Metabolomics analysis I. Selection of biological samples and practical aspects preceding sample preparation

Álvarez-Sánchez, B.; Priego-Capote, Feliciano; Castro, M.D. Luque de

doi:10.1016/j.trac.2009.12.003

Cited by 125 publications

(76 citation statements)

References 59 publications

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“…As amostras coletadas devem ser devidamente armazenadas, para evitar degradação ou ativação de enzimas; isso pode ser alcançado estocando-se as amostras a baixas temperaturas (-80 ºC é usualmente preferida). 12 O preparo de amostra para análise metabolômica, terceira etapa do fluxograma de trabalho, é uma das etapas mais importantes e que demanda maior tempo de trabalho. Isso se deve à complexidade das amostras biológicas, sua heterogeneidade e grande faixa de concentração dos metabólitos presentes.…”

Section: Com O Aumento Dos Estudos Utilizando Abordagens Metabolômicaunclassified

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Metabolômica: Definições, Estado-Da-Arte E Aplicações Representativas

Canuto¹,

Costa²,

Cruz³

et al. 2017

Quim. Nova

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METABOLOMICS: DEFINITIONS, STATE-OF-THE-ART AND REPRESENTATIVE APPLICATIONS.Metabolomics is an emerging and promising omics approach used to understand biological mechanisms. By untargeted and targeted metabolomics analyses, metabolites are determined in biological samples (fluids, cells, tissues, etc.) by comparison of control groups with altered groups, undergoing different therapies, submitted to differing stress levels, dietary modulation, or promoted by a disease, or specific condition, etc., using sophisticated analytical techniques, and advanced data treatment and statistical analyses. In this review, the concepts involved in metabolomics studies were presented, describing in details all steps involved in the metabolomics workflow, for untargeted and targeted strategies. Finally, the potential of metabolomics is illustrated by applications in representative areas: clinical, environmental, food and nutrition, forensic toxicology, microbiology, parasitology, plants, and sports. Relevant reviews were compiled to characterize each of these areas, and a corresponding application of untargeted and targeted metabolomics were described.Keywords: metabolomics; untargeted metabolomics; targeted metabolomics; metabolomics workflow. INTRODUÇÃOAs ciências ômicas buscam o entendimento do funcionamento celular dos organismos e suas alterações biológicas. Fazem parte deste conjunto de ciências, a genômica (estudo da alteração dos genes), a transcriptômica (estudo das alterações dos transcritos), a proteômica (estudo das alterações das proteínas), e a metabolômica (estudo das alterações dos metabólitos). Metabólitos são produtos intermediários ou finais do metabolismo em uma amostra biológica.1 O conjunto de todos os metabólitos de baixa massa molecular (até 1500 Da), presentes ou alterados em um sistema biológico, é chamado de metaboloma (do inglês, metabolome).2 A pesquisa relacionada a metabólitos vem sendo desenvolvida há décadas, mas em 1999, Nicholson et al. definiram a metabonômica (do inglês, metabonomics), como sendo a medida quantitativa da resposta metabólica de um sistema biológico após estímulos fisiopatológicos ou modificações genéticas.3 Já o termo metabolômica (do inglês, metabolomics) foi introduzido em 2001, por Oliver Fiehn, como sendo a análise abrangente e quantitativa do metaboloma de um sistema biológico. 4 Além disso, a literatura nos apresenta outras denominações envolvendo esse campo da ciência, tais como: perfil metabólico (do inglês, metabolic profiling), descrito como sendo a análise de metabólitos previamente selecionados de rotas bioquímicas específicas, 5 a impressão digital metabólica (do inglês, metabolic fingerprinting), que é definida como sendo "uma classificação de amostras de acordo com sua origem ou sua relevância biológica", 3 e, por último, análise footprinting (ainda sem tradução para o português), para se referir aos metabólitos excretados por uma célula em condições controladas. Observa-se, portanto, uma divergência em relação às terminologias aplicadas às análises metabolômic...

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Section: Com O Aumento Dos Estudos Utilizando Abordagens Metabolômicaunclassified

“…Isso se deve à complexidade das amostras biológicas, sua heterogeneidade e grande faixa de concentração dos metabólitos presentes. 12 Sua execução depende da abordagem metabolômica escolhida, tipo de amostra e técnicas de análise que serão de interesse no estudo.…”

Section: Com O Aumento Dos Estudos Utilizando Abordagens Metabolômicaunclassified

Metabolômica: Definições, Estado-Da-Arte E Aplicações Representativas

Canuto¹,

Costa²,

Cruz³

et al. 2017

Quim. Nova

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“…The sample storage conditions will have a direct influence on the sample stabilisation and integrity. [32] Each step in the sample pretreatment has direct consequences on the metabolic fingerprints generated (content and variability). Overall, the repeatability of this sample preparation process appears to be of prime importance and has to be systematically characterized, considering that each particular choice will directly impact on the obtained metabolic profiles and subsequent interpretation.…”

