title: Immunoaffinity in-tube solid phase microextraction coupled with liquid chromatography-mass spectrometry for analysis of fluoxetine in serum samples.
abstract: The inherent selectivity of the antibody was combined with in-tube solid-phase microextraction by immobilization of the antibody into the fused silica capillary. A sensitive, selective, and reproducible immunoaffinity in-tube solid-phase microextraction coupled with liquid chromatography-mass spectrometry (in-tube SPME/LC-MS) method was developed, and validated for fluoxetine analysis in human serum. Important factors in the optimization of in-tube SPME variables, as well as the evaluation of the immunoaffinity capillary capacity are discussed. The in-tube SPME/LC-MS method presented a limit of quantitation of 5.00 ng/mL, and precision intra-assays with RSDs lower than 5%. The response of the in-tube SPME/LC-MS method for fluoxetine was linear over a dynamic range from 5.00 to 50.00 ng/mL, with correlation coefficients better than 0.998. Based on analytical validation it was demonstrated that in-tube SPME/LC-MS method offers high sensitivity, selectivity, and enough reproducibility to permit the quantification of fluoxetine in human serum at therapeutic levels. Thus, the proposed SPME/LC method can be useful tool to determine fluoxetine serum concentrations in patients receiving therapeutic dosages.
authors.affiliation: Departamento de Química, Faculdade de Filosofia Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901 Ribeirao Preto, SP, Brazil. mariaegn@ffclrp.up.br
authors.forename: Maria Eugênia C
authors.initials: ME
authors.lastname: Queiroz
authors.forename: Eduardo B
authors.initials: EB
authors.lastname: Oliveira
authors.forename: François
authors.initials: F
authors.lastname: Breton
authors.forename: Janusz
authors.initials: J
authors.lastname: Pawliszyn
mesh_terms: D000906:Antibodies / Q000276:immunology*; D002853:Chromatography, Liquid; D005473:Fluoxetine / Q000097:blood*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D015203:Reproducibility of Results; D012822:Silicon Dioxide; D052617:Solid Phase Microextraction / Q000379:methods*; D012996:Solutions
chemical_list: D000906:Antibodies; D012996:Solutions; D005473:Fluoxetine; D012822:Silicon Dioxide