title: Biosensor for on-line fluorescent detection of trifluoroperazine based on genetically modified calmodulin.
abstract: This paper describes the development of a novel on-line biosensor based on a fluorescently labeled human calmodulin (CaM), hCaM M124C-mBBr, immobilized on controlled-pore glass (CPG), for the analysis of trifluoroperazine (TFP); a phenothiazine drug in human urine samples. The device was automated by packing hCaM M124C-mBBr-CPG in a continuous-flow microcell connected to a monitoring system, composed of a bifurcated optical fiber coupled to a spectrofluorometer. Operating parameters of the on-line biosensor (flow rate, sample injection volume, and carrier solution and buffer pH) were studied and optimized. Under the optimal conditions, the biosensor provides a detection and a quantification limit of 0.24 and 0.52 μg mL(-1), respectively, and a dynamic range from 0.52 to 61.05 μg mL(-1) TFP (n = 5, correlation coefficient 0.998). The response time (t(100)) was shorter than 42 s (recovery time <4.5 min) and reproducibility and repeatability of the TFP measurements, within the linear response range, were lower than 1.4 and 2.7%, respectively. The device was successfully applied to the analysis of TFP in spiked human urine samples with recoveries ranging between 97 and 101% and with RSDs lower than 5.9%.
authors.affiliation: Departamento de Química Analítica, Facultad de Química, Universidad Complutense, 28040 Madrid, Spain.
authors.forename: Martin
authors.initials: M
authors.lastname: González-Andrade
authors.forename: Elena
authors.initials: E
authors.lastname: Benito-Peña
authors.forename: Rachel
authors.initials: R
authors.lastname: Mata
authors.forename: Maria C
authors.initials: MC
authors.lastname: Moreno-Bondi
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002147:Calmodulin / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods; D011485:Protein Binding; D014268:Trifluoperazine / Q000032:analysis* / Q000652:urine
chemical_list: D002147:Calmodulin; D005456:Fluorescent Dyes; D014268:Trifluoperazine