Forensically Robust Detection of the Presence of Morpholine in Apples—Proof of Principle

Morpholine is a precursor of carcinogenic nitrosamines and although the possibility of their formation in the human stomach after ingestion of morpholine-treated apples is reported as highly unlikely morpholine has not been authorised as a food additive in the EU. Methods for its detection are required since it is permitted in other jurisdictions and may be present on food through direct treatment of fruit with waxes containing the compound, through steam treatment during processing or from packaging. Methods using derivatising agents with the inclusion of UV chromophores such as dansyl chloride yield good separation and high sensitivity but with mass spectrometric fragment ions predominantly originated from the derivatising group rather than the morpholine moiety. An amine acetylation derivatisation method is proposed from which fragment ions originating from the morpholine group are detected using widely available GC–MS. With full validation, a forensically robust confirmation of the presence of morpholine via its N-acetyl derivative is possible in support of regulatory analysis.     

Apparatus and initial results for pressurized planar electrochromatography

Pressurized planar electrochromatography (PPEC) is a new planar chromatographic technique in which the mobile phase is driven by electroosmotic flow, while the sorbent layer is pressurized in a manner that allows heat to flow from the layer through an electrically insulating, thermally conducting, sheet of aluminum nitride ceramic. A prototype apparatus for performing PPEC is described. Separation by PPEC is faster than by conventional TLC, and an example is presented of a 24-fold enhancement in the speed of separation. PPEC was performed on both regular and high-performance C18 layers, and the latter yield substantially faster separation. The sorbent layer requires conditioning at elevated temperature before use, and solute migration velocity increases with this temperature. The flow rate increases in a linear manner with increasing voltage and diminishes in a nonlinear manner with increasing pressure. Both electrical current and Joule heating diminish with increasing pressure, and the diminution of flow at high pressure can be compensated by an increase in voltage. PPEC is more efficient than classical TLC. Theoretical plate heights diminish with increasing Rf and are in the range 29-21 and 55-27 microm for the high-performance and regular plates, respectively. PPEC retains the advantages of classical TLC but has the ability to separate a substantially higher number of samples simultaneously. An example is presented on the separation of nine samples in 1 min on a 2.5 cm x 10 cm sorbent layer.     

Small‐Scale Thermal Studies of Volatile Homemade Explosives

Several homemade or improvised explosive mixtures that either contained volatile components or produced volatile products were examined using standard small‐scale safety and thermal (SSST) testing that employed differential scanning calorimetry (DSC) techniques (constant heating rate and standard sample holders). KClO₃ and KClO₄ mixtures with dodecane exhibited different enthalpy behavior when using a vented sample holder in contrast to a sealed sample holder. The standard configuration produced profiles that exhibited only endothermic transitions. The sealed system produced profiles that exhibited additional exothermic transitions absent in the standard configuration produced profiles. When H₂O₂/fuel mixtures were examined, the volatilization of the peroxide (endothermic) dominated the profiles. When a sealed sample holder was used, the energetic releases of the mixture could be clearly observed. For AN and AN mixtures, the high temperature decomposition appears as an intense endothermic event. Using a nominally sealed sample holder also did not adequately contain the system. Only when a high‐pressure rated sample holder was used the high temperature decomposition of the AN could be detected as an exothermic release. The testing was conducted during a proficiency (or round‐robin type) test that included three U.S. Department of Energy and two U.S. Department of Defense laboratories. In the course of this proficiency test, certain HMEs exhibited thermal behavior that was not adequately accounted for by standard techniques. Further examination of this atypical behavior highlighted issues that may have not been recognized previously because some of these materials are not routinely tested. More importantly, if not recognized, the SSST testing results could lead to inaccurate safety assessments. This study provides examples, where standard techniques can be applied, and results can be obtained, but these results may be misleading in establishing thermal properties.     

Small‐Scale Safety Testing of Ammonium Nitrate and Mixtures

Ammonium nitrate (AN), gunpowder (GP), and an ammonium nitrate gunpowder mixture (AN/GP) were studied for impact sensitivity by four laboratories using the drop hammer apparatus. Bruceton and Neyer methods were used as experimental protocols and for data reduction. The results are presented as 50 % probability of reaction (DH₅₀). For AN, the DH₅₀ values are widely varied among the participants, from sensitive to completely insensitive (limit of the equipment), with no real correlation among results. GP and the AN/GP mixture exhibited much more sensitivity overall and were in some cases within statistical values extrapolated from previous studies of RDX. The variability in results for the AN data is attributed to the difficulty in determining a positive reaction event for AN, as detailed by Neyer experiments and photography during positive reactions. Variability in results for the GP and AN/GP mixtures is attributed to equipment environment and detection criteria. This work was performed by the Integrated Data Collection Analysis (IDCA) program, a multi‐laboratory effort to standardize safety testing of improvised or homemade explosives funded by the Department of Homeland Security.     

