A method for analysing small samples of floral pollen for free and protein-bound amino acids

Pollen provides floral visitors with essential nutrients including proteins, lipids, vitamins and minerals. As an important nutrient resource for pollinators, including honeybees and bumblebees, pollen quality is of growing interest in assessing available nutrition to foraging bees. To date, quantif...

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Main Authors: Stabler, D, Power, E, Borland, A, Barnes, J, Wright, G
Format: Journal article
Language:English
Published: Wiley 2017
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author Stabler, D
Power, E
Borland, A
Barnes, J
Wright, G
author_facet Stabler, D
Power, E
Borland, A
Barnes, J
Wright, G
author_sort Stabler, D
collection OXFORD
description Pollen provides floral visitors with essential nutrients including proteins, lipids, vitamins and minerals. As an important nutrient resource for pollinators, including honeybees and bumblebees, pollen quality is of growing interest in assessing available nutrition to foraging bees. To date, quantifying the protein-bound amino acids in pollen has been difficult and methods rely on large amounts of pollen, typically more than 1 g. More usual is to estimate a crude protein value based on the nitrogen content of pollen, however, such methods provide no information on the distribution of essential and non-essential amino acids constituting the proteins.Here, we describe a method of microwave-assisted acid hydrolysis using low amounts of pollen that allows exploration of amino acid composition, quantified using ultra high performance liquid chromatography (UHPLC), and a back calculation to estimate the crude protein content of pollen.Reliable analysis of protein-bound and free amino acids as well as an estimation of crude protein concentration was obtained from pollen samples as low as 1 mg. Greater variation in both protein-bound and free amino acids was found in pollen sample sizes <1 mg. Due to the variability in recovery of amino acids in smaller sample sizes, we suggest a correction factor to apply to specific sample sizes of pollen in order to estimate total crude protein content.The method described in this paper will allow researchers to explore the composition of amino acids in pollen and will aid research assessing the available nutrition to pollinating animals. This method will be particularly useful in assaying the pollen of wild plants, from which it is difficult to obtain large sample weights.
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spelling oxford-uuid:c91d190c-f26a-404e-8b0e-b55ef3ce3ba82022-03-27T06:56:45ZA method for analysing small samples of floral pollen for free and protein-bound amino acidsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:c91d190c-f26a-404e-8b0e-b55ef3ce3ba8EnglishSymplectic Elements at OxfordWiley2017Stabler, DPower, EBorland, ABarnes, JWright, GPollen provides floral visitors with essential nutrients including proteins, lipids, vitamins and minerals. As an important nutrient resource for pollinators, including honeybees and bumblebees, pollen quality is of growing interest in assessing available nutrition to foraging bees. To date, quantifying the protein-bound amino acids in pollen has been difficult and methods rely on large amounts of pollen, typically more than 1 g. More usual is to estimate a crude protein value based on the nitrogen content of pollen, however, such methods provide no information on the distribution of essential and non-essential amino acids constituting the proteins.Here, we describe a method of microwave-assisted acid hydrolysis using low amounts of pollen that allows exploration of amino acid composition, quantified using ultra high performance liquid chromatography (UHPLC), and a back calculation to estimate the crude protein content of pollen.Reliable analysis of protein-bound and free amino acids as well as an estimation of crude protein concentration was obtained from pollen samples as low as 1 mg. Greater variation in both protein-bound and free amino acids was found in pollen sample sizes <1 mg. Due to the variability in recovery of amino acids in smaller sample sizes, we suggest a correction factor to apply to specific sample sizes of pollen in order to estimate total crude protein content.The method described in this paper will allow researchers to explore the composition of amino acids in pollen and will aid research assessing the available nutrition to pollinating animals. This method will be particularly useful in assaying the pollen of wild plants, from which it is difficult to obtain large sample weights.
spellingShingle Stabler, D
Power, E
Borland, A
Barnes, J
Wright, G
A method for analysing small samples of floral pollen for free and protein-bound amino acids
title A method for analysing small samples of floral pollen for free and protein-bound amino acids
title_full A method for analysing small samples of floral pollen for free and protein-bound amino acids
title_fullStr A method for analysing small samples of floral pollen for free and protein-bound amino acids
title_full_unstemmed A method for analysing small samples of floral pollen for free and protein-bound amino acids
title_short A method for analysing small samples of floral pollen for free and protein-bound amino acids
title_sort method for analysing small samples of floral pollen for free and protein bound amino acids
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