Even though early man did not know the nature of coloring matter in flowers, he used the coloring matter as dye for centuries. There are a variety of reasons for the interest of chemists in natural and artificial coloring matters. One of these is the color pleasure, anothe1· is the commercial importance. The visible color facilitates the experimental work in the search for methods if separation, purification and determination of organic structures. Search for the knowledge of coloring matter goes far back in history. Man has dyed his textiles with the help of mordants from the most ancient times up to the time of Perkin's discovery of mauve. Then a new era in dyestuff chemistry commenced. In the first half of the nineteenth century with the rise and development of the study of Organic chemistry much attention was directed to the extraction and characterization of pure coloring matters from natural sources .... usually the bark, leaves, fruits. or sap of trees or plants. During the twentieth century -- and just a little before that--great progress has been made in the synthesis of many typical natural coloring matters and the improved technique and novel synthetical methods which will bear upon the general progress of chemistry. This research was done with the purpose of isolating and identifying the coloring matter in the flowers of Vinca Major, with the common name of periwinkle. This was done by extracting the pigment of the flowers with methanol, amyl alcohol, and crystallizing the pigment as a chloride. For this pigment, solubility, color reactions, spectra and other physical properties were obtained.

This study concerns the isolation, characterisation and physio-chemical properties of 8,8-methylmethine-(epi)catechin-malvidin-3-glucoside compounds found in red wines. 8,8-Methylmethine-(epi)catechin-malvidin-3-glucoside compounds were isolated via chromatographic methods developed in this study. The compounds were characterised via nuclear magnetic resonance spectrometry which, with the aid of molecular modelling, afforded their possible 3-dimensional structures. Their physio-chemical properties including ionisation and hydration constants, colour parameters and chemical stabilities were determined. The formation of 8,8-methylmethine-flavan-3-ol-malvidin-3-glucoside compounds and other pigments in wines was also studied. 8,8-Methylmethine-(epi)catechin-malvidin-3-glucoside compounds were also synthesised by condensing malvidin-3-glucoside with (epi)catechin in the presence of acetaldehyde. Diastereomers of 8,8-methylmethine-(epi)catechin-malvidin-3-glucoside pigments were isolated from the reaction using size-exclusion liquid chromatography followed by cation-exchange liquid chromatography. The structures of the four 8,8-methylmethine-catechin (and epicatechin)-malvidin-3-glucoside diastereomers were determined using mass spectrometry and one and two-dimensional nuclear magnetic resonance spectroscopy. It was found that for all four compounds, the methylmethine bridge occurs at the 8-positions of malvidin-3-glucoside and (epi)catechin and that the 3-dimensional structural differences between the diastereomers is the positioning of the (epi)catechin moiety with respect to the glucoside group. One diastereomer has the (epi)catechin on the same side, with respect to the malvidin entity whilst it is on the opposite side for the other diastereomer. The proposed structures also afforded the malvidin entity protection from nucleophilic attack via steric hindrance by the (epi)catechin moiety. 8,8-Methylmethine-(epi)catechin-malvidin-3-glucoside pigments have greater colour stability with regards to changes in pH and SO2 bleaching compared to malvidin-3-glucoside providing evidence that little or no hydration in aqueous solutions is occurring for these compounds. Further evidence for little or no hydration occurring is the presence of isosbestic points in the UV-vis spectra observed for the 8,8-methylmethine-(epi)catechin-malvidin-3-glucoside in the pH range 2 to 7. Although the 8,8-methylmethine-(epi)catechin-malvidin-3-glucoside pigments have greater colour stability to pH, SO2 and oxidation, compared to malvidin-3-glucoside, they have lower temporal stabilities and under both aerobic and anaerobic conditions they have significantly higher degradation rate constants than malvidin-3-glucoside. The ionisation constants of the 8,8-methylmethine-(epi)catechin-malvidin-3-glucoside compounds were determined using high voltage paper electrophoresis (HVPE) and UV-visible spectroscopy. The first ionisation constants (pKa1) of the 8,8-methylmethine-(epi)catechin-malvidin-3-glucoside compounds were found to be higher than that of malvidin-3-glucoside whereas the second and third ionisation constants (pKa2 and pKa3) were found to be lower. The correlation of the ionisation constants between HVPE and UV-visible spectroscopy supports the proposal that there is little or no occurrence of hydration for the 8,8-methylmethine-(epi)catechin-malvidin-3-glucoside compounds in the pH range investigated. 8,8-Methylmethine-flavan-3-ol-malvidin-3-glucoside compounds were the major pigments formed during fermentations of chemically defined grape juice media containing malvidin-3-glucoside and various flavan-3-ols. The yeast strain used for fermentation had a major influence on the levels and rates of formation of these pigments during fermentation. The yeast strain used also has an important influence on wine pigment composition, concentration and evolution during maturation thereby affecting the colour density and hue of the resultant wines. The initial formation of 8,8-methylmethine-flavan-3-ol-malvidin-3-glucoside compounds and their subsequent gradual degradation during maturation, allowed a pool of malvidin-3-glucoside to be available for the formation of other colour stable and more temporally stable pigments. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1339479 / Thesis (Ph.D.) - University of Adelaide, School of Agriculture, Food and Wine, 2008

