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Automated Segmented-Flow Analysis - NMR with a Novel Fluoropolymer Flow Cell for High-Throughput Screening.
Bert Wouters1, Paul Miggiels1, Roland Bezemer2
1Metabolomics and Analytics Centre, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 CCLeiden, The Netherlands.
Analytical Chemistry
|October 27, 2022
Summary
A new polychlorotrifluoroethylene (PCTFE) flow cell enables automated, high-throughput Nuclear Magnetic Resonance (NMR) analysis. This cost-effective method offers excellent repeatability and low carryover for complex sample screening.
Area of Science:
- Analytical Chemistry
- Spectroscopy
- Biotechnology
Background:
- High-throughput analysis is crucial in industrial biotechnology, combinatorial chemistry, and life sciences.
- Nuclear Magnetic Resonance (NMR) spectroscopy provides comprehensive molecular data for complex samples.
- Flow NMR offers a time- and cost-efficient approach for large-scale screening.
Purpose of the Study:
- To develop a novel polychlorotrifluoroethylene (PCTFE) flow cell for a segmented-flow analysis (SFA)-NMR automated platform.
- To evaluate the performance, repeatability, and carryover of the new flow cell.
- To demonstrate the platform's applicability for screening complex biological matrices.
Main Methods:
- Development of a 3.0 mm inner diameter PCTFE flow cell for SFA-NMR.
- Utilizing FC-72 fluorinated oil and fluoropolymer components for a fully fluorinated flow path.
- Automated sample transfer from 96-well plates and 1H spectra acquisition on 500/600 MHz NMR spectrometers.
Main Results:
- The PCTFE flow cell demonstrated spectral performance comparable to commercial glass cells.
- Excellent peak area repeatability was achieved (RSD of 0.1-0.5%) with minimal carryover (<0.2%).
- Thermostated transfer lines reduced equilibration time, enhancing throughput.
Conclusions:
- The developed SFA-NMR platform with the novel PCTFE flow cell is highly effective for automated, high-throughput screening.
- The system shows excellent precision and minimal carryover, suitable for complex sample analysis.
- Application in urine sample analysis confirms its utility for screening challenging biological matrices.
