Simultaneous determination of aflatoxins and ochratoxin A in single and mixed spices W.M. Wan Ainiza a, c , S. Jinap a, b, * , M. Sanny a a Food Safety Research Centre (FOSREC), Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia b Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia c National Public Health Laboratory, Ministry of Health, Malaysia article info Article history: Received 27 February 2014 Received in revised form 21 October 2014 Accepted 27 October 2014 Available online 3 November 2014 Keywords: Simultaneous determination Aflatoxins Ochratoxin A Spices abstract Methods for the extraction and high performance liquid chromatography (HPLC) quantification of afla- toxins (AFs) and ochratoxin A (OTA), in a single procedure, from selected individual spices, i.e., coriander, chili, cumin, fennel and turmeric, were developed. The optimized method was then applied to spice mixtures, i.e., curry, kurma and soup powders. The aflatoxins B 1 ,B 2 ,G 1 and G 2 (AFB 1 , AFB 2 , AFG 2 and AFG 2 ) and ochratoxin A were extracted using a multi-mycotoxin immunoaffinity column, and the determination was conducted using reverse-phase HPLC and fluorescence detector (FLD) with a photochemical derivatisation system. The samples, for recovery study, were spiked with 3 different concentrations of AFB 1 and AFG 1 (0.4, 4.0 and 40 mg/kg), AFB 2 and AFG 2 (0.2, 2.0 and 20 mg/kg), and OTA (0.5, 5.0 and 50 mg/kg). Validation was conducted to confirm that the method was suitable for multi-toxin and multi-matrix analysis. The recoveries of AFB 1, AFB 2, AFG 1 and AFG 2 ranged from 72% to 100% for coriander, chili, cumin and all of the spice mixtures, and lower recoveries, ranging from 61% to 80%, were attained for fennel and turmeric. The recovery of OTA ranged from 96% to 113% for coriander and from 73% to 100% for chili, cumin, fennel, turmeric and all of the spice mixtures. The repeatability (RSDr) of the determination of AFB 1, AFB 2, AFG 1, AFG 2 and OTA contained in all of the spices ranged from 2% to 19%, whereas the LOD was 0.1 mg/kg for AFB 1 /AFG 1 , 0.05 mg/kg for AFB 2 /AFG 2 and 0.1 mg/kg for OTA, and the LOQ was 0.4 mg/kg for AFB 1 /AFG 1 , 0.2 mg/kg for AFB 2 /AFG 2 and 0.5 mg/kg for OTA. The results indicated that the method for the simultaneous determination of AFs and OTA can be applied to both single and mixed spices and is suitable for routine laboratory analysis. © 2014 Elsevier Ltd. All rights reserved. 1. Introduction Aflatoxins (AFs: AFB 1 , AFB 2 , AFG 1 and AFG 2 ) are secondary me- tabolites produced by molds, particularly Aspergillus flavus and Aspergillus parasiticus, which were found as contaminant in a wide variety of food matrices (Aydin, Emin Erkan, Baskaya, & Ciftcioglu, 2007; Marasas et al., 2008). The consumption of food with a high level of contamination can cause acute and chronic adverse effects on health which include immunosuppression hepatic disorder and cancer (Abdulkadar, Al-Ali, Al-Kildi, & Al-Jedah, 2004). Aflatoxicosis is a disease caused by AFs; acute toxicity will result in death, whereas chronic aflatoxicosis will result in cancer, immunosup- pression and other pathological conditions (Zain, 2011). AFB 1 is the most potent carcinogen compared with the other AFs, and it has been classified as a Group 1 carcinogen (IARC, 1993; Steyn, 1995). Ochratoxin A (OTA) is a secondary metabolite that is produced by some species of Aspergillus and Penicillium. Commodities from tropical regions are mainly contaminated with Aspergillus ochraceus and Aspergillus carbonarius, whereas those from temperate regions are commonly infected with Penicillium verrucosum (Richard, 2007; Steyn, 1995; Turner, Subrahmanyam, & Piletsky, 2009). OTA is commonly found in cereals, coffee, spices and dried fruits (IARC, 1993; Turner et al., 2009) and is classified as a potential Group 2B carcinogen by the (IARC, 1993). Occurrence of mycotoxins in food has been an increasing concern especially on the safety of spices and herbs; for that reason it has prompted many researchers to conduct more surveys of AFs and OTA contamination in spices. The hot and humid conditions in the fields, drying processes conducted in the open air or on the ground and insufficient training may lead to mycotoxins contamination of spices (Ardic, Karakaya, Atasever, & Durmaz, 2008; Aydin et al., 2007; Bircan, 2005). The occurrence of * Corresponding author. Food Safety Research Centre (FOSREC), Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia. Tel.: þ60 389468393; fax: þ60 389423552. E-mail addresses: sjinap@gmail.com, jinap@upm.edu.my (S. Jinap). Contents lists available at ScienceDirect Food Control journal homepage: www.elsevier.com/locate/foodcont http://dx.doi.org/10.1016/j.foodcont.2014.10.051 0956-7135/© 2014 Elsevier Ltd. All rights reserved. Food Control 50 (2015) 913e918