Selective Oxidations of Organoboron Compounds Catalyzed by Baeyer-Villiger Monooxygenases

The applicability of Baeyer-Villiger monooxygenases (BVMOs) in organoboron chemistry has been explored through testing chemo-and enantioselective oxidations of a variety of boron-containing aromatic and vinylic compounds. Several BVMOs, namely: phenylacetone monooxygenase (PAMO), M446G PAMO mutant, 4-hydroxyacetophenone monooxygenase (HAPMO) and cyclohexanone monooxygenase (CHMO) were used in this study. The degree of chemoselectivity depends on the type of BVMO employed, in which the biocatalysts prefer boron-carbon oxidation over Baeyer-Villiger oxidation or epoxidation. Interestingly, it was discovered that PAMO can be used to perform kinetic resolution of boron-containing compounds with good enantioselectivities. These findings extend the known biocatalytic repertoire of BVMOs by showing a new family of compounds that can be oxidized by these enzymes.

Multivariant optimization, validation, and application of capillary electrophoresis for simultaneous determination of polyphenols and phenolic acids in Brazilian wines

A method for the simultaneous determination of the stilbene resveratrol, four phenolic acids (syringic, coumaric, caffeic, and gallic acids), and five flavonoids (catechin, rutin, kaempferol, myricetin, and quercetin) in wine by CE was developed and validated. The CE electrolyte composition and instrumental conditions were optimized using 2(7-3) factorial design and response surface analysis, showing sodium tetraborate, MeOH, and their interaction as the most influential variables. The optimal electrophoretic conditions, minimizing the chromatographic resolution statistic values, consisted of 17 mmol/L sodium tetraborate with 20% methanol as electrolyte, constant voltage of 25 kV, hydrodynamic injection at 50 mbar for 3 s, and temperature of 25 degrees C. The R(2) values for linearity varied from 0.994 to 0.999; LOD and LOQ were 0.1 to 0.3 mg/L and 0.4 to 0.8 mg/L, respectively. The RSDs for migration time and peak area obtained from ten consecutive injections were less than 2% and recoveries varied from 97 to 102%. The method was applied to 23 samples of inexpensive Brazilian wines, showing wide compositional variation.

Detection of adulterations in processed coffee with cereals and coffee husks using capillary zone electrophoresis

The proposed method for the identification of adulteration was based on the controlled acid hydrolysis of xylan and starch present in some vegetable adulterants, followed by the analysis of the resulting xylose and glucose, which are the monosaccharides that compose, respectively, the two polysaccharides. The acid hydrolysis with HCl increases the ionic strength of the sample, which impairs the electrophoretic separation. Thus, a neutralization step based on anion exchange resin was necessary. The best separations were obtained in NaOH 80 mmol/L, CTAB 0.5 mmol/L, and methanol 30% v/v. Because of the high value of pH, monosaccharides are separated as anionic species in such running electrolyte. The LOQ for both monosaccharides was 0.2 g for 100 g of dry matter, which conforms to the tolerable limits.

Development of a sequential injection chromatography (SIC) method for determination of simazine, atrazine, and propazine

This paper describes the development of a sequential injection chromatography (SIC) procedure for separation and quantification of the herbicides simazine, atrazine, and propazine exploring the low backpressure of a 2.5 cm long monolithic C(18) column. The separation of the three compounds was achieved in less than 90 s with resolution > 1.5 using a mobile phase composed by ACN/1.25 mmol/L acetate buffer (pH 4.5) at the volumetric ratio of 35:65 and flow rate of 40 mu L/s. Detection was made at 223 nm using a flow cell with 40 mm of optical path length. The LOD was 10 mu g/L for the three triazines and the quantification limits were of 30 mu g/L for simazine and propazine and 40 mu g/L for atrazine. The sampling frequency is 27 samples per hour, consuming 1.1 mL of ACN per analysis. The proposed methodology was applied to spiked water samples and no statistically significant differences were observed in comparison to a conventional HPLC-UV method. The major metabolites of atrazine and other herbicides did not interfere in the analysis, being eluted from the column either together with the unretained peak, or at retention times well-resolved from the studied compounds.

Minor sesquiterpenes from the volatile oil from leaves of Guarea guidonia Sleumer (Meliaceae)

The essential oil from leaves of Guarea guidonia was subjected to chromatographic separation procedures to afford nine sesquiterpenes; two of them are new eudesmane derivatives. The chemical structures of the obtained compounds were characterised by spectrometric analysis, mainly mass spectrometry and NMR.

