A LCMS Metabolomic Workflow to Investigate Metabolic Patterns in Human Intestinal Cells Exposed to Hydrolyzed Crab Waste Materials.

Fionn Ó Fearghail,Nathalie Scheers,Patrice Behan,Niklas Engström

doi:10.3389/fbioe.2021.629083

Abstract

We have developed a LCMS metabolomic workflow to investigate metabolic patterns from human intestinal cells treated with simulated gastrointestinal-digested hydrolyzed crab waste materials. This workflow facilitates smart and reproducible comparisons of cell cultures exposed to different treatments. In this case the variable was the hydrolysis methods, also accounting for the GI digestion giving an output of direct correlation between cellular metabolic patterns caused by the treatments. In addition, we used the output from this workflow to select treatments for further evaluation of the Caco-2 cell response in terms of tentative anti-inflammatory activity in the hopes to find value in the crab waste materials to be used for food products. As hypothesized, the treatment identified to change the cellular metabolomic pattern most readily, was also found to cause the greatest effect in the cells, although the response was pro-inflammatory rather than anti-inflammatory, it proves that changes in cellular metabolic patterns are useful predictors of bioactivity. We conclude that the developed workflow allows for cost effective, rapid sample preparation as well as accurate and repeatable LCMS analysis and introduces a data pipeline specifically for probe the novel metabolite patterns created as a means to assess the performing treatments.

Highlights

At present 70% of European shellfish biomass remains as excess material every year
From the enzymatic and bacterial treatments examined in this work, Corolase 800 can be considered the most effective enzymatic treatment when wishing to produce novel bioactive hydrolyzates from commercial crab waste streams
In seeking to add value to commercial crab waste streams by creating new dietary supplements and nutraceuticals, this work demonstrates that this may not be a likely viable option

Summary

Introduction

At present 70% of European shellfish biomass remains as excess material every year This equates to the same mass as sixty fully laden container ships. This 1.5 mega tonnes of material is all of the parts that are not commonly consumed, mainly consisting of shells, tails and unused meat from the most common European shellfish waste streams, especially that of brown crab (Cancer pagurus). At present most of this material ends up going either to landfill, to incinerators or is dumped directly back in to the sea This material is currently treated as a waste product, it is a rich source of a range of useful substances (Bord, 2019; Morris et al, 2019). The minerals that make up the outer surface of the shells present a source of cheap horticultural stimulants for replenishing mineral

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