High-throughput analysis using non-depletive SPME: challenges and applications to the determination of free and total concentrations in small sample volumes

Date

2018-01-18

Author

Boyacı, Ezel
Bojko, Barbara
Reyes-Garces, Nathaly
Poole, Justen J.
Gomez-Rios,Gomez-Rios, German Augusto,German Augusto
Teixeira, Alexandre
Nicol, Beate
Pawliszyn, Janusz

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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

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In vitro high-throughput non-depletive quantitation of chemicals in biofluids is of growing interest in many areas. Some of the challenges facing researchers include the limited volume of biofluids, rapid and high-throughput sampling requirements, and the lack of reliable methods. Coupled to the above, growing interest in the monitoring of kinetics and dynamics of miniaturized biosystems has spurred the demand for development of novel and revolutionary methodologies for analysis of biofluids. The applicability of solid-phase microextraction (SPME) is investigated as a potential technology to fulfill the aforementioned requirements. As analytes with sufficient diversity in their physicochemical features, nicotine, N, N-Diethyl-meta-toluamide, and diclofenac were selected as test compounds for the study. The objective was to develop methodologies that would allow repeated non-depletive sampling from 96-well plates, using 100 mu L of sample. Initially, thin film-SPME was investigated. Results revealed substantial depletion and consequent disruption in the system. Therefore, new ultra-thin coated fibers were developed. The applicability of this device to the described sampling scenario was tested by determining the protein binding of the analytes. Results showed good agreement with rapid equilibrium dialysis. The presented method allows high-throughput analysis using small volumes, enabling fast reliable free and total concentration determinations without disruption of system equilibrium.

Subject Keywords

Multidisciplinary, SOLID-PHASE MICROEXTRACTION, THIN-FILM MICROEXTRACTION, QUANTITATIVE IN-VITRO, PROHIBITED SUBSTANCES, VIVO EXTRAPOLATION, CELL-CULTURES, BINDING, TOOL, ASSAYS, BLOOD

URI

https://hdl.handle.net/11511/43667

Collections

Department of Chemistry, Article