Proteome analysis of human dorsolateral prefrontal cortex using shotgun mass spectrometry

The prefrontal cortex executes important functions such as differentiation of conflicting thoughts, correct social behavior and personality expression, and is directly implicated in different neurodegenerative diseases. We performed a shotgun proteome analysis that included IEF fractionation, RP-LC, and MALDI-TOF/TOF mass spectrometric analysis of tryptic digests from a pool of seven human dorsolateral prefrontal cortex protein extracts. In this report, we present a catalog of 387 proteins expressed in these samples, identified by two or more peptides and high confidence search scores. These proteins are involved in different biological processes such as cell growth and/or maintenance, metabolism/energy pathways, cell communication/signal trarisduction, protein metabolism, transport, regulation of nucleobase, nucleoside, nucleotide and nucleic acid metabolism, and immune response. This analysis contributes to the knowledge of the human brain proteome by adding sample diversity and protein expression data from an alternative technical approach. It will also aid comparative studies of different brain areas and medical conditions, with future applications in basic and clinical research.

Development of a new stir bar sorptive extraction coating and its application for the determination of six pesticides in sugarcane juice

In this article, a novel polydimethylsiloxane/activated carbon (PDMS-ACB) material is proposed as a new polymeric phase for stir bar sorptive extraction (SBSE). The PDMS-ACB stir bar, assembled using a simple Teflon (R)/glass capillary mold, demonstrated remarkable stability and resistance to organic solvents for more than 150 extractions. The SBSE bar has a diameter of 2.36 mm and a length of 2.2 cm and is prepared to contain 92 mu L of polymer coating. This new PDMS-ACB bar was evaluated for its ability to determine the quantity of pesticides in sugarcane juice samples by performing liquid desorption (LD) in 200 mu L of ethyl acetate and analyzing the solvent through gas chromatography coupled with mass spectrometry (GC-MS). A fractional factorial design was used to evaluate the main parameters involved in the extraction procedure. Then, a central composite design with a star configuration was used to optimize the significant extraction parameters. The method used demonstrated a limit of quantification (LOQ) of 0.5-40 mu g/L, depending on the analyte detected; the amount of recovery varied from 0.18 to 49.50%, and the intraday precision ranged from 0.072 to 8.40%. The method was used in the analysis of real sugarcane juice samples commercially available in local markets.

Fluoxetine and norfluoxetine analysis by direct injection of human plasma in a column switching liquid chromatographic system

A column switching LC method is presented for the analysis of fluoxetine (FLU) and norfluoxetine (NFLU) by direct injection of human plasma using a lab-made restricted access media (RAM) column. A RAM-BSA-octadecyl silica (C-18) column (40 min x 4.6 mm, 10 mu m) is evaluated in both backflush and foreflush elution modes and coupled with a C-18 lab-made (50 mm x 4.6 mm, 3 pm) analytical column in order to perform online sample preparation. Direct injection of 100 mu L, of plasma samples is possible with the developed approach. In addition, reduction of sample handling is obtained when compared with traditional liquid-liquid extraction (LLE) and SPE. The total analysis time is around 20 min. A LOQ of 15 ng/mL is achieved in a concentration range of 15-500 ng/mL, allowing the therapeutic drug monitoring of clinical samples. The precision values achieved are lower than 15% for all the evaluated points with adequate recovery and accuracy. Furthermore, no matrix interferences are found in the analysis and the proposed method shows to be an adequate alternative for analysis of FLU in plasma.

Characterization of protein hydrolysates of cosmetic use by CE-MS

Protein hydrolysates have been used as active principles in cosmetic products conferring different properties to the final formulations, which are mostly controlled by the peptide size and its amino acid sequence. In this work, capillary electrophoresis coupled to mass spectrometry analyses were carried out in order to investigate such characteristics of protein hydrolysates. Samples of different origins (milk, soy and rice) were obtained from a local company, and were analyzed without a previous preparation step. The background electrolyte (BGE) and sheath liquid compositions were optimized for each sample. The best BGE composition (860 mmol/L formic acid - pH 1.8 - in 70: 30 v/v water/methanol hydro-organic solvent) was chosen based on the overall peak resolution whereas the best sheath liquid was selected based on increased sensitivity and presented different compositions to each sample (10.9-217 mmol/L formic acid in 75: 25-25: 75 v/v water/methanol hydro-organic solvent). Most of the putative peptides in the hydrolysate samples under investigation presented molecular masses of 1000 Da or less. De novo sequencing was carried out for some of the analytes, revealing the hydrophobicity/polarity of the peptides. Hence, the technique has proved to be an advantageous tool for the quality control of industrial protein hydrolysates.

