title: Validation of a HPLC-MS/MS assay for the determination of total and unbound concentration of temocillin in human serum.
abstract: OBJECTIVE
  The aim of this study was to develop and validate a HPLC-MS/MS assay to determine total and unbound concentrations of temocillin in serum samples.
  
  
  METHODS
  Methanolic protein precipitation and ultrafiltration were used for total and unbound concentration extraction, respectively. Extract was injected into a LC-MS/MS system. Reversed phase chromatography was performed on a phenyl grafted column in gradient mode. Temocillin and internal standard (ticarcillin) were identified in positive electrospray ionization mode using ion transitions of m/z 415.34>339.1 and 385.31>160.3, respectively.
  
  
  RESULTS
  Temocillin total and unbound concentration quantification assays were linear over concentrations ranging from 1 to 500 mg/L and from 0.5 to 300 mg/L, respectively. Both assays presented acceptable intra and inter-assay precision and accuracy <13.9%. Limits of quantification and detection were of 1 and 0.10mg/L, and 0.5 and 0.05 mg/L for total and unbound concentration respectively. Total temocillin concentration recovery ranged from 85.80 to 99.40%. Temocillin ion suppression effect was <36.2 % in both assays.
  
  
  CONCLUSIONS
  The method described is fast, sensitive and selective, with no interferences. This method may be used for both pharmacokinetic studies and therapeutic drug monitoring purposes.
authors.affiliation: Pharmacologie cellulaire et moléculaire, Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium.
authors.forename: Perrin
authors.initials: P
authors.lastname: Ngougni Pokem
authors.affiliation: Pharmacologie cellulaire et moléculaire, Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium; Clinical Pharmacy Research Group, Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium; Center for Clinical Pharmacy, Université catholique de Louvain, Brussels, Belgium.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Miranda Bastos
authors.affiliation: Pharmacologie cellulaire et moléculaire, Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium; Center for Clinical Pharmacy, Université catholique de Louvain, Brussels, Belgium.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Tulkens
authors.affiliation: Clinical Chemistry Department, Cliniques Universitaires St. Luc, Université catholique de Louvain, Brussels, Belgium; Louvain Center for Toxicology and Applied Pharmacology, Université catholique de Louvain, Brussels, Belgium.
authors.forename: Pierre
authors.initials: P
authors.lastname: Wallemacq
authors.affiliation: Pharmacologie cellulaire et moléculaire, Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium; Center for Clinical Pharmacy, Université catholique de Louvain, Brussels, Belgium. Electronic address: francoise.vanbambeke@uclouvain.be.
authors.forename: Françoise
authors.initials: F
authors.lastname: Van Bambeke
authors.affiliation: Clinical Chemistry Department, Cliniques Universitaires St. Luc, Université catholique de Louvain, Brussels, Belgium; Louvain Center for Toxicology and Applied Pharmacology, Université catholique de Louvain, Brussels, Belgium.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Capron
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016903:Drug Monitoring / Q000379:methods*; D006801:Humans; D010406:Penicillins / Q000097:blood*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D010406:Penicillins; C031367:temocillin
keywords: HPLC-MS/MS; Serum concentration; Temocillin; Therapeutic drug monitoring; Unbound fraction