Redução do teor de limoneno e bergapteno do óleo essencial de bergamota adsorvido em sílica gel pelo CO2 - supercrítico

Testou-se a utilização do CO2 supercrítico na extração seletiva de compostos oxigenados (linalol e acetato de linalila) contidos no óleo essencial de bergamota (Citrus bergamia Risso), var. bergamia, adsorvido em sílica gel contendo baixo teor de limoneno e bergapteno. A densidade do dióxido de carbono e a temperatura de extração foram otimizadas, e os extratos obtidos foram analisados por cromatografia. Nas condições estudadas, observou-se uma redução de 72% no teor de limoneno e 92,5% para o bergapteno, sendo extraídos 52% de linalol e 69,4% de acetato de linalila.

Implementação da técnica de extração de dispersão da matriz em fase sólida ("MSPD") para determinação de resíduos de agrotóxicos em laranjas

Um método de extração de multiresíduos baseado na técnica de dispersão da matriz em fase sólida ("MSPD"), foi otimizado e validado para a extração e análise cromatográfica de 27 agrotóxicos (isômeros a, b, g, d do hexaclorociclohexano (HCH), dieldrin, endrin, heptacloro e seu epóxido (HE), a e b-endosulfan, o,p'-DDD, p,p'-DDD, o,p'-DDE, p,p'-DDE, o,p'-DDT, p,p'-DDT, dicofol, metoxicloro, mirex, hexaclorobenzeno (HCB), clorotalonil, parationa metílica, fenitrotiona, malationa, folpete, diazinona, cis e trans-permetrina em laranjas. A amostra macerada é inicialmente homogeneizada com C18, o homogeneizado é transferido para uma coluna de vidro contendo sílica gel onde se adiciona 10mL de acetato de etila como solvente de eluição. O eluente é concentrado, diluído em isooctano e 1mL deste é injetado no cromatógrafo a gás. Para a separação e quantificação dos 27 agrotóxicos, foi utilizado um CG/DCE. A confirmação dos mesmos foi feita por espectrometria de massas. Para a quantificação dos 27 agrotóxicos utilizou-se a padronização externa. As recuperações dos mesmos variaram de 70 a 120%, considerando-se os níveis de adição agrotóxicos/amostra de 0,02 e 2,0mg/kg. Os limites de quantificação variaram de 0,01 a 0,5mg/kg.

Influência de hidrocolóides na textura de gel de amido de milho

O presente trabalho tem como objetivo determinar a influência das interações das gomas xantana e guar com o amido de milho de alto teor de amilose na textura de gel durante o armazenamento. Foram utilizadas amostras de amido milho Hylon VII® (71% de amilose, National Starch, goma guar (Higum 55I®, Rhodia) e goma xantana (Rhodigel 200®, Rhodia). Foram utilizadas diferentes concentrações das gomas guar e xantana, que variaram de 0 a 1%, de acordo com o delineamento experimental central composto rotacional. Em cada tratamento foram utilizadas 50g de amido com alto teor de amilose (Hylon VII), adicionadas das gomas. Estas amostras foram diluídas em água destilada e submetidas à agitação mecânica até completa dissolução. As soluções foram aquecidas até 95°C por 5 minutos para formação dos géis, os quais foram acondicionados em recipientes plásticos de 50mL e mantidos em temperatura de 5-10°C até 120h. Nos tempos T1 (24h), T2 (48h), T3 (72h), T4 (96h) e T5 (120h) de armazenamento foram feitas medidas da força máxima do gel de amilose em texturômetro (Stable Micro-System, Modelo TAX-T2). No período inicial, de 24 horas, a goma guar não apresentou influência positiva na redução da força do gel, sendo que a aplicação de goma xantana entre 0,7 e 1,0% apresentou os menores valores de força do gel de amilose. Após 120h de armazenamento, a força do gel de amilose diminuiu com a adição de 0,5-1,0% de goma xantana e 0-0,15% de goma guar.

