.5-32 % and 3.4-26.8 %. SPH and CPH were fractionated by using RP-HPLC on C18 column. The antioxidant activity of four SPH and CPH fractions (F I, F II, F III, and F IV) was measured by using DPPH radical scavenging assay. For SPH, antioxidant activity of F III (47.7 %) was higher than other fractions at protein concentration of 1 mg/mL and for CPH; F II showed maximum antioxidant activity 27.9 % at protein concentration 1 mg/mL. The results from the SDS-PAGE confirmed the hydrolysis of protein samples. The second part of the study was to measure the impact of high pressure processing (HPP) on the degree of hydrolysis and antioxidant activity of proteins. High pressure processing (HPP) of isolated soybean protein (ISP) and isolated chickpea protein (ICP) was done at 400 MPa and 600 MPa for 5 min and 10 min. The degree of hydrolysis of isolated soybean protein and isolated chickpea protein treated with high pressure processing and with trypsin hydrolysis showed continuous increase from 12.4 to 24.9 % for SPH and 13.6 to 26.2 % for CPH. The DPPH radical scavenging assay showed a more than two fold increase in antioxidant activity of SPH and CPH: 67 % as compared to the 32 % of SPH without HPP and 56.6 % as compared to the 26.8 % of CPH without HPP at concentration 10 mg/mL. These results show that HPP increased the degree of hydrolysis and antioxidant activity of protein hydrolysates.
Le but principal de cette recherche constituait l'analyse du potentiel antioxydant, à diverses concentrations, d'hydrolysats de protéine de soya (HPS) et d'hydrolysats de protéine de pois chiche (HPP). Les hydrolysats de protéine ont été isolés à l'aide de l'enzyme protéolytique trypsine. Les HPS et HPP démontraient respectivement un potentiel antioxydant de 16.5 à 32% et 3.4 à 26.8 % lorsque présents à des concentrations de 2.5 à 10 mg/mL. L'utilisation d'une colonne C18 a permis de séparer, par CLHP-PI, les HPS et HPP en quatre fractions (F I, F II, F III, et F IV) qui furent dosées avec du DPPH (1,1-diphényl-2-picrylhydrazyle) afin de comparer leur pouvoir de scavenging sur les radicaux. Pour les HPS, le potentiel antioxydant de F III (47.7 %) était supérieur à celui des autres échantillons alors que pour les HPP, 27.9 % (F II) était le seuil maximal. Dans les deux cas, les hydrolysats étaient concentrés à 1mg/mL. L'hydrolyse des échantillons de protéine a été confirmée par SDS-page. La deuxième partie de l'étude visait à mesurer l'impact de la pascalisation sur le degré d'hydrolyse et le potentiel antioxydant des protéines. Des isolats de protéine de soya (IPS) et de protéine de pois chiche (IPP) ont été traités à haute pression (400 MPa et 600 MPa) pendant 5 et 10 min. Le degré d'hydrolyse des IPS et IPP soumis à la pascalisation et à la trypsin ont démontré une augmentation constante allant de 12.4 à 24.9 % pour les isolats de protéine de soya et de 13.6 à 26.2 % pour les isolats de protéine de pois chiche. L'analyse au DPPH du pouvoir d'épuration des radicaux a montré que le potentiel antioxydant des hydrolysats a plus que doublé, passant de 32 à 67 % pour les HPS et de 26.8 à 56.6 % pour les HPP, lorsqu'ils étaient traités par hautes pressions. Cela démontre que la pascalisation améliore le degré d'hydrolyse et le potentiel antioxydant des hydrolysats de protéines.

The methanolic crude extract of aerial parts of the plant Scrophularia nodosa was shown to have potent DPPH radical scavenging activity (IC50 = 48.75 ± 7.00 μg/ml). Activity-guided fractionation resulted in the isolation of three principal antioxidant compounds; acteoside, angoroside C and angoroside A. Acteoside (yield = 1.21%, IC50 = 15.2 μM) appeared to be the most abundant and most antioxidant-active. The potent antioxidant activity is in support of the traditional use of the plant for wound healing and anti-inflammatory conditions. The methanolic extract of aerial parts of Tanacetum vulgare has potent DPPH radical scavenging activity (IC50 = 37.00 ± 1.20 μg/ml). Activity-guided fractionation on the methanolic extract of T. vulgare resulted in the isolation of 3,5-di-caffeoylquinic acid (3,5-DCQA), axillarin and luteolin. 3,5-DCQA appeared to be the most abundant and most antioxidant-active compound (yield = 7.28%, IC50 = 9.7 μM). The potent antioxidant activity is in support of the traditional use of the herb for fever, rheumatic conditions and anti-inflammatory conditions. The methanolic crude extract of Cassia auriculata and its fractions were shown to have potent scavenging activity on DPPH, hydroxyl and hydroperoxide radicals, moderate superoxide radical scavenging activity and potent ion(III) reducing power. The activity-directed studies resulted in the isolation of kaempferol-3-0-β-D-rutinoside, kaempferol, luteolin, quercetin and unknown antioxidant inactive compound. The previously reported pharmacological aspects of the above flavonoids and flavonoid glycosides (anti-carcinogenic, anti-inflammatory, hyperglycaemic, antidiabetic) along with the shown antioxidant behaviour explain the traditional medicinal values of the plant. Cassia alata L crude extract and its fractions showed potent radical scavenging activity against formation of lipid peroxide radicals. The activity directed isolations resulted in the isolation of kaempferol along with p-hydroxybenzoic acid. These may contribute towards the traditional medicinal values of the plant as an antidiabetic, anti-microbial and for skin diseases.

The objectives of the studies presented in this thesis were to identify and quantify the major phenolic components of red wine and to assess the contribution of individual compounds to the total antioxidant activity. Red wines were analysed for their phenolic content and antioxidant activity using a range of complementary techniques including HPLC-tandem mass spectrometry, preparative HPLC and HPLC with an on-line antioxidant detection system. HPLC-MS2 revealed the presence of a number of flavoids and phenolic compounds of which 19 were identified, with gallic acid, the flavin-3-ols and anthocyanins being the most abundant. Preparative HPLC was used in an effort to isolate the antioxidant components in red wine and 60 aliquots were collected. Each wine fraction was analysed for total phenolics, catechins and anthocyanins, while antioxidant activity was determined by electron spin resonance spectroscopy (ESR). The preparative HPLC step did into completely separate the compounds in red wine, nonetheless increasing antioxidant activity was highly and significantly associated with total phenolics (r = 0.816, P < 0.001) and total catechins (r = 0.188, p = 0.151). HPLC with an on-line antioxidant detection system was subsequently used to separate and identify red wine phenolics. The findings from this study indicate that gallic acid, (+)-catechin, (-)-epicatechin, and procyanidin dimersB1 and B2 were the major in vitro antioxidants identified in red wine. Collectively, the flavin-3-ols contributed > 50% of the total antioxidant capacity of each wine, while gallic acid contributed between 24-44%. The flavonols and anthocyanins were minor antioxidant components in red wines. By combining HPLC, MS2 and on-line assessment of antioxidant activity, the major phenolic compounds present in red wine were identified, together with their direct contribution to the total antioxidant activity.

Thesis (M.S. in food science)--Washington State University, December 2008.
Title from PDF title page (viewed on Dec. 31, 2008). "School of Food Science and Human Nutrition." Includes bibliographical references.

