A precision apparatus, with solid phase micro-extraction monitoring capability, for incorporation studies of gaseous precursors into insect-derived metabolites

An apparatus is described that facilitates the determination of incorporation levels of isotope labelled, gaseous precursors into volatile insect-derived metabolites. Atmospheres of varying gas compositions can be generated by evacuation of a working chamber followed by admission of the required levels of component gases, using a precision, digitised pressure read-out system. Insects such as fruit-flies are located initially in a small introduction chamber, from which migration can occur downwards into the working chamber. The level of incorporation of labelled precursors is continuously assayed by the Solid Phase Micro Extraction (SPME) technique and GC-MS analyses. Experiments with both Bactrocera species (fruit-flies) and a parasitoid wasp, Megarhyssa nortoni nortoni (Cresson) and oxygen-18 labelled dioxygen illustrate the utility of this system. The isotope effects of oxygen-18 on the carbon-13 NMR spectra of 1,7- dioxaspiro[5,5]undecane are also described.

Analysis of Insect Cuticular Compounds by Non-lethal Solid Phase Micro Extraction with Styrene-Divinylbenzene Copolymers

Insect cuticular hydrocarbons including relatively non-volatile chemicals play important roles in cuticle protection and chemical communication. The conventional procedures for extracting cuticular compounds from insects require toxic solvents, or non-destructive techniques that do not allow storage of subsequent samples, such as the use of SPME fibers. In this study, we describe and tested a non-lethal process for extracting cuticular hydrocarbons with styrene-divinylbenzene copolymers, and illustrate the method with two species of bees and one species of beetle. The results demonstrate that these compounds can be efficiently trapped by ChromosorbA (R) (SUPELCO) and that this method can be used as an alternative to existing methods.

A micro-extraction technique using a new digitally controlled syringe combined with UHPLC for assessment of urinary biomarkers of oxidatively damaged DNA

The formation of reactive oxygen species (ROS) within cells causes damage to biomolecules, including membrane lipids, DNA, proteins and sugars. An important type of oxidative damage is DNA base hydroxylation which leads to the formation of 8-oxo-7,8-dihydro-29-deoxyguanosine (8-oxodG) and 5-hydroxymethyluracil (5-HMUra). Measurement of these biomarkers in urine is challenging, due to the low levels of the analytes and the matrix complexity. In order to simultaneously quantify 8-oxodG and 5-HMUra in human urine, a new, reliable and powerful strategy was optimised and validated. It is based on a semi-automatic microextraction by packed sorbent (MEPS) technique, using a new digitally controlled syringe (eVolH), to enhance the extraction efficiency of the target metabolites, followed by a fast and sensitive ultrahigh pressure liquid chromatography (UHPLC). The optimal methodological conditions involve loading of 250 mL urine sample (1:10 dilution) through a C8 sorbent in a MEPS syringe placed in the semi-automatic eVolH syringe followed by elution using 90 mL of 20% methanol in 0.01% formic acid solution. The obtained extract is directly analysed in the UHPLC system using a binary mobile phase composed of aqueous 0.1% formic acid and methanol in the isocratic elution mode (3.5 min total analysis time). The method was validated in terms of selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), extraction yield, accuracy, precision and matrix effect. Satisfactory results were obtained in terms of linearity (r2 . 0.991) within the established concentration range. The LOD varied from 0.00005 to 0.04 mg mL21 and the LOQ from 0.00023 to 0.13 mg mL21. The extraction yields were between 80.1 and 82.2 %, while inter-day precision (n=3 days) varied between 4.9 and 7.7 % and intra-day precision between 1.0 and 8.3 %. This approach presents as main advantages the ability to easily collect and store urine samples for further processing and the high sensitivity, reproducibility, and robustness of eVolHMEPS combined with UHPLC analysis, thus retrieving a fast and reliable assessment of oxidatively damaged DNA.