Imobilização de enzimas de milho maltado em gel

Este trabalho objetivou a otimização da imobilização das enzimas amilases extraídas do malte do milho, usando alginato de sódio. A concentração do malte no extrato, a porcentagem de alginato de sódio e o pH foram usados como fatores que influenciam na imobilização das enzimas. Os resultados mostraram que as melhores condições de imobilização foram obtidas quando se usou as soluções de malte de milho em duas faixas de concentrações, uma entre 3,75 a 5 g.L-1 e outra entre 15 a 16,25 g.L-1, em pH entre 4,83 a 6,6 e 4% (m/v) de alginato de sódio, condições nas quais se conseguiu imobilizar 100% das enzimas com baixa perda de atividade. Este trabalho mostrou como se obter amilases de malte de milho imobilizadas por oclusão em alginato de sódio e que podem ser usadas em processos industriais de hidrólise de amido.

Chemical and physical properties of aloe vera (Aloe barbadensis Miller) gel stored after high hydrostatic pressure processing

The aim of this study was to evaluate the influence of high hydrostatic pressure (150, 250, 350, 450, and 550 MPa), applied for 5 minutes, on antioxidant capacity, total phenolic content, color, firmness, rehydration ratio, and water holding capacity of aloe vera gel stored for 60 days at 4 °C. The analyzed properties of the pressurized gel showed significant changes after the storage period. The highest value of total phenolic content was found at 550 MPa. However, a decrease in the antioxidant capacity was observed for all pressurized gel samples when compared to the control sample (p < 0.05). The smallest changes in product color were observed at pressure levels between 150 and 250 MP. The application of high hydrostatic pressure resulted in lower gel firmness, and the lowest value was found at 150 MPa (p < 0.05). On the other hand, the untreated sample showed a greater decrease in firmness, indicating that high pressure processing preserves this property. The application of high hydrostatic pressure exhibited modifications in the food matrix, which were evaluated in terms of rehydration ratio and water holding capacity.

ACE-I inhibitory properties of hydrolysates from germinated and ungerminated Phaseolus lunatus proteins

Phaseolus lunatus protein concentrates and the proteases Alcalase(R) and Pepsin-Pancreatin were used for the production of protein hydrolysates that inhibit angiotensin-I converting enzyme (ACE). Protein concentrate obtained from germinated and ungerminated seeds flour was hydrolyzed with Alcalase(R) at enzyme/substrate ratio (E/S) 1/10 and during 0.5 and 2.0 h, respectively. On the other hand, protein concentrate obtained from ungerminated (E/S: 1/10) and germinated (E/S: 1/50) seeds flour was sequentially hydrolyzed with Pepsin-Pancreatin during 1.0 and 3.0 h, respectively. Peptide fractions with ACE inhibitory activity in a range of 0.9 to 3.8 µg/mL were obtained by G-50 gel filtration chromatography and high- performance liquid chromatography C18 reverse phase chromatography. The observed amino acid composition suggests a substantial contribution of hydrophobic residues to the peptides’ inhibitory potency, which potentially acts via blocking of angiotensin II production. These results show that P. lunatus seed proteins are a potential source of ACE inhibitory peptides when hydrolyzed with Alcalase(R) and Pepsin-Pancreatin.

Umidificação de sementes de girassol após ultrassecagem em sílica gel e câmara de secagem

Dentre os processos que precedem a conservação em longo prazo, a secagem tem papel fundamental, uma vez que o conteúdo de água das sementes afeta diretamente a sua longevidade. Os objetivos desse trabalho foram pesquisar o efeito da umidificação prévia das sementes após ultrassecagem em diferentes teores de água sobre a qualidade fisiológica de sementes de girassol. Sementes com conteúdo inicial de água de 4,7% foram embebidas previamente até conteúdo de água de 10,2%, e submetidas à secagem lenta, conduzida em câmara de secagem, e à secagem rápida, em sílica gel, até conteúdos de água de 7,4; 3,2; 2,9% e 5,3; 3,2; 2,1%, respectivamente. Após a secagem rápida ou lenta, as sementes foram submetidas ou não à umidificação em caixas tipo "gerbox" até o teor de água de 15,8 a 17% e então avaliadas quanto à germinação, peso da matéria seca de raiz, parte aérea e total das plântulas e teste de condutividade elétrica. O delineamento experimental foi o inteiramente casualizado em esquema fatorial 5 (teores de água) x 2 (com e sem umidificação). Quando utilizado a secagem rápida, a germinação das sementes não é prejudicada, e a secagem lenta, até conteúdo de água de 2,9%, proporciona uma redução na germinação das sementes. Sementes de girassol podem ser desidratadas tanto em sílica gel quanto em câmara de secagem até teores de água de 3,2%, sem perda de germinação e vigor. O tratamento de umidificação após secagem propicia um melhor desenvolvimento de raiz e menores valores de condutividade elétrica nas sementes desidratadas em sílica gel e em câmara de secagem.

