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Microalgae amino acid extraction and analysis at nanomolar level using electroporation and capillary electrophoresis with laser‐induced fluorescence detection
Author(s) -
Nehmé Reine,
Atieh Carla,
Fayad Syntia,
Claude Bérengère,
Chartier Agnès,
Tannoury Mona,
Elleuch Fatma,
Abdelkafi Slim,
Pichon Chantal,
Morin Philippe
Publication year - 2017
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201601005
Subject(s) - capillary electrophoresis , chromatography , electrophoresis , chemistry , derivatization , extraction (chemistry) , laser induced fluorescence , amino acid , electroporation , fluorescence , analytical chemistry (journal) , mass spectrometry , biochemistry , physics , quantum mechanics , gene
Amino acids play a key role in food analysis, clinical diagnostics, and biochemical research. Capillary electrophoresis with laser‐induced fluorescence detection was used for the analysis of several amino acids. Amino acid labeling with fluorescein isothiocyanate was conducted using microwave‐assisted derivatization at 80°C (680 W) during only 150 s. Good electrophoretic resolution was obtained using a background electrolyte composed of sodium tetraborate buffer (100 mM; pH 9.4) and β‐cyclodextrin (10 mM), and the limits of quantification were 3–30 nM. The developed capillary electrophoresis with laser‐induced fluorescence method was used to analyze amino acids in Dunaliella salina green algae grown under different conditions. A simple extraction technique based on electroporation of the cell membrane was introduced. A home‐made apparatus allowed the application of direct and alternating voltages across the electrochemical compartment containing a suspension of microalgae in distilled water at 2.5 g/L. A direct voltage of 12 V applied for 4 min gave the optimum extraction yield. Results were comparable to those obtained with accelerated‐solvent extraction. The efficiency of electroporation in destroying microalgae membranes was shown by examining the algae surface morphology using scanning electron microscopy. Stress conditions were found to induce the production of amino acids in Dunaliella salina cells.