Detection of Lead Contamination of Water and VOC Contamination of Air Using SOI Micro-Optical Devices

Micro-photonic SOI Mach-Zehnder interferometers were coated with solid-phase micro-extraction materials derived from polydimethylsiloxane to enable sensing of volatile organic compounds of the BTEX class in air. A different coating based on functionalized mesoporous silicates is used to detect lead Pb(II) with a detection limit of <;; 100 ppb in water.

Characterization of volatile organic gunshot residues in fired handgun cartridges by headspace sorptive extraction

In forensic investigation of firearm-related cases, determination of the residual amount of volatile compounds remaining inside a cartridge could be useful in estimating the time since its discharge. Published approaches are based on following the decrease of selected target compounds as a function of time by using solid phase micro-extraction (SPME). Naphthalene, as well as an unidentified decomposition product of nitrocellulose (referred to as "TEA2"), are usually employed for this purpose. However, reliability can be brought into question given their high volatility and the low reproducibility of their extracted quantities. In order to identify alternatives and therefore develop improved dating methods, an extensive study on the composition and variability of volatile residues in nine different types of cartridges was carried out. Analysis was performed using headspace sorptive extraction (HSSE), which is a more exhaustive technique compared to SPME. 166 compounds were identified (several of which for the first time), and it was observed that the final compositional characteristics of each residue were strongly dependent on its source. Variability of single identified compounds within and between different types of cartridge, as well as their evolution over time, was also studied. Many explosion products containing up to 4 aromatic rings were found to be globally present in high proportions amongst residues. 27 of them (excluding naphthalene) also presented detectable decreases during the first 24 h. Therefore, they could be used as complementary target analytes in future dating methods.

Micro-extração em fase sólida (SPME) in situ e comparação de compostos voláteis em variedades de manga (Mangifera indica L.) resistentes e suscetíveis à mosca-das-frutas Anastrepha obliqua (Macquart) (Diptera: Tephritidae)

Pós-graduação em Química - IQ

Analysis of N-nitrosames in water using micellar electrokinetic chromatography (MEKC) and development of online and offline methods of extraction and enrichment of neutral polar analytes

Evaluation of the quality of the environment is essential for human wellness as pollutants in trace amounts can cause serious health problem. Nitrosamines are a group of compounds that are considered potential carcinogens and can be found in drinking water (as disinfection byproducts), foods, beverages and cosmetics. To monitor the level of these compounds to minimize daily intakes, fast and reliable analytical techniques are required. As these compounds are relatively highly polar, extraction and enrichment from environmental samples (aqueous) are challenging. Also, the trend of analytical techniques toward the reduction of sample size and minimization of organic solvent use demands new methods of analysis. In light of fulfilling these requirements, a new method of online preconcentration tailored to an electrokinetic chromatography is introduced. In this method, electroosmotic flow (EOF) was suppressed to increase the interaction time between analyte and micellar phase, therefore the only force to mobilize the neutral analytes is the interaction of analyte with moving micelles. In absence of EOF, polarity of applied potential was switched (negative or positive) to force (anionic or cationic) micelles to move toward the detector. To avoid the excessive band broadening due to longer analysis time caused by slow moving micelles, auxiliary pressure was introduced to boost the micelle movement toward the detector using an in house designed and built apparatus. Applying the external auxiliary pressure significantly reduced the analysis times without compromising separation efficiency. Parameters, such as type of surfactants, composition of background electrolyte (BGE), type of capillary, matrix effect, organic modifiers, etc., were evaluated in optimization of the method. The enrichment factors for targeted analytes were impressive, particularly; cationic surfactants were shown to be suitable for analysis of nitrosamines due to their ability to act as hydrogen bond donors. Ammonium perfluorooctanoate (APFO) also showed remarkable results in term of peak shapes and number of theoretical plates. It was shown that the separation results were best when a high conductivity sample was paired with a BGE of lower conductivity. Using higher surfactant concentrations (up to 200 mM SDS) than usual (50 mM SDS) for micellar electrokinetic chromatography (MEKC) improved the sweeping. A new method for micro-extraction and enrichment of highly polar neutral analytes (N-Nitrosamines in particular) based on three-phase drop micro-extraction was introduced and its performance studied. In this method, a new device using some easy-to-find components was fabricated and its operation and application demonstrated. Compared to conventional extraction methods (liquid-liquid extraction), consumption of organic solvents and operation times were significantly lower.

