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An Improved Method for Measuring Absolute Metabolite Concentrations in Small Biofluid or Tissue Samples.
Fernando, Tharindu; Sawala, Annick; Bailey, Andrew P; Gould, Alex P; Driscoll, Paul C.
Afiliação
  • Fernando T; Physiology and Metabolism Laboratory , The Francis Crick Institute , 1 Midland Road , London NW1 1AT , U.K.
  • Sawala A; Physiology and Metabolism Laboratory , The Francis Crick Institute , 1 Midland Road , London NW1 1AT , U.K.
  • Bailey AP; Physiology and Metabolism Laboratory , The Francis Crick Institute , 1 Midland Road , London NW1 1AT , U.K.
  • Gould AP; Physiology and Metabolism Laboratory , The Francis Crick Institute , 1 Midland Road , London NW1 1AT , U.K.
  • Driscoll PC; Metabolomics Science Technology Platform , The Francis Crick Institute , 1 Midland Road , London NW1 1AT , U.K.
J Proteome Res ; 18(4): 1503-1512, 2019 04 05.
Article em En | MEDLINE | ID: mdl-30757904
ABSTRACT
The measurement of absolute metabolite concentrations in small samples remains a significant analytical challenge. This is particularly the case when the sample volume is only a few microliters or less and cannot be determined accurately via direct measurement. We previously developed volume determination with two standards (VDTS) as a method to address this challenge for biofluids. As a proof-of-principle, we applied VDTS to NMR spectra of polar metabolites in the hemolymph (blood) of the tiny yet powerful genetic model Drosophila melanogaster. This showed that VDTS calculation of absolute metabolite concentrations in fed versus starved Drosophila larvae is more accurate than methods utilizing normalization to total spectral signal. Here, we introduce paired VDTS (pVDTS), an improved VDTS method for biofluids and solid tissues that implements the statistical power of paired control and experimental replicates. pVDTS utilizes new equations that also include a correction for dilution errors introduced by the variable surface wetness of solid samples. We then show that metabolite concentrations in Drosophila larvae are more precisely determined and logically consistent using pVDTS than using the original VDTS method. The refined pVDTS workflow described in this study is applicable to a wide range of different tissues and biofluids.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Metaboloma / Metabolômica Idioma: En Ano de publicação: 2019 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Metaboloma / Metabolômica Idioma: En Ano de publicação: 2019 Tipo de documento: Article