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This research was (partially) funded by X-Omics (NWO Project 184.034.019) and by DSM Biotechnology Delft. The authors acknowledge the contribution of Raphael Zwier from the Fine Mechanics Department at Leiden University for fabricating the PCTFE flow cell, and Manfred Spraul and Ulrich Braumann from Bruker BioSpin for the discussions on the urine analysis and supplying the buffer.

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Automated Segmented-Flow Analysis - NMR with a Novel Fluoropolymer Flow Cell for High-Throughput Screening

Publicado en:ANALYTICAL CHEMISTRY. 94 (44): 15350-15358 - 2022-10-27 94(44), DOI: 10.1021/acs.analchem.2c03038

Autores: Wouters, Bert; Miggiels, Paul; Bezemer, Roland; van der Cruijsen, Elwin A. W.; van Leeuwen, Erik; Gauvin, John; Houben, Klaartje; Gupta, Karthick Babu Sai Sankar; Zuijdwijk, Paul; Harms, Amy; Souza, Adriana Carvalho; Hankemeier, Thomas;

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High-throughput analysis in fields such as industrial biotechnology, combinatorial chemistry, and life sciences is becoming increasingly important. Nuclear magnetic resonance (NMR) spectroscopy is a powerful technique providing exhaustive molecular information on complex samples. Flow NMR in particular is a cost and time-efficient method for large screenings. In this study, we have developed a novel 3.0 mm inner diameter polychlorotrifluoroethylene (PCTFE) flow cell for a segmented-flow analysis (SFA) - NMR automated platform. The platform uses FC-72 fluorinated oil and fluoropolymer components to achieve a fully fluorinated flow path. Samples were repeatably transferred from 96-deepwell plates to the flow cell by displacing a fixed volume of oil, with a transfer time of 42 s. 1H spectra were acquired fully automated with 500 and 600 MHz NMR spectrometers. The spectral performance of the novel PCTFE cell was equal to that of commercial glass cells. Peak area repeatability was excellent with a relative standard deviation of 0.1-0.5% for standard samples, and carryover was below 0.2% without intermediate washing. The sample temperature was conditioned by using a thermostated transfer line in order to reduce the equilibration time in the probe and increase the throughput. Finally, analysis of urine samples demonstrated the applicability of this platform for screening complex matrices.

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