Flavour components of some processed. fish and fishery products of Japan

A study was conducted to examine the flavour components of some processed fish and fishery products of Japan by gas chromatography-mass spectrometry (GC-MS). In brief the method was to absorb the headspace volatiles at 70°C into the fused silica fibre of needle of the solid phase micro extraction fibre. The absorbed components were injected to the GC-MS. The components were identified by computer matching with library database as well as by authentic standard components. In general the number of flavour components were higher in the processed fish and fishery products (except frozen prawn) than that of the raw fish and prawn. The concentration (quantity) of the f1avour components in processed fish and fishery products was much higher than that of the raw fish and prawn. Smoked salmon and baked salmon possessed double number of flavour components than that of the raw salmon. Smoking resulted the highest number of flavour components followed by baking (grilling) and canning, surimi products (kamaboko and chikuwa), drying and lastly salting. However, freezing and frozen storage resulted loss of flavour components in prawn.

Relação do aroma e da espuma dos vinhos espumantes com o potencial enológico das uvas e dos vinhos

Os espumantes produzidos segundo o método Champanhês são obtidos após uma segunda fermentação em garrafa. Quando o vinho é vertido no copo, o CO2 produzido é libertado, sendo a espuma formada o resultado da sua interacção com os constituintes do vinho. A quantidade e a estabilidade da espuma do vinho espumante estão relacionadas com a sua composição química. Para além da espuma, o aroma é também um parâmetro importante de qualidade na apreciação geral de um vinho espumante. O aroma de um vinho espumante provém do contributo das uvas assim como do processo fermentativo. Dependendo do estado de maturação da uva, o contributo dos compostos voláteis para o aroma é diferente. Em virtude da vindima para os vinhos espumantes ser realizada antes da vindima para os vinhos maduros, dependendo da variedade, as uvas poderão não ser colhidas na expressão máxima do seu aroma, podendo verificar-se uma perda significativa do seu potencial varietal volátil. O objectivo desta dissertação é relacionar o aroma e a espuma dos vinhos espumantes com o potencial enológico das uvas e dos vinhos. Para isso, foi estudada a composição volátil das duas castas principais da Bairrada, a casta branca Fernão-Pires (FP) e a casta tinta Baga (BG), sendo estas duas das castas usadas para a produção de espumante. Para estudar a composição volátil das uvas durante a maturação, com vista a avaliar este efeito na expressão máxima de compostos voláteis, foi optimizada para este propósito a metodologia de microextracção em fase sólida em espaço de cabeça (HS-SPME). As uvas foram colhidas semanalmente, em duas vinhas, do pintor à pós-maturidade sendo posteriormente analisadas pela metodologia de HS-SPME seguida de cromatografia de gás acoplada à espectrometria de massa com quadrupolo (GC–qMS). No caso das uvas BG, observou-se um aumento acentuado na expressão máxima de compostos voláteis próximo da maturidade da uva determinada pelo teor em açúcar e acidez titulável, mantendo-se constante durante a pós-maturidade. Na determinação do perfil volátil das uvas ao longo da maturação foram identificados 66 compostos varietais nas uvas provenientes de uma vinha (Pedralvites) e 45 da outra vinha (Colégio). Em ambas as vinhas foram identificados 23 sesquiterpenóides, 13 monoterpenóides, 6 norisoprenóides, 2 álcoois aromáticos e 1 diterpenóide. Os sesquiterpenóides, devido à sua abundância em número e em área cromatográfica, podem ser considerados marcadores da casta BG. As uvas FP apresentaram um comportamento diferente do das uvas BG, sendo a expressão máxima de compostos voláteis expressa durante um curto período de tempo (1 semana), que coincide com a maturidade da uva. Depois de atingido este pico, observa-se uma diminuição drástica logo na semana seguinte. Este comportamento foi observado em ambas as vinhas, onde foram identificados 20 compostos voláteis varietais e 5 pré-fermentativos (álcoois e aldeídos em C6). Estes resultados mostram que quando estas castas são colhidas precocemente (1 semana antes da maturidade) para a produção de espumante, é observada uma redução significativa do potencial volátil que é expresso na