A modified ascending thin layer chromatographic technique has been developed which resolves the major phospholipid and neutral lipid classes of five common fluids and tissues. A one-half milliliter sample is extracted with n-butanol:diisopropylether (40:60 by volume, cholesteryl acetate = 100 ng/ul) for thirty minutes and the organic phase is spotted onto a thin layer chromatography plate and carried through three successive chromatographic developments. The lipids are then visualized either by charring with ammonium bisulfate or staining with phosphomolybdic acid. The use of cholesteryl acetate as an internal standard enables quantitation of the phospholipids and neutral lipid classes. This method may be a very valuable, new technique for research and clinical laboratories.

Call number: LD2668 .T4 1979 L35 / Master of Science

No description available.

Graduation date: 1992

No description available.

Call number: LD2668 .T4 1985 S22 / Master of Science

Graduation date: 1992

Common snook (Centropomus undecimalis) is a relatively new species for aquaculture; considered as a recreational species and not commercial. The aim of this study was to develop common snook larval rearing techniques for stock enhancement. Common snook culture has two main bottlenecks, broodstock management and larval culture. High mortality during the first 6 days is the main limitation for successful larval survival. Broodstock management of common snook is still developing and the only source of common snook eggs is from wild broodstock. Securing a regular supply during the natural spawning was essential to reach the main objective. Finding the optimal spawning sites, as well as optimal spawning time was achieved. Results showed Terra Ceia, Longboat and Cayo Costa to be the best sites for wild broodstock collection. The onset of spawning was triggered by a rise in water temperature. During the 4 years of this study spawning started at the end of May and finished in September. Total capture results and egg quality results, such as fertilization, hatching rate and lipid analysis, indicated June and July as the peak months during the spawning season. Common snook follow a lunar spawning cycle. Results showed that one to three days after the new and full moon were the peak spawning periods and therefore the best days to capture wild stock. Common snook egg lipid composition fits the general marine fish fatty acid composition with saturated fatty acids predominating. On the other hand, the omega 3, omega 6 (n-3/n-6) ratio was lower than the typical marine fish and arachidonic acid values were significantly higher than other marine species. This egg fatty acid profile will be helpful in the future to compare it with captive spawned eggs for egg quality purposes. Description of the common snook embryonic and larval development for the first 14 days was carried out. This has strengthened knowledge for this species’ development, and should provide a helpful tool to identify common snook embryos and larvae in the wild. Novel improvements to existing common snook larval culture protocols were implemented. Larval lipid analysis throughout development, and high mortality around day 6 post hatching, suggested that common snook larvae were dying of starvation. Gross morphological development and ultra-structure findings in the digestive and eye system development during the first three days indicated that day 2 post hatching larvae were capable of capturing and digesting food. Additionally, larval nutritional improvements were made, increasing the larval survival. The most significant ones were: finding a smaller and more nutritional prey (SS type rotifers and copepods), finding an optimal stocking and feeding density and the importance that green water technique has on larval survival. Overall, larval success was improved from a zero percent survival during the first 14 days to a 2% survival rate.