Notice d'une vente... d'un tableau original de Cl. Gelé (dit le Lorrain), et Jean Miel... 30 avril 1839...

[Vente (Art). 1839-04-30. Paris]

Investigation of single tryptophan proteins encapsulated in TEOS-derived sol-gel matrices by fluorescence spectroscopy /

In the work reported here, optically clear, ultrathin TEOS derived sol-gel slides which were suitable for studies of tryptophan (Trp) fluorescence from entrapped proteins were prepared by the sol-gel technique and characterized. The monitoring of intrinsic protein fluorescence provided information about the structure and environment of the entrapped protein, and about the kinetics of the interaction between the entrapped protein and extemal reagents. Initial studies concentrated on the single Trp protein monellin which was entrapped into the sol-gel matrices. Two types of sol-gel slides, termed "wet aged", in which the gels were aged in buffer and "dry-aged", in which the gels were aged in air , were studied in order to compare the effect of the sol-gel matrix on the structure of the protein at different aging stages. Fluorescence results suggested that the mobility of solvent inside the slides was substantially reduced. The interaction of the entrapped protein with both neutral and charged species was examined and indicated response times on the order of minutes. In the case of the neutral species the kinetics were diffusion limited in solution, but were best described by a sum of first order rate constants when the reactions occurred in the glass matrix. For charged species, interactions between the analytes and the negatively charged glass matrix caused the reaction kinetics to become complex, with the overall reaction rate depending on both the type of aging and the charge on the analyte. The stability and conformational flexibility of the entrapped monellin were also studied. These studies indicated that the encapsulation of monellin into dry-aged monoliths caused the thermal unfolding transition to broaden and shift upward by 14°C, and causedthe long-term stability to improve by 12-fold (compared to solution). Chemical stability studies also showed a broader transition for the unfolding of the protein in dry-aged monoliths, and suggested that the protein was present in a distribution of environments. Results indicated that the entrapped proteins had a smaller range of conformational motions compared to proteins in solution, and that entrapped proteins were not able to unfold completely. The restriction of conformational motion, along with the increased structural order of the internal environment of the gels, likely resulted in the improvements in themial and long-term stability that were observed. A second protein which was also studied in this work is the metal binding protein rat oncomodulin. Initially, the unfolding behavior of this protein in aqueous solution was examined. Several single tryptophan mutants of the metal-binding protein rat oncomodulin (OM) were examined; F102W, Y57W, Y65W and the engineered protein CDOM33 which had all 12 residues of the CD loop replaced with a higher affinity binding loop. Both the thermal and the chemical stability were improved upon binding of metal ions with the order apo < Ca^^ < Tb^"^. During thermal denaturation, the transition midpoints (Tun) of Y65W appeared to be the lowest, followed by Y57W and F102W. The placement of the Trp residue in the F-helix in F102W apparently made the protein slightly more thermostable, although the fluorescence response was readily affected by chemical denaturants, which probably acted through the disruption of hydrogen bonds at the Cterminal end of the F-helix. Under both thermal and chemical denaturation, the engineered protein showed the highest stability. This indicated that increasing the number of metal ligating oxygens in the binding site, either by using a metal ion with a higher coordinatenumber (i.e. Tb^*) which binds more carboxylate ligands, or by providing more ligating groups, as in the CDOM33 replacement, produces notable improvements in protein stability. Y57W and CE)OM33 OM were chosen for further studies when encapsulated into sol-gel derived matrices. The kinetics of interaction of terbium with the entrapped proteins, the ability of the entrapped protein to binding terbium, as well as thermal stability of these two entrapped protein were compared with different levels of Ca^"*^ present in the matrix and in solution. Results suggested that for both of the proteins, the response time and the ability to bind terbium could be adjusted by adding excess calcium to the matrix before gelation. However, the less stable protein Y57W only retained at most 45% of its binding ability in solution while the more stable protein CDOM33 was able to retain 100% binding ability. Themially induced denaturation also suggested that CDOM33 showed similar stability to the protein in solution while Y57W was destabilized. All these results suggested that "hard" proteins (i.e. very stable) can easily survive the sol-gel encapsulation process, but "soft" proteins with lower thermodynamic stability may not be able to withstand the sol-gel process. However, it is possible to control many parameters in order to successfully entrap biological molecules into the sol-gel matrices with maxunum retention of activity.

