Microalgae amino acid extraction and analysis at nanomolar level using electroporation and capillary electrophoresis with laser-induced fluorescence detection.
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.
Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans, Orléans, France.
Reine
R
Nehmé
Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans, Orléans, France.
Carla
C
Atieh
Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans, Orléans, France.
Syntia
S
Fayad
Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans, Orléans, France.
Bérengère
B
Claude
Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans, Orléans, France.
Agnès
A
Chartier
Département de Biologie, Faculté des Sciences II, Université Libanaise, Fanar, Liban.
Mona
M
Tannoury
Biotechnologie des algues, Département de Génie biologique, Ecole Nationale d'Ingénieurs de Sfax, University of Sfax, Tunisia.|Centre de Biophysique moléculaire, et Université d'Orléans, France.
Fatma
F
Elleuch
Biotechnologie des algues, Département de Génie biologique, Ecole Nationale d'Ingénieurs de Sfax, University of Sfax, Tunisia.
Slim
S
Abdelkafi
Centre de Biophysique moléculaire, et Université d'Orléans, France.
Chantal
C
Pichon
Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans, Orléans, France.
Philippe
P
Morin
D000596:Amino Acids / Q000032:analysis* / Q000302:isolation & purification*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D000460:Chlorophyta / Q000737:chemistry / Q000378:metabolism; D019075:Electrophoresis, Capillary; D018274:Electroporation; D005453:Fluorescence; D007834:Lasers; D058086:Microalgae / Q000737:chemistry*
D000596:Amino Acids
amino acid extraction; capillary electrophoresis; electroporation; laser-induced fluorescence detection; microwave-assisted derivatization