Targeted serum metabolite profiling and sequential metabolite ratio analysis for colorectal cancer progression monitoring

Abstract

by the separation of true classification models from the random permutation models. Our results 36suggest the potential utility of metabolic profiling for CRC disease monitoring.

Staphylococcus aureus methicillin resistance detected by HPLC-MS/MS targeted metabolic profiling

“…The bacterial cells were harvested by centrifugation at 22024.6g for 10 min, followed by three phosphate buffer saline (PBS, pH = 7.4, which containing NaCl, KCl, Na 2 HPO 4 , KH 2 PO 4 and H 2 O) washes. Intracellular metabolites from each biological replicate were extracted using a cold methanol extraction approach as previously reported [15,16]. Briefly, 250 L methanol was added to the cell pellet and the samples were mixed vigorously for ∼1 min on a vortex machine, then 50 L isotope labeled amino acid mixture was added as internal standards (Cambridge Isotope Laboratories, Inc.) and then mixed one more time.…”

Targeted High Performance Liquid Chromatography Tandem Mass Spectrometry-based Metabolomics differentiates metabolic syndrome from obesity

“…The advantage of our targeted metabolic profiling approach was that it provided broad coverage of relevant metabolites while simultaneously enabling confident metabolite identification, thus avoiding tedious compound database search and confirmation steps required for untargeted metabolic profiling approaches. 5 The metabolite targets were carefully selected from metabolomics studies 10,11,13,14 in which their detectability and measurement reproducibility were validated. Thus, the metabolites reported from the present study are readily interpreted within their biological context to potentially address questions of public health and scientific significance.…”

“…The targeted metabolic profiling was performed in selected-reaction-monitoring (SRM) mode, established by running multiple authentic chemical standards first, and then using the obtained retention time and SRM transition information to identify metabolites from samples. The 221 metabolites were selected according to our published work, [10][11][12] and consistent with previous studies that these represent key metabolites of interest from relevant metabolic pathways. 13,14 The detection parameters for these metabolites are listed in supplementary Table S1.…”

“…10,11 In brief, samples were kept at À80 C until sample preparation steps. Prior to sample preparation, samples were randomized for analysis to ensure no batch effects and prevent experimental bias.…”

“…Targeted metabolomics of plasma samples was performed according to validated procedures, 10,11 with minor modifications. In brief, chromatographic separation and analyte detection was performed using a Thermo Fisher Scientific Dionex Ultimate 3000 HPLC system with inline TSQQuantiva triple quadrupole tandem mass spectrometer (MS/MS) equipped with an electrospray ionization (ESI) source.…”