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Subcellular Fractionation

Subcellular Fractionation

The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
Differential centrifugation is...

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A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample

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Published on: June 1, 2017

High-Frequency Microfluidic Fractionation for Compound-Resolved Bioactivity-Based Metabolomics.

Christian Geibel^1,2, Julian Schubert^1,2, Simon B Knoblauch²

¹Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, 72076 Tübingen, Germany.

Analytical Chemistry

|October 25, 2025

Summary

This summary is machine-generated.

This study introduces a novel metabolomics workflow for rapid drug discovery. The new method efficiently identifies antimicrobial compounds from complex mixtures using liquid chromatography-MS/MS and luminescent bioreporters.

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Multi-step Preparation Technique to Recover Multiple Metabolite Compound Classes for In-depth and Informative Metabolomic Analysis

Multi-step Preparation Technique to Recover Multiple Metabolite Compound Classes for In-depth and Informative Metabolomic Analysis

Published on: July 11, 2014

Area of Science:

Analytical Chemistry
Metabolomics
Drug Discovery

Background:

Specialized metabolites are vital drug candidates, but their identification from extracts is slow.
Nontargeted metabolomics faces bottlenecks in functional assessment, hindering biological understanding.
Urgent need for new drugs, especially antibiotics, necessitates faster discovery methods.

Purpose of the Study:

To develop a compound-resolved bioactivity-based metabolomics workflow.
To accelerate the identification of active agents from complex metabolite samples.
To address the bottleneck in functional assessment within metabolomics.

Main Methods:

Combined nontargeted liquid chromatography tandem mass spectrometry (LC-MS/MS) with high-frequency fractionation.
Utilized microfluidic paper-analytical devices (μPADs) for parallel spotting during MS/MS acquisition.
Employed luminescent bioreporter strains to detect cellular stress and correlate signals with MS data.

Main Results:

Demonstrated high sensitivity (down to 1 ng/spot) for antimicrobial compound detection.
Successfully identified multiple antimicrobial compounds from crude extracts of *Saccharopolyspora erythraea*.
Highlighted the workflow's practicality and high-throughput capability for drug discovery.

Conclusions:

The developed workflow enables rapid, compound-resolved bioactivity-based metabolomics.
This approach significantly speeds up the identification of potential drug candidates from complex samples.
The method advances systematic functional assessment in metabolomics and aids antibiotic discovery.