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Examining concentrations and molecular weights of thiols in microorganism cultures and in Churchill River (Manitoba) using a fluorescent-labeling method coupled to asymmetrical flow field-flow fractionation

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Abstract

In this study, molecular weights of thiols from four laboratory cultures (Scenedesmus obliquus, Chlorella vulgaris, Euglena gracilis, and Attheya septentrionalis) and the Churchill River (Manitoba) were assessed using a fluorescent-labeling method such as monobromotrimethylammoniobimane (qBBr) and asymmetrical flow field-flow fractionation (AF4) coupled to a fluorescence detector. Concentrations of thiols in extracellular fractions ranged from 6.39 ± 3.39 to 39.2 ± 7.43 μmol g−1, and intracellular concentrations ranged from 11.5 ± 4.52 to 41.0 ± 4.1 μmol g−1. In addition, molecular weights (MW) of intracellular thiol ranged from 493 ± 24 to 946 ± 12 Da whereas extracellular thiol MWs varied from 443 ± 36 to 810 ± 174 Da. The novel method of combining AF4 to an on-line fluorometer and the incorporation of the thiol tag provided information regarding thiol concentration and composition of controlled and natural systems. Furthermore, the proposed methods allow for the simultaneous measurement of thiol and DOM MWs produced by microorganisms. By assessing characteristics of naturally produced thiols and lab-grown thiols, information regarding heavy metal complexation can be determined.

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Acknowledgments

The authors acknowledge the financial support by the Natural Sciences and Engineering Research Council of Canada (NSERC), Northern Science Training Program (NSTP), Churchill Northern Science Centre (CNSC), and Earth Rangers Beluga Research Program. Special thanks to Chad Cuss for his AF4 expertise and LeeAnn Fishback and her team at the Churchill Northern Science Centre, Yu Zhu, Jackie Verstege, and Dave Allcorn for their assistance in the field. Final thanks to Yong Xiang Shi, Weibin Chen, and Fan Xu for measuring DOC concentrations and aiding in qBBr troubleshooting.

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Authors and Affiliations

  1. Environmental and Life Sciences Graduate Program, Trent University, 1600 West Bank Drive, Peterborough, ON, K9J 7B8, Canada

    Vaughn Mangal

  2. Chemistry Department, Trent University, 1600 West Bank Drive, Peterborough, ON, K9J 7B8, Canada

    Celine Guéguen

Authors
  1. Vaughn Mangal
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  2. Celine Guéguen
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Correspondence to Celine Guéguen.

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Published in the topical collection Field- and Flow-based Separations with guest editors Gaetane Lespes, Catia Contado, and Bruce Gale.

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Mangal, V., Guéguen, C. Examining concentrations and molecular weights of thiols in microorganism cultures and in Churchill River (Manitoba) using a fluorescent-labeling method coupled to asymmetrical flow field-flow fractionation. Anal Bioanal Chem 407, 4305–4313 (2015). https://doi.org/10.1007/s00216-015-8599-0

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  • DOI: https://doi.org/10.1007/s00216-015-8599-0

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