Journal of Chromatography A, 864 (1999) 89–101 www.elsevier.com / locate / chroma Determination of ochratoxin A in wine by means of immunoaffinity column clean-up and high-performance liquid chromatography * Angelo Visconti , Michelangelo Pascale, Gianluca Centonze Istituto Tossine e Micotossine da Parassiti Vegetali, CNR, Viale L. Einaudi 51, 70125 Bari, Italy Received 1 July 1999; received in revised form 15 September 1999; accepted 15 September 1999 Abstract A new and accurate method to quantify ochratoxin A (OA) in table wine has been developed. The method uses commercial immunoaffinity columns for clean-up and high-performance liquid chromatography (HPLC) with fluorescence detection for quantification of the toxin. Wine was diluted with a solution containing 1% polyethylene glycol (PEG 8000) and 5% sodium hydrogencarbonate, filtered and applied to an OchraTest immunoaffinity column. The column was washed with a solution containing sodium chloride (2.5%) and sodium hydrogencarbonate (0.5%) followed by water. OA was eluted with methanol and quantified by reversed-phase HPLC with fluorometric detection (excitation wavelength 333 nm, emission wavelength 460 nm) using acetonitrile–water–acetic acid (99:99:2) as mobile phase. Average recoveries of OA from white, ´ rose and red wine samples spiked at levels from 0.04 to 10 ng / ml ranged from 88% to 103%, with relative standard deviations (RSDs) between 0.2 and 9.7%. Detection limit was 0.01 ng / ml based on a signal-to-noise ratio of 3:1. The ´ method was applied successfully to 56 samples of red (38), rose (8), white (9) and dessert (1) wine. The levels of OA ´ ranged from ,0.01 to 7.6 ng / ml with red wines more contaminated than rose and white wines. A good correlation ( r50.987) was found by comparative analysis of 20 naturally contaminated samples using this method and the method of Zimmerli and Dick with better recoveries of OA and better performances for the new method. Several advantages of this method with respect to the actually available methods have been pointed out, with particular reference to red wine which appears to be the most difficult to analyze.  1999 Elsevier Science B.V. All rights reserved. Keywords: Wine; Food analysis; Ochratoxin A; Mycotoxins 1. Introduction environmental conditions. OA occurs in various plant products such as cereals (mainly wheat, barley, Ochratoxin A (OA), 7-( L-b-phenylalanyl- maize and oats), beans, groundnuts, spices, dried carbonyl) - carboxyl-5-chloro-8-hydroxy-3,4-dihydro- fruits, coffee, milk, beer and wine, as well as in pig 3R-methylisocumarin, is a widely distributed blood and kidney [1–6]. OA has been shown to be mycotoxin produced mainly by Aspergillus och- nephrotoxic, hepatotoxic, teratogenic and immuno- raceus and Penicillium verrucosum under diverse toxic to several animal species and to cause kidney and liver tumors in mice and rats [1,7]. The IARC (International Agency for Research on Cancer) has *Corresponding author. Tel.: 139-80-5486-013; fax: 139-80- classified OA as a possible carcinogen to humans 5486-063. E-mail address: visconti@area.ba.cnr.it (A. Visconti) (Group 2B) [1]. OA is suspected to be involved in 0021-9673 / 99 / $ – see front matter  1999 Elsevier Science B.V. All rights reserved. PII: S0021-9673(99)00996-6