title: Electrochemical lactate biosensor based upon chitosan/carbon nanotubes modified screen-printed graphite electrodes for the determination of lactate in embryonic cell cultures.
abstract: l-lactate is an essential metabolite present in embryonic cell culture. Changes of this important metabolite during the growth of human embryo reflect the quality and viability of the embryo. In this study, we report a sensitive, stable, and easily manufactured electrochemical biosensor for the detection of lactate within embryonic cell cultures media. Screen-printed disposable electrodes are used as electrochemical sensing platforms for the miniaturization of the lactate biosensor. Chitosan/multi walled carbon nanotubes composite have been employed for the enzymatic immobilization of the lactate oxidase enzyme. This novel electrochemical lactate biosensor analytical efficacy is explored towards the sensing of lactate in model (buffer) solutions and is found to exhibit a linear response towards lactate over the concentration range of 30.4 and 243.9 µM in phosphate buffer solution, with a corresponding limit of detection (based on 3-sigma) of 22.6 µM and exhibits a sensitivity of 3417 ± 131 µAM(-1) according to the reproducibility study. These novel electrochemical lactate biosensors exhibit a high reproducibility, with a relative standard deviation of less than 3.8% and an enzymatic response over 82% after 5 months stored at 4 °C. Furthermore, high performance liquid chromatography technique has been utilized to independently validate the electrochemical lactate biosensor for the determination of lactate in a commercial embryonic cell culture medium providing excellent agreement between the two analytical protocols.
authors.affiliation: Department of Physical Chemistry and Institute of Electrochemistry, Universidad de Alicante, 03080 Alicante, Spain.
authors.forename: Naiara
authors.initials: N
authors.lastname: Hernández-Ibáñez
authors.affiliation: Department of Physical Chemistry and Institute of Electrochemistry, Universidad de Alicante, 03080 Alicante, Spain.
authors.forename: Leticia
authors.initials: L
authors.lastname: García-Cruz
authors.affiliation: Department of Physical Chemistry and Institute of Electrochemistry, Universidad de Alicante, 03080 Alicante, Spain.
authors.forename: Vicente
authors.initials: V
authors.lastname: Montiel
authors.affiliation: Faculty of Science and Engineering, School of Science and the Environment, Division of Chemistry and Environmental Science, Manchester Metropolitan University, Chester Street, Manchester M1 5GD, UK.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Foster
authors.affiliation: Faculty of Science and Engineering, School of Science and the Environment, Division of Chemistry and Environmental Science, Manchester Metropolitan University, Chester Street, Manchester M1 5GD, UK.
authors.forename: Craig E
authors.initials: CE
authors.lastname: Banks
authors.affiliation: Department of Physical Chemistry and Institute of Electrochemistry, Universidad de Alicante, 03080 Alicante, Spain. Electronic address: jesus.iniesta@ua.es.
authors.forename: Jesús
authors.initials: J
authors.lastname: Iniesta
mesh_terms: D015374:Biosensing Techniques; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D003217:Conductometry / Q000295:instrumentation; D004553:Electric Conductivity; D053595:Embryonic Stem Cells / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006108:Graphite / Q000737:chemistry; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000737:chemistry*; D037742:Nanotubes, Carbon / Q000737:chemistry* / Q000648:ultrastructure; D010781:Photography / Q000379:methods
chemical_list: D004800:Enzymes, Immobilized; D037742:Nanotubes, Carbon; D019344:Lactic Acid; D006108:Graphite; D048271:Chitosan; D006899:Mixed Function Oxygenases; C021497:lactate 2-monooxygenase
keywords: Chitosan; Embryo cell culture medium; Lactate; Lactate oxidase; MWCNT; Screen-printed graphite electrode