Abstract

Tissue is considered as a highly informative matrix in omics studies. Most commonly used tissues in metabolomics are fresh-frozen specimens. However, an alternative metabolomics matrix, up to now most often applies in immunohistochemistry or histopathology, is formalin-fixed, paraffin-embedded tissue (FFPE). The main advantages of FFPE tissues over fresh-frozen ones are the possibility to be stored at ambient temperature which is a cost-effective method and, consequently, their higher availability in biobanks. The main aim of the study was to develop and optimize the preparation protocol for FFPE samples in metabolomics, using cancer tissue and corresponding control tissue sections. Several conditions were evaluated: type and volume of deparaffinization solvent, number of purification cycles, solvent used in purification step, extraction solvent, and thickness of FFPE samples. Samples were analyzed by gas chromatography coupled with mass spectrometry (GC–MS). As a result, a sample preparation protocol of FFPE tissues for metabolomics was developed. Moreover, applicability of the proposed method was tested with the use of FFPE tissues obtained from prostate cancer patient and non-cancer controls.

Highlights

  • Metabolomics is a field of study focusing on quantitative and qualitative analysis of products of the metabolic pathways [1]

  • Three different approaches are utilized: metabolic fingerprinting, metabolic profiling, and targeted analysis

  • The main aim of the study was to develop reliable and effective sample preparation protocol for FFPE samples, which could be applied for untargeted metabolomics approach

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Summary

Introduction

Metabolomics is a field of study focusing on quantitative and qualitative analysis of products of the metabolic pathways [1]. The aim of metabolomics is to provide the information about the composition of metabolome and changes within it under various conditions. Three different approaches are utilized: metabolic fingerprinting (determination of global metabolome), metabolic profiling (quantitative analysis of selected set of compounds that are related by the similar function, properties or belong to the same biochemical pathway), and targeted analysis The aim of formalin fixation is to preserve the tissue by the inhibition of proteolytic enzymes as well as the growth of microorganisms [20, 21]. Fixed tissue should remain biologically stable through the time of storage [21]

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