Next generation planar waveguide detection of microcystins in freshwater and cyanobacterial extracts,utilising a novel lysis method for portable sample preparation and analysis

The study details the development of a fully validated, rapid and portable sensor based method for the on-site analysis of microcystins in freshwater samples. The process employs a novel lysis method for the mechanical lysis of cyanobacterial cells, with glass beads and a handheld frother in only 10 min. The assay utilises an innovative planar waveguide device that, via an evanescent wave excites fluorescent probes, for amplification of signal in a competitive immunoassay, using an anti-microcystin monoclonal with cross-reactivity against the most common, and toxic variants. Validation of the assay showed the limit of detection (LOD) to be 0.78 ng mL⁻¹ and the CCβ to be 1 ng mL⁻¹. Robustness of the assay was demonstrated by intra- and inter-assay testing. Intra-assay analysis had % C.V.s between 8 and 26% and recoveries between 73 and 101%, with inter-assay analysis demonstrating % C.V.s between 5 and 14% and recoveries between 78 and 91%. Comparison with LC–MS/MS showed a high correlation (R² = 0.9954) between the calculated concentrations of 5 different Microcystis aeruginosa cultures for total microcystin content. Total microcystin content was ascertained by the individual measurement of free and cell-bound microcystins. Free microcystins can be measured to 1 ng mL⁻¹, and with a 10-fold concentration step in the intracellular microcystin protocol (which brings the sample within the range of the calibration curve), intracellular pools may be determined to 0.1 ng mL⁻¹. This allows the determination of microcystins at and below the World Health Organisation (WHO) guideline value of 1 μg L⁻¹. This sensor represents a major advancement in portable analysis capabilities and has the potential for numerous other applications.     

Strategies to improve the surface plasmon resonance-based immmunodetection of bacterial cells

We have made a comparison of (a) different surface chemistries of SPR sensor chips (such as carboxymethylated dextran and carboxymethylated C1) and (b) of different assay formats (direct, sandwich and subtractive immunoassay) in order to improve the sensitivity of the determination of the model bacteria Acidovorax avenae subsp. citrulli (Aac). The use of the carboxymethylated sensor chip C1 resulted in a better sensitivity than that of carboxymethylated dextran CM5 in all the assay formats. The direct assay format, in turn, exhibits the best sensitivity. Thus, the combination of a carboxymethylated sensor chip C1 with the direct assay format resulted in the highest sensitivity for Aac, with a limit of detection of 1.6?×?10⁶ CFU mL^-1. This SPR immunosensor was applied to the detection of Aac in watermelon leaf extracts spiked with the bacteria, and the lower LOD is 2.2?×?10⁷ CFU mL^?1.

Determining Energies and Cross Sections of Individual Ions Using Higher-Order Harmonics in Fourier Transform Charge Detection Mass Spectrometry (FT-CDMS)

A general method for in situ measurements of the energy of individual ions trapped and weighed using charge detection mass spectrometry (CDMS) is described. Highly charged (>?300 e), individual polyethylene glycol (PEG) ions are trapped and oscillate within an electrostatic trap, producing a time domain signal. A segmented Fourier transform (FT) of this signal yields the temporal evolution of the fundamental and harmonic frequencies of ion motion throughout the 500-ms trap time. The ratio of the fundamental frequency and second harmonic (HAR) depends on the ion energy, which is an essential parameter for measuring ion mass in CDMS. This relationship is calibrated using simulated ion signals, and the calibration is compared to the HAR values measured for PEG ion signals where the ion energy was also determined using an independent method that requires that the ions be highly charged (>?300 e). The mean error of 0.6% between the two measurements indicates that the HAR method is an accurate means of ion energy determination that does not depend on ion size or charge. The HAR is determined dynamically over the entire trapping period, making it possible to observe the change in ion energy that takes place as solvent evaporates from the ion and collisions with background gas occur. This method makes it possible to measure mass changes, either from solvent evaporation or from molecular fragmentation (MSⁿ), as well as the cross sections of ions measured using CDMS.

