Related Experiment Video
Updated: Feb 13, 2026

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
Published on: January 5, 2021
On-Column Dimethylation with Capillary Liquid Chromatography-Tandem Mass Spectrometry for Online Determination of
Rachael E Wilson1, Andrea Jaquins-Gerstl1, Stephen G Weber1
1Department of Chemistry , University of Pittsburgh , Pittsburgh , Pennsylvania 15260 , United States.
Abstract:
We have developed a method for online collection and quantitation of neuropeptides in rat brain microdialysates using on-column dimethylation with capillary liquid chromatography-tandem mass spectrometry (cLC-MS2). This method addresses a number of the challenges of quantifying neuropeptides with cLC-MS. It is also a completely automated and robust method for the preparation of stable isotope labeled-peptide internal standards to correct for matrix effects and thus ensure accurate quantitation. Originally developed for tissue-derived proteomics samples ( Raijmakers et al. Mol. Cell. Proteomics 2008 , 7 , 1755 - 1762 ), the efficacy of on-column dimethylation for native peptides in microdialysate has not been demonstrated until now. We have modified the process to make it more amenable to the time scale of microdialysis sampling and to reduce the accumulation of nonvolatile contaminants on the column and, thus, loss of sensitivity. By decreasing labeling time, we have a temporal resolution of 1 h from sample loading to elution and our peptide detection limits are in the low pM range for 5 μL injections of microdialysate. We have demonstrated the effectiveness of this method by quantifying basal and potassium stimulated concentrations of the neuropeptides leu-enkephalin and met-enkephalin in the rat hippocampus. To our knowledge, this is the first report of quantitation of these peptides in the hippocampus using MS.
More Related Videos
08:56Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
Published on: March 8, 2020
08:14Extraction of Non-Protein Amino Acids from Cyanobacteria for Liquid Chromatography-Tandem Mass Spectrometry Analysis
Published on: December 9, 2022
Related Concept Videos
Tandem Mass Spectrometry
Peptide Identification Using Tandem Mass Spectrometry
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
Gas Chromatography–Mass Spectrometry (GC–MS)
A gas chromatograph consists of a long, narrow capillary column with a polysiloxane coating on the inner wall....
Types Of Column Chromatography
Gel Filtration Chromatography
When the...
Principles Of Column Chromatography
Diffusion on Chromatography Columns
Longitudinal diffusion occurs when the solute molecules in the mobile phase diffuse from the more concentrated center of the chromatographic band to the more dilute regions on either side, both towards and against the flow direction. This...