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Updated: May 16, 2026

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
Published on: February 27, 2020
Direct bioanalytical sample injection with 2D LC-MS
Nm Cassiano1, Jc Barreiro, Rv Oliveira
1Departamento de Química, Universidade Federal de São Carlos, Caixa Postal 676, 13565-905 São Carlos - SP, Brazil.
New analytical platforms enhance complex mixture analysis by increasing sensitivity and throughput. This review covers two-dimensional liquid chromatography (2D LC) methods and their coupling with mass spectrometry (MS) for bioanalysis.
Area of Science:
- Analytical Chemistry
- Chromatography
- Mass Spectrometry
Background:
- Increasing demand for higher sensitivity and throughput in analyzing complex mixtures.
- Limitations of traditional one-dimensional liquid chromatography (1D LC) for intricate samples.
Purpose of the Study:
- To review fundamental principles of target and comprehensive 2D LC method development.
- To discuss applications of LC-LC and LC x LC coupled to MS in bioanalysis.
- To provide rationale for mass analyzer and ionization source selection in 2D LC-MS.
Main Methods:
- Review of target and comprehensive two-dimensional liquid chromatography (2D LC) principles.
- Analysis of online analytical procedures for LC-LC and LC x LC.
- Integration of mass spectrometry (MS) for enhanced detection.
Main Results:
- Detailed overview of 2D LC method development strategies.
- Examples of LC-LC and LC x LC applications in bioanalysis.
- Comparison of various mass analyzers and ionization sources for 2D LC-MS.
Conclusions:
- 2D LC coupled with MS offers significant advantages for complex sample analysis.
- Method development in 2D LC requires careful consideration of separation orthogonality and coupling.
- The choice of MS instrumentation is critical for optimizing bioanalytical outcomes.
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