Uplc Separation
Uplc Separation - Related Videos
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.
07:34
Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)
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Cited by 18 •
2013
Non-targeted metabolite profiling by ultra performance liquid chromatography coupled with mass spectrometry (UPLC-MS) is a powerful technique to investigate metabolism. This article outlines a typical workflow utilized for non-targeted metabolite profiling of serum including sample organization and preparation, data acquisition, data analysis, quality control, and metabolite identification.
03:07
Hydrophilic Interaction Liquid Chromatography: A Technique to Separate Hydrophilic Polar Analytes Using Hydrophilic Beads
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2025
This video demonstrates the principle of separation of hydrophilic polar compounds using hydrophilic interaction liquid chromatography. This technique helps in the purification of various polar analytes, including N-glycans.
08:02
Separation of Mixtures via Precipitation
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2023
Source: Laboratory of Dr. Ana J. García-Sáez — University of Tübingen Most samples of interest are mixtures of many different components. Sample preparation, a key step in the analytical process, removes interferences that may affect the analysis. As such, developing separation techniques is an important endeavor not just in academia, but also in industry. One way to separate mixtures is to use their solubility properties. In this short paper, we will deal with aqueous solutions. The...
07:29
Separating Protein with SDS-PAGE
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2023
Sodium Dodecyl Sulfate Poly-Acrylamide Gel Electrophoresis, or SDS-PAGE, is a widely-used technique for separating mixtures of proteins based on their size and nothing else. SDS, an anionic detergent, is used to produce an even charge across the length of proteins that have been linearized. By first loading them into a gel made of polyacrylamide and then applying an electric field to the gel, SDS-coated proteins are then separated. The electric field acts as the driving force, drawing the SDS...
FAQs
The small particles create a highly packed stationary phase that increases the technique’s resolving power, allowing compounds with similar behaviors to separate more distinctly. Because the mobile phase must pass through this tightly packed column, the system operates at high pressure. This combination supports high-resolution analysis when biological samples contain many chemically different components.
As analytes interact differently with the stationary and mobile phases, they travel through the column at different rates and elute at distinct retention times. Comparing these elution positions helps distinguish compounds within a complex mixture. The separated signals can then support both compound identification and quantification in biological or pharmaceutical analyses.
An analyte’s movement depends on the balance between its interactions with the stationary phase and the mobile phase. Stronger relative interaction with one phase changes how quickly it travels through the column, while different compounds respond differently to the same conditions. These contrasting behaviors produce separation rather than a single combined output from the mixture.
The technique can be applied to samples including blood, cell extracts, and tissue, making it useful across several biological research settings. Depending on the study, researchers may examine metabolites, peptides, proteins, pharmaceuticals, or other biomolecules. Its ability to handle complex biological mixtures supports analysis of both endogenous compounds and drug-related substances.
In biomarker studies, separated compounds can be identified and quantified to characterize molecular patterns in biological samples. During drug development, the same analytical capability helps examine pharmaceuticals alongside biological components. These applications benefit from high resolution, rapid analysis, and low sample requirements, which support biochemical investigations when sample quantity or mixture complexity is limiting.
UPLC separation combines improved resolution with rapid analyses and low sample requirements. Higher resolution helps distinguish components in complex mixtures, while faster analysis can increase the efficiency of experimental workflows. Requiring less sample is particularly useful for blood, cell extracts, or tissue, where available material may be limited. Together, these features strengthen biochemical and biological measurements.