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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

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Pillar Array Mixer for Postcolumn Derivatization Integrated into Liquid Chromatography-Based Microfluidic Device.

Muneki Isokawa¹, Kanki Nakanishi², Takahiro Kanamori¹

¹Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan.

Analytical Chemistry

|June 13, 2024

Summary

A novel microfluidic device integrates a pillar array (PA) mixer for enhanced post-column derivatization in on-chip liquid chromatography (LC). This innovation improves metabolite analysis sensitivity and speed, enabling rapid amino acid detection.

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Area of Science:

Analytical Chemistry
Microfluidics
Biotechnology

Background:

Chemical derivatization is crucial for enhancing sensitivity and selectivity in metabolite analysis using microfluidic devices.
Integrating derivatization units into microfluidic chromatography systems presents significant development challenges.

Purpose of the Study:

To develop a novel microfluidic derivatization unit for on-chip liquid chromatography (LC).
To enhance mixing efficiency for post-column derivatization using a pillar array (PA) mixer.
To demonstrate the device's capability for rapid metabolite analysis.

Main Methods:

Fabrication of microfluidic devices with an integrated LC column and PA mixer on a silicon plate.
Utilized computational fluid dynamics (CFD) for concept validation.
Performed fluid experiments comparing PA mixers with hollow-channel mixers.
Analyzed amino acids using hydrophilic interaction chromatography (HILIC) coupled with post-column derivatization and fluorescence detection.

Main Results:

The PA mixer effectively enhanced transverse mixing of derivatization reagents and analytes.
Analyte peak width was not increased by the PA mixer, preserving separation efficiency.
Achieved rapid analysis of three amino acids within 40 seconds.
Demonstrated successful post-column fluorogenic derivatization using naphthalene-2,3-dicarboxaldehyde.

Conclusions:

The developed PA mixer is effective for post-column derivatization in microfluidic LC systems.
Integrated derivatization units show significant potential for creating advanced micrototal analysis systems (µTAS).
This approach facilitates sensitive and selective bioanalysis with high throughput.