Uhplc-dad-hrms

UHPLC-DAD-HRMS is an integrated analytical platform that separates, detects, and characterizes chemical compounds with high resolution, making it valuable for pharmacological research and drug analysis. Ultra-high-performance liquid chromatography resolves compounds as they pass through a packed column under high pressure, while diode-array detection records ultraviolet-visible absorbance and high-resolution mass spectrometry measures accurate mass-to-charge ratios for molecular identification and elemental-composition assessment. Together, these measurements support the identification and quantification of active pharmaceutical ingredients, metabolites, impurities, and degradation products in complex samples. The approach strengthens bioanalysis, pharmacokinetic studies, quality control, and structural characterization by combining complementary chromatographic, spectroscopic, and mass-spectral information.

Uhplc-dad-hrms - Related Videos

Research

JoVE Journal - Chemistry

Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS

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Cited by 6 •

2017

The present method serves to identify isoform-specific inhibitors of histone deacetylases (HDAC) in HeLa cells by the UHPLC-MS analysis of multiple substrates. This is an antibody-free method developed to reflect HDAC1 and HDAC6 activity in the living cell environment, in contrast to single-isoform cell-free assays.

Education

JoVE Core - Analytical Chemistry

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...

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.

Monitoring the Effects of Illumination on the Structure of Conjugated Polymer Gels Using Neutron Scattering

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Cited by 2 •

2017

A protocol for the analysis of gels formed from the optoelectronic conjugated polymer poly(3-hexylthiophene-2,5-diyl) (P3HT) using small and ultra-small angle neutron scattering in both the presence and absence of illumination is presented.

Research

JoVE Journal - Biology
Free Sample

Automated Modular High Throughput Exopolysaccharide Screening Platform Coupled with Highly Sensitive Carbohydrate Fingerprint Analysis

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Cited by 18 •

2016

We present an automated modular high-throughput-method for the identification and characterization of microbial exopolysaccharides in small scale. This method combines a fast preselection to analyze the total amount of secreted polysaccharides with a detailed carbohydrate fingerprint to enable the fast screening of newly isolated bacterial strains or entire strain collections.

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