Impulse-driven heated-droplet deposition interface for capillary and microbore LC-MALDI MS and MS/MS

An automated off-line liquid chromatography-matrix-assisted laser desorption ionization (LC-MALDI) interface capable of coupling both capillary and microbore LC separations with MALDI mass spectrometry (MS) and tandem mass spectrometry (MS/MS) has been developed. The interface is a combination of two concepts: analyte concentration from heated hanging droplets and impulse-driven droplet deposition of LC fractions onto a MALDI sample plate. At room temperature the interface allows the coupling of capillary LC separations (i.e., flow rate of <5 microL/min) with MALDI MS. With heating, it can be used to combine microbore LC operated at a relatively high flow rate of up to 50 microL/min with MALDI MS. The collected fractions can be analyzed by MALDI MS and MS/MS instruments, such as time-of-flight (TOF) and quadrupole-TOF MS. Performance of the interface was examined using several peptide and protein standards. It was shown that, using MALDI-TOF MS, [GLU1]-fibrinopeptide B could be detected with a total injection amount of 5 fmol to microbore LC. Chromatographic performance was also monitored. A peak width of 12 s at half-height for [GLU1]-fibrinopeptide B showed no evidence of band broadening due to the interface. The ability of the interface to mitigate ion suppression was studied using a mixture of 100 fmol of [GLU1]-fibrinopeptide B and 10 pmol of cytochrome c tryptic digest. Although fully suppressed under direct MALDI conditions, LC-MALDI analysis was able to detect the 100 fmol peptide with 10 s fraction collection. Finally, the ability to inject relatively large sample amounts to improve detectability of low-abundance peptides was illustrated in the analysis of phosphopeptides from alpha-casein tryptic digests. A digest loaded on column to 2.4 microg and analyzed by LC-MALDI MS/MS resulted in 82% sequence coverage and detection of all nine phosphoserine residues. It is concluded that, being able to handle both high- and low-flow LC separations, the impulse-driven heated-droplet interface provides the flexibility to carry out MALDI analysis of peptides and proteins depending on the information sought after, analysis speed, and sample size.     

Inhibitory Effect of Fusarium Mycotoxins on Growth of Brewing Yeasts. 1 Zearalenone and Fumonisin B1*

The Fusarium mycotoxins zearalenone (ZEA) and fumonisin B₁ (FB₁) added to the growth medium in low and high concentrations, were investigated as a possible cause of inhibition of growth of Saccharomyces cerevisiae lager and ale strains. Toxic effects were assessed by measurement of dry weight or growth relative to controls, cell number, viability and conductance changes of the growth medium using indirect and direct methods. The Fusarium mycotoxins studied affected growth on brewing yeasts, but the inhibitory effect was dependent on concentration. Low concentrations (0.1–2 μg/ml) had no significant effect on growth compared to controls. Although high concentrations of both mycotoxins strongly affected growth, the inhibitory effect depended on toxin concentration and type, yeast strain, length of incubation and method used to assess growth. The lowest concentrations of mycotoxin causing significant inhibition on growth of these brewing yeasts were 50 μg/ml ZEA for both yeast strains, and 10 μg/ml FB₁ for the lager strain and 50 μg/ml for the ale strain.     

The density algorithm for pairwise interaction testing

There are many published algorithms for generating interaction test suites for software testing, exemplified by AETG, IPO, TCG, TConfig, simulated annealing and other heuristic search, and combinatorial design techniques. Among these, greedy one‐test‐at‐a‐time methods (such as AETG and TCG) have proven to be a reasonable compromise between the needs for small test suites, fast test‐suite generation, and flexibility to accommodate a variety of testing scenarios. However, such methods suffer from the lack of a worst‐case logarithmic guarantee on test suite size, while methods that provide such a guarantee at present are less efficient or flexible, or do not produce test suites that are competitive in size for practical testing scenarios. In this paper, a new algorithm establishes that efficient, greedy, one‐test‐at‐a‐time methods can indeed produce a logarithmic worst‐case guarantee on the test suite size. In addition, this can be done while still producing test suites that are of competitive size, and in a time that is comparable to the published methods. It is deterministic, guaranteeing reproducibility. It generates only one candidate test at a time, permits users to ‘seed’ the test suite with specified tests, and allows users to specify constraints of combinations that should be avoided. Further, statistical analysis examines the impact of five variables used to tune this density algorithm for execution time and test suite size: weighting of density for factors, scaling of density, tie‐breaking, use of multiple candidates, and multiple repetitions using randomization. Copyright © 2007 John Wiley & Sons, Ltd.     

A density‐based greedy algorithm for higher strength covering arrays

Algorithmic construction of software interaction test suites has focussed on pairwise coverage; less is known about the efficient construction of test suites for t‐way interactions with t≥3. This study extends an efficient density‐based algorithm for pairwise coverage to generate t‐way interaction test suites and shows that it guarantees a logarithmic upper bound on the size of the test suites as a function of the number of factors. To complement this theoretical guarantee, an implementation is outlined and some practical improvements are made. Computational comparisons with other published methods are reported. Many of the results improve upon those in the literature. However, limitations on the ability of one‐test‐at‐a‐time algorithms are also identified. Copyright © 2008 John Wiley & Sons, Ltd.     

Investigation of a simple tensorial permeameter

A device to measure permeability tensors of aligned circular cylinder arrays was designed and tested. Experiments were performed on arrays having solid volume fractions ranging from 0.3 to 0.7, inclusive. Array structures had 60° (hexagonal), 70°, and 90° (square) angles of symmetry. Results for transverse flow across the cylinder arrays were in strong agreement with numerical predictions. Transverse permeability values were also shown to obey tensor transformation rules. Longitudinal test results indicated that transverse flow was present during the experiments, lowering measured permeability values. The influence of longitudinal permeability on the overall tensor was shown to be substantially larger than variations in the transverse permeability values. POLYM. COMPOS., 2010. © 2009 Society of Plastics Engineers     

The Role of Aluminum in the Detonation and Post‐Detonation Expansion of Selected Cast HMX‐Based Explosives

In order to improve understanding of how aluminum contributes in non‐ideal explosive mixtures, cast‐cured formulations have been analyzed in a series of cylinder tests and plate‐pushing experiments. This study describes the contribution of 15 % aluminum (median size of 3.2 μm) vs. lithium fluoride (an inert substitute for aluminum; <5 μm) in cast‐cured HMX formulations in different temporal regimes. Small cylinder tests were performed to analyze the detonation and wall velocities (1–20 μs) for these formulations. Near‐field blast effects of 58 mm diameter spherical charges were measured at 152 mm and 254 mm using steel plate acceleration. Pressure measurements at 1.52 m gave information about free‐field pressure at several milliseconds. While the observed detonation velocities for all formulations were within uncertainty, significantly higher cylinder wall velocities, plate velocities, and pressures were observed for the aluminum formulations at ≥2 μs. Additionally, hydrocode calculations were performed to determine how non‐ideal behavior affected the plate test results. Collectively, this work gives a clearer picture of how aluminum contributes to detonation on timescales from 1 μs to about 2 ms, and how the post‐detonation energy release contributes to wall velocities and blast effects. The experiments indicate that significant aluminum reactions occur after the CJ plane, and continue to contribute to expansion at late times.