Studies were undertaken to identify the basis of differences in seed coat pigmentation between selected yellow- and brown-seeded Brassicaceae, and near-isogenic yellow- and brown-seeded lines of </i>Brassica carinata</i> (PGRC/E 21164). Histochemical analyses of dissected seed coats from six genera of the Brassicaceae revealed condensed tannin (proanthocyanidin) and phlobaphene pigments in dark-seeded species and in scattered spots in most yellow-seeded species. Anthoeyanins were not detected in any seed coat tissue of these species. In leaf tissue, anthocyanin content was lower in yellow-seeded 'B. carinata' at the three- to four-leaf stage grown at 20 to 25°C, compared to similarly grown brown-seeded plants. At 15 to 18°C, both yellow- and brown-seeded lines produced similar amounts of anthocyanin in the seedling leaves. In TLC and HPLC analyses, 't'-cinnamic acid, dihydromyricetin [trace amounts], dihydroquercetin, dihydrokaempferol and flavonols (quercetin, kaempferol) were more abundant in extracts from seed coats of yellow-seeded ' B. carinata' than in extracts from seed coats of brown-seeded ' B. carinata'. Myricetin was not detected in any seed coat extracts. Mass spectra were determined for phenylpropanoid and flavonoid aglycones from the seed coat extracts and for authentic standards. Dihydroflavanol reductase ('DFR') transcripts from developing seed were absent or less abundant at 5, 10, 20 and 30 days after pollination in the yellow-seeded line compared to the brown-seeded line of 'B. carinata'. 'DFR' transcripts in seedling leaves from the yellow-seeded line grown in warm, bright conditions were less abundant compared to similarly grown brown-seeded material. Seedlings grown at 15 to 18°C produced greater amounts of 'DFR' transcript in both yellow- and brown-seeded lines of 'B. carinata' compared to warm-grown plants. Data from these experiments suggest that seed coat pigmentation in the Brassicaceae is due to condensed tannin and phlobaphene accumulation, not anthocyanins, and that seed coat pigment biosynthesis is down-regulated at dihydroflavonol reductase. The regulatory factor controlling 'DFR' expression in developing seed may be temperature sensitive and also affect anthocyanin biosynthesis in seedlings and in related metabolic pathways: thioglycolic lignin concentration was significantly lower in the yellow-seeded Brassicaccae and in the seed coat tissue of yellow-seeded 'B. carinata' compared to dark-seeded samples.

