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Seeking faster, alternative methods for glycolipid biosurfactant characterization and purification

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Abstract

Biosurfactants have been investigated as potential alternatives for synthetic surfactants in several areas, for example, in environmental and pharmaceutical fields. In that regard, extensive research has been carried out with sophorolipids and rhamnolipids that also present various biological properties with therapeutic significance. These biosurfactants are obtained as complex mixtures of slightly different molecules, and thus when studying these microbial glycolipids, the ability to identify and purify the produced compounds is of extreme importance. This study aimed to develop improved methodologies for the identification, separation, and purification of sophorolipids and rhamnolipids. Therefore, an ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) method was modified to ensure faster characterization of both sophorolipids and rhamnolipids, enabling the identification and fragmentation pattern description of 10 and 13 congeners, respectively. The separation and purification of these biosurfactants was achieved with novel reversed-phase solid-phase extraction methods guaranteeing the isolation of different glycolipids, including those considered for their significant biological activity (e.g. antimicrobial, anticancer). It was possible to isolate sophorolipids and rhamnolipids with purity of 94% and 99%, respectively. The methods presented herein can be easily implemented and are expected to make purification of these biosurfactants easier, facilitating the study of their individual properties in further works.

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Acknowledgements

The authors thank Fundação para a Ciência e Tecnologia (FCT) for the financial support under Project PTDC/BTM-SAL/29335/2017, UIDB/04138/2020 and UIDP/04138/2020 (iMed.ULisboa). The authors would also like to acknowledge the financial support from Fundação para a Ciência e Tecnologia (FCT) and Portugal 2020 to the Portuguese Mass Spectrometry Network (Rede Nacional de Espectrometria de Massa RNEM; LISBOA 01 0145 FEDER 402 022125). The authors also acknowledge the Designer Nuno Monge for the artwork and Andreia Bento for the assistance with the UHPLC-MS/MS equipment.

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Authors and Affiliations

  1. Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Avenida Prof. Gama Pinto, 1649-003, Lisbon, Portugal

    Maïssa Dardouri, Rita M. Mendes, Judite Costa & Isabel A. C. Ribeiro

  2. Pharmaceutical Institute, Rheinische Friedrich-Wilhelms-Universität Bonn, An der Immenburg 4, 53121, Bonn, Germany

    Johannes Frenzel

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  1. Maïssa Dardouri
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  2. Rita M. Mendes
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Correspondence to Isabel A. C. Ribeiro.

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Dardouri, M., Mendes, R.M., Frenzel, J. et al. Seeking faster, alternative methods for glycolipid biosurfactant characterization and purification. Anal Bioanal Chem 413, 4311–4320 (2021). https://doi.org/10.1007/s00216-021-03387-4

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