Evaluation of total carotenoids, alpha- and beta-carotene in carrots (Daucus carota L.) during home processing

This study aims to analyze the influence of dehydration and different preparation methods during home processing related toalpha-carotene, beta-carotene and total carotenoids stability in carrots. Vitamin A values were evaluated after different treatments. Thus, carrots were submitted to steam cooking, water cooking with and without pressure, moist/dry cooking and conventional dehydration. Determination of alpha- and beta-carotenes was made by High-Performance Liquid Chromatography (HPLC) (conditions were developed by us) using spectrophotometric detection visible-UV at 470 nm; a RP-18 column and methanol: acetonitrile: ethyl acetate (80: 10: 10) as mobile phase. Total carotenoids quantification was made by 449 nm spectrophotometer. The retention of the analyzed carotenoids ranged from 60.13 to 85.64%. Water cooking without pressure promoted higher retention levels of alpha- and beta-carotene and vitamin A values, while water cooking with pressure promoted higher retention levels of total carotenoids. Dehydration promoted the highest carotenoid losses. The results showed that, among the routinely utilized methods under domestic condition, cooking without pressure, if performed under controlled time and temperature, is the best method as it reduces losses in the amount of alpha- and beta-carotene, the main carotenoids present in the carrots. Despite the significant carotenoid losses, carrots prepared through domestic methods, remain a rich source of provitamin A.

Estudo da produção de beta-galactosidase por fermentação de soro de queijo com Kluyveromyces marxianus

A hidrólise enzimática da lactose por beta-galactosidase desempenha importante papel no processamento de produtos lácteos, como na obtenção de leite com baixo teor de lactose para consumo por indivíduos intolerantes à mesma e na prevenção da cristalização em produtos de laticínio. Neste trabalho, a enzima beta-galactosidase foi produzida pelo cultivo do microrganismo Kluyveromyces marxianus, em meio de cultura à base de soro de queijo em diferentes concentrações iniciais de lactose e extrato de levedura, de acordo com um planejamento fatorial. As fermentações foram conduzidas em incubador rotativo a 150rpm, a 30°C e pH inicial 5,5. A concentração celular inicial foi de 10(7) células/mL. Para a extração da enzima beta-galactosidase, foi realizada autólise das células utilizando como solvente o clorofórmio em tampão fosfato. No meio de cultura enriquecido com (NH4)2SO4, KH2PO4 e MgSO4, nas concentrações iniciais de lactose e de extrato de levedura iguais a 50g/L e 12g/L, respectivamente, obteve-se uma atividade de 28,0UGl/mL e uma concentração celular máxima de 5,3g/L.

Incorporation of conjugated linoleic and alpha linolenic fatty acids into Pimedolus maculatus fillets

The objective of this study was to investigate the fatty acid composition of Pimedolus maculatus fillets of fish fed with different diets. The fish were submitted to an adaptation period (over 30 days) and fed only a control diet (diet I). After this period, the fish were divided into two groups: one group received a diet enriched with flaxseed oil (diet II), and the other received a diet enriched with conjugated linoleic acid (CLA) (diet III). Subsequent collections were undertaken after 25 and 50 days. Compared to the control samples, fish fed diet II showed an increase in concentrations of total fatty acid omega-3 (n-3) in the fillets, with an improvement ratio of n-6/n-3. The incorporation of CLA into fish fed diet III was successful. Supplementing the diet of Pimedolus maculatus with CLA and flaxseed oil led to improvements in nutritional quality and fatty acid profile of the fish fillets.

In-house method validation and occurrence of alpha-, beta-endosulfan, endosulfan sulphate, lambda-cyhalothrin, procymidone and trifluralin residues in strawberry

A method for determination of organohalogen pesticides in strawberry by gas chromatography with electron capture detection was validated and applied in a monitoring program. Linearity, matrix effects, and day effect were evaluated for the analytes alpha-endosulfan, beta-endosulfan, endosulfan sulphate, lambda-cyhalothrin, procymidone, and trifluralin. The linear range varied according to the chromatographic response of the analyte. Significant matrix effects were observed. The mean recoveries ranged from 74.6 to 115.4%, with repeatability standard deviations between 1.6 and 21.0% and intermediate precision between 5.9 and 21.0%. Detection, quantification and decision limit, and detection capacity ranged from 0.003 to 0.007 mg/kg, 0.005 to 0.013 mg/kg; 0.003 to 3.128 mg/kg; and 0.005 to 3.266 mg/kg, respectively. The method was fit for the purpose of monitoring organohalogen residues in strawberries. Residues of these pesticides were detected in 124 of the 186 samples analyzed between 2009 and 2011 in the state of Minas Gerais. Nine of them did not comply with the current legislation requirements; among them, seven (3.8%) had residues of unauthorized pesticide for the culture of strawberry, one (0.5%) had residues above the maximum residue limit, and another one (0.5%) exhibited both non-conformities.

