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d- and l-amino acids in Antarctic lakes: assessment of a very sensitive HPLC-MS method

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Abstract

Amino acids represent a fraction of organic matter in marine and freshwater ecosystems, and a source of carbon, nitrogen and energy. l-Amino acids are the most common enantiomers in nature because these chiral forms are used during the biosynthesis of proteins and peptide. To the contrary, the occurrence of d-amino acids is usually linked to the presence of bacteria. We investigated the distribution of l- and d-amino acids in the lacustrine environment of Terra Nova Bay, Antarctica, in order to define their natural composition in this area and to individuate a possible relationship with primary production. A simultaneous chromatographic separation of 40 l- and d-amino acids was performed using a chiral stationary phase based on teicoplainin aglycone (chirobiotic tag). The chromatographic separation was coupled to two different mass spectrometers—an LTQ-Orbitrap XL (Thermo Fisher Scientific) and an API 4000 (ABSciex)—in order to investigate their quantitative performance. High-performance liquid chromatography coupled with mass spectrometry methods were evaluated through the estimation of their linear ranges, repeatability, accuracy and detection and quantification limits. The high-resolution mass spectrometer LTQ-Orbitrap XL presented detection limits between 0.4 and 7 μg l −1, while the triple quadrupole mass spectrometer API 4000 achieved the best detection limits reported in the literature for the quantification of amino acids (between 4 and 200 ng l −1). The most sensitive method, HPLC-API 4000, was applied to lake water samples.

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Acknowledgements

This work was financially supported by the Italian Programma Nazionale di Ricerche in Antartide (PNRA) through the project “Caratterizzazione Biogeochimica dei laghi sub-glaciali antartici (CaBiLA)” (2009/A2.02). The research was also funded by the National Research Council of Italy (CNR) and by the Early Human Impact ERC Advance Grant of the European Commission’s VII Framework Programme, grant number 267696, contribution no. 13. The authors thank ELGA LabWater for providing the PURE-LAB Option-R and Ultra Analytic, which produced the ultrapure water used in these experiments. We would also like to thank Dr. Daniela Almansi for the revision of our manuscript. In conclusion, we wish to thank Dr. S. Illuminati (Polytechnic University of Marche – Ancona, Italy) for her help and cooperation during the sampling activities in Antarctica.

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Authors and Affiliations

  1. Department of Environmental Sciences, Informatics and Statistics, University of Venice, Ca’ Foscari, Calle Larga Santa Marta 2137, 30123, Venice, Italy

    Elena Barbaro & Andrea Gambaro

  2. Institute for the Dynamics of Environmental Processes CNR, Dorsoduro 2137, 30123, Venice, Italy

    Elena Barbaro, Roberta Zangrando, Marco Vecchiato, Clara Turetta, Carlo Barbante & Andrea Gambaro

  3. Department of Physical Sciences, Earth and Environment, University of Siena, Strada Laterina, 8, 53100, Siena, Italy

    Marco Vecchiato

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  1. Elena Barbaro
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  2. Roberta Zangrando
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Correspondence to Elena Barbaro.

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Barbaro, E., Zangrando, R., Vecchiato, M. et al. d- and l-amino acids in Antarctic lakes: assessment of a very sensitive HPLC-MS method. Anal Bioanal Chem 406, 5259–5270 (2014). https://doi.org/10.1007/s00216-014-7961-y

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