title: Fast and sensitive HPLC method for the determination of neopterin, kynurenine and tryptophan in amniotic fluid, malignant effusions and wound exudates.
abstract: OBJECTIVE
  A new HPLC method for the determination of neopterin, kynurenine and tryptophan using a second-generation monolith stationary phase and high-throughput sample preparation procedure based on microplates was developed and fully validated.
  
  
  METHODS
  As the stationary phase a monolithic C18 Chromolith high-resolution column with dimensions of 4.6 × 100 mm connected to a monolithic 4.6 × 10-mm security guard was used. Separation was achieved using 15 mM phosphate buffer (KH2PO4 +K2HPO4·3H2O at pH 3) and acetonitrile in gradient mode.
  
  
  RESULTS
  Target analytes were determined in 5.5 minutes in amniotic fluid, effusions and wound exudates with a limit of quantification (LOQ) of 1.25 nM for neopterin, 2.5 µM for tryptophan and 0.25 µM for kynurenine.
  
  
  CONCLUSIONS
  The method was applied to real clinical sample measurements, and it will be used to monitor neopterin, kynurenine and tryptophan levels in biological fluids to assess the patient response to therapy and clinical status.
authors.affiliation: Department of Analytical, Chemistry Faculty of Pharmacy, Charles University in Prague, Hradec Kralove, Czech Republic.|3rd Internal Gerontometabolic Clinic, University Hospital, Hradec Kralove, Czech Republic.
authors.forename: Lenka Kujovska
authors.initials: LK
authors.lastname: Krcmova
authors.affiliation: Department of Analytical, Chemistry Faculty of Pharmacy, Charles University in Prague, Hradec Kralove, Czech Republic.|3rd Internal Gerontometabolic Clinic, University Hospital, Hradec Kralove, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Cervinkova
authors.affiliation: 3rd Internal Gerontometabolic Clinic, University Hospital, Hradec Kralove, Czech Republic.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Solichova
authors.affiliation: 3rd Internal Gerontometabolic Clinic, University Hospital, Hradec Kralove, Czech Republic.
authors.forename: Lubos
authors.initials: L
authors.lastname: Sobotka
authors.affiliation: Department of Gynecology & Obstetrics, Faculty of Medicine & Dentistry & University Hospital, Palacky University, Olomouc, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Hansmanova
authors.affiliation: Department of Oncology, Faculty of Medicine & Dentistry & University Hospital, Palacky University, Olomouc, Czech Republic.
authors.forename: Bohuslav
authors.initials: B
authors.lastname: Melichar
authors.affiliation: Department of Analytical, Chemistry Faculty of Pharmacy, Charles University in Prague, Hradec Kralove, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Solich
mesh_terms: D000653:Amniotic Fluid / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005122:Exudates and Transudates / Q000378:metabolism*; D006801:Humans; D007737:Kynurenine / Q000032:analysis*; D019798:Neopterin / Q000032:analysis*; D016066:Pleural Effusion, Malignant / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D014364:Tryptophan / Q000032:analysis*; D014945:Wound Healing
chemical_list: D007737:Kynurenine; D019798:Neopterin; D014364:Tryptophan