Nature-Inspired Chemistry of Complex Alkaloids: Combining Targeted Molecular Networking Approach and Semisynthetic Strategy to Access Rare Communesins in a Marine-Derived Penicillium expansum.

Communesins are rare alkaloids isolated from fungi of the genus Penicillium. In this work, the extract of a marine-derived Penicillium expansum strain was studied using targeted molecular networking approach allowing to detect 65 communesins including 55 new ones. A fragmentation pattern for dimethylvinyl communesins was established and a script was implemented allowing to predict the structure and map all communesins in a global molecular network. A semisynthetic strategy was carried out to obtain some minor congeners from the two isolated communesins A and B. Nine communesins were then synthetised: two of them were already described as produced by the studied strain; four are new natural products which occurrence in the extracts was confirmed; three are new semi-synthetic analogues never described so far. These communesins were evaluated for their cytotoxicity on two human cancer cell lines KB and MCF-7 leading to a preliminary study of their structure-activity relationships.

Untargeted Metabolomics Approach for the Discovery of Environment-Related Pyran-2-ones Chemodiversity in a Marine-Sourced Penicillium restrictum.

Very little is known about chemical interactions between fungi and their mollusc host within marine environments. Here, we investigated the metabolome of a Penicillium restrictum MMS417 strain isolated from the blue mussel Mytilus edulis collected on the Loire estuary, France. Following the OSMAC approach with the use of 14 culture media, the effect of salinity and of a mussel-derived medium on the metabolic expression were analysed using HPLC-UV/DAD-HRMS/MS. An untargeted metabolomics study was performed using principal component analysis (PCA), orthogonal projection to latent structure discriminant analysis (O-PLSDA) and molecular networking (MN). It highlighted some compounds belonging to sterols, macrolides and pyran-2-ones, which were specifically induced in marine conditions. In particular, a high chemical diversity of pyran-2-ones was found to be related to the presence of mussel extract in the culture medium. Mass spectrometry (MS)- and UV-guided purification resulted in the isolation of five new natural fungal pyran-2-one derivatives-5,6-dihydro-6S-hydroxymethyl-4-methoxy-2H-pyran-2-one (1), (6S, 1'R, 2'S)-LL-P880ß (3), 5,6-dihydro-4-methoxy-6S-(1'S, 2'S-dihydroxy pent-3'(E)-enyl)-2H-pyran-2-one (4), 4-methoxy-6-(1'R, 2'S-dihydroxy pent-3'(E)-enyl)-2H-pyran-2-one (6) and 4-methoxy-2H-pyran-2-one (7)-together with the known (6S, 1'S, 2'S)-LL-P880ß (2), (1'R, 2'S)-LL-P880γ (5), 5,6-dihydro-4-methoxy-2H-pyran-2-one (8), (6S, 1'S, 2'R)-LL-P880ß (9), (6S, 1'S)-pestalotin (10), 1'R-dehydropestalotin (11) and 6-pentyl-4-methoxy-2H-pyran-2-one (12) from the mussel-derived culture medium extract. The structures of 1-12 were determined by 1D- and 2D-MMR experiments as well as high-resolution tandem MS, ECD and DP4 calculations. Some of these compounds were evaluated for their cytotoxic, antibacterial, antileishmanial and in-silico PTP1B inhibitory activities. These results illustrate the utility in using host-derived media for the discovery of new natural products.

Detection of ergosterol using liquid chromatography/electrospray ionization mass spectrometry: Investigation of unusual in-source reactions.

RATIONALE: In the field of natural products, de-replication of complex mixtures has become a usual practice to annotate known compounds and avoid their re-isolation. For this purpose, many groups rely on liquid chromatography coupled to high-resolution mass spectrometry (HPLC/MS) to deduce molecular formulae of compounds allowing comparison with public or in-house databases. Electrospray ionization (ESI) is usually considered as the method of choice for investigating a large panel of compounds but, in some cases, it may lead to unusual results as described in this article for ergosterol. METHODS: Ergosterol and other fungal sterols in methanolic solution were analysed using various chromatographic gradients with HPLC/MS using both ion trap time-of-flight MS and Orbitrap MS instruments fitted with an ESI source. Further flow injection analyses were performed to investigate the influence of the solvent composition. MS/MS fragmentation data were acquired to annotate the various ions observed. RESULTS: Contrary to other fungal sterols, ergosterol was found to be highly sensitive to oxidation during ESI. Putative structures were proposed based on MS/MS studies and known oxidation mechanisms of ergosterol by reactive oxygen species that could be formed in the ESI process. The proportion of acetonitrile in the eluent was found to influence this in-source oxidation, with an increased proportion of oxidized sodium adducts with higher proportions of acetonitrile. CONCLUSIONS: While ergosterol is a major sterol found in fungi, this study investigates its ionization by electrospray for the first time. The results reported here will help further detection and annotation of this compound in fungal extracts after HPLC/ESI-MS analyses.

