Simultaneous quantification of succinylacetone and nitisinone for therapeutic drug monitoring in the treatment of Tyrosinemia type 1

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 1072; pp. 259 - 266
• Main Authors: Sundberg, Jonas, Wibrand, Flemming, Lund, Allan Meldgaard, Christensen, Mette
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2018
• Subjects: Bioanalysis; chromatography; derivatization; drugs; hydrazine; hydrochloric acid; Liquid chromatography; Mass spectrometry; monitoring; Nitisinone; Parallelism; patients; storage conditions; Succinylacetone; Tyrosinemia; Tyrosinemia; Bioanalysis; Parallelism; Liquid chromatography; Mass spectrometry; Succinylacetone; Nitisinone
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2017.11.031

[Display omitted] •Simultaneous quantification of succinylacetone and nitisinone in plasma.•LC–MS/MS method with high dynamic range and excellent accuracy and precision.•Single-step derivatization and chromatographic separation using green solvents.•Applicable for screening and therapeutic drug monitoring of Tyrosinemia type 1. We present a straightforward and robust method for simultaneous quantification of succinylacetone and nitisinone in plasma using LC-ESI–MS/MS. The method has been developed for routine therapeutic drug monitoring in hepatorenal tyrosinemia type 1 (HT1) patients undergoing nitisinone treatment. Previous methods are based on separate analyses of succinylacetone and nitisinone, often using the potentially harmful compound hydrazine for derivatization of the former. In the present procedure, succinylacetone is derivatized in a single-step using butanolic HCl. Analyte extraction and sample clean-up is carried out by simple protein precipitation. The linear range for both analytes is 0.1 up to 125μM, covering the vast majority of encountered levels in real-life samples. The sensitivity and limit of quantification allows measurement of succinylacetone in the therapeutical range for HT1 patients. Stability studies show that succinylacetone is highly sensitive to storage conditions, whereas nitisinone shows little to no degradation. Correct sample handling is therefore important for reliable results when monitoring succinylacetone concentrations.