Identification of non-zein proteins in BR473 maize protein bodies by LC-nanoESI-MS/MS

The nutritional value of maize seed is limited due to its high content of storage proteins (zeins), which are deficient in essential amino acids such as lysine and tryptophan. In a previous paper, we showed that protein bodies obtained from BR473 maize variety, developed by Embrapa (Brazilian Agricultural Research Corporation), were mainly constituted by Z27 and a smaller quantity of Z50 gamma-zeins. Besides zein proteins, other not identified protein band in the SDS/PAGE was also observed, which could indicate the presence of non-zein proteins additionally to gamma-zeins. In the present paper, we have demonstrated the presence of non-zein proteins in BR473 maize protein bodies by LC-nanoESI-MS/MS and database searching. This fact could be related to the excellent energetic value and higher protein quality of BR473 maize grains, since high lysine concentration in some maize varieties has been related to the presence of cytoskeleton proteins that are non-zeins. We have identified the following proteins: Brittle-1 protein (chloroplast precursor), Legumin-1, glyceroldehyde-3-phosphate dehydrogenase, and elongation factor 1-alpha.

Coumarins and phenolic fingerprints of oak and Brazilian woods extracted by sugarcane spirit

A total of 25 sugarcane spirit extracts of six different Brazilian woods and oak, commonly used by cooperage industries for aging cachaca, were analyzed for the presence of 14 phenolic compounds (ellagic acid, gallic acid, vanillin, syringaldehyde, synapaldehyde, coniferaldehyde, vanillic acid, syringic acid, quercetin, trans-resveratrol, catechin, epicatechin, eugenol, and myricetin) and two coumarins (scopoletin and coumarin) by HPLC-DAD-fluorescence and HPLC-ESI-MS(n). Furthermore, an HPLC-DAD chromatographic fingerprint was build-up using chemometric analysis based on the chromatographic elution profiles of the extracts monitored at 280 nm. Major components identified and quantified in Brazilian wood extracts were coumarin, ellagic acid, and catechin, whereas oak extracts shown a major contribution of catechin, vanillic acid, and syringaldehyde. The main difference observed among oak and Brazilian woods remains in the concentration of coumarin, catechin, syringaldehyde, and coniferaldehyde. The chemometric analysis of the quantitative profile of the 14 phenolic compounds and two coumarins in the wood extracts provides a differentiation between the Brazilian wood and oak extracts. The chromatographic fingerprint treated by multivariate analysis revealed significant differences among Brazilian woods themselves and oak, clearly defining six groups of wood extracts: (i) oak extracts, (ii) jatoba extracts, (iii) cabreuva-parda extracts, (iv) amendoim extracts, (v) canela-sassafras extracts and (vi) pequi extracts.

Matrix effects observed during pesticides residue analysis in fruits by GC

The influence of the sample matrix in the CC-electron-capture detection analysis of the pesticides dimethoate, diazinon, chlorothalonil.. parathion methyl and fenitrothion in fruits samples has been studied. Experiments have been carried out where the pesticide responses in standard solutions prepared in selected solvent were compared with their response when present in apple, mango, papaya, banana, pineapple and melon extracts. The presence of matrix effects (MEs) and their extent were shown to be simultaneously influenced by several factors (matrix concentration, matrix type, pesticide concentration, analytical range). Pronounced MEs were observed particularly for dimethoate and diazinon in all matrices tested; in lower concentrations, all pesticides presented significant ME. The other pesticides presented variable ME. Higher ME enhancement was detected at lower pesticide concentration levels of and/or at higher matrix concentration solutions. The ME detected for fenitrothion, in the analytical range evaluated, were dependent on matrix type. For each pesticide, solvent and matrix-matched calibrations were compared for all fruit samples, and it could be concluded that quantitation based on standard solutions prepared in blank matrix extract (matrix-matched calibration) should be used to compensate the MEs and to obtain more accurate results for the pesticides studied.

Elevdemokrati i folkhögskola : En undersökning av folkhögskoleelevers uppfattade möjlighet att påverka sin utbildning vid en folkhögskola i mellersta Sverige

