A DNA aptamer with high affinity and specificity to ochratoxin A (OTA) was conjugated to a coupling gel and used as sorbent for the preparation of solid phase extraction (SPE) columns. The SPE columns packed with 300 μl oligosorbent (24 nmol DNA) showed a linear (r = 0.999) behaviour in the range of 0.4-500 ng OTA. After optimisation of the extraction step, SPE columns were used for clean-up of OTA from wheat prior to liquid chromatographic (HPLC) analysis with fluorescence detection (FLD). Average recoveries from wheat samples spiked at levels of 0.5-50 ng/g ranged from 74% to 88% (relative standard deviation <6%) with limits of detection and of quantification of 23 and 77 pg/g, respectively. The comparative HPLC/FLD analyses of 33 naturally contaminated durum wheat samples cleaned-up on both aptamer-SPE and immunoaffinity (IMA) columns showed a good correlation (r = 0.990). Aptamer-SPE columns could be re-used up to five times without any loss of performance.

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Keywords DNA-aptamer, HPLC, Ochratoxin A, Oligosorbent, SPE columns, Wheat
Persistent URL dx.doi.org/10.1016/j.foodchem.2011.01.107
Journal Food Chemistry
Citation
De Girolamo, A. (Annalisa), McKeague, M. (Maureen), Miller, JD, DeRosa, M.C, & Visconti, A. (Angelo). (2011). Determination of ochratoxin A in wheat after clean-up through a DNA aptamer-based solid phase extraction column. Food Chemistry, 127(3), 1378–1384. doi:10.1016/j.foodchem.2011.01.107