Deterioration of the Hanson Logboat: chemical and imaging assessment with removal of polyethylene glycol conserving agent.

The state of preservation of wood in two samples from the Hanson Logboat, currently on display in Derby Museum and Art Gallery, was analysed using elemental analysis (EA), pyrolysis-gas chromatography/flame ionisation detection (Py-GC/FID), pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) and scanning electron microscopy (SEM). The samples were collected in 2003, after the boat had undergone conservation, and in 2011 after the condition of the boat began to deteriorate. Solvent extraction enabled removal of polyethylene glycol, with which the wood had been impregnated during conservation, allowing the degradation of the cellulose and lignin polymeric components of the woods to be assessed. Elemental compositions (C, H, N, O, S), Py-GC/FID, Py-GC/MS and SEM imaging reveal extensive degradation of the wood polymers during the eight year period since conservation.

Mono-, di- and trimethylated homologues of isoprenoid tetraether lipid cores in archaea and environmental samples: mass spectrometric identification and significance.

Higher homologues of widely reported C(86) isoprenoid diglycerol tetraether lipid cores, containing 0-6 cyclopentyl rings, have been identified in (hyper)thermophilic archaea, representing up to 21% of total tetraether lipids in the cells. Liquid chromatography-tandem mass spectrometry confirms that the additional carbon atoms in the C(87-88) homologues are located in the etherified chains. Structures identified include dialkyl and monoalkyl ('H-shaped') tetraethers containing C(40-42) or C(81-82) hydrocarbons, respectively, many representing novel compounds. Gas chromatography-mass spectrometric analysis of hydrocarbons released from the lipid cores by ether cleavage suggests that the C(40) chains are biphytanes and the C(41) chains 13-methylbiphytanes. Multiple isomers, having different chain combinations, were recognised among the dialkyl lipids. Methylated tetraethers are produced by Methanothermobacter thermautotrophicus in varying proportions depending on growth conditions, suggesting that methylation may be an adaptive mechanism to regulate cellular function. The detection of methylated lipids in Pyrobaculum sp. AQ1.S2 and Sulfolobus acidocaldarius represents the first reported occurrences in Crenarchaeota. Soils and aquatic sediments from geographically distinct mesotemperate environments that were screened for homologues contained monomethylated tetraethers, with di- and trimethylated structures being detected occasionally. The structural diversity and range of occurrences of the C(87-89) tetraethers highlight their potential as complementary biomarkers for archaea in natural environments.

Predicting drug candidate victims of drug-drug interactions, using microdosing.

OBJECTIVE: The aim of this crossover human male volunteer study was to investigate the utility of microdosing in the investigation of drug-drug interactions. METHODS: A mixture of midazolam, tolbutamide, caffeine and fexofenadine were administered as a microdose (25 µg each) before and after administration of a combined pharmacological dose of ketoconazole (400 mg) and fluvoxamine (100 mg) to inhibit P-glycoprotein and metabolism by cytochrome P450 (CYP) 1A2, CYP3A4 and CYP2C9. RESULTS: When administered alone, pharmacokinetics for all four microdosed compounds scaled well with those reported for therapeutic doses and with previously performed microdose studies. The pharmacokinetics of each compound administered as a microdose were significantly altered after the administration of ketoconazole and fluvoxamine, showing statistically significant (p < 0.01) 12.8-, 8.1- and 3.2-fold increases in the area under the plasma concentration-time curve from time zero to infinity (AUC(∞)) for midazolam, caffeine and fexofenadine, respectively. A 1.8-fold increase (not statistically significant) in AUC(∞) was observed for tolbutamide. The changes in pharmacokinetics mediated by ketoconazole and fluvoxamine were quantitatively consistent with previously reported, non-microdose, drug-drug interaction data from studies including the same compounds. CONCLUSION: The initial data reported here demonstrate the utility of microdosing to investigate the risk of development drugs being victims of drug-drug interactions.

Novel glycerol dialkanol triols in sediments: transformation products of glycerol dibiphytanyl glycerol tetraether lipids or biosynthetic intermediates?

A glycerol dialkanol triol, similar in structure to glycerol dibiphytanyl glycerol tetraethers but devoid of the carbon atoms of one of the two glycerol termini, has been identified in Messinian sediments (~6 Ma) and characterised using quadrupole ion trap and Fourier transform-ion cyclotron resonance liquid chromatography-tandem mass spectrometry.