Peptides derived from food proteins after enzymatic treatment and/or processing, are known to be bioactive in both biological and food systems; for this reason fish muscle peptides were investigated for their antihypertensive and antioxidant activity. Atlantic mackerel muscle proteins were hydrolysed with pepsin and pancreatine and the resultant hydrolysate was sequentially fractionated on 2 kDa membrane ultrafilters and further by gel filtration, ion exchange and high performance liquid chromatography and the resultant peptide fraction contained the amino acids histidine, proline, tyrosine, methionine, leucine, tryptophan and lysine. For ACE inhibitory activity, the peptide fraction (MFPH-V-JPA) had an inhibitory concentration (IC50) of 0.15 mg/ml and showed competitive inhibition for ACE with an inhibition constant (Ki) of 0.32 mg/ml. In terms of antioxidant activity, the HPLC isolated peptide fraction (LC1-Z) contained the amino acids serine, histidine, tyrosine, phenylalanine, tryptophan and lysine. It inhibited the formation of both peroxides and malonaldehyde in a linoleic acid model emulsion in a dose dependent manner with lipid oxidation inhibitory concentration (IC50) of 1.80 mg/ml. At concentration of 8 mg/ml, the inhibition of linoleic acid oxidation was more than that of 0.01 % butylated hydroxytoluene (BHT) and trolox (p < 0.001). The mechanism of antioxidant activity of the peptide (LC1-Z) was by carbon centered radical scavenging (5.34 %), hydroxyly radical scavenging (IC50 value of 1.60 mg/ml), metal chelating (5.72 %) and reducing ability. In caco-2 cells, 1 mg/ml of the peptide (LC1-Z) was not toxic to the cells seeded at 2 x 104 cells/well. Proxidant tBHP (2.5 mM) reduced cell viability significantly (79.3 %) but this increased to 94.7 % in the presence of the peptide or trolox. The peptide (1 mg/ml) also reduced TBARS formation (33.18 mug/ml) in cells compared to cells treated with tBHP alone (38.18 mug/ml). The activity of caspases-3 and -7, was higher in caco-2 cells treated with tBHP only (157.5 +/- 7.99 %) compared with those treated with the peptide (25.7 + 3.92 %). Morphological modification of the caco-2 cells treated with tBHP was evident as the cells appeared detached from the flask surface compared to those treated with the peptide (LC1-Z) which were healthy and attached to the flask surface. In Ea.hy 926 cells, reactive oxygen species were reduced by 26 % and 39 % in the lucigenin-chemiluminscence and flourescence methods respectively in cells treated with the peptide. In a caco-2 cell monolayer, transepithelial transport of the peptide was observed in both directions with a basolateral to apical apparent permeability of 0.95 +/- 0.12 cm-1 and apical to basolateral flux of 0.74 + 0.20 cm-1. HPLC chromatograms of the buffer solution taken from the apical and basolateral side showed the presence of the peptide in both sides i.e. 11.5% for apical to basolateral flux and 12.2 % for basolateral to apical. These results demonstrate that peptides with antihypertensive and antioxidant activity can be derived from Atlantic mackerel muscle proteins, with potential for neutraceutical applications.

碩士
國立中興大學
食品科學系
89
ABSTRACT The rhizome of the plant Curcuma zedoaria, a topical herb of the Zingiberaceae family native to southern Asia. The Chinese traditional medicine Curcuma zedoaria ROSC. has been clinically used for the treatment of cervical cancer. Many of its components have reputed medicinal properties that have been demonstrated that the extracts and fractions from the dried rhizome of C. zedoaria used in traditional medicine possess anti-inflammatory, antifungal, and antitumor activities. However, the study of its antioxidant activity and antioxidant compounds had not been reported. The objectives of this research were to study the antioxidative activities of extracts from C. zedoaria ROSC simultaneous steam distillation-solvent extraction, different solvent extraction, supercritical fluid extraction and water extraction. This research was also investigate antioxidiative properties of methanolic extracts, water extract, essential oil and supercritical fluid extracts from the rhizome of C. zedoaria, such as reducing power, scavenging effect on 1,1-diphenyl-2-picryl hydrazyl (DPPH●) radical, hydroxyl radical and chelating effect on ferrous ions. C. zedoaria ROSC contained hight carbohydrate contents. The yield of essential oil with simultaneous steam distillation-solvent extraction was very low，but it showed 70-87% inhibition on peroxidation of linoleic acid, and the antioxidative activities of essential oil were better than ascorbic acid. The 36 components of essential oil had been identified by GC-MS, including 17 teperenes, 14 alcohols and 5 ketones. The extracts of supercritical fluid extraction showed the worse antioxidative activities than essential oil, and there were less intense of odor than essential oil. The 31 components of supercritical fluid extracts had been identified by GC-MS, including 15 teperenes, 11 alcohols and 5 ketones. The content of terpene and sesquiterpene might be the main factor to influence antioxidative activities between them. The essential oil was further subjected to silica gel column chromatography eluted with various ratios of solvent′s composites from pentane and ether. The III and IV fractions showed markedly antioxidative activities in 5 separated fractions obtained from silica gel column chromatography. Methanolic and water extracts of C. zedoaria showed 80-94% inhibition on peroxidation of linoleic acid, and the antioxidative activities of them were better than ascorbic acid. Them exhibited the good effect not only in antioxidative activities, but also in other antioxidative properties, such as reducing power, scavenging effect on 1,1-diphenyl-2-picryl hydrazyl (DPPH●) radical, hydroxyl radical and chelating effect on ferrous ions. The antioxidative characteristics of methanolic extracts from C. zedoaria were investigated. The methanolic fractions was further subjected to silica gel column chromatography eluted with various ratios of solvent′s composites from hexane, ethyl acetate and methanol. The VI and VIII fractions showed markedly antioxidative activities in 8 separated fractions obtained from silica gel column chromatography. The antimutageneic characteristics of methanolic extracts from C. zedoaria were investigated. The methanolic and water extracts from C. zedoaria showed markedly inhibition on the mutagenicity of IQ、B[a]P and 4-NQNO to S. typhimurium TA97、TA98、TA100 and TA102. Keyword: Curcuma zedoaria, essential oil, antioxidant properties, antimutagenic properties

碩士
國立嘉義大學
食品科學系碩士班
92
This research used Bupleurum kaio Liu (Chao et Chuang), Gynostemma Pentaphyllum Makino, Codonopsis pilosula Nannf., Panax ginseng C. A. Mey., Lycium chinense Mill. and Momordica grosvenori Swingle to be studied as the compounded herbal tea. The proximate compositions of each herb, antioxidant properties and antioxidant components of water extracts from each herb and the compounded herbal tea were analyzed. The antimutagenic properties and antioxidant properties towards Chang liver cells of the water extracts from each herb and the compounded herbal tea were evaluated. Antioxidant properties of the six Chinese herbs were studied; the water extracts from Bupleurum kaio Liu (Chao et Chuang) and Lycium chinense Mill. showed high antioxidant activity. Free radical scavenging activity of the water extracts from Bupleurum kaio Liu (Chao et Chuang) and Lycium chinense Mill. reached above 87% at 0.75 and 1.25 mg/ml, respectively. Total antioxidant activity of the water extracts from Bupleurum kaio Liu (Chao et Chuang) and Lycium chinense Mill. were 73.9 % and 87.1 % at 0.75 and 1.25 mg/ml, respectively. The elevation in free radical scavenging activity, total antioxidant activity and reducing power of the water extracts from the six Chinese herbs, which were related to the total phenolic content, increased with the increasing of the concentration of water extracts. The ability of scavenging 1,1-diphenyl-2- picrylhydrazyl radical, total antioxidant activity and reducing power of the water extracts from the compounded herbal tea which contained six different Chinese herbs are 91.9%, 95.7% and 1.73 at 3 mg/ml, respectively. Antioxidant activity of the water extracts from compounded herbal tea increased with increasing concentration. Total phenolic compound of six different herbs ranged from 4.57 to 107.10 mg/g, where the highest were found in Bupleurum kaio (107.10 mg/g) and Lycium chinense Mill. (51.15 mg/g). Total phenolic compound of compounded herbal tea contained 37.95 mg/g. The flavonoids content of six different herbs varied from 0.28 to 42.09 mg/g. Once again, Bupleurum kaio contained the highest amount of flavonoids (42.09 mg/g). In addition, the flavonoids content of compounded herbal tea contained 37.95 mg/g. The water extracts from six different herbs and compounded herbal tea which contained the six different Chinese herbs have been tested for their antimutagenic properties against direct-acting mutagen of NQNO (without S9 mix) and indirect-acting mutagen of B[a]P (add S9 mix), using the Salmonella typhimurium tester strains TA98 and TA100. All the tested extracts of six different herbs and compounded herbal tea had no toxicity and mutagenicity. The compounded herbal tea showed best antimutagenicity effect against NQNO and B[a]P toword TA98 and TA100. No toxicity was found in the water extracts of six different herbs and compounded herbal tea towards Chang liver cell. Of the six Chinese herbs, Bupleurum kaio Liu (Chao et Chuang) possessed the highest inhibitory effect of peroxide induced Chang liver cell membrane oxidation. The inhibitory effect of compounded herbal tea was the positive dose-dependent. Thus, the water extracts of compounded herbal tea have protective effect against oxidative damage in Chang liver cells. Based on our data, the six Chinese herbs and compounded herbal tea not only showed negative signs of toxicity or mutagenicity but also possessed the ability to scavenge free radicals and inhibited lipid oxidation in Chang liver cells.

