Using standard additions to improve extraction and quantification of inositol hexakisphosphate in sediment samples by ion chromatography electrospray ionization mass spectrometry.

Several key aspects for the analysis of inositol hexakisphosphate (InsP6) have been investigated in order to establish a suitable method for the study of sediment samples from different aquatic systems. Apparent matrix effects for the ion chromatography electrospray ionization tandem mass spectrometric detection (IC-ESI-MS/MS) method were accounted for with a standard addition approach, which also compensated for variation in extraction efficiency. Several parameters of the extraction method were optimized to improve the extraction efficiency for different sediment types. We observed an improvement in the extraction efficiency between 18% and 720%. Finally, the method was used to gain first insights into the relevance of InsP6 in two aquatic systems located at the German Baltic coastal area. InsP6 was detected in several sediment samples with concentrations between 2.3 and 15.2 µg InsP6-P/g dry weight (DW).

Validation of Arxula Yeast Estrogen Screen assay for detection of estrogenic activity in water samples: Results of an international interlaboratory study.

Endocrine-active substances can adversely impact the aquatic ecosystems. A special emphasis is laid, among others, on the effects of estrogens and estrogen mimicking compounds. Effect-based screening methods like in vitro bioassays are suitable tools to detect and quantify endocrine activities of known and unknown mixtures. This study describes the validation of the Arxula-Yeast Estrogen Screen (A-YES®) assay, an effect-based method for the detection of the estrogenic potential of water and waste water. This reporter gene assay, provided in ready to use format, is based on the activation of the human estrogen receptor alpha. The user-friendly A-YES® enables inexperienced operators to rapidly become competent with the assay. Fourteen laboratories from four countries with different training levels analyzed 17ß-estradiol equivalent concentrations (EEQ) in spiked and unspiked waste water effluent and surface water samples, in waste water influent and spiked salt water samples and in a mixture of three bisphenols. The limit of detection (LOD) for untreated samples was 1.8ng/L 17ß-estradiol (E2). Relative repeatability and reproducibility standard deviation for samples with EEQ above the LOD (mean EEQ values between 6.3 and 20.4ng/L) ranged from 7.5 to 21.4% and 16.6 to 28.0%, respectively. Precision results are comparable to other frequently used analytical methods for estrogens. The A-YES® has been demonstrated to be an accurate, precise and robust bioassay. The results have been included in the ISO draft standard. The assay was shown to be applicable for testing of typical waste water influent, effluent and saline water. Other studies have shown that the assay can be used with enriched samples, which lower the LOD to the pg/L range. The validation of the A-YES® and the development of a corresponding international standard constitute a step further towards harmonized and reliable bioassays for the effect-based analysis of estrogens and estrogen-like compounds in water samples.

TBT and its metabolites in sediments: Survey at a German coastal site and the central Baltic Sea.

Since the 1950s the organotin compound tributyltin (TBT) was intensively used in antifouling paints for marine vessels and it became of concern for the marine environment. Herein, we report on a study from 2015 on TBT and its metabolites monobutyltin (MBT) and dibutyltin (DBT) in sediments from the central Baltic Sea and a Baltic Sea coastal site with strong harbor activities (Warnemünde). Sublayers from a sediment core from the Arkona Basin were analyzed to investigate the long term organotin pressure for the Baltic Sea. For the central Baltic Sea total organotin (MBT+DBT+TBT) ranged from 100 to 500ng/g TOC with distinct areas of high organotin content probably due to historical inputs. For the coastal site total organotin ranged from 10,000 to 60,000ng/g TOC. MBT and DBT were the predominant organotin species detected. Overall, the data obtained indicate the progress of TBT degradation at the investigated sites.

Hexachlorocyclohexane - Long term variability and spatial distribution in the Baltic Sea.

In response to the HELCOM commitment the Leibniz Institute for Baltic Sea Research Warnemuende conducts a monitoring program on listed substances of concern for the Baltic Sea environment which comprises the isomers of technical hexachlorocyclohexane (HCH). After World War II the HCH compounds were applied globally in huge amounts as insecticidal formulation in which only γ-HCH is the effective agent. Monitoring was conducted in water samples from a large Baltic Sea area. Mostly, the HCH compounds were evenly distributed in the Baltic Sea. Data from 1975 to 2015 from the Arkona Basin depict a substantial reduction of the HCH pressure (12,500 to < 400 pg/L). Longest residence is shown for ß-HCH, which is currently the predominant HCH isomer in the Baltic Sea (α/ß/γ:1/2/1). Half lives were determined to be 4-8 years for α- and γ-HCH and 5-20 years for ß-HCH.

Remobilization of Phytol from Chlorophyll Degradation Is Essential for Tocopherol Synthesis and Growth of Arabidopsis.