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Near‐field optical enhancement by lead‐sulfide quantum dots and metallic nanoparticles for SERS

There is a growing interest in using quantum dots (QDs) and metallic nanoparticles (NPs), both for luminescence enhancement and surface‐enhanced Raman scattering (SERS). Here, we study the electromagnetic‐field enhancement that can be generated by lead‐sulfide (PbS) QDs using three‐dimensional finite‐element simulations. We investigate the field enhancement associated with combinations of PbS QDs with metallic NPs and substrates. The results show that high field enhancement can be achieved by combining PbS QDs with metallic NPs of larger sizes. The ideal size for Ag NPs is 25 nm, providing a SERS enhancement factor of ~5*10⁸ for light polarization parallel to the NP dimer axis and a gap of 0.6 nm. For Au NPs, the bigger the size, the higher is the field for the studied diameters, up to 50 nm. The near‐field values for PbS QDs above metallic substrates were found to be lower compared to the case of PbS QD‐metal NP dimers. This study provides the understanding for the design and application of QDs for the enhancement of near‐field phenomena. Copyright © 2013 John Wiley & Sons, Ltd.     

First measurements of the 16O(e, e'pn)14N reaction

This paper reports on the first measurement of the ¹⁶O(e, e'pn)¹⁴N reaction. Data were measured in kinematics centred on a super-parallel geometry at energy and momentum transfers of 215MeV and 316MeV/c. The experimental resolution was sufficient to distinguish groups of states in the residual nucleus but not good enough to separate individual states. The data show a strong dependence on missing momentum and this dependence appears to be different for two groups of states in the residual nucleus. Theoretical calculations of the reaction using the Pavia code do not reproduce the shape or the magnitude of the data. An erratum to this article is available at.     

The influence of cage size on the reactivity of trimetallic nitride metallofullerenes: a mono- and bis-methanoadduct of Gd3N@C80 and a monoadduct of Gd3N@C84

A reactivity study of the higher TNT EMFs of gadolinium is reported here showing that the reactivity substantially decreases when the fullerene cage gets larger.     

Cyclopolymerization. VII. The 13C NMR Spectra of Cyclopolymers Obtained from N,N-Diallyamines

Pulsed Fourier transform ¹³C natural abundance nuclear magnetic resonance spectroscopy has been used to determine the structures of the polymers formed by the radical-induced cyclopolymerization of N-substituted-N, N-diallylamines and N-methyl-N, N-bls(2-alkylallyl)-amines. The polymers of N, N-diallylamines all contain cis-and trans-substituted pyrrolidine rings in the ratio 5:1. The polymers of N-methyl-N, N-bis(2-alkylallyl)amines give complex spectra due to the presence of both cis-and trans-pyrrolidine and -piperidine rings, but the difference in chemical shifts of the N-methyl signals from the different structural types allows the spectra to be analyzed.     

Improved Aldehyde Synthesis: Preparation of 3α, 7α, 12α-Triacetoxy-5β-Cholan-23-al with Ruthenium Tetroxide in Neutral Medium

In relation to our studies on the metabolism of neutral sterols² we have prepared several substituted 24-norcholanic acids. Ruthenium tetroxide was selected as the oxidizing agent^3,4 in the formation of 24-norcholic acid^5,6 from the olefin, 3α, 7α, 12α-triacetoxy-24, 24-diphenyl-5β-chol-23-ene (II), (Fig. 1) since CrO₃ provided multiple products⁵.     

Elucidating Inorganic Nanoscale Species in Solution: Complementary and Corroborative Approaches

The challenge of defining a length on the nanoscale is non‐trivial. For a well‐defined inorganic nanoscale species, a size measurement can describe a number of different dimensions (core, shell, solvation sphere). Often size is reported out of context or even inadvertently misrepresented. Since many of the techniques used to measure size depend on significant and sometimes destructive sample preparation, an additional challenge is defining “what size means” for a nanoscale species in solution. In this Concept, the distinction is made between complementary techniques that can be used together to unveil more information about the material in question, and corroborative techniques, which are used to make multiple measurements of the same property. Additionally, corroborative techniques can be used to measure the same property in and out‐of solution so as to reveal details about solution behaviour. We highlight various approaches to this characterization challenge in the context of three case studies that demonstrate the use of both complementary and corroborative techniques to elucidate the various functional dimensions of different types of inorganic nanoscale species in solution.