A fast atom bombardment mass spectrometry study of tris (3, 6-dioxaheptyl) amine-alkali metal halide complexes and hydrogen bonded complexes

The objective of this thesis was to demonstrate the potential of fast atom bombardment mass spectrometry (FABMS) as a probe of condensed phase systems and its possible uses for the study of hydrogen bonding. FABMS was used to study three different systems. The first study was aimed at investigating the selectivity of the ligand tris(3,6-dioxaheptyl) amine (tdoha) for the alkali metal cations. FABMS results correlated well with infrared and nmr data. Systems where a crown ether competed with tdoha for a given alkali metal cation were also investigated by fast atom bombardment. The results were found to correlate with the cation affinity of tdoha and the ability of the crown ether to bind the cation. In the second and third studies, H-bonded systems were investigated. The imidazole-electron donor complexes were investigated and FABMS results showed the expected H-bond strength of the respective complexes. The effects of concentration, liquid matrix, water content, deuterium exchange, and pre-ionization of the complex were also investigated. In the third system investigated, the abundance of the diphenyl sulfone-ammonium salt complexes (presumably H-bonded) in the FABMS spectrum were found to correlate with qualitative considerations such as steric hindrance and strength of ion pairs.

Fluorometric monitoring of the properties of proteins entrapped in sol-gel derived matrices

The successful development of stable biosensors incorporating entrapped proteins suffers from poor understanding of the properties of the entrapped biomolecules. This thesis reports on the use of fluorescence spectroscopy to investigate the properties of proteins entrapped in sol-gel processed silicate materials. Two different single tryptophan (Trp) proteins were investigated in this thesis, the Ca2 + binding protein cod III parvalbumin (C3P) and the salicylate binding protein human serum albumin (HSA). Furthermore, the reactive single cysteine (Cys) residue within C3P and HSA were labelled with the probes iodoacetoxynitrobenzoxadiazole (C3P) and acrylodan (C3P and HSA) to further examine the structure, stability and function of the free and entrapped proteins. The results show that both C3P and HSA can be successfully entrapped into sol-gelderived matrices with retention of function and conformational flexibility.

Investigations into the extraction and the determination of polycyclic aromatic hydrocarbons (PAHs) from water by solid-phase extraction and capillary gas chromatography/ mass spectrometry

