Preparation of quercetin glucuronides and characterization by HPLC–DAD–ESI/MS

Quercetin is the main flavonol in the human diet, and the most commonly used in studies of biological activity. The major circulating forms of quercetin found in the human plasma after consumption of food containing distinct quercetin glycosides are glucuronides and sulfates. In this work quercetin glucuronides have been obtained from green beans (quercetin 3-glucuronide) and by enzymic synthesis (quercetin 4′-glucuronide) using a modification of the method described by Plumb et al. (Methods in polyphenol analysis, The Royal Society of Chemistry, Cambridge, pp 187, 2003) so as to improve the original low yields of that methodology. The method finally optimised got yields of 19% in the preparation of quercetin 4′-glucuronide, which allows its further isolation for their use in biological assays. In addition, quercetin 3′-glucuronide, 3-glucuronide and a diglucuronide were synthesised with lower yields. The compounds prepared have been employed to perform assays in order to obtain data for their identification by HPLC coupled to photodiode array detection and tandem mass spectrometry. It was observed that the analysis by HPLC–ESI/MS/MS could allow the identification of different quercetin glucuronides based on the presence of some minor key MS² fragments.     

Postharvest storage and cooking techniques affect the stability of glucosinolates and myrosinase activity of Andean mashua tubers (Tropaeolum tuberosum)

The identification and quantification of Glucosinolates (Gls) via UPLC^® MS-MS/PDA/qToF in ten mashua cultivars was carried out. Gls total contents were within the 4.9–54.2 μmol g⁻¹ dry matter range, of which 96–99% corresponded to glucoaubrietin. Other less abundant Gls were glucotropaeolin and tentatively two isomers of hydroxybenzyl Gls. Postharvest refrigeration and shade storage conditions increased the content of Gls up to day 6 in 39.7% and 51.5% respectively. Sun exposure increased the Gls content in 40% up to day 3 but from day 6 considerable losses were attained (92% at day 15). Low correlation levels (R²) between the Gls and myrosinase (MYR) activity of 0.57, 0.28 and 0.39 for the refrigeration, shade and sun exposure treatments were obtained. The cooking regimes tested, boiling, microwaving and baking totally inactivated MYR without affecting the Gls content.     

Synthesis,Structure–Activity,and Structure–Stability Relationships of 2‐Substituted‐N‐(4‐oxo‐3‐oxetanyl) N‐Acylethanolamine Acid Amidase (NAAA) Inhibitors

N‐Acylethanolamine acid amidase (NAAA) is a cysteine amidase that preferentially hydrolyzes saturated or monounsaturated fatty acid ethanolamides (FAEs), such as palmitoylethanolamide (PEA) and oleoylethanolamide (OEA), which are endogenous agonists of nuclear peroxisome proliferator‐activated receptor‐α (PPAR‐α). Compounds that feature an α‐amino‐β‐lactone ring have been identified as potent and selective NAAA inhibitors and have been shown to exert marked anti‐inflammatory effects that are mediated through FAE‐dependent activation of PPAR‐α. We synthesized and tested a series of racemic, diastereomerically pure β‐substituted α‐amino‐β‐lactones, as either carbamate or amide derivatives, investigating the structure–activity and structure–stability relationships (SAR and SSR) following changes in β‐substituent size, relative stereochemistry at the α‐ and β‐positions, and α‐amino functionality. Substituted carbamate derivatives emerged as more active and stable than amide analogues, with the cis configuration being generally preferred for stability. Increased steric bulk at the β‐position negatively affected NAAA inhibitory potency, while improving both chemical and plasma stability.     

Multiple and simultaneous fluorophore detection using fluorescence spectrometry and partial least-squares regression with sample-specific confidence intervals

Fluorescent labeling is widely used in biological and chemical analysis, and the drive for increased throughput is stretching multiplexing capabilities to the limit. The limiting factor in multiplexed analyses is the ability to subsequently deconvolute the signals. Consequently, alternative approaches for interpreting complex data sets are required to allow individual components to be identified. Here we have investigated the application of a novel approach to multiplexed analysis that does not rely on multivariate curve resolution to achieve signal deconvolution. The approach calculates a sample-specific confidence interval for a multivariate (partial least-squares regression (PLSR)) prediction, thereby enabling the estimation of the presence or absence of each fluorophore based on the total spectral signal. This approach could potentially be applied to any multiplexed measurement system and has the advantage over the current algorithm-based methods that the requirement for resolution of spectral peaks is not central to the method. Here, PLSR was used to obtain the concentrations for up to eight dye-labeled oligonucleotides at levels of (0.6-5.3) x 10(-6) M. The sample-specific prediction intervals show good discrimination for the presence/absence of seven of the eight labeled oligonucleotides with efficiencies ranging from approximately 91 to 100%.     

