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Process for detecting Helicobacter pylori using aliphatic amides.
MedLine Citation:
PMID:  21822779     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Helicobacter pylori diagnosis is fundamental in the management of gastrointestinal pathologies, whose current clinical guidelines support a non-invasive 'test-and-treat' strategy. As such, the present work reports the basis of a new, low-cost, specific breath test based on the detection of volatile carboxylic acids resulting from the hydrolysis of short-chain aliphatic amides by H. pylori amidases. Propionamide and butyramide, which are metabolized by amidases to propionic and butyric acids, were elected for this study. Conditions for the extraction of these acids from a vapour phase were optimized concerning the use of solid-phase microextraction (SPME) followed by gas chromatography-quadrupole mass spectrometry (GC-qMS) analysis. SPME-GC-qMS was then used to detect the acids released into a vapour phase upon incubation of a H. pylori reference strain J99 or a clinical specimen with the amides. These experiments have demonstrated that the administration of less than 9 mg of propionamide and/or butyramide to H. pylori cultures, in loads recognized to cause infection (10(6)-10(9) cells), resulted in the formation of detectable and/or quantifiable amounts of propionic and/or butyric acids after 30 min incubation. As such, propionic and butyric acids can be used as biomarkers for H. pylori upon incubation with the corresponding amides. SPME-GC-qMS was also used to verify the hepatic stability of the acids. These experiments were conducted in mouse liver cells and revealed no signs of metabolization that could compromise their bioavailability in future in vivo assays. Moreover, SPME-GC-qMS permitted the detection of both acids in amounts as low as 0.8 μg in systems mimicking exhaled breath, demonstrating the sensitivity of the method for these compounds.
Authors:
José A Ferreira; Elsa Dias; Sílvia M Rocha; Manuel A Coimbra
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-8-6
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  -     ISSN:  1618-2650     ISO Abbreviation:  -     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-8-8     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
QOPNA, Department of Chemistry, University of Aveiro, Campus de Santiago, 3810-193, Aveiro, Portugal.
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