Carbon electrode fabrication from pyrolyzed parylene C

Carbon electrodes coupled with electrochemical detection have been used extensively for the investigation of biogenic amines. Herein we report the fabrication and characterization of carbonaceous electrodes prepared from pyrolyzed parylene C (PPC) films. High-aspect ratio carbonaceous microelectrodes have been prepared by masking PPC coated pipets with an insulating parylene C film. PPC thin film electrodes were characterized electrochemically, spectroscopically, and with electron microscopy. The procedures described here offer a route to fabrication of thin film carbon electrodes that can be patterned and produced in parallel. These electrodes are similar to carbon electrodes derived from pyrolyzed photoresist films but do not require spin-coating or lithography and can readily coat three-dimensional surfaces.     

Detection of C-reactive protein utilizing magnetic permeability detection based immunoassays

A new sensing technology platform integrating magnetic permeability detection and a two-site heterogeneous immunoassay in a one-step analysis is described. As a platform model, measurements of C-reactive protein (CRP), a cardiac and inflammation marker, were performed in a rapid (11.5 min) high-sensitivity (hs) procedure with a low detection limit (0.2 mg/L) and accuracy (CV = 11%). The two-site heterogeneous immunoassay was performed in 1.2-mL disposable reagents vials containing solid phase (polyclonal anti-CRP conjugated silica microparticles), labeling agent (monoclonal anti-CRP conjugated superparamagnetic nanoparticles), and reaction buffer. Whole blood (20 muL) was assayed by introducing the sample into a reagent vial using a glass capillary and mixing its contents by hand for 30 s. After a 11-min sedimentation step, the vial was placed into the coil of the magnetic permeability detector, which measured the enrichment of superparamagnetic nanoparticles in the solid-phase sediment. Magnetic permeability detection and quantification is based on the principle that when paramagnetic materials are placed inside a coil, the inductance of the coil is influenced. Screening of CRP on whole blood patient samples showed good correlation with central hospital measurements for hsCRP (y = 1.018x - 0.021, R(2) = 0.980, n = 103) and normal range CRP (y = 1.02x + 2.53, R(2)= 0.991, n = 33) analyses. The mean differences of the two methods according to the Bland and Altman plots were -0.03 +/- 1.12 mg/L for hsCRP analysis and -3.4 +/- 8.64 mg/L for normal range CRP analysis.     

N-Heterocyclization in Gliotoxin Biosynthesis is Catalyzed by a Distinct Cytochrome P450 Monooxygenase

Gliotoxin and related epidithiodiketopiperazines (ETP) from diverse fungi feature highly functionalized hydroindole scaffolds with an array of medicinally and ecologically relevant activities. Mutation analysis, heterologous reconstitution, and biotransformation experiments revealed that a cytochrome P450 monooxygenase (GliF) from the human-pathogenic fungus Aspergillus fumigatus plays a key role in the formation of the complex heterocycle. In vitro assays using a biosynthetic precursor from a blocked mutant showed that GliF is specific to ETPs and catalyzes an unprecedented heterocyclization reaction that cannot be emulated with current synthetic methods. In silico analyses indicate that this rare biotransformation takes place in related ETP biosynthetic pathways.     

A confessional account of community entry: doing critically reflexive ICT4D work in a deep rural community in South Africa*

ABSTRACT

In this paper I reflect on the community entry phases of doing a critical ethnography in a traditional Zulu community in a deep rural part of South Africa. I present my reflections in the form of confessional narratives on community entry and on how an Information Communication Technology for Development (ICT4D) project was introduced in the community. Drawing on a basic principle of doing social science research, I ask: In what ways should I achieve self-emancipation, in order to ensure the on-going emancipation and empowerment of the people I engage with? I also ask: How should one do critically reflexive community entry and ICT4D project introduction in communities that embrace a people-orientated worldview? The paper is practice-orientated in that it demonstrates critical self-reflexive community entry in a particular ICT4D project situation, how community entry encounters informed follow-up work, and how cultural interpreters empowered me to do community entry successfully. Methodologically, the paper contributes by demonstrating the value of using narrative examples to represent virtuoso expertise of how I developed knowledge of self-reflexivity while excavating hidden social meanings from an ICT4D project situation.