Abstract
BackgroundNodularins are an important class of hepatotoxic cyclic pentapeptides that most current methods are ill-suited to clinically monitor. Therefore, a liquid chromatography–tandem mass spectrometry (LC/MS/MS) assay for the determination of nodularin-R in human plasma was validated and applied to clinical samples in this report. MethodsSample cleanup and enrichment were achieved using solid phase extraction. The concentration of nodularin-R in each sample was calculated using the relative abundance area ratio of nodularin-R and a stable isotope-labeled internal standard. ResultsThe validated calibration range was from 0.50 to 100ng/ml. The intra- and inter-day precision were <6.0 and 9.8%, respectively. The accuracy at the lower limit of quantification (LLOQ) was 2.0%, while for other QC levels, it was <3.4%. The absolute concentrations determined by the LC/MS/MS assay were significantly lower than those measured using a commercial immunoassay. Finally, a variety of demographic factors were evaluated and the findings indicated that the elderly (>50y) and people living close to seashore or lakefront areas (≤10km) were the most susceptible to nodularin-R (p=0.79×10−4 and 2.57×10−6, respectively). ConclusionsThis report was among the first to demonstrate that clinical monitoring of nodularin-R could be achieved using LC/MS/MS.
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