Open AccessLC-MS Quantification of Malondialdehyde-Dansylhydrazine Derivatives in Urine and Serum Samples
Author(s) -
Kostya Kartavenka,
Parinya Panuwet,
Volha Yakimavets,
Churdsak Jaikang,
Kanitarin Thipubon,
Priya Esilda D’Souza,
Dana Boyd Barr,
P. Barry Ryan
Publication year - 2020
Publication title -
journal of analytical toxicology.
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.161
H-Index - 76
eISSN - 1945-2403
pISSN - 0146-4760
DOI - 10.1093/jat/bkz112
Subject(s) - chemistry , chromatography , derivatization , urine , mass spectrometry , quantitative analysis (chemistry) , electrospray ionization , reagent , selected reaction monitoring , tandem mass spectrometry , biochemistry
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.