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Metabolites 2017, 7(1), 1; doi:10.3390/metabo7010001

Fully Automated Trimethylsilyl (TMS) Derivatisation Protocol for Metabolite Profiling by GC-MS

1
School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland 1010, New Zealand
2
The Liggins Institute, University of Auckland, Private Bag 92019, Auckland 1010, New Zealand
3
Lasersan Australasia Pty Ltd., Robina QLD 4226, Australia
4
College of Medicine, Biological Sciences and Psychology, University of Leicester, Leicester LE1 7RH, UK
5
Sustainable Production, The New Zealand Institute for Plant & Food Research Limited, Private Bag 92169, Auckland 1142, New Zealand
*
Author to whom correspondence should be addressed.
Academic Editor: Peter Meikle
Received: 23 September 2016 / Revised: 22 December 2016 / Accepted: 26 December 2016 / Published: 29 December 2016
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Abstract

Gas Chromatography-Mass Spectrometry (GC-MS) has long been used for metabolite profiling of a wide range of biological samples. Many derivatisation protocols are already available and among these, trimethylsilyl (TMS) derivatisation is one of the most widely used in metabolomics. However, most TMS methods rely on off-line derivatisation prior to GC-MS analysis. In the case of manual off-line TMS derivatisation, the derivative created is unstable, so reduction in recoveries occurs over time. Thus, derivatisation is carried out in small batches. Here, we present a fully automated TMS derivatisation protocol using robotic autosamplers and we also evaluate a commercial software, Maestro available from Gerstel GmbH. Because of automation, there was no waiting time of derivatised samples on the autosamplers, thus reducing degradation of unstable metabolites. Moreover, this method allowed us to overlap samples and improved throughputs. We compared data obtained from both manual and automated TMS methods performed on three different matrices, including standard mix, wine, and plasma samples. The automated TMS method showed better reproducibility and higher peak intensity for most of the identified metabolites than the manual derivatisation method. We also validated the automated method using 114 quality control plasma samples. Additionally, we showed that this online method was highly reproducible for most of the metabolites detected and identified (RSD < 20) and specifically achieved excellent results for sugars, sugar alcohols, and some organic acids. To the very best of our knowledge, this is the first time that the automated TMS method has been applied to analyse a large number of complex plasma samples. Furthermore, we found that this method was highly applicable for routine metabolite profiling (both targeted and untargeted) in any metabolomics laboratory. View Full-Text
Keywords: metabolomics; matrix; automation; sample preparation; sugars; amino acids; organic acids metabolomics; matrix; automation; sample preparation; sugars; amino acids; organic acids
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MDPI and ACS Style

Zarate, E.; Boyle, V.; Rupprecht, U.; Green, S.; Villas-Boas, S.G.; Baker, P.; Pinu, F.R. Fully Automated Trimethylsilyl (TMS) Derivatisation Protocol for Metabolite Profiling by GC-MS. Metabolites 2017, 7, 1.

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