The phytochemistry of Ochna serrulata (Hochst.) Walp. was investigated for the first time; two new dimeric chalcones (5-deoxyurundeuvine C and serrulone A) and two new biflavonoid derivatives (4,4’,7-tri-O-methylisocampylospermone A and 4”’-de-Omethylafzelone A) were isolated. These compounds were isolated along with the known compounds lophirone A, afzelone B, campylospermone A, isocampylospermone A, ochnaflavone, 2”,3”-dihydroochnaflavone, lophirone C, psilosin, 3’-O-methylpsilosin, a cyanoglucoside, epicatechin, (2’,4’- dihydroxyphenyl)acetic acid, methyl (2’,4’-dihydroxyphenyl)acetate, irisolone 4’- methyl ether, iriskumaonin 3’-methyl ether, 3',4'-dimethoxy-6,7-methylenedioxyisoflavone, lophirone L, syringaresinol and 16α,17-dihydroxy-entkauran-19-oic acid. The growth inhibitory effect of these compounds was evaluated against three cancer cell line panel of TK 10 (renal), UACC62 (melanoma) and MCF7 (breast) using a sulforhodamine B (SRB) assay. Ochnaflavone and 3’-methoxypsilosin demonstrated selectivity and only inhibited the growth of melanoma cancer cells. However, ochnaflavone showed higher activity by totally inhibiting the growth of melanoma cancer cells at 12.91 μM, whereas, 3’-O-methylpsilosin has this effect at a concentration of 14.11 μM. Lophirone C, a dimeric chalcone, demonstrated the highest cytotoxic activity amongst all isolated compounds against renal, melanoma and breast cancer cells with TGI at 35.63 μM, 11.67 μM and 30.35 μM, respectively. Lophirone A, a rearranged biflavonoid, showed TGI against these cancer cells at 58.96 μM, 26.23 μM and 40.01 μM, respectively. The rest of the compounds showed no significant cytotoxicity against the three cancer cells. The new biflavonoid, 4,4’,7-tri-O-methylisocampylospermone A demonstrated the highest antimalarial activity against chloroquine-resistant strains of Plasmodium falciparum (FCR-3) with IC50 of 11.46 μM, followed by ochnaflavone (17.25 μM). iv Serrulone A (26.52 μM), lophirone A (29.78 μM), 5-deoxyurundeuvine C (31.07 μM), lophirone C (35.31 μM), 4”’-de-O-methylafzelone A (38.43 μM), afzelone B (39.54 μM), irisolone 4’-methyl ether (40.72 μM) and syringaresinol (42.66 μM) were moderately active. The following compounds exhibited the lowest antimalarial activity, 2”,3”-dihydroochnaflavone (61.86 μM), iriskumaonin 3’-O-methyl ether (93.69 μM),3’- O-methylpsilosin (106.35 μM) and16α,17-dihydroxy-ent-kauran-19-oic acid (106.48 μM). Owing to the observed and reported biological/pharmacological activity, ochnaflavone (an ether-linked biflavone consisting of apigenin and luteolin moieties) was selected for synthetic studies. An older method, nucleophilic aromatic substitution (SNAr) was successfully applied in the construction of the diary ether. Oxidative ring cyclization of the ether-linked dimeric chalcone was achieved by using heated pyridine and iodine. The two methods can be extended further in the synthesis of other novel biflavones with ether linkage. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

Oxalis reclinata Jacq., is a dicotyledonous plant. O. reclinata belongs to the family Oxalidaceae. This plant produced callus which accumulated red coloured anthocyanin pigments when cultured in vitro. The levels of anthocyanin accumulated by O. reclinata callus were higher than in the intact plant. The major pigment was isolated and identified as cyanjdin-3-glucoside (CROUCH, VAN STADEN, VAN STADEN, DREWES & MEYER, 1993). In nature, anthocyanins are responsible for orange, red, purple and blue colouration of certain tissues of higher plant s. Due to the toxicity of many synthetic red colouring agents, anthocyanins are regarded as potential substitutes for synthetic food colourants. This research was aimed at investigating mechanisms which induce pigment production as well as to optimize anthocyanin yield from callus cultures of O. reclinata, once anthocyanin production was stimulated. Pigmented and non-pigmented callus lines were generated from O. reclinata (CROUCH & VAN STADEN, 1994) and maintained on MURASHIGE & SKOOG (1962) agar medium (O.8% [w/v], pH 5.7) supplemented with 0.5 mgℓ ¯¹ BA, 5 mgℓ ¯¹ NAA, 30 gℓ ¯¹ sucrose and 0.1 gℓ ¯¹ myo-inositol. Plant tissue culture studies were conducted on red and white lines of O. reclinata to optimize callus yield and anthocyanin production in vitro. This involved manipulating contributory factors of the culture environment (carbohydrates, nitrates, phosphates, phytohormones, light and temperature). In vitro studies showed that, light played an inductive role in anthocyanin production in callus cultures of O. reclinata. The auxin, 2,4- dichlorophenoxyacetic acid (2,4-D) reduced pigment production but increased callus biomass. This hormone probably exerted its effect by reducing the pool of anthocyanin precursors, such as phenylalanine, resulting in increased primary metabolic activity. Suspension cultures were shown to be a viable means of propagating pigmented callus cells of O. reclinata. The growth curves for red and white callus cells were determined using the settled cell volume (SCV) method. Pigmented cell cultures grew for longer periods compared to nonpigmented cells of O. reclinata. White callus cells reached the stationary phase after 18 days. Red callus cells continued growing exponentially for an extra three days compared to white callus cells. The vacuole was identified as the organelle where anthocyanins accumulate using the light microscope. The molecular techniques of two-dimensional electrophoresis and in vitro translation were utilized to analyze differences in gene expression between white and red callus cultures of O. reclinata. Thus far, two-dimensional electrophoresis has shown that the red callus of O. reclinata had more polypeptides compared to the white callus. The level of gene expression was higher in the red callus compared to white callus, as revealed by nonradioactive in vitro translation. With optimization of radioactive in vitro translation, identification of specific structural anthocyanin genes which are under regulatory control should be possible. Future research should aim at acquiring a better understanding about the genetic control of anthocyanin biosynthesis in order to manipulate this pathway effectively. / Thesis (M.Sc.)-Univesity of Natal, Pietermaritzburg, 1996.