碩士
國立中興大學
食品科學系
92
This research used the rhizome of Curcuma aromatica to study its proximate composition, and antioxidant properties and antimutagenic activity of its ethanolic, hot-water extracts, and to evaluate the antioxidant properties and components of essential oils extracts from the rhizome of C. aromatica, C. longa L and C. sichuanensis. In the proximate composition, C. aromatica contained high carbohydrate contents (65.67%). Using the conjugated diene method, the ethanolic and hot-water extracts from the rhizome of C. aromatica showed high antioxidant activities (97.10% and 81.41%) at 1 mg/ml, respectively. At 20 mg/ml, reducing powers were in the order of ethanolic extracts (0.95) > hot-water extracts (0.59). Ethanolic extracts from the rhizome of C. aromatica showed an excellent scavenging ability on 1,1-diphenyl-2- picrylhydrazyl radicals (100.84%) at 5 mg/ml, higher than that of hot-water extracts (68.20%). The scavenging ability of hot-water extracts from the rhizome of C. aromatica on hydroxyl free radicals was the highest (64.72%) at 20 mg/ml. At 5 mg/ml, chelating abilities of the ethanolic and hot-water extracts on ferrous ions were 29.47 and 99.40%, respectively. The hot-water extract showed better chelating effect than ethanolic extract. Total phenols were the major naturally occurring antioxidant components found in ethanolic and hot-water extracts. The ethanolic and hot-water extracts from the rhizome of C. aromatica had been tested for their antimutagenic properties against direct-acting mutagen of NQNO (without S9 mix) and indirect-acting mutagen of B[a]P (add S9 mix), using the Samonella typhimurium strains TA98 and TA100. Six different doses (0.05, 0.1, 0.5, 1.0, 2.0 and 5.0 mg/plate) were used. In hot-water extract, all the tested extracts showed no toxicity and mutagenicity. But, ethanolic extract at 1mg/plate showed toxicity toward TA98. At 2mg/plate, ethanolic extract showed toxicity toward TA100. The ethanolic extracts from the rhizome of C. aromatica showed best antimutagenicity effect. The essential oils of C. aromatica, C. longa and C. sichuanensis obtained by simultaneous steam distillation-solvent extraction, were investigated by GC and GC-MS. The yields of essential oils of C. longa, C. aromatica and C. sichuanensis were 9.85, 7.11 and 3.21 mg/g, respectively. The 23 components of essential oils of C. longa and C. aromatica had been identified by GC-MS, including 10 teperenes, 6 alcohols, 2 ketones and 2 esters. The 21 components of essential oils of C. sichuanensis were identified, but lack of 4-terpineol and caryophyllene. The major constituents of essential oils of C. aromatica, C. longa and C. sichuanensis were curcumol, 1,8-cineole, cis--elemenone, humulene oxide and -cadinene. The higher percentage (35.77%) of curcumol was the essential oil of C. aromatica. The higher percentage (49.03% and 43.52%) of cis--elemenone were the essential oils of C. longa and C. sichuanensis. Using the conjugated diene method, at 20 mg/ml, antioxidant activities of essential oils were in the order of C. longa (99.12%) > C. sichuanensis (95.44%) > C. aromatica (50.34%). At 20 mg/ml, the scavenging ability of essential oils of C. aromatica, C. longa and C. sichuanensis on 1,1-diphenyl-2- picrylhydrazyl radicals were 85.60, 73.78 and 61.16%, respectively.

碩士
國立中興大學
食品科學系
91
Mushroom fruit bodies and mycelia can be used as foods and food flavoring, to produce specifically chemical and medical substances, such as water soluble polysaccharides as well as pharmaceuticals such as antibiotics, anticancer and antidisease drugs. Pleurotus citrinopileatus Sing is a new edible fungi in Taiwan recently. The objectives of this research were to investigate the proximate composition, taste quality, antioxidant properties and antimutagenic effect of Pleurotus citrinopileatus fruit bodies, mycelia and fermented broth. Carbohydrate (30.87-45.82%) and protein (25.99-29.50%) were major components and crude fibers (13.80-18.03%) in fruit bodies and mycelia. In fermented broth, carbohydrate was the major components (78%). In fruit bodies, mannitol was major soluble sugars. Fruit bodies had more succinic acids in orgainic acids assay. Fruit bodies were similar in total amino acid to mycelia higher than fermented broth. Linoleic acid was the major fatty acid in fruit bodies, mycelia and fermented broth. In flavor volatiles, 2-pentyl furan, 1-octanol and 1-octen-3-ol were the major volatile of fruit bodies and mycelia. Fruit bodies might have the highest palatable taste than mycelia and fermented broth in 5''''''''-nucleotides, orgainic acids and free amino acid assay. The antioxidant activity of ethanol extracts were in the order Pleurotus citrinopileatus, fruit bodies (87.91%) > mycelia (79.00%) > fermented broth (64.78%) at 20 mg/ml. The reducing powers of ethanol extracts were 1.06, 0.62 and 1.05 at 10 mg/ml for fruit bodies, mycelia and fermented broth, respectively. The ethanol extracts from fruit bodies (95.85%), mycelia (92.78%) and fermented broth (99.93%) showed an excellent scavenging effect on DPPH than hot extracts at 20 mg/ml. Ethanol extracts didn´t show an excellent scavenging effect on hydroxyl radicals. The chelating effect on ferrous ions of ethanol extracts were in the order Pleurotus citrinopileatus, fruit bodies (75.67%) ≈ mycelia (74.95%) > fermented broth (22.28%)at 20 mg/ml. Hot water extracts were similar in antioxidant properties to cold water extracts. Antioxidant properties of Pleurotus citrinopileatus were studied in the form hot water and cold water extracts, fruit bodies > mycelia > fermented broth. The water extracts from fruit bodies, mycelia and fermented broth showed an excellent scavenging effect on hydroxyl radicals and chelating effect on ferrous ions than extracts from ethanol. Total phenols were the major naturally occurring antioxidant components found in all samples. The antimutagenicity of hot water extraxts of Pleurotus citrinopileatus fruit bodies, mycelia and fermented broth was investigated by Ames test using Salmonella typhimurium TA98 and TA100. No toxicity and mutagenicity were found in hot water extraxts of Pleurotus citrinopileatus at the dose of 0.1-5.0 mg/plate. The hot water extracts from Pleurotus citrinopileatus fruit bodies, mycelia and filtrate showed best antimutagenicity effect to against direct-acting mutagen of NQNO (without S9 mix) and indirect-acting m Pleurotus citrinopileatus utagen of B[a]P (add S9 mix). The hot water extracts from Pleurotus citrinopileatus fermented broth showed excellent effect than fruit bodies, and mycelia to against NQNO and B [a] P toward TA98 and TA100. Considering the studies, in addition their abundant nutrients, Pleurotus citrinopileatus exhibit good antioxidant properties and antimutagenic effects. The basis of developing Pleurotus citrinopileatus health food seems can from our research. Keywords: Pleurotus citrinopileatus, taste quality, antioxidant properties, antimutagenic properties

碩士
國立中興大學
食品科學系
90
The objectives of this research were to study the antioxidant, antimutagenic properties of methanolic and water extracts from L. chinense Mill. , the monosaccharide constituents, and structural analysis of polysaccharides. Their structures were elucidated by spectroscopic analysis and comparison with known compounds. Fruit of L. chinense contained 26.94% of moisture. In dry matter, fibers and carbohydrates were major components, and crude protein (13.15%). Methanolic extracts from L. chinense showed very high antioxidant properties. Antioxidant activity using conjugated diene method, the reducing power, the scavenging effect on 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH) and the chelating effect on ferrous ions of methanolic and water extracts from L. chinense increased with the increased concentration, but water extracts was not as good as expected. Naturally occurring antioxidant compounds, including ascorbic acid, b-carotene, tocopherols, and total phenols were found in methanolic extracts from L. chinense. However b-carotene and tocopherols were not found in water extracts. Total antioxidant components were 6.29, 4.04, 6.58 and 7.57 mg/g for methanolic, cold water, boiling water, heating water to boiling extracts, respectively. The concentration of water extracts from L. chinense used in this study was neither toxicity nor mutagenic to the Salmonella typhimurium TA97, TA98, TA100 and TA102. Methanolic extracts of L. chinense contained antimutagens capable of inhibiting the mutagenicity of direct-acting such as MNNG, NQNO the indirect-acting mutagen, B[a]P. The total sugar contents of boiling alkaline extracts from L. chinense were higher than boiling water extracts. The compositions of neutral monosaccharide in boiling water extracts were ribose, mannose and glucose. Glucose was the major monosaccharide in boiling water and alkaline extracts of polysaccharides from L. chinense. Polysaccharides of L. chinense were separated using gel filtration, and showed a molecular of about 104 Da. X-ray, DSC and NMR elucidated their structure. In DSC and 1H, 13C-NMR spectral comparism showed the polysaccharides crystal of boiling alkaline extracts were more tight and regulation. The DSC and NMR spectrum showed functional group of sugar from water extracts of heating to boiling for 2 hours polysaccharides was lowest, but the absorbance peak markedly. The exothermic behaviors of water extracts of heating to boiling for 2 hours polysaccharides were examined by DSC, so supposition it impure. It could be study their physiological activities, advance analysis. For the application in healthy food, further research on the mechanism of polysaccharide properties is in progress. Considering these studies, the major components in L. chinense were carbohydrates and fibers; methanolic extracts from L. chinense in the antioxidant activity better than water extracts. No toxicity or mutagenicity in S. typhimurium TA97, TA98, TA100 and TA 102. Both boiling water and alkaline extracts contain small molecular polysaccharide, and similar monosaccharide components. Keyword: Lycium chinense, antioxidant properties, antimutagenic properties, polysaccharides, structural analysis

