Optimisation of in-cell accelerated solvent extraction technique for the determination of organochlorine pesticides in river sediments

Organochlorine pesticides (OCPs) are ubiquitous environmental contaminants with adverse impacts on aquatic biota, wildlife and human health even at low concentrations. However, conventional methods for their determination in river sediments are resource intensive. This paper presents an approach that is rapid and also reliable for the detection of OCPs. Accelerated Solvent Extraction (ASE) with in-cell silica gel clean-up followed by Triple Quadrupole Gas Chromatograph Mass Spectrometry (GCMS/MS) was used to recover OCPs from sediment samples. Variables such as temperature, solvent ratio, adsorbent mass and extraction cycle were evaluated and optimised for the extraction. With the exception of Aldrin, which was unaffected by any of the variables evaluated, the recovery of OCPs from sediment samples was largely influenced by solvent ratio and adsorbent mass and, to some extent, the number of cycles and temperature. The optimised conditions for OCPs extraction in sediment with good recoveries were determined to be 4 cycles, 4.5 g of silica gel, 105 ᴼC, and 4:3 v/v DCM: hexane mixture. With the exception of two compounds (α-BHC and Aldrin) whose recoveries were low (59.73 and 47.66 % respectively), the recovery of the other pesticides were in the range 85.35 – 117.97% with precision < 10 % RSD. The method developed significantly reduces sample preparation time, the amount of solvent used, matrix interference, and is highly sensitive and selective.

Accelerated Solvent Extraction for Gas Chromatographic Analysis of Nicotine and Cotinine in Meconium Samples

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)

Geochemistry and sea-surface temperature reconstruction for sediment cores of the Iberian Margin

Biogenic records of the marine palaeoproductivity (carbonates, organic carbon, and C37 alkenones) and the molecular stratigraphy of past sea surface temperatures (SSTs; UK'37) were studied at high resolution in two cores of the Iberian Margin. The comparison of these records indicates that the oceanographic conditions switched abruptly during the past 160 kyr between three kinds of regimes. A first regime with high (17-22°C) SST and low productivity typifies the interglacial periods, marine isotopic stages (MIS) 5 and 1. Several periods during MIS 6, 2, and the terminations II and I are characterised by about 4-5°C colder SST and a higher organic matter accumulation, both of which define the second regime. This anticorrelation between SST and marine productivity suggests that these variations are related to the intensity of the coastal upwelling. By contrast with this upwelling behaviour, extremely low biological productivity and very cold SST (6-12°C) occurred during short phases of glacial MIS 6, 4, and 2, and as abrupt events (~1 kyr or less) during MIS 3. The three oceanographic regimes are consistent with micropalaeontological changes in the same cores based on foraminifera and diatoms. The general trend of these hydrologic changes follows the long-term glacial/interglacial cycle, but the millennium scale variability is clearly related to Heinrich events and Dansgaard-Oeschger cycles. Strengthening of the upwelling corresponds probably to an intensification of the subtropical atmospheric circulation over the North Atlantic which was influenced by the presence of continental ice sheets. However, extreme glacial conditions due to massive discharges of icebergs interrupted the upwelling. Interestingly, both terminations II and I coincided with strong but transient intensification of the upwelling.

Effect of temperature and moisture on high pressure lipid/oil extraction from microalgae

Commercially viable carbon–neutral biodiesel production from microalgae has potential for replacing depleting petroleum diesel. The process of biodiesel production from microalgae involves harvesting, drying and extraction of lipids which are energy- and cost-intensive processes. The development of effective large-scale lipid extraction processes which overcome the complexity of microalgae cell structure is considered one of the most vital requirements for commercial production. Thus the aim of this work was to investigate suitable extraction methods with optimised conditions to progress opportunities for sustainable microalgal biodiesel production. In this study, the green microalgal species consortium, Tarong polyculture was used to investigate lipid extraction with hexane (solvent) under high pressure and variable temperature and biomass moisture conditions using an Accelerated Solvent Extraction (ASE) method. The performance of high pressure solvent extraction was examined over a range of different process and sample conditions (dry biomass to water ratios (DBWRs): 100%, 75%, 50% and 25% and temperatures from 70 to 120 ºC, process time 5–15 min). Maximum total lipid yields were achieved at 50% and 75% sample dryness at temperatures of 90–120 ºC. We show that individual fatty acids (Palmitic acid C16:0; Stearic acid C18:0; Oleic acid C18:1; Linolenic acid C18:3) extraction optima are influenced by temperature and sample dryness, consequently affecting microalgal biodiesel quality parameters. Higher heating values and kinematic viscosity were compliant with biodiesel quality standards under all extraction conditions used. Our results indicate that biodiesel quality can be positively manipulated by selecting process extraction conditions that favour extraction of saturated and mono-unsaturated fatty acids over optimal extraction conditions for polyunsaturated fatty acids, yielding positive effects on cetane number and iodine values. Exceeding biodiesel standards for these two parameters opens blending opportunities with biodiesels that fall outside the minimal cetane and maximal iodine values.

