LC-MS Quantification of Malondialdehyde-Dansylhydrazine Derivatives in Urine and Serum Samples

• Published in: Journal of analytical toxicology Vol. 44; no. 5; pp. 470 - 481
• Main Authors: Kartavenka, Kostya, Panuwet, Parinya, Yakimavets, Volha, Jaikang, Churdsak, Thipubon, Kanitarin, D’Souza, Priya Esilda, Barr, Dana Boyd, Ryan, P Barry
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 02.04.2020
• Subjects: Body Fluids; Chromatography, High Pressure Liquid; Chromatography, Liquid; Dansyl Compounds - metabolism; Humans; Hydrazines - blood; Hydrazines - metabolism; Hydrazines - urine; Limit of Detection; Malondialdehyde - blood; Malondialdehyde - metabolism; Malondialdehyde - urine; Tandem Mass Spectrometry; LC-MS; malondialdehyde; derivatization; serum; urine; marker of lipid peroxidation
• ISSN: 0146-4760; 1945-2403
• DOI: 10.1093/jat/bkz112

Abstract We developed a robust analytical method for quantification of malondialdehyde (MDA) in urine and serum samples using dansylhydrazine (DH) as a derivatizing reagent. The derivatization procedure was partially carried out using an autosampler injection program to minimize errors associated with the low-volume addition of reagents and was optimized to yield a stable hydrazone derivative of MDA and its labeled d2-MDA analogue. The target MDA-DH derivatives were separated on an Agilent Zorbax Eclipse Plus Phenyl-Hexyl (3.0 × 100 mm, 3.5 μm) column. The mass-to-charge ratios of the target derivatives [(M+H)+ of 302 and 304 for MDA-DH and d2-MDA-DH, respectively] were analyzed in single ion monitoring mode using a single quadrupole mass spectrometer operated under positive electrospray ionization. The method limits of quantification were 5.63 nM (or 0.405 ng/mL) for urine analysis and 5.68 nM (or 0.409 ng/mL) for serum analysis. The quantification range for urine analysis was 5.63–500 nM (0.405–36.0 ng/mL) while the quantification range for serum analysis was 5.68–341 nM (0.409–24.6 ng/mL). The method showed good relative recoveries (98–103%), good accuracies (92–98%), and acceptable precisions (relative standard deviations 1.8–7.3% for inter-day precision; 1.8–6.1% for intra-day precision) as observed from the repeat analysis of quality control samples prepared at different concentrations. The method was used to measure MDA in individual urine samples (n = 287) and de-identified archived serum samples (n = 22) to assess the overall performance of the method. The results demonstrated that our method is capable of measuring urinary and serum levels of MDA, allowing its future application in epidemiologic investigations.