Overpressured layer chromatographic determination of aflatoxin B1, B2, G1 and G2 in red paprika Ágnes M. Móricz a, ⁎ , Zsuzsanna Fatér b , Klára H. Otta a , Ernő Tyihák c , Emil Mincsovics d a Department of Chemical Technology and Environmental Chemistry, L. Eötvös University, P.O.B. 32, Budapest 112, H-1518, Hungary b Budapest Institute of the National Public Health and Medical Officer Service, Budapest, Váci út 174, H-1138, Hungary c Plant Protection Institute, Hungarian Academy of Sciences, Budapest, P.O.B. 102, H-1525, Hungary d OPLC-NIT Ltd., Budapest, Andor u. 60, H-1119, Hungary Received 10 January 2006; received in revised form 17 March 2006; accepted 17 March 2006 Abstract Determination of aflatoxin B1 and total aflatoxin (B1 + B2 + G1 + G2) in red paprika powder is described using column chromatographic sample clean-up, overpressured layer chromatography (OPLC) separation and fluorescence densitometric evaluation. Two OPLC methods were developed for separation of the four aflatoxins. The detection limit and quantification limit of aflatoxins in red paprika were 0.5 and 1 μg/kg in both methods, respectively. Recovery experiment was carried out with sample containing 1.74 μg/kg aflatoxin B1 and 3.56 μg/kg total aflatoxins measured by European standard HPLC method. Mean recovery amounted to 78.5% (SD 16.1%, n = 5) for aflatoxin B1 and 81.8% (SD 17.1%, n = 5) for total aflatoxins in the case of method 1. It was 105.3% (SD 10.7%, n = 5) for aflatoxin B1 and 97.4% (SD 18.6%, n = 5) for total aflatoxins using the method 2. Despite of that the Hungarian climate is not proper for the toxin production of moulds high aflatoxin B1 contaminated red paprika purchased from the market was found, which may originate from mixing of imported paprika containing very high level toxin with Hungarian one. © 2006 Elsevier B.V. All rights reserved. Keywords: Aflatoxin; Overpressured layer chromatography (OPLC); Red paprika 1. Introduction More than 300 mycotoxins are known, which are toxic metabolites of various fungi growing in a wide range of food and animal feedstuffs. Among of them, aflatoxins, produced by some Aspergillus moulds such as Aspergillus flavus and Aspergillus parasiticus, represent the main threat world wide owing to their occurrence and toxicity. The aflatoxin B1, B2, G1 and G2 (Fig. 1) may be contamination in the e.g. cereals, oilseeds, coffee and spices like red paprika. They are carcinogenic, mutagenic and teratogenic. Their effects range from acute death (aflatoxicosis) to chronic disease such as tumours [1]. B1, the most potent of them, causes human hepatocellular carcinoma [2]; therefore, it is classified as human carcinogen by WHO-IARC (WHO-International Agency for Research on Cancer) [3]. From these facts it emerges that the aflatoxins are potential risk for human health, therefore, the European Commission set maximum levels for their presence in foodstuffs (Regulation (EC) No. 472/2002). In red paprika, intended for human consumption, the permitted maximum level of aflatoxin B1 is 5 μg/kg and of total aflatoxin (aflatoxin B1, B2, G1 and G2) is 10 μg/kg. Aflatoxins were discovered in 1969 in connection with a disease of turkeys fed with contaminated peanuts. From this time, several methods have been developed for determining them [4], which are mainly based on high-performance liquid chromatography (HPLC) [5–7], thin-layer chromatography (TLC) [8,9] or enzyme-linked immunosorbent assay (ELISA) [10,11]. Overpressured layer chromatography (OPLC) [12] integrates the benefits of HPLC and TLC. The linear OPLC is a forced flow technique, using external pressure on chromatoplate sealed on Microchemical Journal xx (2006) xxx – xxx + MODEL MICROC-00729; No of Pages 5 www.elsevier.com/locate/microc ⁎ Corresponding author. E-mail address: moricz_am@yahoo.com (Á.M. Móricz). 0026-265X/$ - see front matter © 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.microc.2006.03.007 ARTICLE IN PRESS