Reversed-phase LC combined with high-resolution mass spectrometry (HRMS) is one of the most popular methods for cellular metabolomics studies. Due to the difficulties in analyzing a wide range of polarities encountered in the metabolome, 100%-wettable reversed-phase materials are frequently used to maximize metabolome coverage within a single analysis. Packed with silica-based sub-3 μm diameter particles, these columns allow high separation efficiency and offer a reasonable compromise for metabolome coverage within a single analysis. While direct performance comparison can be made using classical chromatographic characterization approaches, a comprehensive assessment of the column's performance for cellular metabolomics requires use of a full LC-HRMS workflow in order to reflect realistic study conditions used for cellular metabolomics. In this study, a comparison of several reversed-phase LC columns for metabolome analysis using such a dedicated workflow is presented. All columns were tested under the same analytical conditions on an LC-TOF-MS platform using a variety of authentic metabolite standards and biotechnologically relevant yeast cell extracts. Data on total workflow performance including retention behavior, peak capacity, coverage, and molecular feature extraction repeatability from these columns are presented with consideration for both nontargeted screening and differential metabolomics workflows using authentic standards and Pichia pastoris cell extract samples.
For the first time, the dual isotope approach was applied to trace the sources of impacts and to identify the governing biogeochemical processes in a river network in the tropical deltaic region of the Red River (Vietnam). Our long term surveys concluded that water in this river network was severely impacted by anthropogenic activities. Analysis has shown strong spatio-temporal variation of nitrate isotopes; ranges of δ(15)N-[Formula: see text] and δ(18)O-[Formula: see text] were from -5 to 15 ‰ and from -10 to 10 ‰, respectively. Average values of δ(15)N-[Formula: see text] and δ(18)O-[Formula: see text] in the dry season, when fertilizer is applied, were 3.54 and 3.15 ‰, respectively. In the rainy season, the values changed to 6.41 and -2.23 ‰, respectively. Denitrification and biological assimilation were active throughout the year, but were especially enhanced during fertilization time. Mineralization of domestic organic matter and consequent nitrification of mineralized [Formula: see text] were the dominant processes, particularly during the rainy period.
The packed column supercritical fluid chromatography has risen as a promising alternative separation technique to the conventional liquid chromatography and gas chromatography. Although the packed column supercritical fluid chromatography has many advantages compared to other chromatographic techniques, its separation mechanism is not fully understood due to the complex combination effects of many chromatographic parameters on separation quality and the lacking of global strategies for studying separation mechanisms. This review aims to provide recent information regarding the chromatographic behaviors and the effects of the parameters on the separation, discuss the results, and point out the remaining bottlenecks in the packed column supercritical fluid chromatography retention mechanism studies.
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