The Open UniversitySkip to content
 

Development and optimization of a gas chromatography/mass spectrometry method for the analysis of thermochemolytic degradation products of phthiocerol dimycocerosate waxes found in Mycobacterium tuberculosis

Nicoara, Simona C.; Minnikin, David E.; Lee, Oona C. Y.; O'Sullivan, Denise M.; McNerney, Ruth; Pillinger, Colin T.; Wright, Ian P. and Morgan, Geraint H. (2013). Development and optimization of a gas chromatography/mass spectrometry method for the analysis of thermochemolytic degradation products of phthiocerol dimycocerosate waxes found in Mycobacterium tuberculosis. Rapid Communications in Mass Spectrometry, 27(21) pp. 2374–2382.

Full text available as:
[img]
Preview
PDF (Version of Record) - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
Download (1MB) | Preview
DOI (Digital Object Identifier) Link: https://doi.org/10.1002/rcm.6694
Google Scholar: Look up in Google Scholar

Abstract

RATIONALE: The phthiocerol dimycocerosates (PDIMs) are certain stable and hydrophobic waxes found in the cell membrane of Mycobacterium tuberculosis, bacteria that cause an infectious disease of growing concern worldwide. Previous studies report the analysis of derivatives of the hydrolysed PDIMs from biological samples, following complex extraction and offline derivatization of PDIMs biomarkers, prior to their analysis by gas chromatography/mass spectrometry (GC/MS).
METHODS: We developed and optimized a GC/MS method based on selected ion monitoring (SIM) to detect the derivatives produced via the thermally assisted hydrolysis and methylation (THM) of the PDIMs from the cell membrane of M. tuberculosis. The extraction of PDIMs from culture is simple, and their thermochemolysis is carried out automatically online, thus avoiding the time-consuming derivatization steps of hydrolysis and esterification, usually performed offline.
RESULTS: For standard PDIMs in petroleum ether, our optimized method gave an excellent linearity (R2 = 0.99) at concentrations between 0.172 and 27.5 ng/mL, a good precision (RSD = 11.42 %), and a limit of detection (LOD) of 100 pg/mL. For the PDIMs extracted from dilutions of M. tuberculosis culture, the method gave good linearity (R2 = 0.9685) and an estimated LOD of 400 CFU/mL (CFU = colony forming units) in sterile distilled water.
CONCLUSIONS: A GC/MS(SIM) method is presented for the rapid and quantitative detection of M. tuberculosis, based on the online thermochemolysis of lipidic biomarkers extracted from the bacterial culture. The method has the potential to be applied in human and veterinary clinical laboratories for the rapid diagnosis of tuberculosis in infected biological samples.

Item Type: Journal Item
Copyright Holders: 2013 The Authors
ISSN: 0951-4198
Academic Unit/School: Faculty of Science, Technology, Engineering and Mathematics (STEM)
Faculty of Science, Technology, Engineering and Mathematics (STEM) > Physical Sciences
Research Group: Space
Item ID: 38788
Depositing User: Astrid Peterkin
Date Deposited: 18 Oct 2013 12:39
Last Modified: 09 Dec 2018 21:22
URI: http://oro.open.ac.uk/id/eprint/38788
Share this page:

Metrics

Altmetrics from Altmetric

Citations from Dimensions

Download history for this item

These details should be considered as only a guide to the number of downloads performed manually. Algorithmic methods have been applied in an attempt to remove automated downloads from the displayed statistics but no guarantee can be made as to the accuracy of the figures.

Actions (login may be required)

Policies | Disclaimer

© The Open University   contact the OU