Visualization of GC/TOF-MS-based metabolomics data for identification of biochemically interesting compounds using OPLS class models

Metabolomics studies generate increasingly complex data tables, which are hard to summarize and visualize without appropriate tools. The use of chemometrics tools, e.g., principal component analysis (PCA), partial least-squares to latent structures (PLS), and orthogonal PLS (OPLS), is therefore of great importance as these include efficient, validated, and robust methods for modeling information-rich chemical and biological data. Here the S-plot is proposed as a tool for visualization and interpretation of multivariate classification models, e.g., OPLS discriminate analysis, having two or more classes. The S-plot visualizes both the covariance and correlation between the metabolites and the modeled class designation. Thereby the S-plot helps identifying statistically significant and potentially biochemically significant metabolites, based both on contributions to the model and their reliability. An extension of the S-plot, the SUS-plot (shared and unique structure), is applied to compare the outcome of multiple classification models compared to a common reference, e.g., control. The used example is a gas chromatography coupled mass spectroscopy based metabolomics study in plant biology where two different transgenic poplar lines are compared to wild type. By using OPLS, an improved visualization and discrimination of interesting metabolites could be demonstrated.     

Olefin Metathesis on a TLC Plate as a Tool for a High‐Throughput Screening of Catalyst‐Substrate Sets

A methodology for screening either various catalysts for a given metathesis reaction, i.e., ring opening‐ring closing alkene metathesis (RO‐RCM) and cross‐metathesis (CM), or various substrates for a given pre‐catalyst on a thin layer chromatography (TLC) plate has been developed. As the substrates elute with the solvent, this TLC‐based system acts as a heterogeneous catalyst bed (“TLC reactor”). Selected promising catalyst candidates were screened on a TLC plate and their initial catalytic potential as observed in the TLC test was later fully confirmed in a classical heterogeneous reaction set‐up using standard commercially available silica (D11‐10). Reacting polyfunctional, natural product‐like substrates in our TLC reactor allows the simultaneous screening of various substrates and the convenient micro‐scale preparation and isolation of potentially biologically active products.     

Profile of fatty acids, muscle structure and shear force of musculus longissimus dorsi (MLD) in growing pigs as affected by energy and protein or protein restriction followed by realimentation

Forty-eight gilts were submitted to a 30% restriction of feed (groups F and F1) or protein intake (group P) from 90 to 118 days of age, followed by realimentation from 119 to 168 days of age. Control pigs (C) were fed during the whole experiment according to a semi ad libitum scale. During realimentation all animals were fed according to semi ad libitum scale except pigs F1 which were fed ad libitum. Six pigs from each group were slaughtered at the end of restriction and realimentation. Restriction decreased the weight of musculus longissimus dorsi (MLD) and increased shear force. Restriction of feed intake depressed MUFA concentration and increased n-6/n-3 ratio while protein restriction decreased n-6/n-3 and PUFA:SFA ratios. Structure of fibers was not affected. After realimentation MLD mass was still lower in all previously restricted pigs, shear force was the lowest in F1 pigs. Only percentage of fast twich oxidative fibers was significantly greater in F1 pigs than in others. Significant correlations between parameters investigated during the study were found.     

Trichloroethylene and tetrachloroethylene elimination from the air by means of a hybrid bioreactor with immobilized biomass

Two-phase bioreactors consisting of bacterial consortium in suspension and sorbents with immobilized biomass were used to treat waste air containing chlorinated ethenes, trichloroethylene (TCE) and tetrachloroethylene (PCE). Synthetic municipal sewage was used as the medium for bacterial growth. The system was operated with loadings in the range 1.48-4.76 gm(-3)h(-1) for TCE and 1.49-5.96 gm(-3)h(-1) for PCE. The efficiency of contaminant elimination was 55-86% in the bioreactor with wood chips and 33-89% in the bioreactor filled with zeolite. The best results were observed 1 week after the pollutant loading was increased. However, in these conditions, the stability of the process was not achieved. In the next 7 days the effectiveness of the system decreased. Contaminant removal efficiency, enzymatic activity and the biomass content were all diminished. The system was working without being supplied with additional hydrocarbons as the growth-supporting substrates. It is assumed that ammonia produced during the transformation of wastewater components induced enzymes for the cometabolic degradation of TCE and PCE. However, the evaluation of nitrogen compound transformations in the system is difficult due to the sorption on carriers and the combined processes of nitrification and the aerobic denitrification. An applied method of air treatment is advantageous from both economic and environmental point of views.     

