title: One-step fabrication of integrated disposable biosensor based on ADH/NAD+/meldola's blue/graphitized mesoporous carbons/chitosan nanobiocomposite for ethanol detection.
abstract: A novel strategy to simplify the dehydrogenase-based electrochemical biosensor fabrication through one-step drop-coating nanobiocomposite on a screen printed electrode (SPE) was developed. The nanobiocomposite was prepared by successively adding graphitized mesoporous carbons (GMCs), meldola's blue (MDB), alcohol dehydrogenase (ADH) and cofactor nicotinamide adenine dinucleotide (NAD(+)) in chitosan (CS) solution. MDB/GMCs/CS film was prepared. Cyclic voltammetry measurements demonstrated that MDB was strongly adsorbed on GMCs. After optimizing the concentration of MDB and the working potential, the MDB/GMCs/CS film presented a fast amperometric response (5s), excellent sensitivity (10.36 nA μM(-1)), wide linear range (10-410 μM) toward NADH and without any other interference signals (such as AA, UA, DA, H2O2 and metal ions). Furthermore, concentrations of ADH and NAD(+) in nanobiocomposite and the detection conditions (temperature and pH) were also optimized. The constructed disposable ethanol biosensor showed an excellent linear response ranged from 0.5 to 15 mM with high sensitivity (67.28 nA mM(-1)) and a low limit of detection (80 μM) and a remarkable long-term stability (40 days). The intra-batch and inter-batch variation coefficients were both less than 5% (n=5). The ethanol recovery test demonstrated that the proposed biosensor offered a remarkable and accurate method for ethanol detection in the real blood samples.
authors.affiliation: Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, No. 1 Yi Xue Yuan Road, Chongqing 400016, PR China.
authors.forename: Erhui
authors.initials: E
authors.lastname: Hua
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Jing
authors.forename: Changtao
authors.initials: C
authors.lastname: Chen
authors.forename: Guoming
authors.initials: G
authors.lastname: Xie
mesh_terms: D000079:Acetaldehyde / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000378:metabolism*; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002244:Carbon / Q000737:chemistry; D002384:Catalysis; D048271:Chitosan / Q000737:chemistry; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods; D006108:Graphite / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009243:NAD / Q000737:chemistry / Q000378:metabolism*; D053761:Nanocomposites / Q000737:chemistry*; D010078:Oxazines / Q000737:chemistry; D015203:Reproducibility of Results; D013696:Temperature; D013997:Time Factors
chemical_list: D010078:Oxazines; D009243:NAD; C005559:Meldola blue; D002244:Carbon; D006108:Graphite; D048271:Chitosan; D000426:Alcohol Dehydrogenase; D000079:Acetaldehyde