Design and Implementation of a Proof-of-Concept Robotic-Microfluidic Interface to Bridge Spatial Comprehensive...

Area of Science:

• Analytical Chemistry
• Chromatography
• Mass Spectrometry

Background:

• Hyphenation of separation techniques to mass spectrometry is crucial for complex sample analysis.
• Spatial comprehensive three-dimensional liquid chromatography (3D-LC) offers enhanced separation power but requires specialized interfaces.

Purpose of the Study:

• To develop and validate a proof-of-concept system for hyphenating spatial 3D-LC with mass spectrometry (MS) detection.
• To demonstrate the capability of the system for high-throughput analysis of intact proteins.

Main Methods:

• A robotic-microfluidic interface was designed, incorporating a 3D fractal microflow distributor with 16 parallel capillary columns.
• An X-Y-Z robotic system was used for microdroplet deposition onto an MS target plate.
• Carryover was minimized by increasing the distance between the capillary and the target plate during deposition.

Main Results:

• System-level variations in flow rates and travel times were assessed and corrected for retention alignment.
• Successful separation of intact proteins was achieved using a 4x4 parallel column configuration.
• MALDI-MS detection was performed after microdroplet spotting on the MS target plate.

Conclusions:

• The developed system provides a robust platform for spatial 3D-LC hyphenated to MS detection.
• The approach enables high-throughput analysis and holds potential for MS imaging applications.
• This technology advances the capabilities for complex biological sample analysis.