Advanced search
Start date
Betweenand
Related content
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Systematic Evaluation of Extraction Methods for Multiplatform-Based Metabotyping: Application to the Fasciola hepatica Metabolome

Full text
Author(s):
Saric, Jasmina [1] ; Want, Elizabeth J. [1] ; Duthaler, Urs [2, 3] ; Lewis, Matthew [1] ; Keiser, Jennifer [2, 3] ; Shockcor, John P. [4] ; Ross, Gordon A. [5] ; Nicholson, Jeremy K. [1] ; Holmes, Elaine [1] ; Tavares, Marina F. M. [1, 6]
Total Authors: 10
Affiliation:
[1] Univ London Imperial Coll Sci Technol & Med, Fac Med, Dept Surg & Canc, London SW7 2AZ - England
[2] Univ Basel, CH-4003 Basel - Switzerland
[3] Swiss Trop & Publ Hlth Inst, Dept Med Parasitol & Infect Biol, CH-4002 Basel - Switzerland
[4] Waters Corp, Pharmaceut Business Operat, Milford, MA 01757 - USA
[5] Agilent Technol Life Sci & Chem Anal, Stockport SK8 3GR, Cheshire - England
[6] Univ Sao Paulo, Inst Chem, BR-05513970 Sao Paulo - Brazil
Total Affiliations: 6
Document type: Journal article
Source: Analytical Chemistry; v. 84, n. 16, p. 6963-6972, AUG 21 2012.
Web of Science Citations: 32
Abstract

Combining data from multiple analytical platforms is essential for comprehensive study of the molecular phenotype (metabotype) of a given biological sample. The metabolite profiles generated are intrinsically dependent on the analytical platforms, each requiring optimization of instrumental parameters, separation conditions, and sample extraction to deliver maximal biological information. An in-depth evaluation of extraction protocols for characterizing the metabolome of the hepatobiliary fluke Fasciola hepatica, using ultra performance liquid chromatography and capillary electrophoresis coupled with mass spectroscopy is presented. The spectrometric methods were characterized by performance, and metrics of merit were established, including precision, mass accuracy, selectivity, sensitivity, and platform stability. Although a core group of molecules was common to all methods, each platform contributed a unique set, whereby 142 metabolites out of 14,724 features were identified. A mixture design revealed that the chloroform:methanol:water proportion of 15:59:26 was globally the best composition for metabolite extraction across UPLC-MS and CE-MS platforms accommodating different columns and ionization modes. Despite the general assumption of the necessity of platform-adapted protocols for achieving effective metabotype characterization, we show that an appropriately designed single extraction procedure is able to fit the requirements of all technologies. This may constitute a paradigm shift in developing efficient protocols for high-throughput metabolite profiling with more-general analytical applicability. (AU)

FAPESP's process: 10/01579-4 - Biomarker transferability from parasite-rodent models into a human sub-Saharan population via targeted UPLC-MS metabolomic analysis
Grantee:Marina Franco Maggi Tavares
Support Opportunities: Scholarships abroad - Research