Review of LCA studies of solid waste management systems--part II: methodological guidance for a better practice.

Life cycle assessment (LCA) is increasingly used in waste management to identify strategies that prevent or minimise negative impacts on ecosystems, human health or natural resources. However, the quality of the provided support to decision- and policy-makers is strongly dependent on a proper conduct of the LCA. How has LCA been applied until now? Are there any inconsistencies in the past practice? To answer these questions, we draw on a critical review of 222 published LCA studies of solid waste management systems. We analyse the past practice against the ISO standard requirements and the ILCD Handbook guidelines for each major step within the goal definition, scope definition, inventory analysis, impact assessment, and interpretation phases of the methodology. Results show that malpractices exist in several aspects of the LCA with large differences across studies. Examples are a frequent neglect of the goal definition, a frequent lack of transparency and precision in the definition of the scope of the study, e.g. an unclear delimitation of the system boundaries, a truncated impact coverage, difficulties in capturing influential local specificities such as representative waste compositions into the inventory, and a frequent lack of essential sensitivity and uncertainty analyses. Many of these aspects are important for the reliability of the results. For each of them, we therefore provide detailed recommendations to practitioners of waste management LCAs.

Review of LCA studies of solid waste management systems--part I: lessons learned and perspectives.

The continuously increasing solid waste generation worldwide calls for management strategies that integrate concerns for environmental sustainability. By quantifying environmental impacts of systems, life cycle assessment (LCA) is a tool, which can contribute to answer that call. But how, where and to which extent has it been applied to solid waste management systems (SWMSs) until now, and which lessons can be learnt from the findings of these LCA applications? To address these questions, we performed a critical review of 222 published LCA studies of SWMS. We first analysed the geographic distribution and found that the published studies have primarily been concentrated in Europe with little application in developing countries. In terms of technological coverage, they have largely overlooked application of LCA to waste prevention activities and to relevant waste types apart from household waste, e.g. construction and demolition waste. Waste management practitioners are thus encouraged to abridge these gaps in future applications of LCA. In addition to this contextual analysis, we also evaluated the findings of selected studies of good quality and found that there is little agreement in the conclusions among them. The strong dependence of each SWMS on local conditions, such as waste composition or energy system, prevents a meaningful generalisation of the LCA results as we find it in the waste hierarchy. We therefore recommend stakeholders in solid waste management to regard LCA as a tool, which, by its ability of capturing the local specific conditions in the modelling of environmental impacts and benefits of a SWMS, allows identifying critical problems and proposing improvement options adapted to the local specificities.

Environmental evaluation of municipal waste prevention.

Waste prevention has been addressed in the literature in terms of the social and behavioural aspects, but very little quantitative assessment exists of the environmental benefits. Our study evaluates the environmental consequences of waste prevention on waste management systems and on the wider society, using life-cycle thinking. The partial prevention of unsolicited mail, beverage packaging and food waste is tested for a "High-tech" waste management system relying on high energy and material recovery and for a "Low-tech" waste management system with less recycling and relying on landfilling. Prevention of 13% of the waste mass entering the waste management system generates a reduction of loads and savings in the waste management system for the different impacts categories; 45% net reduction for nutrient enrichment and 12% reduction for global warming potential. When expanding our system and including avoided production incurred by the prevention measures, large savings are observed (15-fold improvement for nutrient enrichment and 2-fold for global warming potential). Prevention of food waste has the highest environmental impact saving. Prevention generates relatively higher overall relative benefit for "Low-tech" systems depending on landfilling. The paper provides clear evidence of the environmental benefits of waste prevention and has specific relevance in climate change mitigation.

Determination of the concentration of nitrogenous bio-organic compounds using an isotope ratio mass spectrometer operating in continuous flow mode.