The development of soil organic matter in restored biodiverse Jarrah forests of South-Western Australia as determined by ASE and GCMS

Background, aim and scope Soil organic matter (SOM) is known to increase with time as landscapes recover after a major disturbance; however, little is known about the evolution of the chemistry of SOM in reconstructed ecosystems. In this study, we assessed the development of SOM chemistry in a chronosequence (space for time substitution) of restored Jarrah forest sites in Western Australia. Materials and methods Replicated samples were taken at the surface of the mineral soil as well as deeper in the profile at sites of 1, 3, 6, 9, 12, and 17 years of age. A molecular approach was developed to distinguish and quantify numerous individual compounds in SOM. This used accelerated solvent extraction in conjunction with gas chromatography mass spectrometry. A novel multivariate statistical approach was used to assess changes in accelerated solvent extraction (ASE)-gas chromatography-mass spectrometry (GCMS) spectra. This enabled us to track SOM developmental trajectories with restoration time. Results Results showed total carbon concentrations approached that of native forests soils by 17 years of restoration. Using the relate protocol in PRIMER, we demonstrated an overall linear relationship with site age at both depths, indicating that changes in SOM chemistry were occurring. Conclusions The surface soils were seen to approach native molecular compositions while the deeper soil retained a more stable chemical signature, suggesting litter from the developing diverse plant community has altered SOM near the surface. Our new approach for assessing SOM development, combining ASE-GCMS with illuminating multivariate statistical analysis, holds great promise to more fully develop ASE for the characterisation of SOM.

High-anthocyanin strawberries through cultivar selection

BACKGROUND: Diets high in fruit and vegetables are known to have significant health benefits. This is in part due to the presence of phytochemicals, which possess potential protective health benefits. This study focuses on the ability of strawberries to be bred for higher anthocyanin content. This is a major contributor to the characteristic colour and nutritional value of ripe strawberries, together with phenolic acids, ascorbic acid and total antioxidant capacity. RESULTS: Anthocyanins in five commercial strawberry cultivars and three breeding lines were assessed. This led to the discovery of one breeding line (BL 2006-221) as an exceptional source of anthocyanins (approximate to 1 g kg1 fresh weight), with approximately double the levels of current commercial cultivars. Temperature was shown to influence anthocyanin extraction, with 40 degrees C being the best extraction temperature using the accelerated solvent extraction (ASE) method. Hue angle and anthocyanin concentration showed a good correlation (r2 = 0.69). CONCLUSION: The new breeding line BL 2006-221 has the potential to be used in the development of phytochemically rich strawberry cultivars. Using hue angle as a screening tool for total anthocyanin concentration and extraction of anthocyanins from strawberries by ASE at 40 degrees C would support such cultivar development. (c) 2012 Commonwealth of Australia

Bioactive Properties of Mushroom (Agaricus bisporus) Stipe Extracts

The antibacterial activity and total phenolic (TP) content of Agaricus bisporus stipes were assessed using solvent and water extracts to determine its bioactivity. Extraction methods included accelerated solvent extraction (ASE) and hot water followed by membrane concentration. Water extract from ASE had the highest TP of 1.08 gallic acid equivalents (GAE)/g dry weight (DW) followed by ethanol at 0.61 mg GAE/g DW and 0.11 mg GAE/g DW for acetone. Acetone extracts inhibited Escherichia coli and Staphylococcus aureus at less than 50%; ethanol inhibited E. coli at 61.9% and S. aureus at 56.6%; and ASE water inhibited E. coli at 78.6% and S. aureus at 65.4%. The TP content of membrane concentrated extract of mushroom was 17 mg GAE in 100 mL. Membrane concentrated water extracts had a higher percentage inhibition on S. aureus than E. coli. Overall, the results were promising for further application of mushroom stipe extracts as a functional food additive. Practical Applications Mushrooms are known for their health benefits and have been identified as a good source of nutrients. The highly perishable nature of mushrooms warrants further processing and preservation to minimize losses along the supply chain. This study explores the possibility of adding value to mushroom stipes, a by-product of the fresh mushroom industry. The extracts assessed indicate the antibacterial activity and phenolic content, and the potential of using these extracts as functional ingredients in the food industry. This study provides valuable information to the scientific community and to the industries developing novel ingredients to meet the market demand for natural food additives.

