Comparison of plasma and dry blood spots as samples for the determination of nitisinone (NTBC) by high-performance liquid chromatography–tandem mass spectrometry. Study of the stability of the samples at different temperatures

Tyrosinemia is an inborn error of metabolism characterized by the accumulation of tyrosine as well as toxic by-products. NTBC or nitisinone is a drug currently used for the treatment of tyrosinemia that avoids the formation of these toxic substances. This paper presents the determination of NTBC in plasma and dry blood spots by high-performance liquid chromatography (HPLC) coupled to tandem mass spectrometry. The concentration of NTBC in matched plasma-dry blood spots was compared and the study of degradation of NTBC in plasma and dry spots at different temperatures is presented. Method: For sample preparation, plasma proteins were precipitated with acetonitrile and 3-mm discs were extracted with methanol. ESI(+) was used as inozation method and the analytes were detected by multiple reaction monitoring using the transitions 330 > 218 for NTBC and 340 > 228 for mesotrione, used as internal standard. Results: There is good correlation between concentrations obtained in dry blood spots and plasma (r2 = 0.83), although values are 2.4 times higher in plasma samples. NTBC in plasma is stable at least for 45 days frozen at −30 °C and refrigerated at 4 °C. However, it shows slow decomposition at room temperature, approximately 30% after 45 days. The method shows good precision, accuracy and linearity and the detection limit is 50 nmol/L and paper samples are appropriate for the monitorization of NTBC.