Alpha-tocopherol and gamma-tocopherol concentration in vegetable oils

Vegetable oils are the richest dietary sources of vitamin E. Vitamin E determination levels in foods are of great importance to adjust the ingestion of nutrients by the population. The purpose of this paper is to determine the concentration of alpha-tocopherol and gamma-tocopherol in vegetable oils and compare the alpha-tocopherol value to the nutritional requirement of vitamin E. The analysis was performed using High Performance Liquid Chromatography. The values expressed as mg/kg for alpha and gamma-tocopherol were, respectively, 120.3±4.2 and 122.0±7.9 in canola oil; 432.3±86.6 and 92.3±9.5 in sunflower oil; 173.0±82.3 and 259.7±43.8 in corn oil; 71.3±6.4 and 273.3±11.1 in soybean oil. A significant difference was encountered between the alpha-tocopherol concentrations in vegetable oils. Similar results were found for gamma-tocopherol, except for corn and soybean oils. It was concluded that the soybean oil was not considered a source of vitamin E. The canola and corn oils were considered sources, and the sunflower oil was considered an excellent source.

Lactose hydrolysis potential and thermal stability of commercial β-galactosidase in UHT and skimmed milk

Abstract The commercial enzyme (E.C. = 3.2.1.23) from Kluyveromyces lactis (liquid) and Aspergillus oryzae(lyophilized) was investigated for its hydrolysis potential in lactose substrate, UHT milk, and skimmed milk at different concentrations (0.7; 1.0 and 1.5%), pH values (5.0; 6.0; 6.5 and 7.0), and temperature (30; 35; 40 and 55 ºC). High hydrolysis rates were observed for the enzyme from K. lactis at pH 7.0 and 40 ºC, and from A. oryzae at pH 5.0 and 55 ºC. The enzyme from K. lactis showed significantly higher hydrolysis rates when compared to A. oryzae. The effect of temperature and β-galactosidase concentration on the lactose hydrolysis in UHT milk was higher than in skimmed milk, for all temperatures tested. With respect to the thermal stability, a decrease in hydrolysis rate was observed at pH 6.0 at 35 ºC for K. lactisenzyme, and at pH 6.0 at 55 ºC for the enzyme from A. oryzae. This study investigate the hydrolysis of β-galactosidase in UHT and skimmed milk. The knowledge about the characteristics of the β-galactosidase fromK. lactis and A. oryzae enables to use it most efficiently to control the enzyme concentration, temperature, and pH in many industrial processes and product formulations.

Action of gibberellic acid (GA3) on dormancy and activity of alpha-amylase in rice seeds

To evaluate the effectiveness of gibberellic acid (GA3) in breaking rice seed dormancy and the use of alpha-amylase enzyme activity as an indicator of the dormancy level, seed from the intensively dormant irrigated cultivar Urucuia were used. The seeds were submitted to a pre-drying process in a forced air circulation chamber under 40ºC during 7 days and submersed in 30 mL of GA3 solution under 0, 10, 30 and 60 mg/L H2O concentrations, during 2, 24 and 36 hours. After the treatments, the alpha-amylase activity was determined by using the polyacrilamide electrophoresis and spectrophotometry. At the same time, the germination test was made. The results indicated a gain in germination and in alpha-amylase activity in higher concentrations and soaking time of seeds in GA3. These observations support the conclusion that soaking seed in 60 mg GA3/L during 36 hours can be used as a quick and efficient treatment in breaking rice seed dormancy and is equivalent to the forced air circulation chamber at 40ºC during 7 days. The alpha-amylase enzyme activity proved to be as an efficient marker of the seeds dormancy level.

