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Validation data for the determination of ochratoxin A (OTA) in two spice matrices, red paprika and black pepper, were obtained for samples prepared with a simplified single-step clean-up column. Extracts of finely ground samples of red paprika and black pepper were prepared and applied to a Mycosep® 229 Ochra clean-up column. The purified extract was then subjected to HPLC/FLD analysis. The relative standard deviation for repeatability (RSDr) of the method was 11.8% for red paprika and 9.9% for black pepper. The limit of detection (LOD) value (three times the noise) was estimated as corresponding to the response of an extract derived from a blank matrix (previously washed) and spiked at 1.0 µg kg−1. The limit of quantitation (LOQ) (three times LOD) was 3.0 µg kg−1. The performance of the one-step column clean-up procedure appears to be a suitable alternative to commonly used clean-up techniques and allows the precise determination of OTA in two complex matrices. [ABSTRACT FROM AUTHOR]