Development of an untargeted metabolomics method for the analysis of human faecal samples using Cryptosporidium-infected samples

Ng, Josephine; Ryan, Una; Trengove, Robert D.

doi:10.1016/j.molbiopara.2012.08.006

Cited by 73 publications

(84 citation statements)

References 31 publications

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“…Thus far, few studies have investigated optimal sampling conditions for fecal VOC analysis, mostly using the chemical analytical technique “gas chromatography-mass spectrometry” (GC-MS) [23,24,25,26,27]. GC-MS allows for identification of individual VOCs, based on their physiochemical properties, whereas an eNose harbors a chemical array sensor system, allowing for identification of complex gaseous mixtures by deploying pattern recognition algorithms [3,28].…”

Section: Discussionmentioning

confidence: 99%

“…Reade and colleagues demonstrated eight compounds to be detectable at significant higher intensities in samples weighing 450 mg compared to samples with a mass of 100 mg (2,3-butanedione, tetrahydrofurane, ethyl ester propanoic acid, n-propyl acetate, 2-pentenal E, propyl ester propanoic acid, 2-methylpropanal and 1-propanol) [16]. In contrast, results of fecal VOC profile comparison between samples containing higher masses are ambiguous [23,24,26]. Reade and colleagues state in their manuscript that the applied solid phase micro-extraction (SPME) fiber, required for capturing fecal VOCs prior to analysis, reached its limit of absorbance at 0.45 g [24].…”

Section: Discussionmentioning

confidence: 99%

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Effects of Sampling Conditions and Environmental Factors on Fecal Volatile Organic Compound Analysis by an Electronic Nose Device

Berkhout

Benninga

Stein

et al. 2016

Sensors

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Prior to implementation of volatile organic compound (VOC) analysis in clinical practice, substantial challenges, including methodological, biological and analytical difficulties are faced. The aim of this study was to evaluate the influence of several sampling conditions and environmental factors on fecal VOC profiles, analyzed by an electronic nose (eNose). Effects of fecal sample mass, water content, duration of storage at room temperature, fecal sample temperature, number of freeze–thaw cycles and effect of sampling method (rectal swabs vs. fecal samples) on VOC profiles were assessed by analysis of totally 725 fecal samples by means of an eNose (Cyranose320®). Furthermore, fecal VOC profiles of totally 1285 fecal samples from 71 infants born at three different hospitals were compared to assess the influence of center of origin on VOC outcome. We observed that all analyzed variables significantly influenced fecal VOC composition. It was feasible to capture a VOC profile using rectal swabs, although this differed significantly from fecal VOC profiles of similar subjects. In addition, 1285 fecal VOC-profiles could significantly be discriminated based on center of birth. In conclusion, standardization of methodology is necessary before fecal VOC analysis can live up to its potential as diagnostic tool in clinical practice.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Effects of Sampling Conditions and Environmental Factors on Fecal Volatile Organic Compound Analysis by an Electronic Nose Device

Berkhout

Benninga

Stein

et al. 2016

Sensors

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“…In a following study the same group suggested methanol as the optimum organic solvent and a ratio of 1/2 volumes of fecal water to ice-cold methanol [45]. Ng et al, [46] proposed a ratio of 1/5 (sample to methanol volume) as the optimal for GC-MS analysis of human fecal samples.…”

Section: Gc-ms Profiling Datamentioning

confidence: 99%

“…Till now only few studies have dealt with the optimisation of the method used for the sample preparation of feces. In these studies metabolomics analysis was performed by NMR spectroscopy [41][42][43] and GC-MS [44][45][46][47][48] and the investigated parameters included freeze-drying, sonication, filtration, homogenization, sample weight to buffer/solvent ratio, extraction solvent, pH value, extraction duration and repetitions numbers. Up to date there is no study of the optimum treatment conditions of fecal samples for metabolic fingerprinting by LC-MS.…”

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confidence: 99%

Sample preparation optimization in fecal metabolic profiling

Deda

Chatziioannou

Fasoula

et al. 2017

Journal of Chromatography B

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“…A recent preliminary metabolomics study on Cryptosporidium developed a faecal metabolite extraction method for untargeted gas chromatography-mass spectrometry (GC-MS) analysis using Cryptosporidium positive and negative human faecal samples [12]. In that study, faecal metabolite profiles of cryptosporidiosis positive patients could be differentiated from cryptosporidiosis negative patients, suggesting that metabolic homeostasis and intestinal permeability were affected as a result of the infection [12].…”

Section: Introductionmentioning

confidence: 99%

Metabolomic Profiling of Faecal Extracts from Cryptosporidium parvum Infection in Experimental Mouse Models

2013

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Cryptosporidiosis is a gastrointestinal disease in humans and animals caused by infection with the protozoan parasite Cryptosporidium. In healthy individuals, the disease manifests mainly as acute self-limiting diarrhoea, but may be chronic and life threatening for those with compromised immune systems. Control and treatment of the disease is challenged by the lack of sensitive diagnostic tools and broad-spectrum chemotherapy. Metabolomics, or metabolite profiling, is an emerging field of study, which enables characterisation of the end products of regulatory processes in a biological system. Analysis of changes in metabolite patterns reflects changes in biochemical regulation, production and control, and may contribute to understanding the effects of Cryptosporidium infection in the host environment. In the present study, metabolomic analysis of faecal samples from experimentally infected mice was carried out to assess metabolite profiles pertaining to the infection. Gas-chromatography mass spectrometry (GC-MS) carried out on faecal samples from a group of C. parvum infected mice and a group of uninfected control mice detected a mean total of 220 compounds. Multivariate analyses showed distinct differences between the profiles of C. parvum infected mice and uninfected control mice,identifying a total of 40 compounds, or metabolites that contributed most to the variance between the two groups. These metabolites consisted of amino acids (n = 17), carbohydrates (n = 8), lipids (n = 7), organic acids (n = 3) and other various metabolites (n = 5), which showed significant differences in levels of metabolite abundance between the infected and uninfected mice groups (p < 0.05). The metabolites detected in this study as well as the differences in abundance between the C. parvum infected and the uninfected control mice, highlights the effects of the infection on intestinal permeability and the fate of the metabolites as a result of nutrient scavenging by the parasite to supplement its streamlined metabolism.

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Development of an untargeted metabolomics method for the analysis of human faecal samples using Cryptosporidium-infected samples

Cited by 73 publications

References 31 publications

Effects of Sampling Conditions and Environmental Factors on Fecal Volatile Organic Compound Analysis by an Electronic Nose Device

Effects of Sampling Conditions and Environmental Factors on Fecal Volatile Organic Compound Analysis by an Electronic Nose Device

Sample preparation optimization in fecal metabolic profiling

Metabolomic Profiling of Faecal Extracts from Cryptosporidium parvum Infection in Experimental Mouse Models

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