碩士
國立臺灣大學
食品科技研究所
94
In the present study, black soybean kojis were first prepared with Aspergillus awamori、Aspergillus oryzae BCRC30222、Aspergillus sojae BCRC30103、Rhizopus azygosporus BCRC 31158 and Rhizopus sp. No. 2, which are commonly employed as starter organism for production of oriental fermented foods. Changes of tatol phenolic and total anthocyanin contents as well as antimutagenic activity against 4- nitroquinoline- N- oxide (4-NQO) and Benzo[a]pyrene (B[a]P) of black soybean due to fermentation were examined. Besides, effect of cultivation temperature and fermentation period on the tatol phenolic, total anthocyanin content and antimutagenic activity of the A. awamori-prepared black soybean koji were also examined. Additionally, the mechanism of antimutagenic activity of the A. awamori-prepared black soybean koji was also investigated. The result showed tatol phenolic and total anthocyanin contents of black soybean were increased after fermentation. The methanolic extracts of black soybean kojis, regardless of starters used, exhibited a higher antimutagenic activity than that the non-fermented black soybean. While antimutagenicity of black soybean koji varied with the starter used. Among the various kojis examined, koji prepared with A. awamori showed the highest antimutagenic effect against 4-NQO and B[a]P. It also showed the highest tatol phenolic and total anthocyanin contents. In further studies, black soybean koji was prepared with A. awamori at different temperatures (25℃, 30℃ and 35℃) and various fermentation periods (1-5 days). It was found that the methanolic extract of A. awamori koji prepared at 30℃ for 3 days exhibited the highest antimutagenic activity and the highest total contents of phenolic and anthocyanin. Study on the possible antimutagenic mechanism, revealed that the bio-antimutagenic, desmutagenic and blocking effect all contributed to the antimutagenicity of A. awamori-koji against 4-NQO and B[a]P.

碩士
國立中興大學
食品科學系
90
The objestives of this reserch were to investigate the proximate composition, antioxidant properties and antimutagenic effect of methanolic extracts from [Grifola frondosa (Dickson:Fries) Gray], [Morchellaesculenta (L.) Pers.] and [Termitomyces albuminosus (Berkeley & Broome) Heim] mycelia. As a result of our investigation, carbohydrate and crude fat were major components of Grifola frondosa mycelia (32.00% and 30.26%). In Morchella esculenta mycelia, carbohydrate and crude protein were major components (46.67% and 27.56%). In Termitomyces albuminosus mycelia, carbohydrate and crude protein were also major components (30.24% and 40.60%). The antioxidatnt activity, using the conjugated diene methods at 10 mg/ml, was found in the order of T. albuminosus (89.83%) > G. frondosa (78.15%) > M. esculenta (71.3%). The reducing power of methanolc extracts from mycelia increased with increased concentration. Morever, the reducing power of extracts (10 mg/ml) was in the order of G. frondosa (0.62) ~ M. esculenta (0.64) > T. albuminosus (0.43), but they were less than those of DHA and -tocopherol. At 10 mg/ml dose, the scavenging effect on DPPH, M. esculenta (94.06%) was higher than G. frondosa (79.43%) and T. albuminosus (78.82%). Obviously, the scavenging effects of extracts on DPPH also increased with increased concentration. At 5 mg/ml dose, the scavenging effect of samples on hydroxy radicals was in the order of T. albuminosus (5.51%) > M. esculenta (0.91%) > G. frondosa (0.00%). In addition, the chelating effect on ferrous ions was 90~94% at 10 mg/ml for methanolic extracts from these three mycelia, indicating they possessed excellent chelating effect on ferrous ions. The natural antioxidant components found included ascorbic acid, tocopherols, and total phenols. Total phenols were the major components found in all methanolic extracts, which were found in the order M. esculenta (3.63 mg/ml) > G. frondosa (1.80 mg/ml)> T. albuminosus (1.59 mg/ml). However, -carotene was not detected in the above mycelia. No mutagencity in Samonella typhimurium TA97, TA98, TA100, and TA102 was observed at 0.01-500 g/plate of methanolic extracts from G. frondosa, M. esculenta, and T. albuminosus mycelia. At 500 g/plate dose, the methanolic extracts from three mycelia showed markedly antimutagenicity effect of Benzo [a] pyrene, N-methyl-N''-nitrosoguanidine, and 4-nitro-quinoline-N-oxide to Salmonella typhimurium TA97, TA98, TA100 and TA102. Summarily, in addition to their abundant nutrients, G. frondosa, M. esculenta, and T. albuminosus mycelia exhibit good antioxidant properties and antimutagenic effects. Therefore, the results of this study could provide valuable information for application agriculture, health foods and other related industries to prepare and formulate nutrient foods or as their ingredient.

碩士
國立屏東科技大學
食品科學系
88
The objectives of this study were to investigate the antimutagenic activity of 50% alcohol extracted Anoectochilus formosanus Hayata, Vitis thunbergii Sieb. & Zucc. var. taiwaniana Lu., Chrysanthemum indicum L., Glossogyne tenuifolia (Labill) Cass, Lycium chinense Mill, Rhinacanthus nasutus (L.) Kurz, Boehmeria nivea (L.) Gaud. var. nivea, Agastache rugosa (Fisch et Mey) O. Kuntze, Ganoderma lucidum (Leyss. ex. Fr.) Karst, and water extracted Anoectochilus formosanus Hayata and Morus australis Poir. The studies were investigated by Ames test using Salmonella typhimurium TA98、TA100、TA1535 and immediate mutagen of NQNO (without S9 mix) and indirect mutagen of B[a]P (add S9 mix) to these ten plants. Five different dosage were used, which were 0.5, 1.0, 2.0, 2.5, 5.0 mg/plate. The toxicity were found at Ganoderma lucidum (Leyss. ex. Fr.) Karst, Agastache rugosa (Fisch et Mey) O. Kuntze, Chrysanthemum indicum L., Glossogyne tenuifolia (Labill) Cass, Lycium chinense Mill, Rhinacanthus nasutus (L.) Kurz, Vitis thunbergii Sieb. & Zucc. var. taiwaniana Lu., Anoectochilus formosanus Hayata (water extracted), Morus australis Poir without S9 mix, Glossogyne tenuifolia (Labill) Cass, Anoectochilus formosanus Hayata (alcohol and water extracted) with S9 mix for TA 98, and Vitis thunbergii Sieb. & Zucc. var. taiwaniana Lu. without S9 mix, Vitis thunbergii Sieb. & Zucc. var. taiwaniana Lu. and Lycium chinense Mill with S9 mix at the dosage of 5 mg/plate. But at the reduced dosage level of 2.5 mg/plate, the toxicity was not found. The mutagenicity was not found at every dosage tested for all the samples. In the antimutagenic activity, effect against B[a]P toward TA 1535 were found for all samples, others were dependent on tested samples. Anoectochilus formosanus Hayata (alcohol extracted) had to effect against NQNO toward TA 98. Vitis thunbergii Sieb. & Zucc. var. taiwaniana Lu. showed effect gainst NQNO toward TA 98. Chrysanthemum indicum L. exhibited effect to against NQNO toward TA 100. Glossogyne tenuifolia (Labill) Cass reveled effect against NQNO toward TA 1535. Morus australis Poir had effect to against B[a]P toward TA 98 and NQNO toward TA 100.Lycium chinense Mill showed effect gainst B[a]P toward TA 98. Rhinacanthus nasutus (L.) Kurz had effect against NQNO toward TA 98. Boehmeria nivea (L.) Gaud. var. nivea had effect against NQNO toward TA 98 and TA 100. Agastache rugosa (Fisch et Mey) O. Kuntze had effect against NQNO toward TA 1535. Ganoderma lucidum (Leyss. ex. Fr.) Karst had effect against NQNO toward TA 98 and TA 100. Significantly antimutagenic activities were found for Anoectochilus formosanus Hayata (extracted alcohol), Vitis thunbergii Sieb. & Zucc. var. taiwaniana Lu., Rhinacanthus nasutus (L.) Kurz, Boehmeria nivea (L.) Gaud. var. nivea and Ganoderma lucidum (Leyss. ex. Fr.) Karst against NONO, and Morus australis Poir and Lycium chinense Mill against B[a]P toward TA 98, Chrysanthemum indicum L., Morus australis Poir, Boehmeria nivea (L.) Gaud. var. nivea and Ganoderma lucidum (Leyss. ex. Fr.) Karst against NONO, but showed no effect against B[a]P toward TA 100, Glossogyne tenuifolia (Labill) Cass and Agastache rugosa (Fisch et Mey) O. Kuntze against NONO.