The major lipid cores of the archaeon Ignisphaera aggregans: implications for the phylogeny and biosynthesis of glycerol monoalkyl glycerol tetraether isoprenoid lipids.

The lipid cores from Ignisphaera aggregans, a hyperthermophilic Crenarchaeon recently isolated from New Zealand hot springs, have been profiled by liquid chromatography-tandem mass spectrometry. The distribution revealed includes relatively high proportions of monoalkyl (also known as H-shaped) tetraether cores which have previously been implicated as kingdom-specific biomarkers for the Euryarchaeota. Such high expression of monoalkyl tetraether lipids is unusual in the archaeal domain and may indicate that formation of these components is an adaptive mechanism that allows I. aggregans to regulate membrane behaviour at high temperatures. The observed dialkyl tetraether and monoalkyl tetraether lipid distributions are similar but not fully concordant, showing differences in the average number of incorporated rings. The similarity supports a biosynthetic route to the ring-containing dialkyl and monoalkyl tetraether lipids via a dialkyl tetraether core containing zero rings, or a closely related structural relative, as an intermediate. Currently, however, the precise nature of the biosynthetic route to these lipids cannot be deduced.

An RNA-aptamer-based assay for the detection and analysis of malachite green and leucomalachite green residues in fish tissue.

A robust screening assay employing solid phase extraction (SPE) followed by a novel aptamer-based procedure is presented for the rapid detection and semiquantitation of the triphenylmethane dye, Malachite Green (MG) and its primary metabolite Leucomalachite Green (LMG) in fish tissue. To the authors' knowledge, this is the first reported use of an RNA aptamer for the development of a diagnostic assay for the detection of chemical residues in food. The aptamer based screening assay is found to be highly specific for MG; but has negligible affinity for the LMG metabolite. However, because the LMG metabolite is lipophilic and known to be highly persistent in tissues, an oxidation step has been incorporated within the sample cleanup procedure to ensure that all LMG residues are converted to MG prior to measurement. This article provides evidence that an oligonucleotide aptamer can be used as an alternative recognition element to conventional antibodies with application to the detection of residues in food. Furthermore, this finding has the future potential to reduce the number of animals currently being used in the production of antibodies for immunodiagnostic kits.

A rapid microbial inhibition-based screening strategy for fluoroquinolone and quinolone residues in foods of animal origin.

A rapid, high-throughput antimicrobial screening assay for the detection of fluoroquinolone and 4-quinolone residues in foods of animal origin has been developed in ampoule format. The assay employs a single Escherichia coli species sensitive to those Gram-negative inhibitiory antimicrobial compounds and is presented in a comparable format to the existing commercially available Premi Test and Delvotest ampoule-based microbial inhibition tests (DSM, Delft, The Netherlands). In the novel E. coli assay the microorganism, in vegetative state, is inoculated into a nutrient agar pellet containing a pH sensitive acid-base indicator dye. A simple extraction protocol that is selective for fluoroquinolone and quinolone compounds was developed to recover, cleanup and concentrate the target analyte(s) from a variety of tissue types and matrices prior to screening analysis. The method detected 16 target compounds at concentrations equal to or below the maximum residue limits (where applicable). The method has been validated using the prototype assay in accordance with the 2002/657/EC guidelines for the validation of qualitative screening assays. False positive and false negative responses rates for the procedure have been determined as less than 5%. The stability of a selection of representative target analytes has been demonstrated for a 20-week period under a variety of storage conditions both in tissue and in extract.

Rapid discrimination of archaeal tetraether lipid cores by liquid chromatography-tandem mass spectrometry.

Atmospheric pressure chemical ionization liquid chromatography-tandem mass spectrometry (APCI LC-MS/MS) of tetraether lipid cores of archaeal origin reveals distinct dissociation pathways for three classes of core lipid extracted from Methanobacter thermautotrophicus. Within these classes, two isobaric tetraether lipids, one a scarcely reported lipid constituent of M. thermautotrophicus and the other an artefact formed during extraction from cultured cells, were identified and distinguished via their MS(2) spectra. APCI LC-MS/MS discriminates different tetraether core lipid types and isobaric species and reveals the mass of the constituent biphytanyl chains within the tetraether cores, albeit without full elucidation of their structures. Furthermore, the method allows direct estimation of the relative proportions of tetraether core lipids from chromatographic peak area measurement, allowing rapid profiling of these compounds in microbiological and environmental extracts.

Antioxidant activity of oregano, parsley, and olive mill wastewaters in bulk oils and oil-in-water emulsions enriched in fish oil.

