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

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

2041-210X
430-438
Stabler, Daniel
b275ba93-2cd8-460a-b5dc-b527a268f351
Power, Eileen F
a406c2de-f8fd-4535-95d6-d6095f08d2c2
Borland, Anne M
f67cc093-98cb-4d2e-b537-e6f13ad6ea7b
Barnes, Jeremy D
a5b7a763-dc14-48a1-ad83-b12a773298a7
Wright, Geraldine A
f1dac0e1-1920-43ce-802a-c9056c817eb0
Stabler, Daniel
b275ba93-2cd8-460a-b5dc-b527a268f351
Power, Eileen F
a406c2de-f8fd-4535-95d6-d6095f08d2c2
Borland, Anne M
f67cc093-98cb-4d2e-b537-e6f13ad6ea7b
Barnes, Jeremy D
a5b7a763-dc14-48a1-ad83-b12a773298a7
Wright, Geraldine A
f1dac0e1-1920-43ce-802a-c9056c817eb0

Stabler, Daniel, Power, Eileen F, Borland, Anne M, Barnes, Jeremy D and Wright, Geraldine A (2018) A method for analysing small samples of floral pollen for free and protein-bound amino acids. Methods in Ecology and Evolution, 9 (2), 430-438. (doi:10.1111/2041-210X.12867).

Record type: Article

Abstract

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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Published date: 1 February 2018

Identifiers

Local EPrints ID: 477086
URI: http://eprints.soton.ac.uk/id/eprint/477086
ISSN: 2041-210X
PURE UUID: 594fa48f-b455-42da-a657-a30d20b54557

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Date deposited: 25 May 2023 16:49
Last modified: 10 May 2024 17:01

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Contributors

Author: Daniel Stabler
Author: Eileen F Power
Author: Anne M Borland
Author: Jeremy D Barnes
Author: Geraldine A Wright

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