Bioactive Properties of Mushroom (Agaricus bisporus) Stipe Extracts

The antibacterial activity and total phenolic (TP) content of Agaricus bisporus stipes were assessed using solvent and water extracts to determine its bioactivity. Extraction methods included accelerated solvent extraction (ASE) and hot water followed by membrane concentration. Water extract from ASE had the highest TP of 1.08 gallic acid equivalents (GAE)/g dry weight (DW) followed by ethanol at 0.61 mg GAE/g DW and 0.11 mg GAE/g DW for acetone. Acetone extracts inhibited Escherichia coli and Staphylococcus aureus at less than 50%; ethanol inhibited E. coli at 61.9% and S. aureus at 56.6%; and ASE water inhibited E. coli at 78.6% and S. aureus at 65.4%. The TP content of membrane concentrated extract of mushroom was 17 mg GAE in 100 mL. Membrane concentrated water extracts had a higher percentage inhibition on S. aureus than E. coli. Overall, the results were promising for further application of mushroom stipe extracts as a functional food additive. Practical Applications Mushrooms are known for their health benefits and have been identified as a good source of nutrients. The highly perishable nature of mushrooms warrants further processing and preservation to minimize losses along the supply chain. This study explores the possibility of adding value to mushroom stipes, a by-product of the fresh mushroom industry. The extracts assessed indicate the antibacterial activity and phenolic content, and the potential of using these extracts as functional ingredients in the food industry. This study provides valuable information to the scientific community and to the industries developing novel ingredients to meet the market demand for natural food additives.

Estudo em Escala Piloto de Parâmetros de Monitoramento e Operação de Biopilhas no Tratamento de Solos Contaminados por Hidrocarbonetos de Petróleo

O objetivo do trabalho foi identificar ferramentas e indicadores adequados ao monitoramento e à otimização de processos de biorremediação, incluindo parâmetros físicos, químicos e microbiológicos definidos em estudos de tratabilidade de solos contaminados por óleo cru em escala de laboratório e comparar estratégias de biorremediação, tais como bioestímulo e bioaumento conduzidas em simulações de biopilhas dinâmicas ou estáticas. Quando três métodos de extração de hidrocarbonetos de petróleo de solo arenoso e franco-argiloso para análise cromatográfica (Soxhlet-SOX, microondas-MARS e extração acelerada por solvente-ASE) foram comparados entre si, concluiu-se que a técnica que promove a melhor recuperação depende da fração de interesse (n-alcanos, HRP, MCNR, HPA), das características texturais do solo (teores de areia, silte e argila) e da idade da contaminação. Dentre os indicadores de densidade populacional microbiana (microrganismos heterotróficos totais-PHT, população de fungos-PF e população microbiana degradadora de óleo (PDO) passíveis de utilização para indicar a taxa de degradação de compostos orgânicos presentes no solo tais como os hidrocarbonetos de petróleo, o PDO mostrou-se o mais adequado em conjunto com a produção de CO2 aferida pelo método respirométrico. Quando a estratégia de biorremediação de solo franco-argiloso contaminado com óleo cru a 3% (m m-1) utilizando bioestímulo (ajuste de pH, umidade e taxa C:N:P) foi comparada ao bioaumento (bioestímulo e adição de inóculo de microrganismos extraídos, enriquecidos e aclimatizados ao óleo cru como fonte de carbono), em sistemas de bancada simulando biopilha dinâmica (microcosmo M) e biopilha estática com aeração forçada (reator B), o tratamento que apresentou melhor remoção (32%) de HTP após 121 dias foi o bioaumento em biopilha estática. Para HPA, o tratamento que alcançou a melhor remoção (33%) foi com bioestímulo também em biopilha estática. A avaliação da taxa de mortalidade (%) de Eisenia andrei exposta tanto a solos recém-contaminados por óleo cru e preparados para bioestímulo (BIOS) e bioaumento (BIOA) a serem tratados em biopilhas dinâmicas e estáticas em escala de laboratório mostrou que após 56 dias de exposição da E. andrei, todos os solos produziram letalidade de 100%, quer fossem os solos recém-contaminados e preparados para os diferentes tratamentos (BIOS M, BIOS B, BIOA M, BIOA B) ou após 121 dias de tratamento. Tal resultado confirma que a biorremediação foi incipiente também do ponto de vista de remoção da ecotoxicidade. Em linhas gerais, a biorremediação de solo franco-argiloso contaminado por óleo cru, contendo tanto contaminação antiga quanto recente, reúne os maiores desafios à biorremediação, tanto do ponto de vista da composição textural do solo quanto da natureza do contaminante. Os processos são aparentemente lentos e requerem ferramentas auxiliares para aceleração dos mesmos. Recomenda-se no futuro, condução de experimentos com o uso de diferentes surfactantes, com ênfase em biosurfactantes