The antioxidant activity of oregano, parsley, olive mill wastewaters (OMWW), Trolox, and ethylenediaminetetraacetic acid (EDTA) was evaluated in bulk oils and oil-in-water (o/w) emulsions enriched with 5% tuna oil by monitoring the formation of hydroperoxides, hexanal, and t-t-2,4-heptadienal in samples stored at 37 degrees C for 14 days. In bulk oil, the order of antioxidant activity was, in decreasing order (p < 0.05), OMWW > oregano > parsley > EDTA > Trolox. The antioxidant activity in o/w emulsion followed the same order except that EDTA was as efficient an antioxidant as OMWW. In addition, the total phenolic content, the radical scavenging properties, the reducing capacity, and the iron chelating activity of OMWW, parsley, and oregano extracts were determined by the Folin-Ciocalteau, oxygen radical absorbance capacity, ferric reducing antioxidant power, and iron(II) chelating activity assays, respectively. The antioxidant activity of OMWW, parsley, and oregano in food systems was related to their total phenolic content and radical scavenging capacity but not to their ability to chelate iron in vitro. OMWW was identified as a promising source of antioxidants to retard lipid oxidation in fish oil-enriched food products.

High-throughput methods to assess lipophilic and hydrophilic antioxidant capacity of food extracts in vitro.

Assays comprising three probes for different mechanisms of antioxidant activity in food products have been modified to allow better comparison of the contributions of the different mechanisms to antioxidant capacity (AOC). Incorporation of a common format for oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), and iron(II) chelating activity (ICA) assays using 96-well microplates provides a comprehensive and high-throughput assessment of the antioxidant capacity of food extracts. The methods have been optimized for aqueous extracts and validated in terms of limit of quantification (LoQ), linearity, and precision (repeatability and intermediate reproducibility). In addition, FRAP and ORAC assays have been validated to assess AOC for lipophilic extracts. The relative standard deviation of repeatability of the methods ranges from 1.2 to 6.9%, which is generally considered to be acceptable for analytical measurement of AOC by in vitro methods. Radical scavenging capacity, reducing capacity, and iron chelating properties of olive mill wastewaters (OMWW), oregano, and parsley were assessed using the validated methods. OMWW showed the highest radical scavenging and reducing capacities, determined by ORAC and FRAP assays, respectively, followed by oregano and parsley. The ability to chelate Fe (2+) was, in decreasing order of activity ( p > 0.05) parsley congruent with oregano > OMWW. Total phenol content, determined by the Folin-Ciocalteu method, correlated to the radical scavenging and reducing capacities of the samples but not to their chelating properties. Results showed that the optimized high-throughput methods provided a comprehensive and precise determination of the AOC of lipophilic and hydrophilic food extracts in vitro.

Quantification and screening of pesticide residues in food by gas chromatography-exact mass time-of-flight mass spectrometry.

A gas chromatography-exact mass time-of-flight mass spectrometry (GC-TOF-MS) method has been developed for the quantification of approximately one hundred pesticides in baby food, pear and lettuce samples. Prior to analysis, co-extractives were removed from acetonitrile:toluene (80:20) extracts using dispersive solid-phase extraction with primary secondary amine (50mg) and carbon sorbent (50mg). The concentration of pesticides in the extracts was measured using an extracted mass chromatogram window of 0.05 Th. Samples spiked with pesticides at 0.01 and 0.1 mgkg(-1) yielded average recoveries in the range of 70-109% with relative standard deviations less than 26% and displayed good linearity for the majority of the analytes. The method was applied to the screening of pear and lettuce samples for pesticide residues. Targeted quantification and exact mass peak detection, deconvolution and library searching packages were used successfully to detect and identify incurred residues present in the samples at concentrations above 0.01 mgkg(-1). The new feature dynamic range enhancement, improved mass accuracy, and hence detection and quantification of the analytes across the concentration range studied.

Adduction of solvent molecules by ions isolated within an ion trap mass spectrometer under atmospheric pressure ionisation conditions.

