LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate Reagents

This study provides a basis for a new and straightforward method for LC/MS/MS-based screening of N-terminal protein adducts. This procedure is denoted the “FIRE procedure” as fluorescein isothiocyanate (FITC) gave superior sensitivity by LC/MS/MS when measuring adducts (R) of electrophilic compounds with a modified Edman procedure. The principles of the FIRE-procedure are that adducts to N-terminal amino acids selectively are detached and measured from of proteins after derivatisation by isothiocyanate Edman reagents. In this study, FITC, 4-N,N-dimethylaminoazobenzene 4′-isothiocyanate (DABITC) and 4-dimethylamino-1-naphthyl isothiocyanate (DNITC) were used to synthesize thiohydantoin analytes from valine and N-methylvaline. The sensitivity by LC/MS/MS was enhanced by up to three orders of magnitude as compared to phenyl isothiocyanate and higher as compared to pentafluorophenyl isothiocyanate. The FITC reagent will enable measurements of low background adduct levels. Synthesized analytes were characterised with, for example, 1H NMR, 13C NMR, LC/MS/MS, and UV

In vitro inhalation bioaccessibility of phthalate esters and alternative plasticisers present in indoor dust using artificial lung fluids

Phthalate esters (PEs) are plasticiser additives imparting durability, elasticity and flexibility to consumer products. The low migration stability of PEs along with their ubiquitous character and adverse health effects to humans and especially children has resulted in their classification as major indoor contaminants. This study assesses inhalation exposure to PEs via indoor dust using an in vitro inhalation bioaccessibility test (i.e. uptake) for of dimethyl phthalate (DMP), diethyl phthalate (DEP) and di-(2-ethylhexyl) phthalate (DEHP) and the alternative non phthalate plasticisers bis(2-ethylhexyl) terephthalate (DEHT) and cyclohexane-1,2-dicarboxylic acid diisononyl ester (DINCH), exposure. Using artificial lung fluids, which mimicktwo distinctively different pulmonary environments, namely artificial lysosomal fluid (ALF, pH = 4.5) representing the fluid that inhaled particles would contact after phagocytosis by alveolar and interstitial macrophages within the lung and Gamble’s solution (pH = 7.4), the fluid for deep dust deposition within the pulmonary environment. Low molecular weight (MW) PEs such as DMP and DEP were highly bioaccessible (> 75 %) in both artificial pulmonary media, whereas highly hydrophobic compounds such as DEHP, DINCH and DEHT were < 5 % bioaccessible via the lung. Our findings show that the in vitro pulmonary uptake of PEs is primarily governed by their hydrophobicity and water solubility, highlighting thus the need for the establishment of a unified and biologically relevant inhalation bioaccessibility test format, employed within the risk assessment framework for volatile and semi-volatile organic pollutants

Methodologies to assess the fate of polar organic compounds in aquatic environments

Polar organic compounds (POCs) are chemicals with polar functional groups in their structure. The functional groups make the compounds hydrophilic and less prone to partition with biota. However, the knowledge of their fate is limited due to difficulties associated with their measurements. Although, the persistence of POCs in the environment is generally low, they are considered to be semi-persistent compounds due to their continuous introduction to the environment via wastewater. Studies have shown that complex mixtures of POCs of different classes may have synergistic toxic effects on biota at environmental concentration levels. Therefore, it is important to develop analytical methods in order to establish the occurrence and fate of POCs in aquatic environments. In Study I, a positive correlation between the sorption of a novel poly(ethylene-co-vinyl acetate-co-carbon monoxide) (PEVAC) material and the theoretical logarithmic dissociation partition coefficient (Log D) for seven POCs was observed. The PEVAC material showed an enhanced sorption of the POCs compared to the silicone material. Study II, demonstrated that the PEVAC sampler assess the freely dissolved concentration of POCs in aquatic environments. The results showed that the PEVAC polymer is an attractive alternative to silicone for mimicing the biological uptake of POCs in aquatic environments. Additionally, Study II showed that total extraction is appropriate for determination of the freely dissolved concentration of uncharged POCs with Log KOW &lt; 2.67 in natural water. In study III, a novel bag-solid phase extraction (bag-SPE) technique was compared to a conventional SPE-technique. Despite that the extraction efficiencies for POCs in wastewater were lower using the bag-SPE method, the two methods showed similar detection limits due to the lower ion-suppression experienced with the bag-SPE. In study IV the bag-SPE method was further developed with the aim of lowering the detection limits for POCs. Detection limits (LOD) below 13 ng/L showed that the bag-SPE method was suitable for determination of POCs in surface sea water.  This research was financially supported by European Union (European Commission, FP6 Contract No. 003956) “Novel Methods for Integrated Risk Assessment of Cumulative Stressors in the Environment” (NoMiracle) and by the Swedish research council Formas

