Comparison of sample preparation methods for analysis of isoflavones in foodstuffs

Due to the lack of one universally applicable and commonly used reference method, sample preparation in isoflavone (IF) analysis has been performed by many different methods which renders comparison and quality assessment of published IF contents in foodstuffs difficult.In the present work, the impact of different experimental parameters on the IF concentrations determined in soybeans, tofu, soy drink and textured vegetable protein by different extraction and hydrolysis methods was assessed and IF contents obtained by optimized orthogonal methods were compared. Chromatographic analysis was performed by HPLC-UV-ESI-MS. If possible sources of error - which are also pointed out in this work - are avoided, IF contents obtained by extraction, acid-, base- and enzymatic hydrolysis are similar. However, these sample preparation methods differ in the amount of time, standard compounds and instruments required, ruggedness, and in their applicability to analysis of complex composite samples containing soy as minor ingredient. Enzymatic hydrolysis with Helix pomatia juice after extraction by sonication with first 50, then 80% aqueous acetonitrile in the presence of zinc sulfate heptahydrate and after adjustment to ≤10% organic solvent turned out to be the method of choice if only aglucone equivalent contents are required. The advantages of this method are short chromatographic run times, smallest danger of coelution, lowest achievable limits of quantitation and therefore best suitability for work-up of complex composite samples and that only aglucone standards are needed for quantitation.     

Development of a broad toxicological screening technique for urine using ultra-performance liquid chromatography and time-of-flight mass spectrometry

Withdrawal of the support for the REMEDi HS drug profiling system has necessitated its replacement within our laboratories with an alternative broad toxicological screening technique. To this end, a novel method, based on ultra-performance liquid chromatography (UPLC) and time-of-flight (TOF) mass spectrometry, was developed for the routine analysis of urine samples. Identification was achieved by comparison of acquired data to libraries containing more than 300 common drugs and metabolites, and was based on a combination of retention time, exact mass and fragmentation patterns. Validation data for the method is presented and comprised an evaluation of the following parameters: precision; transferability of the methodology between the six collaborating laboratories; specificity; extraction recovery and stability of processed samples; matrix effects and sensitivity.This paper presents the benefits of supplementary fragmentation data with particular regard to increasing specificity and confidence of identification and its usefulness with overdosed samples. The utility of the method was assessed by the parallel analysis of 30 authentic urine samples using the REMEDi HS and UPLC-TOF. The latter provided enhanced detection, leading to the identification of twice as many drugs. Furthermore it did not miss any compounds that were identified by REMEDi HS. The UPLC-TOF findings were further verified by a combination of data from three other conventional screening techniques, i.e., GC-MS, HPLC-DAD and UPLC-MS/MS.     

Asymptotic random graph intuition for the biased connectivity game

We study biased Maker/Breaker games on the edges of the complete graph, as introduced by Chvátal and Erd?s. We show that Maker, occupying one edge in each of his turns, can build a spanning tree, even if Breaker occupies b ≤ (1 ? o(1)) · edges in each turn. This improves a result of Beck, and is asymptotically best possible as witnessed by the Breaker‐strategy of Chvátal and Erd?s. We also give a strategy for Maker to occupy a graph with minimum degree c (where c = c(n) is a slowly growing function of n) while playing against a Breaker who takes b ≤ (1 ? o(1)) · edges in each turn. This result improves earlier bounds by Krivelevich and Szabó. Both of our results support the surprising random graph intuition: the threshold bias is asymptotically the same for the game played by two “clever” players and the game played by two “random” players. © 2009 Wiley Periodicals, Inc. Random Struct. Alg., 2009     

Gold-catalyzed rearrangement of substituted allyl aryl ethers

Triphenylphosphinegold(I) complexes catalyze the Claisen-type rearrangement of aryl allyl ethers to the corresponding branched and linear products. The product distribution depends on the olefin geometry of the allylic ether. Stereochemical transfer experiments support an ionic mechanism.     

Proficiency testing in analytical chemistry, microbiology, and laboratory medicine – working group discussions on current status, problems, and future directions

Working group (WG) discussions on proficiency testing (PT) held at the joint Eurachem/ EQALM workshop, Borås, Sweden, 24–26 September 2000 are summarized. The discussions focused on aspects of PT and accreditation (WG 1), general aspects of PT in analytical chemistry (WG 2), microbiology (WG 3), and laboratory medicine (WG 4), incorporation of measurement uncertainty into PT schemes (WG 5), international harmonization of PT schemes (WG 6), and the role of PT in the international structure of chemical measurement (WG 7). Current status, problems and future directions are identified. Each WG contained a majority of participants experienced in the subject being covered by that WG, and a few participants with different expertise. This was done to promote cross-fertilization of ideas between sectors, a key objective of the workshop. The WG issues reflected the content of the keynote lectures and some issues were covered from different perspectives by more than one group.     

Stable Galerkin reduced order models for linearized compressible flow

The Galerkin projection procedure for construction of reduced order models of compressible flow is examined as an alternative discretization of the governing differential equations. The numerical stability of Galerkin models is shown to depend on the choice of inner product for the projection. For the linearized Euler equations, a symmetry transformation leads to a stable formulation for the inner product. Boundary conditions for compressible flow that preserve stability of the reduced order model are constructed. Preservation of stability for the discrete implementation of the Galerkin projection is made possible using a piecewise-smooth finite element basis. Stability of the reduced order model using this approach is demonstrated on several model problems, where a suitable approximation basis is generated using proper orthogonal decomposition of a transient computational fluid dynamics simulation.     

Cyclodextrin-modified micellar electrokinetic chromatography for the analysis of Esterom,a topical product consisting of hydrolyzed benzoylecgonine in propylene glycol

Esterom, a new drug currently in human clinical trials, is a mixture of compounds in a propylene glycol vehicle. It is being evaluated as a topical treatment to aid in the relief of muscle pain and to increase range of motion. Benzoylecgonine is the major component of Esterom and there are at least nine other minor constituents, including four hydroxypropyl esters that have multiple diasteriomers. The aim of the study was to develop a capillary electrophoresis method for the simultaneous separation of the main components in Esterom, including the multiple proposed diastereomers of the esters. Due to the complex sample composition, the use of micelles and cyclodextrins as buffer modifiers was evaluated. A cyclodextrin-modified micellar electrokinetic chromatography method was able to determine 7 of the 8 UV-active Esterom components, with baseline separation of 7 of the 10 diastereomers of the hydroxypropyl esters.     

Designing cytokine variants by phage-display

Cytokines are important mediators of many cellular functions including coordination of the immune system and regulation of regenerative processes. Therefore, cytokines can be exploited for therapeutic strategies. Cytokines can be altered in a way that their biologic activity is enhanced or antagonized. This can be accomplished by changing the interaction of cytokines with their cognate cytokine receptor complexes. Therefore, many research groups tried to design cytokines, which bind with higher affinity to their receptors. Alternatively, cytokine variants have been created which do bind to their receptors but do not elicit a signal. Such strategies have been followed using high throughput techniques like error-prone polymerase chain reaction and phage display. Designer cytokines can be used to specifically inhibit cytokine functions. Moreover, peptides have been generated with the help of phage display techniques, which exhibit cytokine activity. Surprisingly, such mimetic peptides do not show any sequence similarity to the parental cytokines. Such peptide mimetics can be used as lead structures for the generation of non-peptidic chemical compounds with cytokine activity.