Benzylpyridine and papaverine, an alkyl quinoline, both produce product ions containing an azepinium ring during atmospheric pressure chemical ionisation or electrospray multistage mass spectrometry. By controlling the trapping conditions, an isolated azepinium ion was held within the trap for an extended period of time without excitation. A subsequent analytical scan revealed a mass spectrum containing ions at two mass-to-charge (m/z) ratios, the first at the m/z of the isolated product ion and the second at an m/z ratio corresponding to the adduction of a molecule of solvent. Isolation and resonance excitation of the adduct ion remove the solvent molecule, resulting in recovery of the azepinium ion at the same signal intensity as the adduct ion. Isolating and trapping the ion for a further period allowed the solvent adduct ion to be re-formed. Modulation of the solvent flowing into the source while the ion was trapped allowed variation in the solvent molecule adducted to the trapped ion. The proportion of the ion current due to the adduct ion depends on the nature of the isolated ion, the proton affinity of the solvent and the length of time for which the ion was trapped. Adduct ion formation, deliberately maximised in this study, can occur to a significant extent under standard ion trap operating conditions, reducing the ion current of product ions of interest and, ultimately, the response in tandem mass spectrometric assays.

Ultra-performance liquid chromatography for the determination of pesticide residues in foods by tandem quadrupole mass spectrometry with polarity switching.

An ultra-performance liquid chromatographic (UPLC) electrospray ionisation tandem quadrupole mass spectrometric method has been developed for the determination of 52 pesticides in cereal-based baby foods, oranges and potatoes. The fast polarity switching capability of the mass spectrometer used enabled the determination of 44 of the compounds in the positive ionisation mode and 8 of the compounds in the negative ionisation mode in a single run. Prior to analysis, co-extractives were removed from acetonitrile extracts using dispersive solid-phase extraction with primary secondary amine (50 mg). The UPLC method separates all of the pesticides, resolves structural isomers (e.g. butocarboxim sulfoxide and aldicarb sulfoxide) and has a short (7 min) cycle time. Extracts spiked with pesticides at 0.10 and 0.01 mg kg(-1) yielded average recoveries in the range of 66-124% with relative standard deviations less than 19% for the majority of the analytes.

The impact of coral bleaching on the pigment profile of the symbiotic alga, Symbiodinium.

Bleaching of corals by loss of symbiotic dinoflagellate algae and/or photosynthetic pigments is commonly triggered by elevated temperatures coupled with high irradiance, and is a first-order threat to coral reef communities. In this study, a high-resolution high-performance liquid chromatography method integrated with mass spectrometry was applied to obtain the first definitive identification of chlorophyll and carotenoid pigments of three clades of symbiotic dinoflagellate algae (Symbiodinium) in corals, and their response to experimentally elevated temperature and irradiance. The carotenoids peridinin, dinoxanthin, diadinoxanthin (Dn), diatoxanthin (Dt) and beta-carotene were detected, together with chlorophylls a and c2, and phaeophytin a, in all three algal clades in unstressed corals. On exposure to elevated temperature and irradiance, three coral species (Montastrea franksi and Favia fragum with clade B algae, and Montastrea cavernosa with clade C) bleached by loss of 50-80% of their algal cells, with no significant impact to chlorophyll a or c2, or peridinin in retained algal cells. One species (Agaricia sp. with clade C) showed no significant reduction in algal cells at elevated temperature and irradiance, but lost substantial amounts of chlorophyll a and carotenoid pigments, presumably through photo-oxidative processes. Two coral species (Porites astreoides and Porites porites both bearing clade A algae) did not bleach. The impact of elevated temperature and irradiance on the levels of the photoprotective xanthophylls (Dn + Dt) and beta-carotene varied among the corals, both in pool size and xanthophyll cycling, and was not correlated to coral bleaching resistance.

Analysis of androgenic steroid Girard P hydrazones using multistage tandem mass spectrometry.

Seven androgenic steroids have been converted into steroid hydrazones using Girard P hydrazine and analysed by electrospray ionisation multistage tandem mass spectrometry. The cationic derivatives 17alpha-testosterone hydrazone, 17beta-nortestosterone hydrazone, 17beta-bolasterone hydrazone, 17alpha-boldenone hydrazone, 17beta-fluoxymesterone hydrazone, 17alpha-trenbolone hydrazone and 4-chloroandrosten-3,17-dione hydrazone show good response in positive ion mode with enhancements for the method of up to 33 times relative to the native species. Detailed characterisation of fragmentation pathways reveals structurally specific ions formed by fragmentation of the hydrazine moiety. Comparison of structural similarities among the androgenic steroids allows recognition of common ions/fragmentation processes as well as analyte-specific transitions. The suitability of the derivatisation approach in the screening of heifer urine for the presence of administered hormones has been demonstrated using partially purified urine spiked with a mixture of androgenic steroids.

Semiautomated determination of pesticides in water using solid phase extraction disks and gas chromatography-mass spectrometry.