No description available.

Phytoplankton Chlorophyll a (CHLa), total protein, colloidal carbohydrates, storage carbohydrates and taxonomic pigment relationships were studied in two cyanophytes (Microcystis aeruginosa and Synnechococcus elongatus), two chlorophytes (Dunaliella tertiolecta and Scenedesmus quadricauda), one cryptophyte (Rhodomonas salina), two diatoms (Cyclotella meneghiniana and Thalassiosira weissflogii) and one dinophyte (Amphidinium carterae) to assess if algal biomass could be expressed in other indices than just chlorophyll a alone. Protein and carbohydrates are more useful currencies for expressing algal biomass, with respect to energy flow amongst trophic levels. These phytoplankton were grown at low light (LL = 37 (So(Bmol photons m-2 s-1), medium light (ML = 70-75 (So(Bmol photons m-2 s-1), and high light (HL= 200 (So(Bmol photons m-2 s-1). / Even though pigment per cell increased with increasing light intensity, statistically light had very little effect on the CHL a : taxonomic marker pigment ratios, as they covaried in the same way. Protein, colloidal carbohydrates and storage carbohydrates per cell all increased with increasing light intensity, but they did not covary with CHLa. Statistical data showed that light intensity had a more noticeable effect on protein: CHL a, colloidal carbohydrate: CHLa, storage CHO: CHLa, therefore a general mathematical expression for these relationships cannot be generated. This study showed that light intensity does have an influence on these biomass indices, therefore, seasonal and latitudinal formulas may be required for meaningful algal biomass estimation. However, more studies are needed if that goal is to be realized. / While studying the effects of light intensity on algal pigment content and concentration, a new pigment was isolated from a cyanophyte (Scytonema hofmanii) growing between 300-1800 (So(Bmol photons¨m-2¨s-1 and from samples collected in areas of the Florida Everglades. This pigment was characterized and structurally determined to possess indolic and phenolic subunits that are characteristic of scytonemin and its derivatives. In addition, the pigment has a ketamine functionality which gives it its unique polarity and spectral properties. Based on the ultra violet/visible absorbance data, this pigment was postulated to be protecting the chlorophyll a and cytochrome Soret bands as well as a and (Sb (Bbands of the cytochromes (e.g. cytc-562) in the photosynthetic unit. / by Cidya Grant. / Thesis (Ph.D.)--Florida Atlantic University, 2011. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2011. Mode of access: World Wide Web.

No description available.

