1. Determination of ochratoxin A in wine by means of immunoaffinity column clean-up and high-performance liquid chromatography Ochratoxin A (OTA) Toxicity  OTA is hepatotoxic, nephrotoxic, teratogenic and carcinogenic to animals  OTA was classified by the International Agency for Research on Cancer (IARC) as a possible human carcinogen (group 2B) Regulatory limits set by the European Union COMMISSION REGULATION (EC) No 123/2005  2,0 ppb for wine (red, rose and white)

2.  The clean-up procedure for ochratoxin A involves generally IAC, SPE  Immunoaffinity chromatography (IAC) clean-up is based on the binding interaction of ochratoxin A-specific antibodies and has been set as European Standard (SR EN 14133) for ochratoxin A analysis in wine and beer Method Immunoaffinity column design containing solid extraction phase made from antibody-modified support beads binding ochratoxin A  Chromatographic determination  Sample preparation and clean-up

3. Sample preparation and immunoaffinity clean-up

4. Chromatogram of OTA contaminated red wine Chromatographic conditions:  OTA was quantified by reverse phase high–performance liquid chromatography (HPLC) with fluorescence detection (excitation wavelength 330nm, emission wavelength 450nm).  mobile phase consisted of a mixture of acetonitrile–water–acetic acid (99:99:2)  analytical column were reverse phase, C-18 with 5 μm particles  flow-rate 1 mL/min  Injection 20 yL HPLC determination of ochratoxin A