Carbohydrate‐based poly(ester‐urethane)s: A comparative study regarding cyclic alditols extenders and polymerization procedures

A set of segmented poly(ester‐urethane)s were prepared from diisocyanates HDI or MDI and using 1,4‐butanediol and D ‐glucose‐derived cyclic diols (1,4 : 3,6‐dianhydro‐D ‐glucitol (isosorbide) or 2,4;3,5‐di‐O‐methylidene‐D ‐glucitol (gludioxol) or mixtures of them) as extenders. Hydroxyl end‐capped polycaprolactone with a molecular weight of 3000 g·mol^?1 was used as soft segment. Two polymerization methods, in solution and in bulk, were applied for the synthesis of these poly(ester‐urethane)s. The influence of the preparation procedure and composition in cyclic extender on synthesis results, structure, and properties of the novel poly(ester‐urethane)s was comparatively evaluated and discussed. The effect of replacement of 1,4‐butanediol by isosorbide or gludioxol on hydrodegradability was also assessed; the hydrolysis rate increased noticeably with the presence of glucitol derived units, although degradation of the polymers took place essentially by hydrolysis of the polyester soft segment. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Process integrated wire-bond quality control by means of cytokine-Formal Immune Networks

Ultrasonic wire bonding is one of the most frequently used techniques in semiconductor production to establish electrical interconnections. Improper bonding process parameters, wire or substrate contamination or low substrate quality are some of the causes of failed bonds. Process integrated wire-bond quality control techniques compare process feedback signals to a reference for monitoring online the quality of a bond. The feedback signals sampled at high frequencies, constitute high dimensional vectors representing the bonding process characteristics. In the area of online bond failure detection, dimensionality reduction of the input signals and feature extraction of the characteristics of the process are very demanding. Cytokine-Formal Immune Network (cFIN) is a procedure for pattern recognition which presents a low recognition failure rate and a fast recognition due to the reduction of dimensions and feature extraction of the training pattern data set produced in the learning phase. We use cytokine-Formal Immune Networks for recognizing faults present during the wire bonding process. The recognition methodology is intended to be applied into a process integrated quality control system. Further an automated optimization procedure has been developed to find optimal cFIN training parameters. Very promising results for two wire bonding process setups are shown in this paper.     

Improved creep performance of melt-extruded polycaprolactone/organo-bentonite nanocomposites

In this work, biodegradable nanocomposites based on polycaprolactone reinforced with pristine and organo-modified bentonites are prepared by melt extrusion. Bentonite is exchanged with benzalkonium chloride (CBK) in a pilot plant scale reactor. The influence of clay type and loading on morphology, rheology, mechanical properties, and creep performance of the resulting materials is analyzed. Besides, several theoretical models then applied to experimental creep data and master curves are used to relate time and temperature with the compliance of the materials. The morphology characterization of the nanocomposites show that the organo-modification of the clay greatly improves its dispersion in the polymer matrix. As a consequence, it is demonstrated that reinforcement of PCL with 3 wt% loading of organoclay produces the strongest improvement in creep resistance. The instantaneous creep strain and the experimental creep rate decrease more than 9% and 27%, respectively, in the range of temperatures analyzed. Moreover, the experimental values are used to adequately fit theoretical creep models for different clay loadings. On the other hand, the material with optimal creep behavior also shows the greatest improvements in tensile mechanical properties.     

Phytoplankton size-distribution and community structure in relation to mucilage occurrence in the northern Adriatic Sea

The spatial and temporal variations of pico-, nano- and microphytoplankton abundance and composition were investigated over a 37 month period, focusing on the ecological role of different size classes of phytoplankton, and on the changes of the community structure that might occur during periods when large mucilage macroaggregates appear. Samples were collected monthly from June 1999 to July 2002 at 11 stations, along three transects covering the northern Adriatic basin. Highest abundances were observed in late-winter/spring for microphytoplankton (mainly diatoms), in spring-summer for nanophytoplankton, and in summer for picophytoplankton. The autotrophic component was more abundant in the summers of 2000 and 2002 (when large mucilage aggregates occurred) than in the summers of 1999 and 2001 (when a massive phenomenon was not observed). This increase was statistically significant for pico-, nano- and, among microphytoplankton, only for dinoflagellates. Blooms of picophytoplankton were often observed at the bottom layer during mucilage summers. The microphytoplankton community during mucilage phenomena was characterized by a species composition (Chaetoceros spp., Cerataulina pelagica, Pseudo-nitzschia delicatissima, P. pseudodelicatissima, Cylindrotheca closterium, Dactyliosolen fragilissimus) comparable to that observed in summers without extensive mucilage occurrence. However, some species appeared with significantly higher densities in the summers of 2000 and 2002: Ceratium furca, C. closterium, Oxytoxum spp., Hemiaulus hauckii and Gonyaulax fragilis. Microscopic observation of aggregates revealed that the microphytoplankton species composition inside the aggregates was comparable to that observed in the water column, with an enrichment of opportunistic species such as C. closterium and P. delicatissima. The presence of mucilage aggregates affects the phytoplankton populations in the water column, even when aggregates are at early stages. It seems that there is a mutual relationship between phytoplankton and aggregates, i.e., several diatom and dinoflagellate species may contribute to the aggregate formation and enlargement, but mucilage aggregates themselves may also affect the phytoplankton populations, allowing the development of a rich diatom community and in general enhancing nanophytoplankton growth.