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METHOD ABSTRACTS203.1 / MULTI-RESIDUE MYCOTOXIN ANALYSISSINGLE RUN ANALYSIS OF DEOXYNIVALENOL, AFLATOXINS, OCHRATOXIN A,ZEARALENONE AND FUMONISIN BY HPLC AND POST-COLUMN DERIVATIZATIONMaria Otserova, PhD, Sareeta Nerkar, PhD, Michael Pickering, PhDAlthough Aspergillus (Aatoxins, Ochratoxin A) are generally associated with peanuts and Fusarium (Deoxynivalenol,Zearalenone) with wheat, these fungi and those that produce other toxins are not host selective and so can cross plant species.This situation is complicated by the fact that the microscopic mold may not be visible to the naked eye. Also, when infectedgrains are processed, any visible mold is lost but the toxic metabolites carry over into the nished products. Thus, multi-residueanalytical screens for toxins in grain and nished goods are a wiser choice than single-family protocols. We present a singlescreen to cover ve families of toxins. This method is suitable for analyzing beverages, grains and feeds.Sample Extraction and Clean-up25 g of nely grounded sample is extracted with 150 mL of water/Methanol mixture (30/70). 20 mL of ltered extractis diluted with 70 mL of Phosphate Buffered Saline (PBS). Aatoxins, Zearealenone and Ochratoxin A are isolated usingAOZ Immunoafnity column (Vicam, USA) according to the procedure from the column manufacture. Toxins are elutedwith 2 x 2 mL of Methanol. Fumonisins are isolated using FumoniTest Immunoafnity column (Vicam, USA) according tothe procedure from the column manufacture. Toxins are eluted with 2 x 1.5 mL of Methanol.To isolate DON 3 mL of ltered extract is mixed with 6 mL of Acetonitrile and cleaned with MycoSep 277 column(Romer Labs, USA) according to the manufacture’s instructions. The cleaned solution is ltered and 5.5 mL of it is combinedwith eluants from AOZ and FumoniTest columns. The solution is evaporated to 0.5 mL and nal volume is adjustedto 1 mL with Methanol.METHODAnalytical ConditionsColumn: MYCOTOX reversed-phase C 18,4.6x250 mm Catalog No. 1612124Temperature: 40 ºCFlow Rate: 1 mL/minMobile Phase: Sodium Phosphate buffer, pH 3.5Catalog No 1700-1108/MeOH/ACNHPLC PROGRAMTIME (Min) 1700-1108 ,% METHANOL, % ACETONITRILE, %0.0 85 0 155.0 85 0 155.1 57 28 1520.0 57 28 1523.0 40 60 040.0 40 60 050.0 20 0 8060.0 20 0 80REFERENCES:Ofi tserova, M., Nerkar, S., Pickering, M., Torma, L., Thiex, N.,Multiresidue Mycotoxin Analysis in Corn Grain by ColumnHigh-performance Liquid Chromatography with Post-columnPhotochemical and Chemical Derivatization: Single-LaboratoryValidation., (2009), J AOAC Int., 92, 15-25Post-column ConditionsPost-column System: Pinnacle PCXReactor Volume: 1.4 mLTemperature: 60 CReagent: OPA, Thiouor, Brij 35® in GA104Photochemical Reactor: UVEDetection:FluorescenceAatoxins (photochemical derivatization) ex= 365 nm; em= 455 nmFumonisins (post-column derivatization with OPA) ex= 330 nm; em= 465 nmOchratoxin A ex= 335 nm; em= 455 nmZearalenone ex= 275 nm; em= 455 nmUV/VisDeoxynivalenol=218 nm