Factors affecting the detennination of PAHs by capillary GC/MS were studied. The effect of the initial column temperature and the injection solvent on the peak areas and heights of sixteen PAHs, considered as priority pollutants, USillg crosslinked methyl silicone (DB!) and 5% diphenyl, 94% dimethyl, 1% vinyl polysiloxane (DBS) columns was examined. The possibility of using high boiling point alcohols especially butanol, pentanol, cyclopentanol, and hexanol as injection solvents was investigated. Studies were carried out to optimize the initial column temperature for each of the alcohols. It was found that the optimum initial column temperature is dependent on the solvent employed. The peak areas and heights of the PAHs are enhanced when the initial column temperature is 10-20 c above the boiling point of the solvent using DB5 column, and the same or 10 C above the boiling point of the solvent using DB1 column. Comparing the peak signals of the PAHs using the alcohols, p-xylene, n-octane, and nonane as injection solvents, hexanol gave the greatest peak areas and heights of the PAHs particularly the late-eluted peaks. The detection limits were at low pg levels, ranging from 6.0 pg for fluorene t9 83.6 pg for benzo(a)pyrene. The effect of the initial column temperature on the peak shape and the separation efficiency of the PARs was also studied using DB1 and DB5 columns. Fronting or splitting of the peaks was obseIVed at very low initial column temperature. When high initial column temperature was used, tailing of the peaks appeared. Great difference between DB! and.DB5 columns in the range of the initial column temperature in which symmetrical.peaks of PAHs can be obtained is observed. Wider ranges were shown using DB5 column. Resolution of the closely-eluted PAHs was also affected by the initial column temperature depending on the stationary phase employed. In the case of DB5, only the earlyeluted PAHs were affected; whereas, with DB1, all PAHs were affected. An analytical procedure utilizing solid phase extraction with bonded phase silica (C8) cartridges combined with GC/MS was developed to analyze PAHs in water as an alternative method to those based on the extraction with organic solvent. This simple procedure involved passing a 50 ml of spiked water sample through C8 bonded phase silica cartridges at 10 ml/min, dried by passing a gentle flow of nitrogen at 20 ml/min for 30 sec, and eluting the trapped PAHs with 500 Jll of p-xylene at 0.3 ml/min. The recoveries of PAHs were greater than 80%, with less than 10% relative standard deviations of nine determinations. No major contaminants were present that could interfere with the recognition of PAHs. It was also found that these bonded phase silica cartridges can be re-used for the extraction of PAHs from water.

Sol-gel routes to supported Friedel-Crafts alkylation catalysts

Aluminosilicate catalysts containing supported ZnCl2 and metal fluoride salts have been prepared using a sol-gel based route, tested and characterized. The activities of these ZnCl2 + metal fluoride catalysts, while greater than "Clayzic" (ZnCI2 supported on montmorillonite KIO) are not as good as supported ZnCl2 only supported on aluminosilicate. Alumina supports have also been prepared via a sol-gel route using various chemical additives to generate a mesoporous structure, loaded with ZnCl2 and tested for activity. The activities for these alumina-supported catalysts are also significantly higher than that of "Clayzic", an effective Friedel-Crafts catalyst. Characterizations of these two types of catalysts were done by magic angle spinning (MAS) NMR, diffuse reflectance infrared (DRIFT) spectroscopy and additionally for the alumina nitrogen adsorption studies were done. Supported aluminum trichloride was also investigated as an alternative to the traditional use of aluminum trichloride.

Transmission measurements using a new far-infrared set-up

A system comprised of a Martin-Puplett type polarizing interferometer and a Helium-3 cryostat was developed to study the transmission of materials in the very-far-infrared region of the spectrum. This region is of significant interest due to the low-energy excitations which many materials exhibit. The experimental transmission spectrum contains information concerning the optical properties of the material. The set-up of this system is described in detail along with the adaptations and improvements which have been made to the system to ensure the best results. Transmission experiments carried out with this new set-up for two different varieties of materials: superconducting thin films of lead and biological proteins, are discussed. Several thin films of lead deposited on fused silica quartz substrates were studied. From the ratio of the transmission in the superconducting state to that in the normal state the superconducting energy gap was determined to be approximately 25 cm-1 which corresponds to 2~/kBTc rv 5 in agreement with literature data. Furthermore, in agreement with theoretical predictions, the maximum in the transmission ratio was observed to increase as the film thickness was increased. These results provide verification of the system's ability to accurately measure the optical properties of thin low-Tc superconducting films. Transmission measurements were carried out on double deionized water, and a variety of different concentrations by weight of the globular protein, Bovine Serum Albumin, in the sol, gel and crystalline forms. The results of the water study agree well with literature values and thus further illustrate the reproducibility of the system. The results of the protein experiments, although preliminary, indicate that as the concentration increases the samples become more transparent. Some weak structure in the frequency dependent absorption coefficient, which is more prominent in crystalline samples, may be due to low frequency vibrations of the protein molecules.