Quantitation of Alpha-Amanitin, T-2, HT-2, DAS and DON in Human Urine
Wed 3:00 PM - Track 3: Emergency Response
Rudolph Johnson
*Rudy Johnson, #Chris Pittman, *Leigh Swaim, *Yingtao Zhou

*Centers for Disease Control and Prevention
#Battelle Memorial Institute
Alpha-Amanitin is a mushroom toxin that causes liver failure within days of consumption. Human urine was spiked with low ng/mL concentrations of alpha-Amanitin, and a synthetic internal standard analogue. The mixture was prepared using a styrene divinylbenzene solid phase extraction sorbent, in a 96-well format. Extracts were pre-concentrated through evaporation, reconstituted and then analyzed. A reversed-phase gradient HPLC method was applied here prior to electrospray ionization, followed by either low or high resolution mass spectrometric analysis. This method was combined with an existing method for measuring human exposure to ricin and abrin by monitoring the copurified components on those toxins, ricinine and abrine, respectively.

A separate method for the measurement of mycotoxins was also developed. Mycotoxins are known to contaminate up to 25% of the world’s grain supply, and are considered a biothreat. T-2, HT-2, DAS and DON were analyzed in a separate, similar method to alpha-amanitin, and analyzed using high resolution orbitrap mass spectrometry only. The method was validated for concentrations between 5 and 200 ng/mL using 20 analytical batches. Each analytical batch consisted of a blank, calibrators and two quality control materials. Statistical evaluation of the method, and analyte stability testing will be discussed here.