maturidade. Para a análise da composição volátil dos vinhos espumantes foi optimizada uma metodologia de microextracção que permite usar uma maior quantidade de fase estacionária, a extracção sorptiva em barra de agitação (SBSE). O método foi optimizado usando 10 padrões de compostos voláteis representativos das principais famílias químicas presentes no vinho, nomeadamente, ésteres, monoterpenóides, sesquiterpenóides, norisoprenóides em C13 e álcoois. O método proposto apresenta uma boa linearidade (r2 > 0,982) e a reprodutibilidade varia entre 8,9 e 17,8%. Os limites de detecção para a maioria dos compostos é bastante baixo, entre 0,05 e 9,09 μg L-1. O método foi aplicado para a análise da composição volátil dos vinhos espumantes. Dentro dos vinhos espumantes analisados, foi estudada a influência da casta, do tipo de solo e do estado de maturação das uvas na sua composição volátil. A casta FP pode dar origem a vinhos com maior potencial de aroma do que a casta BG. Relativamente à avaliação dos diferentes estados de maturação, verificou-se que as uvas da maturidade e as da colheita tardia (uma semana depois da maturidade) deram origem aos vinhos com maior quantidade de compostos voláteis. Para os três tipos de solo estudados (arenoso, argiloso e argilo-calcário), o vinho obtido a partir de uvas colhidas no solo argilo-calcário foi o que mostrou a maior concentração de compostos voláteis varietais. A espuma destes vinhos espumantes foi também avaliada quanto à sua quantidade máxima (HM) e tempo de estabilidade (TS). O vinho espumante que apresentou um maior TS foi o vinho produzido a partir da casta FP proveniente de uma colheita tardia e solo argiloso. Os vinhos provenientes dos solos arenosos e argilo-calcários são os que apresentaram valores mais baixos de TS. Com vista a avaliar quais os conjuntos de moléculas do vinho que estão relacionados com as propriedades da espuma e possíveis sinergismos entre eles, para cada vinho espumante foi separada a fracção hidrofóbica de baixo peso molecular (MeLMW), a fracção de elevado peso molecular (HMW) e duas fracções de peso molecular intermédio (AqIMW e MeIMW). As propriedades da espuma dos vinhos modelo, reconstituídos com estas fracções e suas misturas, foram avaliadas. A combinação da fracção HMW com a MeLMW aumentou o TS 2,7 vezes quando comparado com o observado para a fracção HMW isoladamente, produzindo um efeito sinergético. Este aumento do TS ainda foi maior quando se combinou a fracção HMW com as subfracções obtidas a partir da fracção MeLMW, principalmente para as fracções menos apolares. A subfracção hidrofóbica menos apolar foi caracterizada por espectrometria de massa de ionização por electrospray (ESI-MS/MS) tendo sido identificada uma série de oligómeros de polietileno glicol e um potencial composto tensioactivo, o 8-hidroxi-tridecanoato de dietilenoglicolglicerilacetato. A fracção MeLMW foi também isolada da espuma do vinho espumante e caracterizada por ESI-MS/MS, permitindo identificar vários compostos potenciais tensioactivos, nomeadamente, dois monoacilgliceróis e quatro derivados de ácidos gordos com gliceriletilenoglicol. Estes resultados confirmam que estes compostos relacionados com a estabilidade da espuma existem em maior número na espuma do que no vinho. O vinho foi ainda fraccionado em 12 grupos de moléculas: 3 fracções de manoproteínas, 3 de arabinogalactanas, 3 de misturas de polissacarídeos, proteínas e compostos fenólicos e 3 fracções de peso molecular intermédio e baixo, compostas por uma mistura de hidratos de carbono, peptídeos e compostos fenólicos. Foram usados vinhos modelo reconstituídos com cada uma das fracções isoladas na concentração em que estas se encontraram no vinho. Foram também efectuados ensaios com soluções modelo dez vezes mais concentradas e com misturas de algumas das fracções. Todas as soluções formadas foram avaliadas quanto às propriedades da espuma. O aumento da concentração para dez vezes faz com que a solução contendo a fracção rica em manoproteínas (MP1) aumente para mais do dobro a HM e 7,4 vezes mais o TS. A combinação entre a fracção MP1 e a MeLMW produziu um aumento significativo nos parâmetros de HM e TS. A combinação da fracção HMW (manoproteínas com baixo teor em proteína) com a MeLMW (tensioactivos derivados de ácidos gordos com gliceriletilenoglicol) contém os compostos chave de um vinho espumante para se obter uma maior quantidade e estabilidade da espuma.