Caracterização de alfa-galactosidase em embrião e cotilédones de sementes de Platymiscium pubescens Micheli, var. pubecens (tamboril-da-mata)

Sementes de Platymiscium pubescens foram colocadas para embeber em água, sendo retiradas amostras para as caracterizações bioquímica e cinética da enzima alfa-galactosidase do eixo embrionário e dos cotilédones. A atividade específica no eixo embrionário aumenta de zero até o tempo de 96 horas de embebição, estabilizando em seguida. A atividade da enzima dos cotilédones mostrou pequeno incremento durante esse mesmo tempo. A alfa-galactosidase do eixo embrionário apresentou atividade máxima no intervalo de pH de 4,5 a 6,0. Por outro lado, para a enzima proveniente dos cotilédones, a maior atividade foi detectada na faixa de 4,0 a 6,0. A temperatura de 55ºC foi a que mais estimulou as atividades da alfa-galactosidase do eixo embrionário e dos cotilédones. As enzimas do eixo embrionário e dos cotilédones mostraram-se termotolerantes, não se alcançando a meia vida na temperatura de 40ºC, no período de 1.500 minutos. A atividade da alfa-galactosidase do eixo embrionário foi inibida por melibiose, CuSO4 e SDS, enquanto a dos cotilédones foi por todos os efetores, exceto com SDS, CuSO4 e galactose que tiveram efeito neutro sobre a atividade da alfa-galactosidase dos cotilédones. Os valores de K M para as alfa-galactosidases do embrião e para o cotilédone foram 3,37 e 0,26 mM, respectivamente.

Alterações nas atividades das enzimas alfa-galactosidase e poligalacturonase e nas reservas de carboidratos de sementes de Schizolobium parahyba (vell.) Blake (guapuruvú) durante a germinação

A retomada do metabolismo do embrião durante a germinação é realizada por processos metabólicos que culminam na protrusão da radícula e no fornecimento de energia para o desenvolvimento inicial da plântula. O objetivo deste trabalho foi estudar as variações nas atividades das enzimas alfa-galactosidase e poligalacturonase e nas reservas de mono e oligossacarídeos em sementes de guapuruvú durante a germinação. Para tanto, as sementes foram colocadas para germinar e as reservas do eixo embrionário e cotilédones, avaliadas periodicamente. Os teores de galactose no eixo embrionário diferiram significativamente somente entre a testemunha e o oitavo dia, muito embora houvesse aumento contínuo até o quarto dia. Somente no sexto dia de germinação houve aumento no teor de galactose nos cotilédones. Houve tendência de aumento nos teores de arabinose, manose e glicose no eixo embrionário, não sendo detectada a presença de xilose no oitavo dia. Nos cotilédones os mesmos açúcares não foram originalmente detectados no tempo zero, mas apresentaram valores mais altos nas amostras do oitavo dia. Os teores de galactose oscilam tanto no eixo embrionário, quanto nos cotilédones durante o período de germinação de sementes de guapuruvu. Os teores de sacarose aumentam e os de rafinose decrescem nos cotilédones e no eixo embrionário. Os teores de estaquiose permanecem aproximadamente estáveis no eixo e nos cotilédones, com decréscimo no eixo, no oitavo dia. A enzima alfa-galactosidase é pré-formada, tendo clara redução na sua atividade específica, no segundo dia, permanecendo constante até o oitavo. A atividade nos cotilédones apresenta aumento no quarto dia, decrescendo posteriormente. A enzima polygalacturonase é tambem pré-formada, com maior atividade inicial no eixo. A atividade nos cotilédones aumenta até o sexto dia, alcançando maiores valores que o do eixo embrionário e, em seguida, decresce para valores menores do que aqueles.

Intramolecular carbenoid insertions : the reactions of [alpha]-diazoketones derived from furanyl, thienyl, benzofuranyl and benzothienyl acetic acids with rhodium (II) acetate /

A number of synthetically useful ring systems can be prepared via the intramolecular insertion of a metal-stabilized carbenoid into a heteroaromatic systems. The chemical outcome of these reactions are dependent not only on the nature of the heteroatom but also on the length of the aliphatic tether linking the carbenoid moiety with the aromatic fragment. Our work with furanyl and thienyl systems containing a single methylene tether have allowed for some rather atypical chemistry. For example, treatment of l-diazo-3-(2-thienyl)-2-propanone (6) with catalytic rhodium (II) acetate yields 5,6- dihydro-4^-cyclopenta[Z>]thiophen-5-one (3) while, the isomeric l-diazo-3-(3-thienyl)-2- propanone(15) gives a spiro-disulphide (20). Novel chemistry was also exhibited in the analogous furanyl systems. While treatment of l-diazo-3-(3-furanyl)-2-propanone (52) with Rh2(OAc)4 resulted in the expected 2-(4-Oxo-2-cyclopentenyliden)acetaldehyde (54), isomeric l-diazo-3-(2- furanyl)-2-propanone (8) undergoes vinylogous Wolff rearrangement to give a mixture of 6a-methyl-2,3,3a,6a-tetrahydrofuro[2,i-^>]furan-2-one (44) and 2-(2-methyl-3-furyl)acetic acid (43). Rhodium acetate catalyzed decomposition of l-diazo-3-(3-benzofuranyl)-2- propanone (84) and l-diazo-3-(2-benzofuranyl)-2-propanone (69)also allows for vinylogous Wolff rearrangement, a chemistry unseen in benzofuranyl systems with longer tethers. A number of interesting products were isolated from the trapping of intermediate ketenes. Decomposition of l-diazo-3-(3-benzothienyl)-2-propanone (100) resulted in the formation of 2,3-dihydro-l//-benzo[^]cyclopenta[^thiophen-2-one (102). However, in addition to (102), a dimer was also generated from the decomposition of l-diazo-3-(2- benzothienyl)-2-propanone (109). The insight into the mechanistic underpinnings of the above reactions are provided by molecular modeling at a PM3 level.

