| HORKÁ, Marie, Pavel KARÁSEK, Jiří ŠALPLACHTA, Filip RŮŽIČKA, Marie VYKYDALOVÁ, Anna KUBESOVÁ, Vladimír DRÁB, Michal ROTH a Karel ŠLAIS. Capillary isoelectric focusing of probiotic bacteria from cow's milk in tapered fused silica capillary with off-line matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification. Analytica Chimica Acta. AMSTERDAM: ELSEVIER SCIENCE BV, 2013, roč. 788, 25 July 2013, s. 193-199. ISSN 0003-2670. doi:10.1016/j.aca.2013.05.059. |
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@article{1138468,
author = {Horká, Marie and Karásek, Pavel and Šalplachta, Jiří and Růžička, Filip and Vykydalová, Marie and Kubesová, Anna and Dráb, Vladimír and Roth, Michal and Šlais, Karel},
article_location = {AMSTERDAM},
article_number = {25 July 2013},
doi = {http://dx.doi.org/10.1016/j.aca.2013.05.059},
keywords = {Capillary isoelectric focusing; Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; Tapered fused silica capillary; Lactic acid bacteria; Milk},
language = {eng},
issn = {0003-2670},
journal = {Analytica Chimica Acta},
title = {Capillary isoelectric focusing of probiotic bacteria from cow's milk in tapered fused silica capillary with off-line matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification},
volume = {788},
year = {2013}
}
TY - JOUR
ID - 1138468
AU - Horká, Marie - Karásek, Pavel - Šalplachta, Jiří - Růžička, Filip - Vykydalová, Marie - Kubesová, Anna - Dráb, Vladimír - Roth, Michal - Šlais, Karel
PY - 2013
TI - Capillary isoelectric focusing of probiotic bacteria from cow's milk in tapered fused silica capillary with off-line matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification
JF - Analytica Chimica Acta
VL - 788
IS - 25 July 2013
SP - 193-199
EP - 193-199
PB - ELSEVIER SCIENCE BV
SN - 00032670
KW - Capillary isoelectric focusing
KW - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
KW - Tapered fused silica capillary
KW - Lactic acid bacteria
KW - Milk
N2 - In this study, combination of capillary isoelectric focusing (CIEF) in tapered fused silica (FS) capillary with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is presented as an efficient approach for unambiguous identification of probiotic bacteria in real sample. For this purpose, bacteria within genus Lactobacillus were selected as model bioanalytes and cow's milk was selected as a biological sample. CIEF analysis of both the cultivated bacteria and the bacteria in the milk was optimized and isoelectric points characterizing the examined bacteria were subsequently determined independently of the bacterial sample origin. The use of tapered FS capillary significantly enhanced the separation capacity and efficiency of the CIEF analyses performed. In addition, the cell number injected into the tapered FS capillary was quantified and an excellent linearity of the calibration curves was achieved which enabled quantitative analysis of the bacteria by CIEF with UV detection. The minimum detectable number of bacterial cells was 2 x 10(6) mL(-1). Finally, cow's milk spiked with the selected bacterium was analyzed by CIEF in tapered FS capillary, the focused and detected bacterial cells were collected from the capillary, deposited onto the cultivation medium, and identified using MALDI-TOF MS afterward. Our results have revealed that the proposed procedure can be advantageously used for unambiguous identification of probiotic bacteria in a real sample.
ER -
HORKÁ, Marie, Pavel KARÁSEK, Jiří ŠALPLACHTA, Filip RŮŽIČKA, Marie VYKYDALOVÁ, Anna KUBESOVÁ, Vladimír DRÁB, Michal ROTH a Karel ŠLAIS. Capillary isoelectric focusing of probiotic bacteria from cow's milk in tapered fused silica capillary with off-line matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification. \textit{Analytica Chimica Acta}. AMSTERDAM: ELSEVIER SCIENCE BV, 2013, roč.~788, 25 July 2013, s.~193-199. ISSN~0003-2670. doi:10.1016/j.aca.2013.05.059.
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