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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
•We found method for rapid separation of Lactobacillus in tapered FS capillary.•CIEF separation capacity and efficiency was increased.•CIEF was coupled with off-line MALDI-TOF MS identification.•Method was tested on spiked milk. In this study, combination of capillary isoelectric focusing (CIEF) in...
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Published in: | Analytica chimica acta 2013-07, Vol.788, p.193-199 |
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creator | Horká, Marie Karásek, Pavel Šalplachta, Jiří Růžička, Filip Vykydalová, Marie Kubesová, Anna Dráb, Vladimír Roth, Michal Šlais, Karel |
description | •We found method for rapid separation of Lactobacillus in tapered FS capillary.•CIEF separation capacity and efficiency was increased.•CIEF was coupled with off-line MALDI-TOF MS identification.•Method was tested on spiked milk.
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 2×106mL−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. |
doi_str_mv | 10.1016/j.aca.2013.05.059 |
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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 2×106mL−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.</description><identifier>ISSN: 0003-2670</identifier><identifier>EISSN: 1873-4324</identifier><identifier>DOI: 10.1016/j.aca.2013.05.059</identifier><identifier>PMID: 23845500</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Bacteria ; Calibration ; Capillarity ; Capillary isoelectric focusing ; Cattle ; Electrophoresis, Capillary - methods ; Food Analysis - methods ; Hydrogen-Ion Concentration ; Ionization ; Isoelectric focusing ; Isoelectric Focusing - instrumentation ; Isoelectric Focusing - methods ; Lactic acid bacteria ; Lactobacillus ; Lactobacillus - isolation & purification ; Lasers ; Mass spectrometry ; Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry ; Milk ; Milk - microbiology ; Pasteurization ; Probiotics ; Probiotics - analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods ; Tapered fused silica capillary</subject><ispartof>Analytica chimica acta, 2013-07, Vol.788, p.193-199</ispartof><rights>2013 Elsevier B.V.</rights><rights>Copyright © 2013 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c485t-b4d12bca1de3045f1861cbca30a79cef3e7d8db59168a2119230d392670e96653</citedby><cites>FETCH-LOGICAL-c485t-b4d12bca1de3045f1861cbca30a79cef3e7d8db59168a2119230d392670e96653</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23845500$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Horká, Marie</creatorcontrib><creatorcontrib>Karásek, Pavel</creatorcontrib><creatorcontrib>Šalplachta, Jiří</creatorcontrib><creatorcontrib>Růžička, Filip</creatorcontrib><creatorcontrib>Vykydalová, Marie</creatorcontrib><creatorcontrib>Kubesová, Anna</creatorcontrib><creatorcontrib>Dráb, Vladimír</creatorcontrib><creatorcontrib>Roth, Michal</creatorcontrib><creatorcontrib>Šlais, Karel</creatorcontrib><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</title><title>Analytica chimica acta</title><addtitle>Anal Chim Acta</addtitle><description>•We found method for rapid separation of Lactobacillus in tapered FS capillary.•CIEF separation capacity and efficiency was increased.•CIEF was coupled with off-line MALDI-TOF MS identification.•Method was tested on spiked milk.
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 2×106mL−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.</description><subject>Animals</subject><subject>Bacteria</subject><subject>Calibration</subject><subject>Capillarity</subject><subject>Capillary isoelectric focusing</subject><subject>Cattle</subject><subject>Electrophoresis, Capillary - methods</subject><subject>Food Analysis - methods</subject><subject>Hydrogen-Ion Concentration</subject><subject>Ionization</subject><subject>Isoelectric focusing</subject><subject>Isoelectric Focusing - instrumentation</subject><subject>Isoelectric Focusing - methods</subject><subject>Lactic acid bacteria</subject><subject>Lactobacillus</subject><subject>Lactobacillus - isolation & purification</subject><subject>Lasers</subject><subject>Mass spectrometry</subject><subject>Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry</subject><subject>Milk</subject><subject>Milk - microbiology</subject><subject>Pasteurization</subject><subject>Probiotics</subject><subject>Probiotics - analysis</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</subject><subject>Tapered fused silica