Orientador: Admar Costa de Oliveira / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-04T15:40:31Z (GMT). No. of bitstreams: 1 Botelho_AdrianaPrais_M.pdf: 512202 bytes, checksum: bcf5ef975b16eb94dafdbac80cdd5ce6 (MD5) Previous issue date: 2005 / Resumo: O ácido linoléico conjugado (CLA), um conjunto de isômeros de posição e geométricos do ácido linoléico com duplas ligações conjugadas, ocorre em pequenas quantidades em uma grande variedade de alimentos. O CLA pode ser originado no rúmen por meio da biohidrogenação incompleta de ácidos graxos poliinsaturados provenientes da dieta e também, pela dessaturação do ácido graxo C18:1 trans-11. Dessa maneira, concentrações significativas de CLA podem ser encontradas nas carnes, no leite e seus produtos derivados. O objetivo deste trabalho foi verificar o efeito da suplementação com ácido linoléico conjugadosobre o perfil lipídico e a composição corporal de ratos Wistar saudáveis em crescimento. Foram realizados dois ensaios biológicos: um em que variou a quantidade de CLA suplementada, e outro em que variaram as misturas comerciais de CLA utilizadas. Para o primeiro ensaio biológico, foram utilizados 40 ratos albinos, machos, recém-desmamados, distribuídos aleatoriamente em 4 grupos com 10 animais cada, de acordo com a quantidade de suplemento administrada. Os animais foram suplementados diariamente durante 21 dias com AdvantEdge® CLA (EASTM) nas concentrações 1, 2 e 4 % sobre o consumo diário de dieta, constituindo respectivamente os grupos AE1, AE2 e AE4, e com ácido linoléico na concentração de 2 % sobre o consumo diário de dieta, constituindo o grupo controle ©. Com este ensaio procurava-se identificar qual a quantidade de suplemento mais adequada para reduzir a gordura corporal dos ratos. Para o segundo ensaio biológico, foram utilizados 30 ratos albinos, machos, recém-desmamados, da linhagem Wistar, divididos aleatoriamente em 3 grupos com 10 animais cada, de acordo com a marca de suplemento administrada. Utilizando-se a quantidade de suplemento identificada no primeiro ensaio, os animais receberam diariamente, durante 42 dias, as misturas comerciais AdvantEdge® CLA (EASTM) e CLA One® (Pharmanutrients), constituindo os grupos AE e CO, respectivamente, e ácido linoléico, constituindo o grupo controle ©, na concentração de 2 % sobre o consumo diário de dieta. Durante os dois períodos experimentais os animais tiveram o peso e consumo de dieta monitorados a cada dois dias. Ao final de cada experimento, os animais foram mortos por deslocamento cervical sob anestesia (pentobarbital sódico . 46 mg/kg), sendo o sangue utilizado para as determinações séricas de triacilgliceróis, colesterol total e leptina e a carcaça empregada para a determinação da composição corporal centesimal. Para esta avaliação, foi removido todo o conteúdo intestinal para obtenção da carcaça vazia. Em seguida a carcaça foi congelada em nitrogênio líquido, fatiada, liofilizada, moída e armazenada a - 80 °C até o momento das determinações de umidade, cinzas, proteína bruta e gordura. A eficiência alimentar dos ratos não foi alterada com a suplementação de CLA em ambos os ensaios biológicos. Com relação aos valores séricos de triacilgliceróis, estes não apresentaram diferença significativa (p > 0,05) após a suplementação com CLA. Quanto aos teores de colesterol total no primeiro ensaio, estes demonstraram uma redução dose dependente após 21 dias de tratamento, tomando-se em conta as suplementações. No entanto, no segundo ensaio biológico, aos 42 dias de tratamento, a administração de CLA aumentou os teores de colesterol total dos animais. No tocante à composição corporal, constatou-se uma redução média de 18,0 % dos teores de gordura corporal dos grupos AE2 (11,2 %) e AE4 (11,6 %), quando comparados ao teor do grupo controle (13,9 %). A mesma redução foi observada no segundo ensaio biológico nos grupos AE e CO, em relação ao controle (18,1 %, 16,7 % e 21,2 %, respectivamente). Após 42 dias de suplementação com CLA, os animais dos grupos AE e CO, no segundo ensaio biológico, obtiveram aumento de 7,5 % nos teores de cinzas e diminuição de 22,4 % da concentração sérica de leptina. Tendo em vista os resultados encontrados, pôde-se concluir que a suplementação com ácido linoléico conjugado na concentração de 2 % sobre o consumo médio diário de dieta reduziu a gordura corporal e aumentou os teores de cinzas em ratos / Abstract: Conjugated linoleic acid (CLA), a group of positional and geometric isomers of linoleic acid with conjugated double bonds, occurs in small quantities in a wide variety of foods. CLA can originate in the rumen by biohydrogenation