Cyclolinopeptides (CLs) are hydrophobic cyclic peptides found in flaxseed. They show immunosuppressive activity, but the biological function of these compounds is largely unknown. This thesis presents the results of studies that were conducted to determine whether CLs could act as antioxidants. In the first study, flaxseed oil was passed over a silica adsorbent column to remove polar compounds. The polar compounds were then eluted from the silica absorbant using a series of increasingly polar solvents. Individual polar fractions were then added back to the silica-treated flaxseed oil and the oxidative stability index of these samples was determined at 100 °C. A polar fraction containing mainly CLA, β/γ- and δ-tocopherol increased the induction time of silica-treated flaxseed oil from 2.3 ± 0.28 h to 3.2 ± 0.41 h. A positive effect of the polar fraction containing a mixture of CLA and CLD-CLG on the oxidative stability of oil was also observed. The antioxidant mechanism of CLs was investigated in several model systems using electron spin resonance spectroscopy. The concentration of radicals in a DMPO (5,5-dimethyl-1-pyrroline-N-oxide) radical-CLs reaction mixture was monitored. All CLs exhibited dose dependent scavenging activities. CLA–CLC reactions with DMPO-OH at a concentration of 5 mM resulted in a 24–30% decrease in electron paramagnetic resonance (EPR) signal intensity. The reaction of CLs and the stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•) revealed a more complex interaction than simple radical scavenging. Peptides (CLG and CLG") that contained both tryptophan and methionine showed stronger radical scavenging activity than did CLs containing methionine or methionine sulfoxide but not tryptophan (CLB and CLC). Irradiation of the reaction mixture of DPPH• and peptide with UV light also affected the radical scavenging behaviour. Scavenging activities of DPPH• by CLB, CLC and CLA were enhanced by light, whereas scavenging of DPPH• by the tryptophan containing peptides CLG and CLG″ was not affected. High-performance liquid chromatography with mass spectrometry (HPLC-MS) analysis of the reaction mixtures after a radical scavenging reaction was used to determine the impact of radical scavenging on the peptides. These reactions revealed new masses that were identified and characterized. It was established that DPPH• reacted with the methionine of CLB and with tryptophan in CLG and CLG, by formation of a new covalently-bonded species. Covalent linkages between these amino acids (alone or in peptides) and DPPH• have not been reported previously.

博士
國立中興大學
食品科學系
91
Glycosaminoglycan (GAG), recognized as a functional component with antioxidant, antimutagenic and hypolipidemic activities, was isolated and identified from body wall of sea cucumber Metriatyla scabra. Antioxidative and antimutagenic activities were investigated by using intestine 407 cell model system and additing of 4NQO (4-nitroquinoline-1-oxide) and MNNG （N-methyl-N-nitroso-guanidine）as mutagenic agents. Hypolipidemic effects and mechanism of the GAG were also evaluated through cholesterol supplemented diet with oral administration of GAG to rats. The results are: 1. Compositions of dried sea cucumber were: moisture 3%, protein 88%, carbohydrate 4%, lipid 2%, ash 3%; after rehydration they were 95%, 4%, 0.2%, 0.1%, 0.1%, respectively. Extraction in the order of water, alkali solution, acidic solution, and hot water indicates that major collagen contains in sea cucumber body wall were water-soluble (42%) and acid-soluble (48%), i.e., Type I and Type II collagens. 2. Two peaks (peak-1 and peak-2) were observed for the sea cucumber hydrolyzed with papain and followed by ion-exchange and gel-filtration chromatography. Peak-1 M.W. 200-500kDa, contained mainly GAG as hexuronic acid and hexosamine, while peak-2 M.W. 40-200kDa, contained mostly free glycan as fucose with little hexuronic acid or hexosamine. The peak-1 fraction was used to evaluate hypolipidemic effects. 3. Single cell gel electrophoresis (comet assay) shows that crude proteoglycan (crude PG), GAG (peak-1) and free glycan exhibited DNA damage, which may result from good antimutagenic effects, inducted by 4NQO and MNNG. GAG exhibited antioxidant capacity including scavenging of DPPH free radical, H2O2, superoxide anion and chelating Fe++ and Cu++. No ability for reduction of nitrate further verified the inhibition and antimutagenic activity. 4. Oral administration of GAG to male Wister rats shows that plasma levels of total cholesterol, LDL-cholesterol and atherogenic index, SOD, GSH-Px activity in plasma were significantly decreased, while HDL-cholesterol was significantly increased, and plasma and hepatric tissue MDA concentraction, although these effects of the GAG were only dose-dependent at dose higher than 20mg/kg b.w. Similarly, the GAG significantly prevented the increase (p<0.05) in hepatic contents of triglyceride, cholesterol and phospholipid. 5. GAG increased serum GSH levels. Howerever, the relationship between antioxidant activities and GSH levels was not obvious. Furthermore, the GAG did not significantly prevent the increase of bile acid in the fecal. Sea cucumber GAG probably was identified to attain the reduction of plasma and hepatic cholesterol.

碩士
國立中興大學
食品暨應用生物科技學系
94
Tremella flava Chee-Jen Chen, a fungus isolated from Taiwan, was used to ferment soy bean and black soy bean milk. During the cultivation, the growth and β-glucosidase activity of Tremella flava, the hydrolysis of conjugated isoflavones in soymilk fermentation, the antimutagenic and antioxidant properties of fermented soymilk and fermented black-soymilk with T. flava were evaluated. The growth of T. flava was found to reach 8.69 and 8.25 log CFU/mL, respectively after the 36 h of incubation in soybean and black soybean milk. After 48 h of fermentation, the pH of fermented soymilk and black-soymilk was found ranging from 6.4 to 6.7. The concentrations of isoflavones genistein and daidzein in soymilk after 48 hr of cultivated with T. flava were 24.00 μg/mL and 14.22μg/mL, respectively. On the other hand, 159.14 μg/mL and 55.17 μg/mL of genistein and daidzein were detected in black soybean milk, respectively after fermentation. The β-glucosidase activity in soymilk and black-soymilk fermented with T. flava reached 48.56 mU/mL and 95.71 mU/mL, respectively after 36 h of incubation. It was also noted that as the fermentation time extended, the contents of aglycones and activity of β-glucosidase in soymilk and black-soymilk raised In regard of antioxidant ability, the reducing power of both soymilk and black-soymilk increased significantly after T. flava fermentation. In addition, fermented black soybean milk demonstrated a greater reducing power than fermented soymilk. After 36 h of cultivation, the 1, 1-diphenyl- 2-picrylhydrazyl (DPPH) radical-scavenging activities of fermented black-soymilk was 42.47%, which was significantly higher than those of unfermented black soymilk. The Trolox Equivalent Antioxidant Capacity (TEAC) of the fermented soymilk increased to 0.46 mM after 36 h of incubation. However, no such pattern was observed in fermented black soybean milk. Soymilk and black-soymilk fermented with T. flava did not alter the ferrous ion chelating ability The antimutagenic activity of the fermented soymilk and black soybean milk was determined by means of the Salmonella mutagenicity assay. Both fermented soymilk and fermented black soymilk demonstrated significantly greater antimutagenic activity than unfermented ones against 4-nitroquinoline-N-oxide (4-NQO).

The use of natural antioxidants to improve the oxidative stability of food lipids has received special attention because of the worldwide trend to avoid the use of synthetic food additives. A wide range of natural sources has been shown to contain antioxidant properties, these include plant extracts, herbs and spices, citrus fruits, oilseeds and legumes. Some antioxidants have been found to be fonned during the heat processing of foods, including the Maillard reaction products that are formed by the reaction of amino acids, peptides and proteins with reducing carbohydrates. A study was undertaken to investigate the antioxidant activity of Maillard reaction products fonned during extrusion of soyabeans. A preliminary oxidation study carried out to identify a suitable substrate revealed that sunflower oil stripped of antioxidants was a suitable substrate with a low induction period of 15 minutes via the Rancimat Method and 4.5 hours via the method of Ross and de Muelenaere. Methyllinoleate was found to be sensitive to oxidation, but not readily available and costly. Storage test of antioxidant stripped sunflower oil under various headspace conditions showed that the substrate stability was best at 4°C under nitrogen or vacuum. Under such conditions the product could be stored for a period of 136 days. Nitrogen was chosen as the most suitable for this exercise as it was not easy to remove all residual air from the samples by vacuum. Furthermore with nitrogen headspace residual 02 could be measured based on Ni02 ratio changes. Hexane solvent was found to be able to remove all lipids from soyabeans. Under the experimental conditions practised it was found that the induction periods for extruded and unextruded soya flour hexane extracted lipids were very similar. Addition of glucose or fructose to the extrusion mixture increased induction period of hexane extracted lipids by 37.5% and 1.5% respectively as measured by the Ross and de Muelenaere method and by 50% and 6.5% respectively as measured by the Rancimat Method. Available lysine of glucose containing extrudate was reduced by 69% while that of the fructose containing extrudate was reduced by 23%. Residual glucose and fructose analysis of extrudates showed that 66% of glucose was utilized in the formation of the Maillard reaction products while only 21% of fructose was utilized during extrusion processing. Comparison of induction periods of soya glucose and soya fructose extrudates to induction period of TBHQ antioxidants (200ppm) in antioxidant stripped sunflower oil gave antioxidant activity of 86ppm and 9ppm for soya glucose extrudates and soya fructose extrudates respectively. The observed antioxidant activity of Maillard reaction products could be utilized with success in different types of processed foods without the need for extensive testing as required for synthetic antioxidants but supplementation of lysine may be required to maintain nutritional balance.
Thesis (M.Sc.)-University of Natal, 1994.