Self-declarations of environmental classification at Fass.se

Since 2005 Sweden has a unique environmental classification system for pharmaceutical products. It is a self-declaration system where each pharmaceutical company is responsible for their own environmental information. The environmental risk assessments are published on the web based portal www.Fass.se, which is open to the public. Prior to publication the environmental risk assessments are reviewed by IVL as an independent, external part to make sure that the classifications are based on a scientifically acceptable interpretation of the guidance for the pharmaceutical companies. The review of pre-published environmental risk assessments and system evaluation is an on-going task and the present report describes the experiences from the review process during the year 2014. With its iterative process, IVL gives feedback to LIF regarding the system as such, both from a scientific perspective as well as from a quality assurance perspective. In 2015, 400 environmental risk assessments were sent in for review. Of these 41% received the comment no remarks and were recommended to be published. Another 41% received the assessment remark and were recommended to be corrected before publication and 19% needed to be corrected and sent in for another review before publication. The number of times a document is re-sent in for revision has decreased during the last three years. One of the reasons is the development and implementation of the assessment tool, which has improved the review process and clarified the comments sent to the companies. The statistical calculations of the environmental risk assessments in this report are based on documents that were published at Fass.se during 2015. The total number of unique substances was 391 and 19% of these were classified regarding environmental risk. 50% were exempted from classification and 31% were reviewed, but no classification could be made due to lack of data. The work of improving the review system is an on-going process. Den här rapporten finns endast på engelska. Svensk sammanfattning finns i rapporten.Since 2005 Sweden has a unique environmental classification system for pharmaceutical products. It is a self-declaration system where each pharmaceutical company is responsible for their own environmental information. The environmental risk assessments are published on the web based portal www.Fass.se, which is open to the public. Prior to publication the environmental risk assessments are reviewed by IVL as an independent, external part to make sure that the classifications are based on a scientifically acceptable interpretation of the guidance for the pharmaceutical companies. The review of pre-published environmental risk assessments and system evaluation is an on-going task and the present report describes the experiences from the review process during the year 2014. With its iterative process, IVL gives feedback to LIF regarding the system as such, both from a scientific perspective as well as from a quality assurance perspective. In 2015, 400 environmental risk assessments were sent in for review. Of these 41% received the comment no remarks and were recommended to be published. Another 41% received the assessment remark and were recommended to be corrected before publication and 19% needed to be corrected and sent in for another review before publication. The number of times a document is re-sent in for revision has decreased during the last three years. One of the reasons is the development and implementation of the assessment tool, which has improved the review process and clarified the comments sent to the companies. The statistical calculations of the environmental risk assessments in this report are based on documents that were published at Fass.se during 2015. The total number of unique substances was 391 and 19% of these were classified regarding environmental risk. 50% were exempted from classification and 31% were reviewed, but no classification could be made due to lack of data. The work of improving the review system is an on-going process.This report describes the experiences gained during the review process in year 2015 of the system of self-declarations of environmental classification at www.Fass.se, and has been prepared with the aim to achieve transparency by explaining the role and the experiences of the reviewer, which may be useful in future development of the system. The main target groups are LIF and its member companies, as well as users of the environmental classifications, e.g. county councils and researchers

Self-declarations of environmental classification in www.fass.se

In parallel to the implementation of the environmental classification system at www.fass.se, IVL has conducted a study of the review process with the aim to identify and address the pitfalls of the system. With experience from the reviewing process, IVL took part in a discussion, led by LIF, with the pharmaceutical companies about how to improve the implementation of the guidelines for environmental risk assessment developed by LIF and their expert group on environment. The goal of this reviewing process was to establish a common praxis for the implementation of the guideline among the different companies and to provide feedback from the experience of the self-declaration process to the system owners, LIF. The review of pre-published environmental risk assessments and system evaluation is an on-going task and the present report describes the experiences from the review process during the year 2012.A

