Analysis of pseudomonas quinolone signal and other bacterial signalling molecules using capillaries coated with highly charged polyelectrolyte monolayers and boron doped diamond electrode

Download
  1. Get@NRC: Analysis of pseudomonas quinolone signal and other bacterial signalling molecules using capillaries coated with highly charged polyelectrolyte monolayers and boron doped diamond electrode (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1016/j.chroma.2012.06.064
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titleJournal of Chromatography A
ISSN0021-9673
Volume1251
Pages169175
SubjectCapillary electrophoresis; PQS; HHQ; Biomarker detection
AbstractCoated capillary electrophoresis equipped with a boron doped diamond (BDD) electrode was developed for analysis of chemically synthesised 2-heptyl-3-hydroxy-4-quinolone (HHQ), 2-heptyl-3-hydroxy-4-quinolone (PQS), and 2-methyl analogues. Detection was then extended to biological samples. PQS and its biological precursor, HHQ, are two key regulators of bacterial cooperative behaviour known as quorum sensing in the nosocomial pathogen Pseudomonas aeruginosa. The fused silica capillary was coated with a thin layer of poly (diallyldimethylammonium) chloride to reverse the electroosmosis, allowing fast migration of PQS and HHQ with improved selectivity. The four model compounds were baseline resolved using a 50 mM H3PO4–Tris, pH 2.0 buffer with 20% (v/v) acetonitrile as buffer additive. With an injection time of 3 s, the detection limits of four analytes ranging from 60 to 100 nM (S/N = 3) were observed when the BDD electrode was poised at +1.5 V vs. 3 M Ag/AgCl. As expected, no PQS or HHQ was detected from the supernatant of the P. aeruginosa (pqsA) mutant. A concentration of HHQ of 247 μM was detected from the supernatant of the pqsH mutant, which catalyses the conversion of HHQ to PQS in the presence of molecular oxygen by monooxygenase. The separation and detection scheme was applicable to follow the conversion of HHQ to PQS in P. aeruginosa when entering the stationary phase of growth. The results obtained by coated capillary electrophoresis with BDD detection were validated and compared well with LC–MS data.
Publication date
LanguageEnglish
AffiliationAquatic and Crop Resource Development; National Research Council Canada
Peer reviewedYes
IdentifierS0021967312009582
NPARC number21268698
Export citationExport as RIS
Report a correctionReport a correction
Record identifierdd247f8f-f88c-4bd4-af58-b067f8d83b58
Record created2013-11-07
Record modified2016-05-09
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)