Phytol from chlorophyll degradation can be phosphorylated to phytyl-phosphate and phytyl-diphosphate, the substrate for tocopherol (vitamin E) synthesis. A candidate for the phytyl-phosphate kinase from Arabidopsis thaliana (At1g78620) was identified via a phylogeny-based approach. This gene was designated VITAMIN E DEFICIENT6 (VTE6) because the leaves of the Arabidopsis vte6 mutants are tocopherol deficient. The vte6 mutant plants are incapable of photoautotrophic growth. Phytol and phytyl-phosphate accumulate, and the phytyl-diphosphate content is strongly decreased in vte6 leaves. Phytol feeding and enzyme assays with Arabidopsis and recombinant Escherichia coli cells demonstrated that VTE6 has phytyl-P kinase activity. Overexpression of VTE6 resulted in increased phytyl-diphosphate and tocopherol contents in seeds, indicating that VTE6 encodes phytyl-phosphate kinase. The severe growth retardation of vte6 mutants was partially rescued by introducing the phytol kinase mutation vte5. Double mutant plants (vte5 vte6) are tocopherol deficient and contain more chlorophyll, but reduced amounts of phytol and phytyl-phosphate compared with vte6 mutants, suggesting that phytol or phytyl-phosphate are detrimental to plant growth. Therefore, VTE6 represents the missing phytyl-phosphate kinase, linking phytol release from chlorophyll with tocopherol synthesis. Moreover, tocopherol synthesis in leaves depends on phytol derived from chlorophyll, not on de novo synthesis of phytyl-diphosphate from geranylgeranyl-diphosphate.

Innovative methods in soil phosphorus research: A review.

Phosphorus (P) is an indispensable element for all life on Earth and, during the past decade, concerns about the future of its global supply have stimulated much research on soil P and method development. This review provides an overview of advanced state-of-the-art methods currently used in soil P research. These involve bulk and spatially resolved spectroscopic and spectrometric P speciation methods (1 and 2D NMR, IR, Raman, Q-TOF MS/MS, high resolution-MS, NanoSIMS, XRF, XPS, (µ)XAS) as well as methods for assessing soil P reactions (sorption isotherms, quantum-chemical modeling, microbial biomass P, enzymes activity, DGT, 33P isotopic exchange, 18O isotope ratios). Required experimental set-ups and the potentials and limitations of individual methods present a guide for the selection of most suitable methods or combinations.

Fatty acid phytyl ester synthesis in chloroplasts of Arabidopsis.

During stress or senescence, thylakoid membranes in chloroplasts are disintegrated, and chlorophyll and galactolipid are broken down, resulting in the accumulation of toxic intermediates, i.e., tetrapyrroles, free phytol, and free fatty acids. Chlorophyll degradation has been studied in detail, but the catabolic pathways for phytol and fatty acids remain unclear. A large proportion of phytol and fatty acids is converted into fatty acid phytyl esters and triacylglycerol during stress or senescence in chloroplasts. We isolated two genes (PHYTYL ESTER SYNTHASE1 [PES1] and PES2) of the esterase/lipase/thioesterase family of acyltransferases from Arabidopsis thaliana that are involved in fatty acid phytyl ester synthesis in chloroplasts. The two proteins are highly expressed during senescence and nitrogen deprivation. Heterologous expression in yeast revealed that PES1 and PES2 have phytyl ester synthesis and diacylglycerol acyltransferase activities. The enzymes show broad substrate specificities and can employ acyl-CoAs, acyl carrier proteins, and galactolipids as acyl donors. Double mutant plants (pes1 pes2) grow normally but show reduced phytyl ester and triacylglycerol accumulation. These results demonstrate that PES1 and PES2 are involved in the deposition of free phytol and free fatty acids in the form of phytyl esters in chloroplasts, a process involved in maintaining the integrity of the photosynthetic membrane during abiotic stress and senescence.

Plastid lipid droplets at the crossroads of prenylquinone metabolism.

Lipid droplets called plastoglobules (PGs) exist in most plant tissues and plastid types. In chloroplasts, the polar lipid monolayer surrounding these low-density lipoprotein particles is continuous with the outer lipid leaflet of the thylakoid membrane. Often small clusters of two or three PGs, only one of them directly connected to thylakoids, are present. Structural proteins (known as plastid-lipid associated proteins/fibrillins or plastoglobulins) together with lipid metabolic enzymes coat the PGs. The hydrophobic core of PGs contains a range of neutral lipids including the prenylquinones [tocopherols (vitamin E), phylloquinone (vitamin K(1)), and plastoquinone (PQ-9)]. In this review the function of PGs and their associated enzymes in prenylquinone metabolism will be discussed.