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MEDI: Macronutrient Extraction and Determination from invertebrates, a rapid, cheap and streamlined protocol

Lookup NU author(s): Dr Jordan CuffORCiD



This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Macronutrients, comprising carbohydrates, proteins and lipids, underpin many ecological processes, but their quantification in ecological studies is often inaccurate and laborious, requiring large investments of time and bulk samples, which make individual-level studies impossible. This study presents Macronutrient Extraction and Determination from Invertebrates (MEDI), a protocol for the direct, rapid and relatively low-cost determination of macronutrient content from single small macroinvertebrates.Macronutrients were extracted by a sequential process of soaking in 1:12 chloroform:methanol solution to remove lipid and then solubilising tissue in 0.1 M NaOH. Proteins, carbohydrates and lipids were determined by colorimetric assays from the same individual specimens.The limits of detection of MEDI with the equipment and conditions used were 0.067, 0.065 and 0.006 mg/ml for proteins, carbohydrates and lipids respectively. Adjusting the volume of reagents used for extraction and determination can broaden the range of concentrations that can be detected. MEDI successfully identified taxonomic differences in macronutrient content between five insect species.Macronutrient Extraction and Determination from Invertebrates can directly and rapidly determine macronutrient content in tiny (dry mass ~3 mg) and much larger individual invertebrates. Using MEDI, the total macronutrient content of over 50 macroinvertebrates can be determined within around 3 days of collection at a cost of ~$1.35 per sample.

Publication metadata

Author(s): Cuff JP, Wilder SM, Tercel MPTG, Hunt R, Oluwaseun S, Morley PS, Badell-Grau RA, Vaughan IP, Bell JR, Orozco-terWengel P, Symondson WOC, Müller CT

Publication type: Article

Publication status: Published

Journal: Methods in Ecology and Evolution

Year: 2021

Volume: 12

Issue: 4

Pages: 593-601

Print publication date: 01/04/2021

Online publication date: 17/01/2021

Acceptance date: 04/12/2020

Date deposited: 29/10/2021

ISSN (electronic): 2041-210X

Publisher: Wiley-Blackwell Publishing Ltd.


DOI: 10.1111/2041-210X.13551


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Funder referenceFunder name
Biotechnology and Biological Sciences Research Council. Grant Number: BB/M009122/1
BBSRC's Core Capability. Grant Number: BBS/E/C/000J0200
Natural Environment Research Council. Grant Number: NE/L002434/1
US National Science Foundation. Grant Number: DEB1838988