Lc-hrms Analysis
Lc-hrms Analysis - Related Videos
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High-Resolution Mass Spectrometry (HRMS)
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2024
The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For example, the mass of helium...
11:00
Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)
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Cited by 33 •
2013
Untargeted metabolomics provides a hypothesis generating snapshot of a metabolic profile. This protocol will demonstrate the extraction and analysis of metabolites from cells, serum, or tissue. A range of metabolites are surveyed using liquid-liquid phase extraction, microflow ultraperformance liquid chromatography/high-resolution mass spectrometry (UPLC-HRMS) coupled to differential analysis software.
15:04
A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity
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Cited by 46 •
2009
This article describes a novel protocol and reagent set designed for sensitive measurement of neurotoxic effects of compounds and treatments on co-cultures of neurons and astrocytes using high content analysis. Results demonstrate that high content analysis represents an exciting novel technology for neurotoxicity assessment.
10:39
Automated Analysis of Intracellular Phenotypes of Salmonella Using ImageJ
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Cited by 3 •
2022
Salmonella invades and replicates inside intestinal epithelial cells both in Salmonella-specific vacuoles and free in the cytosol (hyper-replication). A high-throughput fluorescence microscopy-based protocol is described here to quantify the intracellular phenotypes of Salmonella by two complementary image analyses through ImageJ, reaching single-cell resolution and scoring.
11:24
High-throughput Imaging and Analysis Workflow for Evaluating Skin Cell Phenotypes and Proliferation States in Tissue Samples
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Cited by 1 •
2025
The combination of iterative-bleaching-extends-multiplexity (IBEX) and a commercial nucleotide labeling assay (Click-iT EdU) enables the detection and categorization of dividing cell types in highly dynamic processes in fixed frozen murine tissue sections. Furthermore, a novel open-source image processing pipeline provides high-throughput image acquisition and analysis.
FAQs
The two instrument stages contribute different kinds of selectivity. Liquid chromatography separates compounds before they enter the mass spectrometer, reducing overlap among analytes in a complex biological sample. High-resolution measurement then distinguishes ions by accurate mass-to-charge ratio. Together, these steps improve confidence in compound detection and support subsequent molecular-formula assignment or structural characterization.
Accurate mass-to-charge measurement provides more than a signal indicating that an ion is present. In LC-HRMS analysis, the measured value can support assignment of a molecular formula and contribute to structural identification. This information helps biochemists interpret detected compounds and relate their presence to metabolites, lipids, peptides, or other biochemical constituents.
Retention behavior and polarity influence when compounds emerge from liquid chromatography. Compounds with different properties can therefore be resolved before mass analysis, which is especially important for complex biological samples containing many chemical species. This separation affects how clearly individual analytes can be detected and characterized, while the mass spectrometer supplies complementary mass-to-charge information.
A basic workflow begins with a complex biological sample, followed by liquid-chromatographic separation of its analytes. The separated compounds are transferred into the gas phase by ionization, and the mass spectrometer measures their mass-to-charge ratios. Researchers can then use the accurate-mass information for compound characterization, biochemical profiling, or quantitative comparison of changes in the sample.
Targeted and untargeted metabolomics address different biochemical questions. Targeted work focuses on selected metabolites, whereas untargeted work surveys detected chemical features more broadly. The same LC-HRMS analysis platform can also support lipid profiling and peptide characterization, allowing investigators to choose an analytical scope that matches the molecules and biological question under study.
In biochemistry, LC-HRMS analysis supports biomarker research and the examination of biochemical changes associated with health and disease. Its sensitivity, selectivity, and mass accuracy help investigators detect and quantify molecular differences, then use those patterns to investigate affected pathways. The approach connects measurements of metabolites, lipids, or peptides with broader biological interpretation.