香菇是一種重要的藥用蘑菇。數千年來，香菇一直被人們作為食物和藥物來使用。許多研究表明，香菇的提取物具有抗氧化活性，而且他們的抗氧化活性與他們的酚類化合物的含量相關。然而，到目前為止該研究大多集中在對香菇子實體的研究，對香菇菌絲體分泌物的研究就少見報導。
在本課題研究中，使用不同的體外抗氧化測定方法和酚類化合物含量測定方法來研究兩種香菇菌絲體分泌物（1358DE 和L5458DE）。實驗結果表明， 在不同的體外抗氧化實驗中1358DE 和L5458DE均具有明顯不同的抗氧化活性。在清除DPPH自由基，清除氫氧根離子，清除超氧陽離子，清除過氧化氫離子，螯合亞鐵離子，還原能力，抑制老鼠紅細胞溶血和抑制脂質過氧化的實驗中，1358DE 和L5458DE的IC50 分別為3.3和132.6; 44.5和 > 1000; 26.9和53.7; 153.6和 >175.0; 176.0和521.0; 26.7和746.4; 47.8和736.9; 3.1和 > 1000 μg/ml。他們的多酚化合物的含量分別為237.33 and 24.08 mg (GAE)/g of DE。實驗資料表明，1358DE的抗氧化活性高於L5458DE，其原因可能是1358DE的酚類化合物含量較高。
由於1358DE具有較好的抗氧化活性，採用有機溶劑萃取的方法將其分成水溶性部位和乙酸乙酯部位。體外抗氧化實驗表明，水溶性部位的抗氧化活性與1358DE相近，而乙酸乙酯部位則沒有表現出抗氧化活性。因此，使用聚醯胺柱色譜（可以將多酚類化合物從其他成分中分離出來）對水溶性部位進行進一步的分離，可以得到兩個聚醯胺洗脫部位（P-1和P-2）。與原來的水溶性部位比較，P-1的糖的含量明顯增加，而多酚化合物含量明顯減少，抗氧化活性也明顯降低；相反，P-2的糖的含量明顯減少，而多酚化合物含量明顯增加，抗氧化活性也明顯增加。該實驗結果表明，糖對抗氧化活性的貢獻遠不及多酚化合物。因此，多酚化合物是1358DE的抗氧化活性成分。基質輔助鐳射解吸電離飛行時間質譜和三氯化鐵試劑測定結果表明，P-2是一類水溶性多酚低聚物（WSP），它的分子量在600~1200Da之間。
水溶性多酚低聚物（WSP）是1358DE的主要抗氧化活性成分。採用過氧化氫引導細胞毒性的細胞(V79-4)模型來進一步研究WSP的抗氧化活性。在細胞毒性試驗中，在所有測試濃度，WSP在濃度6.25~50 μg/ml均能明顯抑制過氧化氫引致的細胞毒性。此外，WSP還能明顯抑制由過氧化氫引起的丙二醛（MDA）增加和抗氧化酶（SOD，CAT，GSH-Px）的減少。
許多抗氧化劑被報導具有抗血管增生活性，該活性與其抗氧化活性相關。由於WSP具有非常好的抗氧化活性，因此，採用斑馬魚模型來研究WSP的抗血管增生活性。在內源性鹼性磷酸酶測定實驗結果表明，WSP在濃度50，100，150，200，250 μg/ml，斑馬魚（野生型）的血管生成明顯分別減低為87.2, 85.6, 74.8, 69.4, and 62.8%（與空白對照組相比）。此外，在螢光顯微鏡下可觀察到WSP在濃度為250μg/ml能明顯抑制螢光斑馬魚（fli1a:EGFP）的節間血管形成。
本研究表明，水溶性多酚低聚物（WSP）是香菇菌絲體分泌物的抗氧化成分，WSP不僅具有抗氧化活性，同時還具有抗血管增生活性。此外，本研究結果表明香菇菌絲體分泌物是很好的天然抗氧化劑的來源。
Shiitake mushroom (Lentinus edodes), known in China as Xiang-gu, is one of the most valuable medicinal mushrooms, and has been used for thousands of years both as food and medicine. Shiitake mushroom extracts have also been found to have antioxidant properties and their antioxidant ability is positively correlated with their phenolic content. However, thus far, investigation of the antioxidant ability of shiitake mushroom has mainly focused on the fruiting body, and the antioxidant properties of its mycelial exudates are rarely reported.
In this study, exudates (DE) secreted from two shiitake mushroom mycelia (strains 1358 and L5458) were evaluated for their antioxidative properties and phenolic content. 1358DE and L5458DE showed distinct antioxidant activity in different in vitro assays, including scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, hydroxyl radical, superoxide anions and hydrogen peroxide; the ability to chelate ferrous ions; reducing power; hemolysis inhibition activity in rat erythrocyte; and lipid peroxidation inhibition (IC₅₀ values of 1358DE and L5458DE were 3.3 and 132.6; 44.5 and > 1000; 26.9 and 53.7; 153.6 and >175.0; 176.0 and 521.0; 26.7 and 746.4; 47.8 and 736.9; and 3.1 and > 1000 μg/mL, respectively). Their total phenolic content was 237.33 and 24.08 mg gallic acid equivalent (GAE)/g of dry DE, respectively. Overall, these results show that 1358DE generally possesses better antioxidant properties than L5458DE, possibly due to its larger total phenolic content.
1358DE were selected to further investigate for its better antioxidant effect. 1358DE was fractionated using water-solvent partition and two fractions [water soluble fraction (WF) and ethyl acetate fraction (EF)] were obtained. The antioxidant effects of WF were similar to those of the original 1358DE, while EF did not possess any antioxidant activities. The WF was further isolated with polyamide column, which can separated the polyphenols from other components, and two sub-fraction (P-1 and P-2) were obtained. After the WF passing through the polyamide column, carbohydrate content in the sub-fraction 1 (P-1) was significantly increased, while its total phenolic content reduced dramatically, and its antioxidant activity decreased. However, the sub-fraction 2 (P-2) was the opposite. Carbohydrate content in P-2 was significantly reduced, while its total phenolic content increased dramatically, and its antioxidant activity increased. Apparently, carbohydrate contributed little to the antioxidant effect than that of the phenolic compounds as shown from this investigation. These results suggest that the antioxidant effect in 1358DE was contributed by the presence of polyphenols. Besides, results from Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and ferric trichloride reaction suggested that P-2 was oligomers of water soluble polyphenols (WSP) with the molecular weight of about 600~1200 Da.
The water soluble polyphones (WSP) were the potent antioxidant components in 1358DE and further study its protective effect against the hydrogen peroxide which induced cytotoxicity in V79-4 cells. In the cell viability experiments, pretreatment of WSP at the concentrations of 6.25~50 μg/ml increased the cell viability significantly more than at the presence of H₂O₂ only. Besides, the pretreatment of cells with WSP significantly inhibited the increase of Malondialdehyde (MDA, which is a by-product of lipid peroxidation) production and the decrease of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) activities induced by H₂O₂.
Quite a few antioxidant compounds have been reported that a causative relationship may exist between the anti-angiogenic activity and antioxidant effect. Therefore, a zaebrafish model was using to investigate the anti-angiogenic activity of the WSP because of its excellent antioxidant activity. In quantitative of endogenous alkaline phosphatase (EAP) assay, after the embryos treated with WSP at final concentrations of 50, 100, 150, 200, 250 μg/ml, and the vessel formation were significantly (p < 0.05) reduced to 87.2, 85.6, 74.8, 69.4, and 62.8% of the control value, respectively. Moreover, from the microscope, compare to the control, WSP at the concentration of 250 μg/ml also showed potent inhibition on the intersegmental blood vessels (ISVs) formation in Tg(fli1a:EGFP)y1 zebrafish embryos. Thus, the finding indicated that WSP could inhibit vessel formation in zeabrafish.
Overall, this study revealed that water soluble polyphenols (WSP) was the active components of 1358DE. Besides of the antioxidant effect, the WSP could inhibit vessel formation significantly in zebrafish. The findings indicate that exudates of shiitake mushroom mycelia have good potential as a source of natural antioxidants.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Huang, Weihuan.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2012.
Includes bibliographical references (leaves 90-103).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.
Acknowledgements --- p.i
Abstract --- p.ii
摘要 --- p.v
List of Abbreviations --- p.vii
List of Figures --- p.ix
List of Tables --- p.xi
Chapter Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- Oxidation and antioxidant --- p.1
Chapter 1.1.1 --- Reactive oxygen species (ROS) --- p.1
Chapter 1.1.2 --- Sources of ROS --- p.1
Chapter 1.1.3 --- The role of ROS in normal physiology --- p.2
Chapter 1.1.4 --- Oxidative damage to DNA, lipids and proteins --- p.2
Chapter 1.1.5 --- Antioxidant defense systems in vivo --- p.4
Chapter 1.1.6 --- Sources of antioxidants --- p.6
Chapter 1.2 --- Assessment of antioxidant capacity in vitro and in vivo (Antioxidant methodology) --- p.9
Chapter 1.2.1 --- Assessment of Free Radical Scavenging Capacity in vitro --- p.9
Chapter 1.2.2 --- Antioxidant capacity in cultured Cells --- p.10
Chapter 1.2.3 --- Antioxidant capacity in vivo --- p.11
Chapter 1.3 --- Mushrooms --- p.12
Chapter 1.3.1 --- Mushroom life cycle --- p.12
Chapter 1.3.2 --- Nutritional and medicinal values of mushroom --- p.14
Chapter 1.4 --- Shiitake mushroom (Lentinus edodes) --- p.15
Chapter 1.5 --- Objectives --- p.17
Chapter Chapter 2 --- Antioxidant activity and total phenolic content in Shiitake mycelial exudates --- p.18
Chapter 2.1 --- Introduction --- p.18
Chapter 2.2 --- Materials and methods --- p.19
Chapter 2.2.1 --- Materials --- p.19
Chapter 2.2.2 --- Sample preparation --- p.20
Chapter 2.2.3 --- In vitro antioxidant activity assays --- p.20
Chapter 2.2.4 --- Determination of total phenolic content --- p.25
Chapter 2.2.5 --- Statistical analysis --- p.25
Chapter 2.3 --- Results and discussion --- p.25
Chapter 2.3.1 --- Antioxidant activity --- p.25
Chapter 2.3.2 --- Total phenolic content --- p.37
Chapter 2.3.3 --- Antioxidant activity (IC₅₀ values) and phenolic content --- p.37
Chapter 2.4 --- Conclusion --- p.39
Chapter Chapter 3 --- The antioxidant components of Shiitake mycelial exudates (1358DE) --- p.42
Chapter 3.1 --- Introduction --- p.42
Chapter 3.2 --- Materials and methods --- p.42
Chapter 3.2.1 --- Materials --- p.42
Chapter 3.2.2 --- Sample preparation --- p.43
Chapter 3.2.3 --- HPLC analytical condition --- p.43
Chapter 3.2.4 --- Sample fractionated using solvent-water partition and polyamide column chromatographic method guided by in vitro antioxidant assays --- p.43
Chapter 3.2.5 --- Determination of total phenolic content --- p.44
Chapter 3.2.6 --- Determination of the contents of carbohydrate --- p.45
Chapter 3.2.7 --- MALDI-TOF MS analysis --- p.45
Chapter 3.2.8 --- Statistical analysis --- p.46
Chapter 3.3 --- Results and discussions --- p.46
Chapter 3.3.1 --- HPLC analytical results --- p.46
Chapter 3.3.2 --- Samples fractionation using solvent-water partition --- p.47
Chapter 3.3.3 --- Water soluble fraction (WF) was further isolated using polyamide column chromatographic --- p.50
Chapter 3.3.4 --- Molecular weight determination of P-2 --- p.54
Chapter 3.4 --- Conclusion --- p.56
Chapter Chapter 4 --- Antioxidative effect of water soluble polyphenols (WSP) in Shiitake mycelial exudates (1358DE) against H2O2-induced cytotoxicity in V79-4 cells --- p.57
Chapter 4.1 --- Introduction --- p.57
Chapter 4.2 --- Sample preparation, materials and methods --- p.58
Chapter 4.2.1 --- Preparation of the water soluble polyphenols (WSP) --- p.58
Chapter 4.2.2 --- Materials --- p.58
Chapter 4.2.3 --- Cell culture and treatment --- p.59
Chapter 4.2.4 --- MTT assay --- p.60
Chapter 4.2.5 --- Lactate dehydrogenase (LDH) release assay --- p.60
Chapter 4.2.6 --- Assay for lipid peroxidation measuring the malondialdehyde (MDA) --- p.61
Chapter 4.2.7 --- Assay for antioxidant enzymes --- p.62
Chapter 4.2.8 --- Protein determination --- p.62
Chapter 4.2.9 --- Statistical analysis --- p.63
Chapter 4.3 --- Results and discussion --- p.63
Chapter 4.3.1 --- Cytotoxicty of WSP in V79-4 cells --- p.63
Chapter 4.3.2 --- Determined the time of WSP pretreatment in V79-4 cell against H₂O₂-induced cytotoxicity --- p.63
Chapter 4.3.3 --- Protective effect of WSP treated cell against H₂O₂-induced cytotoxicity --- p.64
Chapter 4.3.3 --- Inhibition of WSP on lipid peroxidation --- p.66
Chapter 4.3.4 --- Effects of WSP on antioxidant enzyme activities --- p.66
Chapter 4.4 --- Conclusion --- p.72
Chapter Chapter 5 --- Anti-angiogenic property of water soluble polyphenols (WSP) in Shiitake mycelial exudates (1358DE) --- p.73
Chapter 5.1 --- Introduction --- p.73
Chapter 5.1.1 --- Angiogenesis --- p.73
Chapter 5.1.2 --- Angiogenesis as a therapeutic target --- p.73
Chapter 5.1.3 --- Tumors angiogenesis --- p.75
Chapter 5.1.4 --- Reactive oxygen species (ROS) and tumor angiogenesis --- p.75
Chapter 5.1.5 --- Anti-angiogenic effects of polyphenols --- p.76
Chapter 5.1.6 --- Experimental model for studying anti-angiogenic agents --- p.76
Chapter 5.2 --- Sample preparation, materials and methods --- p.78
Chapter 5.2.1 --- Preparation of the water soluble polyphenols (WSP) --- p.78
Chapter 5.2.2 --- Materials --- p.78
Chapter 5.2.3 --- Methods --- p.79
Chapter 5.2.4 --- Statistical analysis --- p.80
Chapter 5.3 --- Results --- p.81
Chapter 5.3.1 --- Anti-angiogenic effect of WSP on zebrafish model --- p.81
Chapter 5.3.2 --- Microscopic imaging --- p.82
Chapter 5.4 --- Discussion and conclusion --- p.82
Chapter Chapter6 --- Conclusions --- p.84
Chapter 6.1 --- Conclusion --- p.86
Chapter 6.2 --- Future works --- p.88
References --- p.89
Chapter Appendix 1 --- Publication --- p.104