Traditional algarvian distillats and liqueurs historic scientific aspects

All the evidence indicates that distillation and liqueurs preparation began in Monchique mountain because this place was pointed as a possible capital of the oldest population of Algarve and an important Arabic village (Barreto, 1972: 19). It was possible to find lots of vestiges like the alembic produced by Arabic population near the X century (Telo, 1988: 77). Traditionally the Algarvian people produce the Arbutus unedo L., fig, carob, grape distillates. At the same time they produce liqueur-using maceration of parts of plants or fruits in some kinds of distillates. Most of the work about Algarvian distillates started by studying the basic compounds of Arbutus unedo spirits by gas chromatography (GC) and mass spectrometry (MS) as well as other physical-chemical properties. In a second phase aged distillates were studied by their phenolic compounds evolution using high resolution liquid chromatography (HPLC). Volatile compounds of traditional liqueurs were identified by head space micro extraction solid phase (HE-SPME) and also analysed by gas chromatography mass spectrometry (GC-MS) and when possible confirmed with standards. Total phenols were determined by Folin-Ciocalteur method. Flavenoids were studied by high performance liquid chromatography (HPLC). Sensorial analysis was also done in every drink studies. The results showed that the arbutus distillate doesn’t present a high level of methanol according to the current legislation. The excesses of acidity or ethyl acetate present normal values when the fermentation is well done (Galego, et al. 1995: 341; Galego, et al. 1995: 685). During the aging process, the colour of spirits tend to become darker, the colour changes occurred more rapidly in the arbutus spirits located in cellars with higher temperatures (Galego, et al. 2001: 432). In the sensory evaluation of samples aged during 12 months into 50 L medium toasting level oak wood barrels, panellists considered that samples of arbutus spirit had too much wood flavour and they were not able to detect the characteristic aroma of arbutus fruit (Galego, et al., 2001: 183). Differences in liqueurs were observed using HS-SPME-GC, HS-SPME-GC-MS or HPLC analysis and this observation was confirmed by a sensorial panel (Galego, et al. 2003: 60).

Influence of abiotic stress factors on VOCs emission from Portuguese rice paddy fields: relation with increased climate change

Dissertação para obtenção do Grau de Mestre em Engenharia do Ambiente Perfil de Gestão de Sistemas Ambientais

Flavour characterization of fresh and processed pennywort (Centella asiatica L.) juices

The flavour characteristics of fresh and processed pennywort juices treated by pasteurization, sterilization and high pressure processing (HPP) were investigated by using solid-phase micro-extraction combined with gas chromatography-mass spectrometry. Sesquiterpene hydrocarbons comprised the major class of volatile components present and the juices had a characteristic smell due to the presence of volatile compounds including β-caryophyllene, humulene, E-β-farnesene, α-copaene, alloaromadendrene and β-elemene. All processing operations caused a reduction in the total volatile concentration, but HPP caused more volatile acyclic alcohols, aldehydes and oxygenated monoterpenoids to be retained than pasteurization and sterilization. Ketones were not present in fresh pennywort juice, but 2-butanone and 3-nonen-2-one were generated in all processed juices, and 2-nonanone and 2-hexanone were present in pasteurized and sterilized juices. Other chemical changes including isomerization were also reduced by HPP compared to pasteurization, and sterilization.