Den här uppsatsen är en fallstudie av elevinflytande vid en folkhögskola i mellersta Sverige. Jag har undersökt hur elevinflytandet som fenomen tar sig uttryck vid en folkhögskola. Undersökningen utgår ifrån hur elever uppfattar sin möjlighet att påverka sin utbildning. I vilket jag har undersökt förhållandet mellan informellt och formellt inflytande, och hur förhållandet dem emellan ser ut. Vidare har jag undersökt om det finns någon skillnad mellan de manliga och kvinnliga elevgrupperna i fråga om synen på deras inflytande över sin utbildning och sina studier. Informellt elevinflytande är det inflytande elever har över den rena undervisningssituationen och formellt inflytande är det inflytande vari eleverna som grupp har representanter i skolråd och liknande, samt ett elevråd.Undersökningen bygger på en enkätundersökning, som tagit avstamp i olika kategorier av informellt och formellt inflytande, vari eleverna tagit ställning till vilken position dom anser att deras åsikter har i de olika kategorierna. I det formella inflytandet har eleverna svarat utifrån kategorierna: Skolans fysiska utformning; inköp av material; skolans kurs- och programutbud; avstängning av studerande; avstängning av lärare; anställning av lärare. Och i det informella inflytandet: Val av temaområden; enskilda lektioners utformning; val av arbetsformen; val av läromedel/kunskapskälla; val av examinationsform. I dessa kategorier har jag gjort en indelning av olika nivåer av vilken ställning elevernas åsikter anses ha utifrån den svarandes syn. Nivåerna är en omarbetning av Kjell Jormfeldts modell gällande elevinflytande. Mina inflytandenivåer utgår från elevernas åsikters ställning inför fattande av beslut i de specifika kategoriernas frågor. Dessa nivåer är: För informellt inflytande obefintligt, läraren bestämmer alternativ, läraren tar intryck, avgörande. För formellt inflytande: obefintligt, de lyssnar, de tar intryck, avgörande.I den undersökta skolan har det visat sig att eleverna har en hög nivå av inflytande. Till synes så är det informella inflytandet dock mer utvecklat och ligger på en högre nivå än det formella. I samtliga kategorier av det formella inflytandet så visade det sig att elevernas åsikter, till största delen var i nivån av att de lyssnades på. I kategorierna gällande anställande och avstängande av lärare samt avstängning av elever så finns det klara brister då eleverna i en väldigt stor utsträckning ansåg att deras åsikter hade en obefintlig ställning i dessa frågor. I det informella inflytandet var det väldigt få elever som ansåg att deras inflytande var obefintligt. Tonvikten hos elevernas åsikt, i samtliga kategorier, var att lärarna inte bara lyssnar på elevernas åsikter utan även att de tar intryck av dessa!I jämförelsen mellan de manliga och kvinnliga eleverna har jag inte kunna finna några helhetstäckande mönster, i det informella inflytandet. De få saker jag fann var till exempel att det var en större procent hos den manliga än den kvinnliga gruppen som ansåg att inflytandet var obefintligt gällande kategorierna inom informellt inflytande. Men i det stora hela var det i vissa kategorier som kvinnorna åsikter pekade på ett högre inflytande än hos männen, och i andra kategorier var det helt tvärtom. Gällande formellt inflytande var det dock tydligt att det fanns större andelar kvinnor som ansåg att inflytandet var obefintligt i jämförelse med männen, och på ett liknande sätt ansåg färre av kvinnorna att deras åsikter hade en avgörande ställning i formella frågor jämfört med männen. Jag har därför konstaterat att kvinnorna som grupp anser att inflytandet är lägre i jämförelse med vad männen anser att det är, ifråga om formellt inflytande!

A paixão proibida no Hipólito de Eurípedes e em Fedra de Sêneca

Este estudo visa a analisar a peça Hipólito de Eurípides e a Fedra de Sêneca, a partir da intertextualidade, termo criado e difundido por Julia Kristeva no âmbito dos estudos da Literatura Comparada. Procuramos investigar o tema “a paixão proibida” em ambos os textos, destacando o diálogo mútuo que envolvem tais obras. Nessa pesquisa, estudamos essencialmente o discurso feminino, o desenvolvimento da paixão em seus diversos estágios, o ódio masculino contra as mulheres, o pensamento estóico subjacente nas falas das personagens de Sêneca. Outros aspectos também foram investigados como a caracterização das personagens e o enredo. Sêneca apropriou-se de alguns elementos do texto de Eurípides e reformulou-os em um amplo processo de reescrita o que culminou em uma nova obra estética com traços da sua cultura romana.

Indústria de transformação brasileira: análise do progresso tecnológico e dos determinantes de sua disseminação

Este estudo utiliza o modelo de fronteira estocástica paramétrica com dados em painel para estimação das funções de produção asso- ciadas aos grupos da indústria de transfomação brasileira no período de 1996-2005. Um ponto importante dessa técnica é a possibilidade de utilização de um conjunto de variáveis explicativas para o com- ponente de ine ciência. Com o objetivo de captar as dinâmicas da fronteira tecnológica e do nível de ine ciência foram introduzidos com- ponentes de tendência linear e quadrática. Veri cou-se para o período a ocorrência de aumento do nível de ine ciência técnica e de progresso tecnológico a taxas constantes. Observou-se ainda que o nível de ine - ciência relaciona-se negativamente com os salários médios do pessoal direta e indiretamente ligado à produção, com o percentual de tributos sobre a receita total ,com o grau de terceirização e com o percentual da receita total originário da atividade principal da indústria.