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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, food and veterinary sciences |
| UTAS Author: | Phung, SC (Ms Sui Phung) |
| UTAS Author: | Nai, YH (Mr Ryan Nai) |
| UTAS Author: | Powell, SM (Dr Shane Powell) |
| UTAS Author: | Macka, M (Professor Mirek Macka) |
| UTAS Author: | Breadmore, MC (Professor Michael Breadmore) |
| ID Code: | 79331 |
| Year Published: | 2013 |
| Web of Science® Times Cited: | 17 |
| Deposited By: | Tasmanian Institute of Agriculture |
| Deposited On: | 2012-09-01 |
| Last Modified: | 2014-05-16 |
| Downloads: | 0 |
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