Results from the Swedish Screening programme 2012

IVL Svenska Miljöinstitutet har på uppdrag av Naturvårdsverket genomfört en screening avseende pyritioner och deras viktigaste nedbrytningsprodukter under 2012/2013. Zink pyrition (ZnPT) är en bredspektrumbiocid mot bakterier, svampar och alger. ZnPT används kommersiellt som antimjällmedel i schampon och som konserveringsmedel i kosmetika. ZnPTs löslighet i vatten gör ämnet lämpligt som tillsatts i båt- och utomhusfärger. Natrium pyrition (HPT) har samma användningsområden som ZnPT och förekommer som konserveringsmedel i kosmetika och i vätskor som används inom metallindustrin. Syftet med studien var att bestämma halter och att bidra till förståelsen av pyritioners omvandling genom att även analysera 2-pyridinsulfonsyra (PSA) och andra nedbrytnings­produkter i prover från reningsverk och småbåtshamnar. ZnPT och/eller kopparpyrition (CuPT) uppmättes i 79 % av inkommande och i 7 % av utgående avloppsvatten från kommunala reningsverk. Minskningen av halten ZnPT och/eller CuPT mellan inkommande och utgående avloppsvatten visar att nedbrytning kan ske i reningsverksprocessen. Att ZnPT och/eller CuPT inte kunde detekteras i slam tyder på att nedbrytning av dessa ämnen sker i kontakt med slam. Nedbrytningsprodukten PSA var den mest frekvent förekommande föreningen och uppmättes i de flesta avloppsvatten samt i samtliga slamprover. Koncentrationen av PSA i inkommande vatten var 73 – 480 (median 240) ng/L (n=14), i utgående vatten <2 - 330 (median 59) ng/L (n=28) och i slam 25 - 280 (median 110) µg/kg TS (n=21). Minskningen i halten PSA mellan inkommande och utgående avloppsvatten tyder på att PSA kan brytas ner eller adsorberas till slam. I en småbåtshamn där ZnPT troligen emitteras från båtbottenfärger, och också kan omvandlas till den ännu mer toxiska formen CuPT, kunde ingen av dessa former påvisas i vattenfasen eller sedimentet. Däremot kunde PSA detekteras i sedimentet. I sediment där belastningen av ZnPT är mer diffus (Göta älv, gradient från Göteborg), kunde ingen ackumulering av PSA påvisas. Eftersom den akuta toxiciteten för pyritioner minskar med ökad nedbrytning och PSA är den minst giftiga av de kända nedbrytningsprodukterna indikerar resultatet att en process pågår i sedimentet som leder till minskande toxicitet. Avsaknaden av kroniska tester av PSA i den vetenskapliga litteraturen gör det inte möjligt att uppskatta långsiktig påverkan på den marina miljön.As an assignment from the Swedish Environmental Protection Agency, a screening study of pyrithiones and their major degradation products has been performed by IVL during 2012/2013. Zinc pyrithione (ZnPT) is a broad-spectrum biocide effective against bacteria, fungi and algae. ZnPT is commercially used as anti-dandruff agent in shampoos and as preservatives in cosmetics. Due to ZnPTs solubility in water it is suitable for use in outdoor and marine paints. Sodium pyrithione (HPT) is used as a preservative in cosmetics and as protective agent for fluids used in the metal industries. The objective of the present screening was to determine concentrations and to illuminate the fate of pyrithiones in sewage treatment plants and in small boat marinas by broadening the chemical analysis also to 2-pyridinesulfonic acid (PSA) and other pyrithione residues. ZnPT and/or cupper pyrithione (CuPT) were detected in 79% of influent and in 7% of effluent wastewaters from sewage treatment plants (STPs). A decrease in concentration from influent to effluent indicates that ZnPT and CuPT are degraded in the STPs. The absence of ZnPT and/or CuPT in sludge suggests that degradation will take place in contact with sludge. The degradation product PSA was the most abundant compound in the study. PSA was present in the majority of the wastewater samples and in all the sludge samples analysed. The concentration range of PSA in influents were 73 - 480 (median 240) ng/L (n=14), in effluents <2 - 330 (median 59) ng/L (n=28) and in sludge 25 - 280 (median 110) µg/kg dw (n=21). The decrease in concentration of PSA from influent to effluent wastewater shows that PSA may be further degraded or adsorbed to sludge in the STP. In a small boat marina where ZnPT is probably constantly emitted from anti-fouling paints, (and could be trans-chelated to the even more toxic CuPT) none of these species could be detected in the water or in the sediment. PSA, however, was detected in the sediment. In the sediments from Gothenburg, where the ZnPT load is of a more diffuse character, accumulation of PSA was not detected. As the acute toxicity of pyrithiones decreases with increased degradation and PSA is the least toxic of the known pyrithione residues, the result indicates that a process leading to less toxic sediments is taking place. However, the absence of chronic tests of PSA in the scientific literature makes it impossible to estimate the long-term effect on the marine environment

Årsrapport 2010 för projektet RE-PATH. Mätningar av PFAS i närområdet till Stockholm Arlanda Airport och Göteborg Landvetter Airport

