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3 results on '"Ezel Boyacı"'

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

Author

Ezel Boyacı, Barbara Bojko, Nathaly Reyes-Garcés, Justen J. Poole, Germán Augusto Gómez-Ríos, Alexandre Teixeira, Beate Nicol, and Janusz Pawliszyn

Subjects
Medicine, Science
Abstract

Abstract 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 µL 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.

Published
2018
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2. The use of solid phase microextraction for metabolomic analysis of non-small cell lung carcinoma cell line (A549) after administration of combretastatin A4

Author

Ezel Boyacı, Janusz Pawliszyn, Karol Jaroch, and Barbara Bojko

Subjects
0301 basic medicine, Lung Neoplasms, Metabolite, lcsh:Medicine, Solid-phase microextraction, Article, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Metabolomics, Carcinoma, Non-Small-Cell Lung, Stilbenes, Extracellular, Humans, Solid phase extraction, lcsh:Science, A549 cell, Multidisciplinary, Chromatography, Solid Phase Extraction, lcsh:R, Biological activity, 030104 developmental biology, chemistry, A549 Cells, Cell culture, lcsh:Q, 030217 neurology & neurosurgery
Abstract

Use of solid phase microextraction (SPME) for cell culture metabolomic analysis allows for the attainment of more sophisticated data from in vitro cell cultures. Moreover, considering that SPME allows the implementation of multiple extractions from the same sample due to its non/low-depletive nature, time course studies using the same set of samples are thus facilitated via this method. Such an approach results in a reduction in the number of samples needed for analysis thus eliminates inter-batch variability related to biological variation occurring during cell culturing. The current work aims to demonstrate the capability of SPME for measurements of combretastatin A4 (CA4) effectiveness on non-small cell cancer cell line. A cultivation protocol was established in the 96-well plate, and a fiber format of SPME was selected for metabolite extraction. The extracellular metabolic pattern of cells was changed after administration of the tested drug. This suggests pharmacological activity of the administered compound towards the studied cell line model. Results support that the use of direct immersion SPME for analysis of cell cultures does not affect cells growth or contaminate sample. Consequently, SPME allows the attainment of accurate information regarding drug uptake, metabolism, and metabolomic changes in the studied cells induced by exposure to the drug simultaneously in a single experiment.

Published
2019
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3. Quantitative analysis of biofluid spots by coated blade spray mass spectrometry, a new approach to rapid screening

Author

Nathaly Reyes-Garcés, Janusz Pawliszyn, Germán Augusto Gómez-Ríos, Justen J. Poole, Marcos Tascon, and Ezel Boyacı

Subjects
Bioanalysis, Analyte, lcsh:Medicine, engineering.material, 010402 general chemistry, Mass spectrometry, 01 natural sciences, Mass Spectrometry, Article, Matrix (chemical analysis), Plasma, Coating, Tandem Mass Spectrometry, Humans, lcsh:Science, Multidisciplinary, Chromatography, Spots, Chemistry, 010401 analytical chemistry, lcsh:R, Reference Standards, Pain management, 0104 chemical sciences, engineering, lcsh:Q, Quantitative analysis (chemistry), Blood Chemical Analysis
Abstract

This study demonstrates the quantitative capabilities of coated blade spray (CBS) mass spectrometry (MS) for the concomitant analysis of multiple target substances in biofluid spots. In CBS-MS the analytes present in a given sample are first isolated and enriched in the thin coating of the CBS device. After a quick rinsing of the blade surface, as to remove remaining matrix, the analytes are quickly desorbed with the help of a solvent and then directly electrosprayed into the MS analyzer. Diverse pain management drugs, controlled substances, and therapeutic medications were successfully determined using only 10 µL of biofluid, with limits of quantitation in the low/sub ng·mL−1 level attained within 7 minutes.

Published
2017
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