Analysis of low numbers of bacterial cells by CE and MALDI-TOF MS utilizing roughened capillary

0577800 - ÚIACH 2024 CZ eng A - Abstract
Šalplachta, Jiří - Kubesová, Anna - Karásek, Pavel - Růžička, F. - Roth, Michal
Analysis of low numbers of bacterial cells by CE and MALDI-TOF MS utilizing roughened capillary.
CECE 2023. 18th International Interdisciplinary Meeting on Bioanalysis. Brno: Ústav analytické chemie AV ČR, v. v. i., 2023. s. 69-69. ISBN 978-80-908154-0-7.
[CECE 2023. International Interdisciplinary Meeting on Bioanalysis /18./. 23.10.2023-25.10.2023, Brno]
R&D Projects: GA MZd(CZ) NU22-05-00110
Institutional support: RVO:68081715
Keywords : time-of-flight mass spectrometry (MALDI-TOF MS) * capillary electrophoresis (CE) * biological samples
OECD category: Analytical chemistry
https://www.ce-ce.org/user_uploads/CECE_2023/CECE%202023%20Proceedings.pdf

Detection and identification of low amount of pathogens in the biological samples is of increased interest in recent years. This study presents rapid and sensitive method for identification of pathogenic bacteria in biological samples based on a combination of capillary electrophoresis (CE) with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). A part of single-piece fused silica capillary was etched with supercritical water with the aim of using it for adhesion (static or dynamic) of bacterial cells from large sample volumes. Adhered bacterial cells were desorbed and concentrated from the roughened capillary surface by transient isotachophoretic stacking. Charged cells were separated in micellar electrokinetic chromatography. Concentrated and separated bacteria were collected from the capillary and off-line analyzed by MALDI-TOF MS. Dynamic adhesion allowed concentration of bacteria from large sample volumes with the limits of detection in the range from 1.8 × 102 to 1.7 × 103 cells mL-1, depending on the sample origin. Nevertheless, the limits of detection were the same for all the examined bacteria. The recovery of the method was about 83 % and it was independent on the sample matrix. A combination of CE with MALDI-TOF MS required at least 4×103 cells mL-1 to obtain reliable results.
Permanent Link: https://hdl.handle.net/11104/0346894