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Rapid and sensitive microbial analysis by capillary isotachophoresis with continuous electrokinetic injection under field amplified conditions

Citation

Phung, SC and Nai, YH and Powell, SM and Macka, M and Breadmore, MC, Rapid and sensitive microbial analysis by capillary isotachophoresis with continuous electrokinetic injection under field amplified conditions, Electrophoresis, 34, (11) pp. 1657-1662. ISSN 0173-0835 (2013) [Refereed Article]

Copyright Statement

Copyright 2013 Wiley-VCH Verlag GmbH & Co.

DOI: doi:10.1002/elps.201200479

Abstract

A highly sensitive capillary isotachophoresis method with LIF detection for microbial analysis was developed. This allowed the reliable analysis of Escherichia coli bacteria with a LOD of 14 cells in a sample volume of 100 μL, or 1.35 102 cell/mL, which is 47 times lower than reported byCE-LIF and 148 times lower thanCE-UV with on-line concentration. A leading electrolyte of 50 mM Tris-HCl was used wh le the cells were diluted in 5 mM Tris HEPES as the terminator. To facilitate detection, cells were stained with the universal nucleic acid fluorophore SYTO 9. Continuous electrokinetic injection of the cells from the terminator under field amplified conditions concentrated cells into a single peak at the leader/terminator boundary allowing quantitation by measurement of peak height. The method was applied to water collected from two local streams, with only filtration through a 5-μm syringe filter to remove large particulate matter followed by a ten times dilution in terminator, with total analysis time approximately 40 min. The detected cell numbers in the water samples by the isotachophoresis method were 3.70 105 cell/mL and 2.62 104 cell/mL, which were slightly higher than the 9.50 104 cell/mL and 1.96 104 cell/mL obtained by conventional microbiological plate counting.

Item Details

Item Type:Refereed Article
Keywords:capillary isotachophoresis, electrokinetic injection, field amplified, microbial
Research Division:Chemical Sciences
Research Group:Analytical Chemistry
Research Field:Analytical Chemistry not elsewhere classified
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Agricultural and Veterinary Sciences
Author:Phung, SC (Ms Sui Phung)
Author:Nai, YH (Mr Ryan Nai)
Author:Powell, SM (Dr Shane Powell)
Author:Macka, M (Professor Mirek Macka)
Author:Breadmore, MC (Professor Michael Breadmore)
ID Code:79331
Year Published:2013
Web of Science® Times Cited:15
Deposited By:Tasmanian Institute of Agriculture
Deposited On:2012-09-01
Last Modified:2014-05-16
Downloads:0

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