A method based on semiautomated solid phase extraction using octadecyl-bonded silica disks and gas chromatography-mass spectrometry, operated in selected ion monitoring mode, allows detection and quantification of approximately 100 pesticides and transformation products in drinking water. Samples (500 mL) were passed through the disk, and the retained pesticides were eluted with acetone and ethyl acetate. Typical recoveries for pesticides at 0.1 microg L(-1) in water were in the range of 72-120% with relative standard deviations less than 20%. Calibration curves were linear over the range of 0.025-0.5 microg mL(-1) (equivalent to a concentration range in drinking water of 0.05-1.0 microg L(-1)).

Application of programmable temperature vaporisation injection with resistive heating-gas chromatography flame photometric detection for the determination of organophosphorus pesticides.

The combination of a programmable temperature vaporisation (PTV) injector with resistive heating GC (RH-GC), a form of fast GC, has been applied to the analysis of organophosphorus (OP) pesticides. The PTV injector was optimised in the 'at-once' solvent vent mode for the injection of ethyl acetate (10-40 microL) or ACN (10 microL). The short RH-GC column (5 m x 0.25 mm ID) with fast temperature ramps (up to 153 degrees C/ min) allowed the separation of a total of 20 OP pesticides in less than 6 min. Average recoveries between 67 and 119% were obtained for pesticides spiked at 0.01 mg/kg into apple and pear matrix. Extraction of orange juice with ACN provided higher recoveries (92-104%) for methamidophos, acephate and omethoate compared to ethyl acetate (62-73%). Results for analysis of OP pesticides in samples containing incurred residues were in good agreement with those obtained using GC-MS. The overall method was rapid, allowing 20 samples to be analysed in 4 h.

Atmospheric pressure ionisation mass spectrometric fragmentation pathways of noscapine and papaverine revealed by multistage mass spectrometry and in-source deuterium labelling.

Two opium alkaloids, noscapine and papaverine, show good response as [M+H]+ ions in positive ion electrospray mass spectrometry and atmospheric pressure chemical ionisation mass spectrometry. The two compounds exhibit markedly different fragmentation pathways and behaviour under multistage mass spectrometry (MSn), with papaverine displaying a wealth of ions in MS2 and noscapine providing a single dominant ion at each stage of MSn prior to MS4. Elucidation of the fragmentation pathways using the MSn capability of the ion trap was aided by spraying the analytes in 2H2O to incorporate an isotopic label. Simplex optimisation allowed optimum trapping and fragmentation parameters to be determined, leading to a six-fold improvement in response for one transition and a seven-fold improvement for one transition sequence.

Comparison of ultra-performance liquid chromatography and high-performance liquid chromatography for the determination of priority pesticides in baby foods by tandem quadrupole mass spectrometry.

Determination of 16 priority pesticides and transformation products specified in the EU Baby Food Directive 2003/13/EC has been compared using high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC) coupled to tandem quadrupole mass spectrometry (MS/MS). Prior to analysis, co-extractives were removed from acetonitrile extracts using dispersive solid-phase extraction (SPE) with primary secondary amine (50 mg). Extracts spiked with pesticides at 1 microg kg(-1) yielded average recoveries in the range 85-119%, with relative standard deviations less than 17%. The HPLC-MS/MS and UPLC-MS/MS multi-residue methods developed are simple, rapid and suitable for the quantification and confirmation of the 16 priority pesticides in fruit-, potato- and cereal-based baby food at 1 microg kg(-1). The major advantages of UPLC, using 1.7 microm particles, over HPLC are the speed of analysis, the narrower peaks (giving increased signal-to-noise ratio) and improved confirmation for the targeted pesticides in the analyses of baby foods.

Determination of priority pesticides in baby foods by gas chromatography tandem quadrupole mass spectrometry.

A gas chromatography-tandem quadrupole mass spectrometry (GC-MS/MS) method for the determination of twelve priority pesticides, and transformation products (e.g. metabolites) specified in the EU Baby Food Directive 2003/13/EC is described. Prior to GC-MS/MS analysis, co-extractives were removed from acetonitrile extracts using dispersive solid phase extraction with octadecyl (200 mg) and primary secondary amine (50 mg) sorbents. The clean up proved essential for the satisfactory long-term chromatographic performance during the analysis of a range of representative commercially pre-prepared baby food samples. Extracts spiked with pesticides at 1-8 microg kg(-1), yielded average recoveries in the range 60-113% with relative standard deviations less than 28%.