Projektet RE-PATH syftar till att undersöka effekter och spridning av PFAS-ämnen (främst PFOS) från brandskum (AFFF) i området runt Göteborg Landvetter Airport och Stockholm Arlanda Airport. PFOS (perfluoroktansulfonat) klassas som ett PBT-ämne. Det bryts inte ned i naturen, bioackumuleras i näringskedjan och är giftigt för vattenlevande organismer.Denna rapport sammanfattar andra årets mätningar som bekräftar förra årets resultat att koncentrationerna av PFOS var förhöjda i vatten och fisk från lokaler i nära anslutning till flygplatsområdena. I ytvatten föreligger en stark korrelation mellan PFOS-halt och avståndet till källan som är respektive brandövningsplats och provtagningslokal. Vid Göteborg Landvetter Airport och området nedströms uppmättes höga halter av PFOS i dammarna på flygplatsområdet och koncentrationerna är fortsatt höga längs med avrinningsområdet söderut. Halterna i fisk från Lilla Issjön belägen direkt söder om flygplatsen är många gånger högre jämfört med fisk från Västra Ingsjön som är belägen längre nedströms. Vid Stockholm Arlanda Airport är PFOS-halten i vatten och fisk från Halmsjön kraftigt förhöjd, jämfört med en närbelägen sjö utanför avrinningsområdet. De höga koncentrationerna i vatten från Halmsjön späds ut i Märstaån men halten vid mynningen i Mälaren är fortfarande förhöjd, liksom halterna i fångad fisk. Halterna av PFOS i abborre och mört korrelerar med PFOS-koncentrationen i vattnet. Inga skillnader i halter i fisk har kunnat kopplas till kön eller ålder. Abborrhonor där flera organ har analyserats visar högst anrikning av PFOS i lever och hjärta/blod följt av gälar och gonad, det senare visar på en möjlig utsöndringsväg av PFOS för honor när de leker.Effektstudier på yngel från åkergroda och abborre visar att PFOS-fritt brandskum är något högre toxicitet jämfört med AFFF. Detta indikerar att det är andra komponenter i skummet som ger det största bidraget till den uppmätta toxiciteten mot vattenlevande organismer. PFAS-ämnen förekommer i för låg halt i AFFF för att bidra till en betydande toxicitet

Self-declarations of environmental classification in www.fass.se

In 2005 environmental information was published for the first two groups of products in www.fass.se, to test a new model for classification, developed on an initiative from LIF (The Research-Based Pharmaceutical Industry in Sweden). The initiative was a response to an increasing public demand for environmental information on pharmaceuticals and an attempt to develop a model accepted by Swedish stakeholders, but also by the global pharmaceutical industry. In 2010, all groups of pharmaceuticals (ATC codes) on the Swedish market had been the subject of an environmental risk assessment. During the implementation of the environmental classification system IVL Swedish Environmental Research Institute (IVL) performed a project with the aim to identify and address the pitfalls of the system. This project was financed by LIF and the Foundation for IVL Swedish Environmental Research Institute (SIVL). IVL reviewed the pre-published data and took part in a discussion, led by LIF, with the pharmaceutical companies about how to implement the guideline for environmental risk assessment, developed by LIF (LIF Expert Group Environment) in cooperation with stakeholders and the international industry. The goal of this reviewing process was to establish a common praxis for the implementation of the guideline among the different companies and to feed back the experience from the self-declaration process to the system owners, LIF. The review of pre-published environmental risk assessments and system evaluation is an on-going task and the present report describes the experiences from the review process during the year 2013. With its iterative process, IVL gives feedback to LIF regarding the system as such, both from a scientific perspective as well as from a quality assurance perspective. Den här rapporten finns endast på engelska. Svensk sammanfattning finns i rapporten.Report on how the work to implement environmental information in www.fass.se will be achieved through a common practice

Self-declarations of environmental classification in www.fass.se

In parallel to the implementation of the environmental classification system at www.fass.se, IVL has conducted a study of the review process with the aim to identify and address the pitfalls of the system. With experience from the reviewing process, IVL took part in a discussion, led by LIF, with the pharmaceutical companies about how to improve the implementation of the guidelines for environmental risk assessment developed by LIF and their expert group on environment. The goal of this reviewing process was to establish a common praxis for the implementation of the guideline among the different companies and to provide feedback from the experience of the self-declaration process to the system owners, LIF. The review of pre-published environmental risk assessments and system evaluation is an on-going task and the present report describes the experiences from the review process during the year 2012.A