A New Competitive Fluorescence Assay for the Detection of Patulin Toxin

Abstract

Patulin is a toxic secondary metabolite of a number of fungal species belonging to the genera Penicillum and Aspergillus. It has been mainly isolated from apples and apple products contaminated with the common storage-rot fungus of apples, Penicillum expansum, but it has also been extracted from rotten fruits, moldy feeds, and stored cheese. Human exposure to patulin can lead to serious health problems, and according to a long-term investigation in rats, the World Health Organization has set a tolerable weekly intake of 7 ppb body weight. The content of patulin in foods has been restricted to 50 ppb in many countries. Conventional analytical detection methods involve chromatographic analyses, such as HPLC, GC, and, more recently, techniques such as LC/MS and GC/MS. However, extensive protocols of sample cleanup are required prior to the analysis, and to accomplish it, expensive analytical instrumentation is necessary. An immunochemical analytical method, based on highly specific antigen-antibody interactions, would be desirable, offering several advantages compared to conventional techniques, i.e., low cost per sample, high selectivity, high sensitivity, and high throughput. In this paper, the synthesis of two new derivatives of patulin is described, along with their conjugation to the bovine serum albumin for the production of polyclonal antibodies. Finally, a fluorescence competitive immunoassay was developed for the on-line detection of patulin.

A near-infrared fluorescence assay method to detect patulin in food

“…Thus, we adopted the follow strategy: a PAT derivative, whose synthesis is described in the literature [19], was covalently attached to a protein carrier (BSA).…”

“…These characteristics represent a large advance in terms of cost and time execution of analyses with respect to the described analytical methods [19].…”

“…A high stable patulin derivate lacking the highly reactive C7-C7a double bond, PAT-Sat, was synthesized from L-arabinose and conjugated to BSA as described by De Champdorè and coworkers [19]. In brief, a solution of PAT-Sat (1.…”

“…Both assays were based on the use of specific antibodies against novel chemically synthesized PAT derivative compounds [19,20].…”

A novel fluorescence polarization assay for determination of penicillin G in milk

“…The affinity column was prepared according to (De Champdoré et al, 2007). Briefly, the resin was washed sequentially with H 2 O at pH 4.5 (20 mL), NaCl (0.5 mol/L, 20 mL), and again with H 2 O at pH 4.5 (20 mL) and finally suspended in 2.0 mL of H 2 O.…”