The quality of the determination of compound-specific isotopic content at natural abundance by gas chromatography-isotope ratio measurement-mass spectrometry (GC-irm-MS) relies on the stability of the voltage generated by the ion detector Faraday cages. The application of GC-irm-MS to the determination of δ(13)C (‰) and δ(15)N (‰) is now routine. However, for numerous applications, it is necessary to determine both the isotope content (δ(15)N) and the quantity (in micromoles) of analyte present. We now show that it is possible for nitrogen-containing compounds to measure how much analyte is present with an irm mass spectrometer linked to a GC by exploiting the integrated N(2) total ion current intensity (Vs) generated by measuring the (15)N/(14)N isotope ratio. The method is validated over a range of concentration (2-70 mmol/L) and δ(15)N (-70 to +50‰) values for six molecules of diverse chemical nature and functionality (nortropine, norpseudotropine, nortropinone, cysteine, taurine, glutathione). It is shown that once the ion current is calibrated, the quantitative values are of a comparable quality to those obtained from GC with flame ionization detection (GC-FID). In addition, it is demonstrated that over a definable range, the δ(15)N (‰) value is independent of the quantity of analyte introduced, confirming the validity of this method.

Models for waste life cycle assessment: review of technical assumptions.

A number of waste life cycle assessment (LCA) models have been gradually developed since the early 1990 s, in a number of countries, usually independently from each other. Large discrepancies in results have been observed among different waste LCA models, although it has also been shown that results from different LCA studies can be consistent. This paper is an attempt to identify, review and analyse methodologies and technical assumptions used in various parts of selected waste LCA models. Several criteria were identified, which could have significant impacts on the results, such as the functional unit, system boundaries, waste composition and energy modelling. The modelling assumptions of waste management processes, ranging from collection, transportation, intermediate facilities, recycling, thermal treatment, biological treatment, and landfilling, are obviously critical when comparing waste LCA models. This review infers that some of the differences in waste LCA models are inherent to the time they were developed. It is expected that models developed later, benefit from past modelling assumptions and knowledge and issues. Models developed in different countries furthermore rely on geographic specificities that have an impact on the results of waste LCA models. The review concludes that more effort should be employed to harmonise and validate non-geographic assumptions to strengthen waste LCA modelling.

Determination of the natural abundance delta15N of nortropane alkaloids by gas chromatography-isotope ratio mass spectrometry of their ethylcarbamate esters.

An important route for the detoxification of tropane alkaloids involves N-demethylation to the nor-compounds followed by further degradation. In order to study the mechanisms of the pertinent reactions, a suitable means to determine the isotope ratios of the substrates and products is required. However, the polarity and functionality of the nortropane compounds makes their analysis as free bases difficult. A method is described which allows both the quantification of nortropane alkaloids and the determination of their natural abundance delta(15)N values. The protocol exploits the derivatisation of the alkaloids by reaction with ethyl chloroformate in aqueous medium and the quantitative extraction of the ensuing ethylcarbamate esters. The improved chromatographic properties of these derivatives gives ample separation of the isomeric nortropine and norpseudotropine for measurement of their delta(15)N (per thousand) values by isotope ratio mass spectrometry interfaced to gas chromatography. Adequate separation could not be achieved with the underivatised compounds. Repeatability and precision are sufficient to allow differences in the delta(15)N values (Deltadelta(15)N) > 0.8 per thousand to be measured, with a standard deviation routinely approximately 0.3 per thousand. The methodology has been tested by determining the changes in the delta(15)N values of nortropine and norpseudotropine during degradation by cell suspension cultures of a Pseudomonas strain expressing a specific capacity for tropine catabolism. The precision and reproducibility are shown sufficient to allow the evolution of the delta(15)N values to be followed during the fermentation.

Greenhouse gas accounting and waste management.

Accounting of emissions of greenhouse gas (GHG) is a major focus within waste management. This paper analyses and compares the four main types of GHG accounting in waste management including their special features and approaches: the national accounting, with reference to the Intergovernmental Panel on Climate Change (IPCC), the corporate level, as part of the annual reporting on environmental issues and social responsibility, life-cycle assessment (LCA), as an environmental basis for assessing waste management systems and technologies, and finally, the carbon trading methodology, and more specifically, the clean development mechanism (CDM) methodology, introduced to support cost-effective reduction in GHG emissions. These types of GHG accounting, in principle, have a common starting point in technical data on GHG emissions from specific waste technologies and plants, but the limited availability of data and, moreover, the different scopes of the accounting lead to many ways of quantifying emissions and producing the accounts. The importance of transparency in GHG accounting is emphasised regarding waste type, waste composition, time period considered, GHGs included, global warming potential (GWP) assigned to the GHGs, counting of biogenic carbon dioxide, choice of system boundaries, interactions with the energy system, and generic emissions factors. In order to enhance transparency and consistency, a format called the upstream-operating-downstream framework (UOD) is proposed for reporting basic technology-related data regarding GHG issues including a clear distinction between direct emissions from waste management technologies, indirect upstream (use of energy and materials) and indirect downstream (production of energy, delivery of secondary materials) activities.