Ecotoxicidade aguda, crônica e genotoxicidade de solo contaminado por óleo lubrificante usado e biorremediado para Eisenia andrei.

O objetivo do presente deste trabalho foi avaliar a toxicidade aguda, crônica e a genotoxicidade sobre E. andrei causadas por solo recém-contaminado com óleo lubrificante usado e após biorremediação por diferentes estratégias, após 22 meses, e paralelamente ao estudo de ecotoxicidade, foi conduzida uma investigação comparativa de três métodos de extração de HTP e HPA de solos para análise cromatográfica. A comparação das técnicas de extração evidenciou que para HTP, a técnica de extração acelerada por solvente-ASE foi a que melhor recuperou n-alcanos; já para as frações HRP e MCNR as técnicas soxhlet e micro-ondas-MARS não apresentaram diferenças significativas e foram melhores que ASE. Para HPA, a técnica de extração por soxhlet foi a que apresentou melhor recuperação em todos os solos. O teste de mortalidade apresentou, aos 14 dias, taxas crescentes de mortalidade de 10 6%, 20 0%, 73 25%, 93 12% e 100 0% para amostras de CONT (solo controle, sem contaminação artificial), BIOS (solo contaminado com 5% de OLU e biorremediado por bioestimulo), BIOA1 (solo contaminado com 5% de OLU e biorremediado por bioestimulo + bioaumento com adição de 10% de RSU maturado), e BIOA2 (solo contaminado com 5% de OLU e biorremediado por bioestimulo + bioaumento com adição de 10% de RSU semi-maturado) e OLU (solo contaminado com 5% de OLU), respectivamente. Aos 28 dias, entretanto, BIOS e OLU apresentaram taxas de mortalidade de 97 % 6 % e de 100 % 0 % respectivamente, valores estes significativamente superiores ao CONT. Foram observadas deformações anatômicas nos indivíduos mantidos em BIOS e OLU, assim como diminuição da biomassa em todas as amostras, evidenciando efeitos crônicos. O teste de reprodução, aos 28 dias, foram observadas grandes quantidades de indivíduos jovens nos solos biorremediados e recém-contaminado. No entanto, aos 56 dias houve uma diminuição dessas formas e o controle (CONT) exibiu uma quantidade maior de formas juvenis. O teste de densidade e viabilidade celular mostrou ser indicador sensível para toxicidade crônica apresentando queda nos solos BIOS e OLU em relação ao CONT com diferenças significativas (p <0.05). Não foram observados micronúcleos nos solos em estudo. Tal observação reforça a necessidade de testes de ecotoxicidade para avaliar a real eficácia de tecnologias de tratamento.

Ecotoxicidade aguda, crônica e genotoxicidade de solo contaminado por óleo lubrificante usado e biorremediado para Eisenia andrei.