An application of scanning electron microscopy combined with roentgen microanalysis (SEM-EDS) in canine urolithiasis

Urolithiasis is a common diagnostic and therapeutic problem in small-animal veterinary practice. The traditional diagnostic approach usually consists of clinical, radiological and ultrasonographic examination of the patient. The main diagnostic material is still urine sediment, ignoring the fact that presence of crystalluria is not always of pathological significance. In order to establish the most effective therapeutic and preventative strategies, especially in the case of multicomponent stone, it is crucial to define the exact elemental composition of the given stone including crystallization nidus chemical contents. In the course of the research, the usefulness of scanning electron microscopy combined with X-ray-dispersive spectrometry in analysis of canine mixed and compound stones was investigated. The obtained results indicated that the tested method allows one to trace the dynamics of the crystallization process, including crystallization nucleus detection, and concurrently and quantitatively assess the elemental composition of the given urinary concrement. Moreover, the conducted research showed epidemiological data of urolithiasis occurrence in a population of dogs coming from the southern part of Poland.     

Application of various methods for determination of the color of cocoa beans roasted under variable process parameters

Cocoa beans of Forastero variety from Togo were subjected to roasting under either constant or variable process parameters. The variable process parameters were roasting air flow rate, temperature and relative humidity. The color of roasted cocoa beans was determined by pigment extraction under various conditions followed by either their spectrophotometric assays or CIE L*a*b measurements. Also the Maillard compounds index and total polyphenols content correlated with progress of the browning of beans were determined. It was found that an increase in the roasting air relative humidity stimulated formation of brown pigments, while elevated temperature caused worsening of color parameters of roasted cocoa beans. The most suitable method of color characterization of roasted cocoa beans was found to be pigments extraction combined with either separation of their fractions or CIE L*a*b measurements. These assays revealed that cocoa beans roasted under variable roasting air flow rate were characterized by improved color parameters. The relatively simple and inexpensive CIE L*a*b measurements ensured fast analysis of color parameters, while total polyphenols in roasted cocoa beans were quickly estimated by using F–C reagent. Furthermore, quantification of melanoid pigments in roasted cocoa beans can be based on determination of the index of nonenzymatic browning products content, which is relatively simple and inexpensive.     

The application of d-SPE in the QuEChERS method for the determination of PAHs in food of animal origin with GC–MS detection

This paper reports the evaluation of the Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) method for the determination of polycyclic aromatic hydrocarbons (PAHs) in food of animal origin with GC–MS detection. Although in the available literature, there is a lot of information about sample preparation method for PAHs determination in food samples, but the QuEChERS method application for PAHs determination in food of animal origin has not been reported as yet. The results showed that the best recovery ratios 72.4–110.8 % with relative standard deviation lower than 10 % for all determined compounds were received for the method with ethyl acetate as an extraction solvent, primary–secondary amine and C₁₈ sorbents and evaporation to dryness and dissolving the residues in the hexane. The limit of quantification ranged from 0.0003 to 0.0030 mg kg^?1 for pyrene and benzo[a]anthracene, respectively. This method was also used for the determination of PAHs in 15 samples of pork ham. In 8 of 15 samples selected, PAHs were identified. It was observed that in 6 cooked ham and one smoked and cooked samples, any PAHs were found. In other samples, which were smoked and roasted, some low concentration of PAHs was detected. In one sample benzo[a]pyrene (0.0015 mg kg^?1), in one sample benzo[b]fluoranthene (0.0015 mg kg^?1) and in one sample chrysene (0.0024 mg kg^?1) were detected. A number of other less harmful PAHs were also determined. There were no exceedances of maximum levels (according to Commission Regulation (EU) No 835/2011) for determined PAHs in any of the analysed samples.