Intramolecular carbenoid insertion : reactions of [alpha] -diazoketones derived from benzothienyl, pyrolyl and indolyl alkanoic acids with rhodium (II) acetate

Recent studies have shown that the rhodium (II) acetate decomposition chemistry observed for a-diazoketones tethered to thienyl, furanyl, and benzofuranyl moieties is dependent not only on the nature of the heteroatom but also on the length of the aliphatic tether linking the diazoketone moiety with the aromatic fragment. The present thesis expands on these results and focuses on a-diazoketones tethered to benzothiophenes, pyrroles and indoles by a methylene linker. In the case of benzothiophenes, it was shown that the rhodium catalyst decomposition of I-diazo-4-(3-benzothienyl)-2-butanone (146) and 1-diazo-4-(3benzothienyl)- 2-butanone (152) allow for the isolation of 1,2,3a,3b-tetrahydro-3Hbenzo[ b]cyclopenta[1,3]cyclopropa- [1 ,2-d]thiophen-3-one (147) and 1,2,3a,3btetrahydro- 3H-benzo[b]cyclopenta[1,3]cyclopropa[1,2-d]thiophen-3-one (153). However treatment of 1-diazo-3-(3-Benzothienyl)-2-Propanone (165) with Rh(II) acetate results in the formation of 2,3-Dihydro-1H-benzo[b]cyclopenta[d]thiophen-2-one (159), while 1diazo- 3-(2-Benzothienyl)-2-Propanone with the same condition gives 5,5-bis( 1benzothiophen- 2-ylmethyl)-2(5H)-furanone (166) along with the tricycle 159. The chemistry of the pyrrolyl and the indolyl moieties linked to terminal adiazoketone systems was also investigated. The decomposition of I-diazo-(2-pyrrolyl)-2propanone (173) results in the formation of two products; the N-H insertion product IHpyrrolizin- 2(3H)-one (176) and the alkylation product 4,6-dihydrocyclopenta[b]pyrrol5( 1 H)-one (180). When 1-Diazo-3-(3-indoly)-3-propanone (194) is treated with catalytic amount of Rh (II) 3,4-dihydrocyclopenta[b]indol-2(1H)-one (193) is isolated quantitatively. The later reaction when monitored using IH NMR the intermediate 200 can be seen whose structure was confirmed by the comparison to series of model compounds. The mechanisms underlying these reactions as well as their synthetic utility is discussed.

alpha-Tocopherol's Antioxidant Role: A Biophysical Perspective

I present evidence of an antioxidant mechanism for vitamin E that correlates strongly with its physical location in a model lipid bilayer. These data address the overlooked problem of the physical distance between the vitamin's reducing hydrogen and lipid acyl chain radicals. The combined data from neutron diffraction, NMR and UV spectroscopy experiments, all suggest that reduction of reactive oxygen species and lipid radicals occurs specifically at the membrane's hydrophobic-hydrophilic interface. The latter is possible when the acyl chain adopts conformations in which they snorkel to the interface from the hydrocarbon matrix. Moreover, not all model lipids are equal in this regard, as indicated by the small differences in the vitamin's location. The present result is a clear example of the importance of lipid diversity in controlling the dynamic structural properties of biological membranes. Importantly, these results suggest that measurements of alpha-tocopherol oxidation kinetics, and its products, should be revisited by taking into consideration the physical properties of the membrane in which the vitamin resides.

Evaluación de características morfofisiológicas y de productividad de papa (Solanum tuberosum L.) variedad Alpha proveniente de diferentes condiciones agrotecnológicas [por] Gloria Estela Terán Sarabia

Tesis (Maestría en Ciencias con Especialidad en Botánica) U.A.N.L.