O objetivo do presente deste trabalho foi avaliar a toxicidade aguda, crônica e a genotoxicidade sobre E. andrei causadas por solo recém-contaminado com óleo lubrificante usado e após biorremediação por diferentes estratégias, após 22 meses, e paralelamente ao estudo de ecotoxicidade, foi conduzida uma investigação comparativa de três métodos de extração de HTP e HPA de solos para análise cromatográfica. A comparação das técnicas de extração evidenciou que para HTP, a técnica de extração acelerada por solvente-ASE foi a que melhor recuperou n-alcanos; já para as frações HRP e MCNR as técnicas soxhlet e micro-ondas-MARS não apresentaram diferenças significativas e foram melhores que ASE. Para HPA, a técnica de extração por soxhlet foi a que apresentou melhor recuperação em todos os solos. O teste de mortalidade apresentou, aos 14 dias, taxas crescentes de mortalidade de 10 6%, 20 0%, 73 25%, 93 12% e 100 0% para amostras de CONT (solo controle, sem contaminação artificial), BIOS (solo contaminado com 5% de OLU e biorremediado por bioestimulo), BIOA1 (solo contaminado com 5% de OLU e biorremediado por bioestimulo + bioaumento com adição de 10% de RSU maturado), e BIOA2 (solo contaminado com 5% de OLU e biorremediado por bioestimulo + bioaumento com adição de 10% de RSU semi-maturado) e OLU (solo contaminado com 5% de OLU), respectivamente. Aos 28 dias, entretanto, BIOS e OLU apresentaram taxas de mortalidade de 97 % 6 % e de 100 % 0 % respectivamente, valores estes significativamente superiores ao CONT. Foram observadas deformações anatômicas nos indivíduos mantidos em BIOS e OLU, assim como diminuição da biomassa em todas as amostras, evidenciando efeitos crônicos. O teste de reprodução, aos 28 dias, foram observadas grandes quantidades de indivíduos jovens nos solos biorremediados e recém-contaminado. No entanto, aos 56 dias houve uma diminuição dessas formas e o controle (CONT) exibiu uma quantidade maior de formas juvenis. O teste de densidade e viabilidade celular mostrou ser indicador sensível para toxicidade crônica apresentando queda nos solos BIOS e OLU em relação ao CONT com diferenças significativas (p <0.05). Não foram observados micronúcleos nos solos em estudo. Tal observação reforça a necessidade de testes de ecotoxicidade para avaliar a real eficácia de tecnologias de tratamento.

Cellular and in-vitro models to assess antioxidant activities of seaweed extracts and the potential use of the extracts as ingredients

Seaweeds contain a range of antioxidant compounds such as polyphenols, carotenoids, sulphated polysaccharides and vitamins and have the potential to be used as ingredients in neutraceuticals. The antioxidant activity of crude 60% methanol extracts prepared from five Irish seaweeds, Ascophyllum nodosum, Laminaria hyperborea, Pelvetia canaliculata, Fucus vesiculosus and Fucus serratus were examined using in-vitro assays and a cell model system to determine the antioxidant activity of the extracts and their ability to protect against H2O2 and tert-BOOH-induced DNA damage and alterations in cellular antioxidant status in the human adenocarcinoma, Caco-2 cell line. To optimise the extraction of antioxidant compounds from seaweeds, an accelerated solvent extraction (ASE®) was used in combination with food grade solvents. The antioxidant activity of these extracts against H2O2 and tert-BOOH-induced DNA damage and alterations in cellular antioxidant status was also assessed. Extracts that exhibited the highest antioxidant activity, A. nodosum (100% water and 80% ethanol extracts) and F. vesiculosus (60% ethanol extract) were selected as ingredients for incorporation into fluid milk and yogurt at concentrations of 0.25% and 0.5%. The addition of the seaweed extracts to milk and yogurt did not affect the pH or shelf-life properties of the products. Seaweed addition did however significantly influence the colour properties of the milk and yogurt. Yellowness values were significantly higher in yogurts containing F. vesiculosus at both concentrations and A. nodosum (80% ethanol) at the 0.5% concentration. In milk, the F. vesiculosus (60% ethanol) and A. nodosum (80% ethanol) at both the 0.25% and the 0.5% concentrations had higher greenness and yellowness values than the milk containing A. nodosum (100% water). Sensory analysis revealed that appearance and flavour governed the overall acceptability of yogurts with the control yogurt, and yogurts containing A. nodosum (100% water) were the most preferred samples by panellists. However, in the milk trial the perception of a fishy taste was the determining factor in the negative perception of milk. The unsupplemented control and the milk containing A. nodosum (100% water) at a concentration of 0.5% were the most overall accepted milk samples by the sensory panellists. The antioxidant activity of the extracts in milk and yogurt remained stable during storage as determined by the in-vitro assays. Seaweed supplemented milk and yogurt were also subjected to an in-vitro digestion procedure which mimics the human digestive system. The milk and yogurt samples and their digestates were added to Caco-2 cells to investigate their antioxidant potential however neither the undigested or digested samples protected against H2O2-induced DNA damage in Caco-2 cells.