碩士
國立屏東科技大學
熱帶農業暨國際合作系所
99
Chainey root (Smilax spinosa Mill.) is popularly used in Nicaragua’s traditional medicine for its believed anti-bacterial, anti-toxin and antioxidant properties. Despite the widespread use of this plant in Nicaragua, especially on the Atlantic coast, literature contains few reports on the antioxidant activity and chemical composition of S. spinosa. Scientific evidence is required to verify its medicinal effects. Therefore, the aims of this study were to investigate the antioxidant properties of S. spinosa’s root methanolic (ME) extract and its ethyl acetate (EA), n-butanol and water soluble fractions. The total phenolic and flavonoid content, 1,1-Diphenyl-2-Picrylhydrazyl (DPPH) free radical scavenging capacity and Iron (II) chelating activity were investigated. The cytotoxic effect of the EA fraction on human liver hepatoma (HepG2) cells and its cytoprotective properties against H2O2 induced oxidative stress on healthy human liver (FL83B) cells were established. Results demonstrated the antioxidant activity of S. spinosa root to be concentration dependant. Phenolics and flavonoids were found in the ME extract as well as in the EA, n-butanol and water soluble fractions. EA fraction revealed the highest gallic acid equivalence (GAE) of 71.81± 0.36 mg/g DW. Quercetin equivalent was also highest for the EA fraction (45.27± 31.27 mg/g). On the other hand, lowest phenolics and flavonoids were observed in the water soluble fraction. The DPPH scavenging capacity and Iron (II) chelating activity of all four fractions were above 70% at 0.1 mg/ mL. However, none of the fractions were better iron chelators than positive control EDTA. EA showed the highest DPPH scavenging percentage (93%) at 0.2 mg/mL. Among all test samples, the EC50 value of EA soluble fraction was the lowest (0.033 ± 0.002 mg/mL), indicating stronger antioxidant activity than those of positive controls ascorbic acid (0.067 ± 0.0002 mg/mL) and gallic acid (0.068 ± 0.0004 mg/mL). Because of its outstanding antioxidant activity, the EA fraction was selected for further assessment of cytotoxic and cytoprotective effects on the viability of human liver cells. Cytotoxic effect of S. spinosa root EA fraction on the proliferation of human liver hepatoma (HepG2) cells was assessed with the 2-(4-Iodophenyl)-3 - (4-nitrophenyl-5 (2, 4-disulfophenyl) - 2H-tetrazolium (WST-1) assay. S. spinosa EA fraction was capable of interrupting the development and proliferation of HepG2 cells at a concentration dependant manner. At 1500 µg/mL the scavenging of 100% cancerous cells was accomplish by the EA fraction. The Protective effect of the EA fraction on healthy human liver (FL83B) cells viability was assessed by treating the cells with EA for 1 hr prior to the addition of H2O2. The relative cell survival was determined by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. At a concentration of 12.5 µg/mL, the EA fraction enhanced a protective effect on the cells viability allowing 77% cells survival. Results provided a scientific support to the antioxidant properties of S. spinosa Mill. root extract in vitro. Further evaluation of its antioxidative properties in vivo is needed to ensure functionality and long term safety.