Section: Sample Preparationmentioning

confidence: 99%

Metabolomics in food analysis: application to the control of forbidden substances

Dervilly

Courant

Chéreau

et al. 2012

Drug Testing and Analysis

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Metabolomics is a science of interest in food analysis to describe and predict properties of food products and processes. It includes the development of analytical methods with the ultimate goal being the identification of so-called 'quality markers', (i.e. sets of metabolites that correlate with, for example, quality, safety, taste, or fragrance of foodstuffs). In turn, these metabolites are influenced by factors as genetic differences of the raw food ingredients (such as animal breed or crop species differences), growth conditions (such as climate, irrigation strategy, or feeding) or production conditions (such as temperature, acidity, or pressure). In cases where the routine-based measurement of a food property faces some limitations such as the lack of knowledge regarding the target compounds to monitor, monitoring based on a limited set of crucial biomarkers is a good alternative, which is of great interest for food safety purposes regarding growth promoting practices. Such an approach may be more efficient than using a classic approach based on a limited set of known metabolites of anabolic compounds. In this context, screening strategies allowing detection of the physiological response resulting from anabolic compound administration are promising approaches to detect their misuse. The global metabolomics workflow implemented for such studies is presented and illustrated through various examples of biological matrices profiling (tissue, blood, urine) and for different classes of anabolic compounds (steroids, b-agonists and somatotropin).

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“…Recently, proteomics and metabolomics tools have been applied for the analysis of proteome and metabolome of CSF [3,4]. However, metabolome analysis of CSF remains to be an analytical challenge because of low concentrations of many metabolites present in CSF and the availability of only small volumes of samples.…”

Section: Introductionmentioning

confidence: 99%

Qualitative Metabolome Analysis of Human Cerebrospinal Fluid by¹³C-/¹²C-Isotope Dansylation Labeling Combined with Liquid Chromatography Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Guo

Bamforth

2011

J. Am. Soc. Mass Spectrom.

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Metabolome analysis of human cerebrospinal fluid (CSF) is challenging because of low abundance of metabolites present in a small volume of sample. We describe and apply a sensitive isotope labeling LC-MS technique for qualitative analysis of the CSF metabolome. After a CSF sample is divided into two aliquots, they are labeled by 13 C-dansyl and 12 C-dansyl chloride, respectively. The differentially labeled aliquots are then mixed and subjected to LC-MS using Fourier-transform ion cyclotron resonance mass spectrometry (FTICR MS). Dansylation offers significant improvement in the performance of chromatography separation and detection sensitivity. Moreover, peaks detected in the mass spectra can be readily analyzed for ion pair recognition and database search based on accurate mass and/or retention time information. It is shown that about 14,000 features can be detected in a 25-min LC-FTICR MS run of a dansyl-labeled CSF sample, from which about 500 metabolites can be profiled. Results from four CSF samples are compared to gauge the detectability of metabolites by this method. About 261 metabolites are commonly detected in replicate runs of four samples. In total, 1132 unique metabolite ion pairs are detected and 347 pairs (31%) matched with at least one metabolite in the Human Metabolome Database. We also report a dansylation library of 220 standard compounds and, using this library, about 85 metabolites can be positively identified. Among them, 21 metabolites have never been reported to be associated with CSF. These results illustrate that the dansylation LC-FTICR MS method can be used to analyze the CSF metabolome in a more comprehensive manner.

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Metabolomics analysis I. Selection of biological samples and practical aspects preceding sample preparation

Cited by 125 publications

References 59 publications

Metabolômica: Definições, Estado-Da-Arte E Aplicações Representativas

Metabolômica: Definições, Estado-Da-Arte E Aplicações Representativas

Metabolomics in food analysis: application to the control of forbidden substances

Qualitative Metabolome Analysis of Human Cerebrospinal Fluid by¹³C-/¹²C-Isotope Dansylation Labeling Combined with Liquid Chromatography Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

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Metabolomics analysis I. Selection of biological samples and practical aspects preceding sample preparation

Cited by 125 publications

References 59 publications

Metabolômica: Definições, Estado-Da-Arte E Aplicações Representativas

Metabolômica: Definições, Estado-Da-Arte E Aplicações Representativas

Metabolomics in food analysis: application to the control of forbidden substances

Qualitative Metabolome Analysis of Human Cerebrospinal Fluid by13C-/12C-Isotope Dansylation Labeling Combined with Liquid Chromatography Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Contact Info

Product

Resources

About

Qualitative Metabolome Analysis of Human Cerebrospinal Fluid by¹³C-/¹²C-Isotope Dansylation Labeling Combined with Liquid Chromatography Fourier Transform Ion Cyclotron Resonance Mass Spectrometry