Internal Validation of Rapid and Performance Method for Patulin Determination in Apple Cider by High-Performance Liquid Chromatography

Abstract

http://www.revistadechimie.ro 3921 Mycotoxins are toxic secondary metabolites produced by fungi that pose a potential hazard for food safety. Patulin is a mycotoxin produced by more than 60 species of fungi belonging to greater than 30 genera [1, 2]. Patulin is produced mainly by Penicillium expansum, but and other fungi, including P. claviforme, P. urticae, P. patulum, Aspergillus clavatus, A. giganteus, Byssoclamys fulva, B. nivea, and Alternaria alternata [3]. Frequently found in the rotten tissues of apples, patulin accumulation correlates with apparent symptoms of blue mold. Apple blue mold usually starts with the invasion of P. expansum spores on the wounds of fresh apples. Such stem punctures, insect injuries, and bruises are created during the picking and handling operations in the apple orchard, until the final processing steps of products [4]. Apple juice and other derived fruit products achieved from apples infected by Penicillium expansum, are the main source of patulin. Tannous et al. [5] developed and validated a mathematical model for predicting the P. expansum growth under natural conditions on apples, with an acceptable precision. This model could be used by apple growers and fruit juice producers to anticipate growth of P. expansum during storage and apple processing [5]. Long-term consumption of patulin-contaminated fruit juice can cause serious health problems because this mycotoxin is thought to have mutagenic, neurotoxic, genotoxic, and immunotoxic effects to animals [6, 7]. Initially identified as an antimicrobial agent, patulin was reclassified as mycotoxin in the 1960s because of its acute toxicity into human body [7]. Joint FAO/WHO Expert Committee on Food Additives established in 1995 a provisional maximum tolerable daily intake of patulin at 0.4 μg/kg body weight/day [8]. The LD50 of patulin ranges from 15 to 5 mg/kg, being dependent on the mode of exposure to this contaminant and the route of ingestion [9]. The amount of patulin in apple products is a quality indicator of them concerning food safety. In case of apple juice and apple cider the maximum level of patulin, established by the European Commission [10] is 50 μg/L. Worldwide, the incidence of patulin contamination is a problem, especially in case of producers of apples and apple-based products, such as China, the EU, and the USA [11]. In a study, Harris et al. [12] have evaluated the concentrations of patulin in apple cider and apple juice produced and marketed by Michigan apple cider mills during 2002-2006 (n=493 samples), using highperformance chromatography. Patulin was detected (> or =4 μg/L) in 18.7% of all cider mill samples, and 11 samples (2.2%) having patulin concentrations of > or =50 μg/L. In case of cider samples obtained from mills using thermal pasteurization, patulin was detected in 28.4% of samples, and in the case of those taken from mills using UV light radiation, in 13.5% of samples. Among retail grocery store samples (n=159), in 23% of apple juice and cider samples, patulin was detected, and in case of 18 samples (11.3%) concentration of patulin was > or =50 μg/L. This paper presents the internal validation of a rapid and performance method for patulin determination in apple cider by high-performance liquid chromatography (HPLCDAD). Moreover, assessing the patulin content of apple cider samples (purchased from commerce), was performed.