Liquid chromatography tandem mass spectrometry method for the quantitation of NTBC (2-(nitro-4-trifluoromethylbenzoyl)1,3-cyclohexanedione) in plasma of tyrosinemia type 1 patients.
In this study, we describe a bioanalytical method for quantification of NTBC in plasma of patients with hereditary tyrosinemia type 1 (HT-1) using high-performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). After protein precipitation with acetonitrile including Mesotrione as internal standard, separation of NTBC was achieved by RP-HPLC. Detection was performed by positive ion electrospray ionization (ESI) in selected reaction monitoring (SRM) mode. NTBC recovery in the developed method was found to be more than 90%. The lower limit of quantification was calculated to be 0.35 microM. The intra-day and inter-day precision of three different quality control samples (measured as RSD%) was less than 10% and 15%, respectively. The standard calibration curves showed good linearity within the range of 2.5-40 microM and the determined correlation coefficients were r(2)>or=0.995. This presented method is rapid, sensitive, specific and suitable for clinical practice and research.
Department of General Paediatrics, University Children's Hospital, Heinrich-Heine University Düsseldorf, Germany.
Diran
D
Herebian
Ute
U
Spiekerkötter
Marc
M
Lamshöft
Eva
E
Thimm
Maurice
M
Laryea
Ertan
E
Mayatepek
D002853:Chromatography, Liquid / Q000379:methods*; D003512:Cyclohexanones / Q000097:blood*; D006801:Humans; D009579:Nitrobenzoates / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D020176:Tyrosinemias / Q000097:blood*
D003512:Cyclohexanones; D009579:Nitrobenzoates; C077073:nitisinone