of fatty acids from ingested food, and by the desaturation of the trans-11 C18:1 fatty acid. Thus, significant concentrations of CLA are found in beef, milk and dairy products. The purpose of this study was to assess the effect of conjugated linoleic acid supplementation on lipid profile and body composition of healthy growing Wistar rats. Two biological assays were performed: one varying CLA supplement concentration in the diet, and another varying the commercial brands of CLA used. For the first assay, 40 albino male, weaning rats were distributed at random in 4 groups of 10 animals each, according to the amount of supplement to be administered. Animals in groups AE1, AE2 and AE4 were supplemented daily for 21 days with the commercial product AdvantEdge® CLA (EASTM) at 1, 2 and 4 % of food intake respectively, and those in group C (control) with linoleic acid at 2% of food intake. The aim of this first assay was to find the optimum amount of supplement for the purpose of body fat reduction. In the second assay, 30 albino male, weaning Wistar rats were distributed at random in 3 groups of 10 animals each, according to the brand of supplement. Animals were supplemented daily for 42 days at a concentration of 2 %, chosen on the basis of results in the previous assay. Group AE received AdvantEdge® CLA (EASTM); group CO was fed CLA One® (Pharmanutrients); and group C (control) was given linoleic acid at 2 % of food intake. Throughout the experimental period animals had their weight and food intake controlled every 2 days. At the end of each experiment, the animals were killed by cervical displacement under anesthesia (sodium pentobarbital . 46 mg/kg). The blood was used for the determinations of serum triacylglycerols, total cholesterol and leptin; and the carcass was used for determining body composition. Gut contents were removed to obtain empty carcass weight. The carcass was then frozen in liquid nitrogen, chopped, dried, ground and stored at - 80 °C until determinations of water, ash, protein and fat were performed. Feeding efficiency of the rats was not altered by CLA supplementation in either of the assays. No significant difference (p > 0.05) was observed in the serum levels of triacylglycerols after supplementation with CLA. Total cholesterol values, as measured in the first essay after 21 days of treatment, presented a dose-dependent reduction. In the second assay, however, CLA supplementation was found to increase total cholesterol after 42 days. An average reduction of 18.0 % on body fat percentage was found in groups AE2 (11.2 %) and AE4 (11.6 %), compared to the control (13.9 %). Body fat percentage was also reduced by 18.0 % in the second assay in groups AE and CO, compared to the control (18.1 %, 16.7 % e 21.2 %, respectively). After 42 days of CLA supplementation, animals in groups AE and CO, in the second assay, displayed an increase of 7.5 % in ash content and a decrease of 22.4 % in the serum leptin concentration. Considering the results obtained it can be concluded that the conjugated linoleic acid supplementation at a concentration of 2 % of food intake reduced the body fat and increased the ash content of rats / Mestrado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Mestre em Alimentos e Nutrição

The capability to characterize disease states by way of determining novel biomarkers has led to a high demand of single cell and organelle analytical methodologies due to the unexpected heterogeneity present in cells of the same type. Lipids are of particular interest in the search for biomarkers due to their active roles in cellular metabolism and energy storage. Analyzing localized lipid chemistry from individual cells and organelles is challenging however, due to low analyte volume, limited discriminate instrumentation, and common requirements of separation procedures and expenditure of cell sample. Using nanomanipulation in combination with mass spectrometry, individual cells and organelles can be extracted from tissues and cultures in vitro to determine if heterogeneity at the cellular level is present. The discriminate extraction of a single cell or organelle allows the remainder of cell culture or tissue to remain intact, while the high sensitivity and chemical specificity of mass spectrometry provides structural information for limited volumes without the need for chromatographic separation. Mass analysis of lipids extracted from individual cells can be carried out in multiple mass spectrometry platforms through direct-inject mass spectrometry using nanoelectrospray-ionization and through matrix-assisted laser/desorption ionization.