Metabolomics analysis II. Preparation of biological samples prior to detection

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

doi:10.1016/j.trac.2009.12.004

Cited by 135 publications

(83 citation statements)

References 53 publications

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“…Sample preparation is critical for optimal metabolite recovery [17,18] and should yield a reasonable number of metabolites with high coverage and reproducible chromatographic data. Various extraction methods have been developed for different types of samples for metabolomic analyses.…”

Section: Results (A) Quantitative Analysis Of Metabolite Extractionmentioning

confidence: 99%

“…Figure 4e is a pNLC that extracted the neutral loss of 141.018 Da from MS/MS spectra of all selected precursors in the MS/MS analysis. The loss of fragment 141.018 Da is a characteristic neutral loss in MS/MS fragmentation of PE species and this identification method has been described previously [18]. Therefore, the pNLC basically shows the intensity and RT distribution of the whole PE species in the LC-MS/MS analysis.…”

Section: (D) Temporal Trend Of All-trans-retinal and N-retinyl-n-retimentioning

confidence: 94%

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Quantitative metabolomics of photoreceptor degeneration and the effects of stem cell-derived retinal pigment epithelium transplantation

Wang

Westenskow

Fang

et al. 2016

Phil. Trans. R. Soc. A.

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metabolomics was further used to determine the temporal effects of the therapeutic intervention of grafting stem cell-derived retinal pigment epithelium (RPE) in dystrophic retinas, which significantly prevented photoreceptor atrophy in our previous studies. The result revealed that lipid levels such as phosphatidylethanolamine in eyes were restored in those animals receiving the RPE grafts. In conclusion, this study provides insight into the metabolomics of retinal degeneration, and further understanding of the efficacy of RPE transplantation.This article is part of the themed issue 'Quantitative mass spectrometry'. BackgroundThe leading cause of central vision loss in industrialized countries is age-related macular degeneration (AMD), which could be caused by ageing, smoking and obesity [1][2][3]. AMD is caused by dysfunction or loss of a neighbouring transporting epithelium, i.e. the retinal pigment epithelium (RPE). RPE cells physically contact the tips of photoreceptors and provide critical metabolic support, recycling light-sensitive pigments required for vision. The photoreceptors are the most numerous and metabolically demanding cells in the retina; the RPE and photoreceptors are considered to be one functional unit due to their co-dependency [4]. The risk of AMD can be reduced through low-glucose and omega-6-enriched diets. Numerous small molecules such as polyunsaturated fatty acids (PUFAs), fatty acid primary amines and amides, amino acids, vitamin A analogues, phospholipids, oxidized species and lipofuscin pigments are enriched in the outer retina. Therefore, the correct balance of these metabolites may be critical for maintaining retinal homeostasis. A more complete understanding of metabolic pathways of retinal development and pathogenesis may be very useful for developing therapeutic interventions to prevent or slow disease progression. To date, no cure for the most common form of AMD, geographical atrophy or 'dry' AMD, has been identified, but it is possible that RPE transplantation could offer a promising therapeutic approach. We and others have shown that transplantation of healthy RPE cells is effective in preclinical studies to significantly, albeit transiently [5], promote photoreceptor survival in the face of RPE-mediated retinal degeneration [6][7][8][9][10]; clinical trials are underway to test the safety and efficacy of stem cell-derived RPE grafts in different types of retinal degeneration [11]. Whether the grafted RPE cells provide essential functional or trophic (or both) support remains unresolved. We have previously examined some of the functional effects of grafted RPE, but changes to the metabolome post transplantation have not been investigated. As retinal metabolism is greatly understudied, and the metabolic pathways that are most essential for vision and retinal homeostasis are not completely understood, we set out to perform a comprehensive metabolomic analysis of the retinal degeneration metabolome. Identification of important classes of metabolites that are important for pho...

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Section: Results (A) Quantitative Analysis Of Metabolite Extractionmentioning

confidence: 99%

Section: (D) Temporal Trend Of All-trans-retinal and N-retinyl-n-retimentioning

confidence: 94%

Quantitative metabolomics of photoreceptor degeneration and the effects of stem cell-derived retinal pigment epithelium transplantation

Wang

Westenskow

Fang

et al. 2016

Phil. Trans. R. Soc. A.

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“…Different mobile phase compositions and analysis time were evaluated using a modified central composite design. Gradient elution mode has been used in complex matrices because of better resolution effects and wider selectivity [15,20,43]. However, CCD has been applied only for optimization in isocratic elution mode limiting the use of dependent variables such as mobile phase composition [21][22][23][24][25]30].…”

Section: Influence Of Organic Phase Composition and Analysis Time In mentioning

confidence: 99%

“…Nevertheless, organic-solvent-based extractions remain as the best choice due to its efficiency and compatibility with major analytical platforms such as GC-MS, LC-MS, and NMR spectroscopy [14][15][16][17][18][19][20]. Although advances in GC derivatization protocols have allowed simultaneous polar and nonpolar compounds analysis, thermostability problems, and low versatility in terms of instrumental coupling hinder its application in metabolomics [12][13][14][15].…”

Section: Introductionmentioning

confidence: 99%

Partial least squares model and design of experiments toward the analysis of the metabolome of Jatropha gossypifolia leaves: Extraction and chromatographic fingerprint optimization

Pilon

Neto

Freire

et al. 2016

J of Separation Science

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A major challenge in metabolomic studies is how to extract and analyze an entire metabolome. So far, no single method was able to clearly complete this task in an efficient and reproducible way. In this work we proposed a sequential strategy for the extraction and chromatographic separation of metabolites from leaves Jatropha gossypifolia using a design of experiments and partial least square model. The effect of 14 different solvents on extraction process was evaluated and an optimized separation condition on liquid chromatography was estimated considering mobile phase composition and analysis time. The initial conditions of extraction using methanol and separation in 30 min between 5 and 100% water/methanol (1:1 v/v) with 0.1% of acetic acid, 20 μL sample volume, 3.0 mL min(-1) flow rate and 25°C column temperature led to 107 chromatographic peaks. After the optimization strategy using i-propanol/chloroform (1:1 v/v) for extraction, linear gradient elution of 60 min between 5 and 100% water/(acetonitrile/methanol 68:32 v/v with 0.1% of acetic acid), 30 μL sample volume, 2.0 mL min(-1) flow rate, and 30°C column temperature, we detected 140 chromatographic peaks, 30.84% more peaks compared to initial method. This is a reliable strategy using a limited number of experiments for metabolomics protocols.

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“…methanol, acetonitrile, ether, acetone, hexane, cyclohexane) which leads not surprisingly to the production of different and complementary fractions of the studied metabolome. [30] Subsequent purification steps may then be employed (e.g. liquid-liquid partitioning, solid-phase extraction) depending on the degree of selectivity wished to be reached for the particular application.…”

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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Metabolomics analysis II. Preparation of biological samples prior to detection

Cited by 135 publications

References 53 publications

Quantitative metabolomics of photoreceptor degeneration and the effects of stem cell-derived retinal pigment epithelium transplantation

Quantitative metabolomics of photoreceptor degeneration and the effects of stem cell-derived retinal pigment epithelium transplantation

Partial least squares model and design of experiments toward the analysis of the metabolome of Jatropha gossypifolia leaves: Extraction and chromatographic fingerprint optimization

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

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