Abstract
BackgroundThe amino acid profile of plants is an important parameter in assessments of their growth potential, resource-use efficiency and/or quality as food and feed. Screening studies may involve large number of samples but the classical amino acid analysis is limited by the fact that it is very time consuming with typical chromatographic run times of 70 min or more.ResultsWe have here developed a high-throughput method for analysis of amino acid profiles in plant materials. The method combines classical protein hydrolysis and derivatization with fast separation by UHPLC and detection by a single quadrupole (QDa) mass spectrometer. The chromatographic run time is reduced to 10 min and the precision, accuracy and sensitivity of the method are in line with other recent methods utilizing advanced and more expensive mass spectrometers. The sensitivity of the method is at least a factor 10 better than that of methods relying on detection by fluorescence or UV. It is possible to downscale sample size to 20 mg without compromising reproducibility, which makes the method ideal for analysis of very small sample amounts.ConclusionThe developed method allows high-throughput analysis of amino acid profiles in plant materials. The analysis is robust and accurate as well as compatible with both free amino acids and protein hydrolysates. The QDa detector offers high sensitivity and accuracy, while at the same time being relatively simple to operate and cheap to purchase, thus significantly reducing the overall analytical costs compared to methods based on more advanced mass spectrometers.
Highlights
The amino acid profile of plants is an important parameter in assessments of their growth potential, resource-use efficiency and/or quality as food and feed
The purpose of the present study was to develop a high-throughput amino acid analysis exploiting the advantages of single quadrupole MS for detection in combination with stable isotope labelled amino acids as internal standards
The only exception was the isobaric amino acids isoleucine/leucine, which appeared in the same channel and had a retention of about 7.5 min (Fig. 1; Table 1)
Summary
The amino acid profile of plants is an important parameter in assessments of their growth potential, resource-use efficiency and/or quality as food and feed. A large number of research questions in basic plant biology and plant breeding require analysis of the amino acid profiles of proteins in single plant tissues or whole plants This is required in order to improve parameters associated with plant growth, tolerance towards environmental stress and resource-use efficiency. The current societal focus on biorefining and bio-based economy has prompted a booming interest and spurred new research activities on how to exploit proteins extracted from green biomass [2,3,4] These studies have often been performed without assessing the nutritional values of the obtained proteins by amino acid analysis [5, 6]. Plant-derived proteins are believed to have an enormous untapped potential for animal feed, food, food ingredients and as raw materials for other valuable products in future biorefinery contexts [7]
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