Global warming factor of municipal solid waste management in Europe.

The global warming factor (GWF; CO(2)-eq. tonne(-1) waste) performance of municipal waste management has been investigated for six representative European Member States: Denmark, France, Germany, Greece, Poland and the United Kingdom. The study integrated European waste statistical data for 2007 in a life-cycle assessment modelling perspective. It is shown that significant GWF benefit was achieved due to the high level of energy and material recovery substituting fossil energy and raw materials production, especially in Denmark and Germany. The study showed that, despite strong regulation of waste management at European level, there are major differences in GWF performance among the member states, due to the relative differences of waste composition, type of waste management technologies available nationally, and the average performance of these technologies. It has been demonstrated through a number of sensitivity analyses that, within the national framework, key waste management technology parameters can influence drastically the national GWF performance of waste management.

Greenhouse gases, radiative forcing, global warming potential and waste management--an introduction.

Management of post-consumer solid waste contributes to emission of greenhouse gases (GHGs) representing about 3% of global anthropogenic GHG emissions. Most GHG reporting initiatives around the world utilize two metrics proposed by the Intergovernmental Panel on Climate Change (IPCC): radiative forcing (RF) and global warming potential (GWP). This paper provides a general introduction of the factors that define a GHG and explains the scientific background for estimating RF and GWP, thereby exposing the lay reader to a brief overview of the methods for calculating the effects of GHGs on climate change. An objective of this paper is to increase awareness that the GWP of GHGs has been re-adjusted as the concentration and relative proportion of these GHGs has changed with time (e.g., the GWP of methane has changed from 21 to 25 CO(2)-eq). Improved understanding of the indirect effects of GHGs has also led to a modification in the methodology for calculating GWP. Following a presentation of theory behind GHG, RF and GWP concepts, the paper briefly describes the most important GHG sources and sinks in the context of the waste management industry. The paper serves as a primer for more detailed research publications presented in this special issue of Waste Management & Research providing a technology-based assessment of quantitative GHG emissions from different waste management technologies.

C balance, carbon dioxide emissions and global warming potentials in LCA-modelling of waste management systems.

Global warming potential (GWP) is an important impact category in life-cycle-assessment modelling of waste management systems. However, accounting of biogenic CO(2) emissions and sequestered biogenic carbon in landfills and in soils, amended with compost, is carried out in different ways in reported studies. A simplified model of carbon flows is presented for the waste management system and the surrounding industries, represented by the pulp and paper manufacturing industry, the forestry industry and the energy industry. The model calculated the load of C to the atmosphere, under ideal conditions, for 14 different waste management scenarios under a range of system boundary conditions and a constant consumption of C-product (here assumed to be paper) and energy production within the combined system. Five sets of criteria for assigning GWP indices to waste management systems were applied to the same 14 scenarios and tested for their ability to rank the waste management alternatives reflecting the resulting CO(2) load to the atmosphere. Two complete criteria sets were identified yielding fully consistent results; one set considers biogenic CO(2) as neutral, the other one did not. The results showed that criteria for assigning global warming contributions are partly linked to the system boundary conditions. While the boundary to the paper industry and the energy industry usually is specified in LCA studies, the boundary to the forestry industry and the interaction between forestry and the energy industry should also be specified and accounted for.

Natural-abundance isotope ratio mass spectrometry as a means of evaluating carbon redistribution during glucose-citrate cofermentation by Lactococcus lactis.

The cometabolism of citrate and glucose by growing Lactococcus lactis ssp. lactis bv. diacetylactis was studied using a natural-abundance stable isotope technique. By a judicious choice of substrates differing slightly in their 13C/12C ratios, the simultaneous metabolism of citrate and glucose to a range of compounds was analysed. These end-products include lactate, acetate, formate, diacetyl and acetoin. All these products have pyruvate as a common intermediate. With the objective of estimating the degree to which glucose and citrate metabolism through pyruvate may be differentially regulated, the delta13C values of the products accumulated over a wide range of concentrations of citrate and glucose were compared. It was found that, whereas the relative accumulation of different products responds to both the substrate concentration and the ratio between the substrates, the delta13C values of the products primarily reflect the availability of the two substrates over the entire range examined. It can be concluded that in actively growing L. lactis the maintenance of pyruvate homeostasis takes precedence over the redox status of the cells as a regulatory factor.