capillary</subject><issn>0003-2670</issn><issn>1873-4324</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><recordid>eNqNkktv1DAQxyMEokvhA3BBvtFLtn7GiTihFS-pEhc4W44zbmdJ4mB7KfRT8pFwtKVHFsmvsX7zn7Fnquolo1tGWXO531pnt5wysaWqjO5RtWGtFrUUXD6uNpRSUfNG07PqWUr7YnJG5dPqjItWKkXppvq9swuOo42_CKYAI7gc0REf3CHhfE2CJ0sMPYZcbnvrMkS0xMcwERduXycy4fiN4EyyXSDCQPwhlTXhiM4S9yB-i_mmiPl6xBnIZEuUn7VNCVMu-GgTRDJACnHJGObLMvHOrkeScYI6-NqPeH2Ti2tKJC1rnmGCvOY9wJzRl3gr_7x64u2Y4MX9fl59ff_uy-5jffX5w6fd26vayVblupcD472zbABBpfKsbZgrtqBWdw68AD20Q6861rSWM9ZxQQfRrZ8JXdMocV5dHHXL93w_QMpmwuSgvHaGcEiGadrpVvOmOY02mimlZEtPo4pT0WpN5WlUNnJVlf-RgOi6TnCteUHZEXUxpBTBmyXiVCpoGDVr05m9KU1n1qYzVJXRFZ9X9_KHfoLhweNvlxXgzRGAUpAfCNEkhzA7GDCWQpoh4D_k_wA2pOud</recordid><startdate>20130725</startdate><enddate>20130725</enddate><creator>Horká, Marie</creator><creator>Karásek, Pavel</creator><creator>Šalplachta, Jiří</creator><creator>Růžička, Filip</creator><creator>Vykydalová, Marie</creator><creator>Kubesová, Anna</creator><creator>Dráb, Vladimír</creator><creator>Roth, Michal</creator><creator>Šlais, Karel</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7QL</scope><scope>C1K</scope><scope>7SP</scope><scope>7U5</scope><scope>8FD</scope><scope>L7M</scope></search><sort><creationdate>20130725</creationdate><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</title><author>Horká, Marie ; Karásek, Pavel ; Šalplachta, Jiří ; Růžička, Filip ; Vykydalová, Marie ; Kubesová, Anna ; Dráb, Vladimír ; Roth, Michal ; Šlais, Karel</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c485t-b4d12bca1de3045f1861cbca30a79cef3e7d8db59168a2119230d392670e96653</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animals</topic><topic>Bacteria</topic><topic>Calibration</topic><topic>Capillarity</topic><topic>Capillary isoelectric focusing</topic><topic>Cattle</topic><topic>Electrophoresis, Capillary - methods</topic><topic>Food Analysis - methods</topic><topic>Hydrogen-Ion Concentration</topic><topic>Ionization</topic><topic>Isoelectric focusing</topic><topic>Isoelectric Focusing - instrumentation</topic><topic>Isoelectric Focusing - methods</topic><topic>Lactic acid bacteria</topic><topic>Lactobacillus</topic><topic>Lactobacillus - isolation & purification</topic><topic>Lasers</topic><topic>Mass spectrometry</topic><topic>Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry</topic><topic>Milk</topic><topic>Milk - microbiology</topic><topic>Pasteurization</topic><topic>Probiotics</topic><topic>Probiotics - analysis</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</topic><topic>Tapered fused silica capillary</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Horká, Marie</creatorcontrib><creatorcontrib>Karásek, Pavel</creatorcontrib><creatorcontrib>Šalplachta, Jiří</creatorcontrib><creatorcontrib>Růžička, Filip</creatorcontrib><creatorcontrib>Vykydalová, Marie</creatorcontrib><creatorcontrib>Kubesová, Anna</creatorcontrib><creatorcontrib>Dráb, Vladimír</creatorcontrib><creatorcontrib>Roth, Michal</creatorcontrib><creatorcontrib>Šlais, Karel</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Electronics & Communications Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><jtitle>Analytica chimica acta</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Horká, Marie</au><au>Karásek, Pavel</au><au>Šalplachta, Jiří</au><au>Růžička, Filip</au><au>Vykydalová, Marie</au><au>Kubesová, Anna</au><au>Dráb, Vladimír</au><au>Roth, Michal</au><au>Šlais, Karel</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>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</atitle><jtitle>Analytica chimica acta</jtitle><addtitle>Anal Chim Acta</addtitle><date>2013-07-25</date><risdate>2013</risdate><volume>788</volume><spage>193</spage><epage>199</epage><pages>193-199</pages><issn>0003-2670</issn><eissn>1873-4324</eissn><abstract>•We found method for rapid separation of Lactobacillus in tapered FS capillary.•CIEF separation capacity and efficiency was increased.•CIEF was coupled with off-line MALDI-TOF MS identification.•Method was tested on spiked milk.
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 2×106mL−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.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>23845500</pmid><doi>10.1016/j.aca.2013.05.059</doi><tpages>7</tpages></addata></record> |
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subjects | Animals Bacteria Calibration Capillarity Capillary isoelectric focusing Cattle Electrophoresis, Capillary - methods Food Analysis - methods Hydrogen-Ion Concentration Ionization Isoelectric focusing Isoelectric Focusing - instrumentation Isoelectric Focusing - methods Lactic acid bacteria Lactobacillus Lactobacillus - isolation & purification Lasers Mass spectrometry Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry Milk Milk - microbiology Pasteurization Probiotics Probiotics - analysis Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods Tapered fused silica capillary |
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 |
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