碩士
國立中興大學
生命科學院碩士在職專班
105
Tea is a healthy way at home and abroad. It is considered a magical effect of herbs in ancient time, has become the daily drink for everyone. Studies have shown that tea has a significant effect on human health. The main organic compounds in tea are tea polyphenols. Polyphenols are strong antioxidant, with physiological activity and immune function in the body. Catechin is one of the important ingredients of polyphenols. It universal exist in tea, foods. Green tea is rich in catechins and also a source of bitterness in tea. The four main structures of catechins are (-)-epicatechin (EC), (-)-epicatechin-3-gallate (ECG), (-)-epigallocatechin (EGC) and (-)-epigallocatechin-3-gallate (EGCG). Most of the previous studies were discussed for EGCG, relatively less than the other three structures. So this study will evaluate that antioxidant activity in a tube and cell test in vitro for EC, ECG, EGC, EGCG. Compare with the difference between the four in antioxidant activities and the degree of influence in the cell. At antioxidant activities, the results showed that the ability of scavenging DPPH free radicals of catechins four kinds of structures was not very different, and the low concentration had a good scavenging effect. In the superoxide anion scavenging project, results are expressed EGCG has the best clearance ability, while EC is the worst. ABTS free radical scavenging results show that ECG and EGCG had a similar clearance effect, EGC and EC are slightly worse. Cell test section, an intracellular ROS production test used high dose and reaction 24 hours condition. The results shown ECG, EGCG has the effect of significantly reducing ROS generation. EC and EGC have no difference. In cell viability, ECG and high concentration of EGC, EGCG were significantly different, representing three structures with the ability to kill cancer cells. Therefore, catechin effectively remove ROS and kill cancer cells to achieve anti-cancer effect. Overall, ECG has an advantage in scavenging oxides and free radicals. EC effectively removes free radicals, but on ROS generation was not affected. The antioxidant activity of four kinds of catechins was ECG > EGCG > EGC > EC. The four structures of catechins have high antioxidant activity, but have different scavenging effects of different classes of free radicals, especially in cell tests. For a single structure catechin antioxidant mechanism is necessary to further study, look forward to catechins in the treatment can be used with a single structures catechins to achieve more efficient use.

碩士
環球科技大學
生物技術研究所
101
Hedychium coronarium belongs to Zingiberaceae family and is widely distributed throughout tropical and subtropical regions. The aim of the present study was designed to investigate the antioxidative activity and tyrosinase activity of Hedychium coronarium. First, we use of DNA sequence by similarity analysis to compare the naming of molecular identification and morphological identification. And then, the dried leaf, stem, rhizome and flower of Hedychium coronarium were extracted by hot water and ethanol for 180 mins. The antioxidant activities were examined by DPPH, reducing power, ferrous chelating ability, inhibition of lipid peroxidation, scavenging of hydrogen peroxide and superoxide anion scavenging activity. We also measured the amount of trolox equivalent antioxidant capacity, total flavonoids and total polyphenolic in the Hedychium coronarium. Among the tested, water extract of freeze dried leaf is found to be the best free radical scavenging activities. The EC50 of the freeze dried leaf extract for scavenging DPPH free radicals is 1.34 mg/ml. The EC50 of the freeze dried leaf water extract for chelating ferrous ion activity is 3.92 mg/ml. In the concentration of 3.125 µg/ml, the inhibition of lipid peroxidation is 83.3%. The Trolox equivalent antioxidant capacity is 1.44 mmol TE/mg for freeze dried leaf water extract. In the concentration of 500 μg/ml, the ability for scavenging superoxide anion radicals is 87%. Total phenol contents of water extract are 66.4 mg of GAE/g and total flavonoids contents are 88.7µg of QE/mg. In addition, at concentration of 1 mg/ml for ethanol extract of freeze dried leaf exhibited 88.3% tyrosinase inhibitory activity. The phenolic and total flavonoids contents are all correlated with these activities. Based on the assays presented here, it can be concluded that Hedychium coronarium is an accessible source of natural antioxidants that provides the expected health benefits.

碩士
大葉大學
生物產業科技學系
101
Red job’s tears (Taichung 3) produced in Er-lin were used as material to prepare adlay wine products by mixing dehulled alday with glutinous rice through both cooking and direct fermentation processes. The processes for three wine items including (1)yellow alday wine with 10-12% in alcohol content; (2)adlay spirit with 40% in alcohol content; and (3) a type of light wine with about 3% in alcohol content and specific adlay flavor were confirmed and their functions were also investigated. The results obtained were as follows. 1. According to sensory evaluation: (1)The best fermented alday wine was made from entire alday that was cooked. More specific alday flavor with moderate acidity was the feature of this product; (2)The best adlay spirit was made from entire alday that was not cooked. This product was quite different from common rice spirit or kaoliang spirit due to more obvious alday flavor; (3)A light alday wine with good quality was made by adjusting entire alday wine with cooked method. The product suits for the female sex or groups they do not accept products with higher alcohol content. 2. Functionality analyses showed that DPPH-scavenging capability reached 83% for the fermented alday wine made from entire uncooked alday, while 55% for that of from cooked. On the other hand, similar reducing activities to 200 ppm of butylated hydroxyl anisole(BHA) were observed for both fermented wines made from cooked or uncooked alday. 3. About 5% adlay oil contained in red job’s tears. Nineteen fatty acids and various functional components including phytols, phenols, and coixol are present in the oil. HPLC analysis showed 11.5 mg/mL of coixol that is the special physiological functional ingredient. GC analysis indicated that methyl oleate (C19H36O2) with 6.2 mg/mL was the primary composition among fatty acids. Methyl linoleate(C19H34O2)with 1.97 mg/mL and methyl hexadecanoate (C17H34O2) with 0.29 mg/mL came the next. 4. The alday wine made from cooked entire alday only contained 0.73 mg/mL of alday oil. Further studies for increasing alday oil content and preventing from oxidation of the wine was necessary in the future.

碩士
國立高雄海洋科技大學
水產食品科學研究所
101
The objective of present work is to extract and isolate the antioxidative substances from banana leaves, and to study their antioxidative activity. The banana leaves were dried and milled into powder. The proximate composition of banana leaf powder was listed as follows: moisture content, 29.6%; carbohydrate, 49.6%; ash, 19.5%; crude fat, 0.3%; crude protein, 0.9%. Efficiency of solvent to extract antioxidative substance from banana leaf powder was in the following: boiling water> cooling water > 50% ethanol. Those extracted by 95% ethanol, methanol, ethyl acetate, hexane, acetone showed less efficiency. The optimal extracting conditions were boiling water for 30 min. Crude extract of banana leaves was further fractionated by using XAD column separation, the 4th fraction (named XAD-4) was obtained with IC50 287.76(μg/mL) and purification fold 1.36. The antioxidative substances from XAD-4 were possessed of ability to prevent peroxidation of linoleic acid, DPPH and hydroxyl radical scavenging activity, reducing power, Fe (II) chelating activity. Total phenol content of crude extract from banana leaf was 1.34 ± 0.06 mg/g; flavonoid content, 2.14 ± 0.06 mg/g. Total phenol content of XAD-4 from XAD column separation was 2.44± 0.06mg/g; flavonoid content, 3.48 ± 0.06 mg/g.