Department of Chemistry / MSc (Chemistry) / See the attached abstract below

\'Piraroxa\' e \'Vanda\' são cultivares de alface cujos comportamentos pós-colheita ainda não foram caracterizados. Um dos diferenciais destas cultivares pode estar relacionado ao seu alto potencial antioxidante como resultado da ação de seus compostos fenólicos. A fim de avaliar os parâmetros pós-colheita e a viabilidade do uso destas cultivares como fontes de antioxidantes, o presente trabalho teve como objetivo investigar suas respostas fisiológicas e bioquímicas, comparando-as entre si em diferentes temperaturas de armazenamento e em épocas de colheita distintas. Tanto no verão quanto no inverno, as hortaliças foram armazenadas por 28 dias à 0ºC e 95-97%UR e por quatro dias em condição ambiente monitorada. Em todas as condições de armazenamento e períodos de colheita, não houve diferença entre as cultivares e tendências decrescentes foram observadas para a produção de etileno e as taxas respiratórias. O armazenamento refrigerado proporcionou acentuado declínio destas variáveis. Já em condição ambiente, na colheita de verão, a cv. Vanda apresentou produção superior de etileno e, na colheita de inverno, a cv. Piraroxa apresentou taxas respiratórias mais elevadas. Os parâmetros de coloração permaneceram constantes. As cultivares apresentaram tendências similares entre si, tanto para a perda de massa fresca quanto para o conteúdo de clorofila e carotenoides, e o armazenamento refrigerado e as altas umidades relativas mostraram-se eficazes para o controle da perda de massa. O conteúdo de antocianinas foi influenciado pela temperatura de armazenamento, mas não pela época de colheita, e não demonstrou estar diretamente relacionado ao potencial antioxidante da cv. Piraroxa. As características de qualidade representadas pela aparência das hortaliças foram preservadas, em armazenamento refrigerado, até o 28º dia e, em condição ambiente, até o primeiro e segundo dias. A atividade da polifenoloxidase (PPO) e o potencial antioxidante das hortaliças foram influenciados pela época de colheita, mas não pela temperatura de armazenamento. A cv. Vanda apresentou atividade da PPO superior na colheita de verão e a cv. Piraroxa apresentou potencial antioxidante superior em todas as condições de armazenamento e épocas de colheita. O teor de compostos fenólicos e a atividade sequestrante do radical livre DPPH apresentaram valores superiores na colheita de verão para ambas as cultivares, com tendências à constância de valores, enquanto o sistema-modelo ?-caroteno/ácido linoleico demonstrou tendências crescentes, com elevações significativas a partir do segundo dia de armazenamento em condição ambiente e a partir do 21º dia de armazenamento refrigerado. Os parâmetros fisiológicos analisados evidenciaram grande similaridade em relação ao comportamento pós-colheita das duas cultivares e a durabilidade e manutenção das características qualitativas das hortaliças durante o armazenamento refrigerado foi extremamente satisfatória. Em contrapartida, o armazenamento, tanto em condição ambiente monitorada quanto em refrigeração, não demonstrou constituir-se como técnica válida para a maximização do potencial antioxidante das hortaliças, embora a alface constitua fonte dietética importante de antioxidantes. / \'Piraroxa\' and \'Vanda\' are lettuce cultivars whose postharvest behavior has not yet been characterized. One of the differentials of these cultivars may be related to its high antioxidant potential as a result of its phenolic compounds action. In order to evaluate the postharvest parameters and feasibility of using these cultivars as sources of antioxidants, this study aimed to investigate their physiological and biochemical responses, comparing them to each other at different storage temperatures and at different harvest times. Vegetables were stored for 28 days at 0ºC and 95-97%RH and for four days at ambient monitored condition regardless of summer or winter seasons. In all storage conditions and harvest periods there was no difference between cultivars and decreasing trends were observed for ethylene production and respiratory rates. Cold storage provided dramatic decline of these variables. At ambient condition, however, cv. Vanda showed higher production of ethylene in the summer harvest and cv. Piraroxa had higher respiratory rates in the winter harvest. Color parameters remained constant. Cultivars showed similar trends among one another for both weight loss and the content of chlorophyll and carotenoids. Also, cold storage and high relative humidity were effective for the control of weight loss. Anthocyanins content was influenced by storage temperature, not by the harvest season, and did not show to be directly related to the antioxidant potential of cv. Piraroxa. Vegetables quality appeared to be preserved on cold storage until day 28, and at ambient condition until the first and second days. Polyphenol oxidase (PPO) activity and antioxidant potential of vegetables were influenced by harvest time, but not by the storage temperature. \'Vanda\' showed higher PPO activity in the summer harvest and \'Piraroxa\' showed higher antioxidant potential in all storage conditions and harvest times. Phenolic content and free radical scavenging activity of DPPH showed higher values in summer harvest for both cultivars, with constant value tendencies, while the model system ?-carotene/linoleic acid showed increasing trends, with significant increases in the second day of storage at ambient condition and after 21 days of refrigerated storage. The physiological parameters analyzed showed great similarity to postharvest behavior of the two cultivars, and the durability and maintenance of vegetables quality during refrigerated storage were extremely satisfactory. In contrast, both ambient and refrigerated monitored storages did not show as valid techniques for maximizing the antioxidant potential of vegetables, although lettuce constitutes an important source of dietary antioxidants.