Bruker (Canada)

Abstract High throughput proteomics is gaining increasing traction as it facilitates screening of large sample cohorts required in clinical research and systems biology studies. Recent developments in mass spectrometry-based proteomics resulted in improved hardware and software providing deep proteome coverage, robustness, and scale accessible to a wide range of laboratories. Here, we benchmark dia-PASEF, a data-independent acquisition scheme that integrates trapped ion mobility with high scan speed, with a high-resolution time-of-flight mass analyzer (timsTOF HT) for the deep proteome analysis of a human cell line applying short 5-minute gradients. To show intra-and interlaboratory reproducibility, we performed a multi-laboratory study including 11 sites. We demonstrate that on average 7,072 protein groups and 99,835 peptides were identified in human chronic myelogenous leukemia cells on the timsTOF HT with low variation. Our results underline that dia-PASEF data acquisition combined with reproducible chromatography enables high robustness and data consistency across instruments and laboratories, which is a prerequisite for translational biomedical insights.

Development of analytical methods to streamline real-time measurements during process development is a priority for the evolution of bioprocess manufacturing. Current methods to measure downstream processes are limited

According to CODEX, moisture and volatile matter are olive oil quality parameters and the development of a rapid screening method for the determination of moisture is of interest. We recently demonstrated for the first time that the weak near-infrared (NIR) band near 5260 cm-1 is primarily attributed to a water O-H combination band. To determine the intensity of this band, we measured the peak-to-peak (p-p) height of its first derivative and generated exponential calibration curves for p-p height versus gravimetrically determined concentrations of spiked water in olive oils that had been purged of their initial moisture contents. To further optimize this univariate calibration method, calibration curves were generated in the present study based on plotting the moisture band first derivative p-p heights for neat olive oils (that were neither purged nor spiked) versus the moisture concentrations obtained by the Karl-Fischer (KF) primary reference method. To enhance the speed of FT-NIR data collection, measurements were carried in the transmission mode using disposable glass tubes. We also developed and compared a multivariate partial least squares approach to the univariate one. All the spectra were collected in two separate laboratories using two FT-NIR spectrometers of the same brand and model and no significant difference (p > 0.05) was found between the two laboratory determinations and the KF reference values at a 95% confidence interval. High accuracies were found with the two FT-NIR instruments used, as indicated by the low root mean squared error (RMSE, %) for predicted values obtained with the univariate procedure (RMSE = 0.008% and 0.010%) and the multivariate one, which yielded an even lower value (RMSE= 0.007% for both instruments). These results suggest that, once validated, the FT-NIR approach could potentially be a rapid substitute for the KF method.

Microcoil nuclear magnetic resonance (NMR) has been interfaced with digital microfluidics (DMF) and is applied to monitor organic reactions in organic solvents as a proof of concept. DMF permits droplets to be moved and mixed inside the NMR spectrometer to initiate reactions while using sub-microliter volumes of reagent, opening up the potential to follow the reactions of scarce or expensive reagents. By setting up the spectrometer shims on a reagent droplet, data acquisition can be started immediately upon droplet mixing and is only limited by the rate at which NMR data can be collected, allowing the monitoring of fast reactions. Here we report a cyclohexene carbonate hydrolysis in dimethylformamide and a Knoevenagel condensation in methanol/water. This is to our knowledge the first time rapid organic reactions in organic solvents have been monitored by high field DMF-NMR. The study represents a key first step towards larger DMF-NMR arrays that could in future serve as discovery platforms, where computer controlled DMF automates mixing/titration of chemical libraries and NMR is used to study the structures formed and kinetics in real time.

Abstract Microcoil nuclear magnetic resonance (NMR) has been interfaced with digital microfluidics (DMF) and is applied to monitor organic reactions in organic solvents as a proof of concept. DMF permits droplets to be moved and mixed inside the NMR spectrometer to initiate reactions while using sub‐microliter volumes of reagent, opening up the potential to follow the reactions of scarce or expensive reagents. By setting up the spectrometer shims on a reagent droplet, data acquisition can be started immediately upon droplet mixing and is only limited by the rate at which NMR data can be collected, allowing the monitoring of fast reactions. Here we report a cyclohexene carbonate hydrolysis in dimethylformamide and a Knoevenagel condensation in methanol/water. This is to our knowledge the first time rapid organic reactions in organic solvents have been monitored by high field DMF‐NMR. The study represents a key first step towards larger DMF‐NMR arrays that could in future serve as discovery platforms, where computer controlled DMF automates mixing/titration of chemical libraries and NMR is used to study the structures formed and kinetics in real time.

The combination of FT-IR spectroscopy with electrochemistry offers insight in the molecular change during electrochemical reactions in an addition to the electrochemical response of the studied media. This method can be applied for investigations of electrolytes or reactions at electrode surfaces. New accessories for the spectroelectrochemical studies will be presented. Accessories can be configured for the reflection-absorption or for the attenuated total reflectance (ATR) measurements. IR reflection-absorbance spectroscopy (IRRAS) set-up is used for the studies of the electrolyte and the electrode surface while ATR configuration is used to analyze the electrode surface without strong influence of the electrolyte. Optimization of spectroelectrochemical setup and details of communication between the potentiostat and the FTIR spectrometer will be discussed. Example of applications will include electro oxidation of metal-organic complexes, alcohols, and glycerol.

In the present study, we propose a characterization technique to determine the amount of calcium phosphate (CP) precipitate formed on a titanium substrate. The quantitative analysis of the CP precipitate on a metallic substrate is significant for researchers of metallic biomaterials because CP that spontaneously precipitates in a simulated-body fluid gives information on the bioactivity of the metallic biomaterials. We focused on X-ray fluorescence (XRF) analysis and adopted the thin-film fundamental parameter method (thin-film FP method) because it allows direct (non-pretreatment) and rapid quantitative analysis without any reference materials. We show that XRF analysis using the thin-film FP method can be adequately applied to the quantitative analysis of the CP precipitate in the simulated-body fluid immersion test. We also show that the density of the CP-precipitate layer can be estimated by combining the XRF results with images of cross-sectional scanning electron microscopy (SEM). Consequently, XRF analysis combined with the thin-film FP method can provide a convenient means to evaluate the CP precipitate on a titanium substrate, which improves the accuracy and accessibility of the simulated-body fluid immersion test.

Seeds are complex entities composed of liquids, gels, and solids. NMR spectroscopy is a powerful tool for studying molecular structure but has evolved into two fields, solution and solid state. Comprehensive multiphase (CMP) NMR spectroscopy is capable of liquid-, gel-, and solid-state experiments for studying intact samples where all organic components are studied and differentiated in situ. Herein, intact (13)C-labeled seeds were studied by a variety of 1D/2D (1)H/(13)C experiments. In the mobile phase, an assortment of metabolites in a single (13)C-labeled wheat seed were identified; the gel phase was dominated by triacylglycerides; the semisolid phase was composed largely of carbohydrate biopolymers, and the solid phase was greatly influenced by starchy endosperm signals. Subsequently, the seeds were compared and relative similarities and differences between seed types discussed. This study represents the first application of CMP-NMR to food chemistry and demonstrates its general utility and feasibility for studying intact heterogeneous samples.

We describe a conceptually novel “accelerated aging” approach for the synthesis of metal–organic materials.