Selective Extraction and Cleanup of Patulin Mycotoxin from Apple Juice Samples

Introduction

Patulin (Figure 1) is a mycotoxin produced by a number of fungal species. The mold Penicillium Expansum, in particular, is known as the main source of patulin contamination.¹ Patulin occurrence is found most often in rotting apples and products made from spoiled apples. Included among these products is apple juice and apple puree.²

Figure 1. Structure of Patulin

Because studies have shown that patulin possesses immunotoxic and genotoxic properties, regulatory limits for patulin have been set in 50 countries, including USA and European Union. In Europe, the maximum limit recommended for patulin in fruit juices is 50 μg/kg. The maximum patulin level permitted by the US FDA in apple juice is 50 μg/L.2-4 Historically, analytical methods for patulin have employed liquidliquid
extraction (LLE) followed by HPLC separation with UV detection at 276 nm.1,5 Researchers have highlighted problems with these methodologies, including:

• Tedious sample preparation associated with liquid-liquid extraction
• Patulin instability in alkaline conditions resulting from sodium carbonate cleanup
• The requirement of extra cleanup or chromatographic method development to prevent the coelution of patulin and interfering matrix component 5-hydroxymethylfurfural (HMF).1,6

Therefore, a quick, simple and robust sample preparation method for patulin analysis is needed.

Molecularly Imprinted Polymers

SupelMIP^® SPE – Patulin, a sample cleanup product used in this application, is made using the technology of molecularly imprinted polymers (MIPs). During MIP synthesis, a polymerization takes place around a template molecule that is similar to the analyte of interest. This template guides the formation of specific cavities within the polymer that are sterically and chemically complimentary to the target analyte. The subsequent removal of the template results in a polymer that is selectively retentive for the specific analyte of interest. During sample preparation, the MIP phase extracts the target analyte with a high degree of selectivity. The use of vigorous washes during the SPE procedure results in superior cleanup, producing a highly pure, concentrated sample for analysis. In this application, the use of SupelMIP SPE – Patulin is demonstrated for the cleanup of patulin in apple juice. Please note that other apple-containing products and matrices can be analyzed using similar methodology.

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