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Optimization and Validation of a Method Without Alkaline Clean-Up for Patulin Analysis on Apple Puree Agar Medium (APAM) and Apple Products

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Abstract

A sensitive high-performance liquid chromatography-UV (HPLC-UV) method, based on the Association of Analytical Communities (AOAC) Official method 2000.02, was developed and validated for the high-throughput analysis of patulin in in vitro experiments on apple puree agar medium (APAM). The importance of repeating the ethyl acetate extraction step at liquid-liquid extraction (LLE) was examined, as well as the extent of patulin degradation during the sodium carbonate clean-up. In addition to this alkaline clean-up, the efficiency of using an Oasis HLB or C18 cartridge as solid-phase extraction (SPE) clean-up was compared. This resulted in a two-step ethyl acetate LLE, followed by an Oasis HLB SPE clean-up, without alkaline clean-up conditions. The method was fully validated for APAM, cloudy apple juice, and apple puree. Average patulin recoveries at levels of 100, 500, and 1000 μg kg−1 of APAM varied between 95 and 113 % over 3 independent days, with an interday precision (RSDR) of 5 to 10 %. Recovery experiments carried out with the spiked apple juice (at 50 μg kg−1) and apple puree (10 μg kg−1) showed average recovery rates laying between 80–101 % (RSDR = 12 %) and 77–100 % (RSDR = 9 %), respectively. This method offered a detection limit of 3–4 μg kg−1 and a quantification limit of 5–8 μg kg−1 for APAM, apple juice, and puree.

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Acknowledgments

This research was funded by the Belgian Federal Public Service (FPS) of Health, Food Chain Safety and Environment (RF6198 PATULINE). We are grateful to Martine Merchiers and Petra De Neve for their excellent technical assistance.

Conflict of Interest

Nikki De Clercq declares that she has no conflict of interest. Els Van Pamel declares that she has no conflict of interest. Els Van Coillie declares that she has no conflict of interest. Geertrui Vlaemynck declares that she has no conflict of interest. Frank Devlieghere declares that he has no conflict of interest. Bruno De Meulenaer declares that he has no conflict of interest. Els Daeseleir declares that she has no conflict of interest.

Ethical Statement

This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. Institute for Agricultural and Fisheries Research (ILVO), Member of Food2Know, Brusselsesteenweg 370, 9090, Melle, Belgium

    Nikki De Clercq, Els Van Pamel, Els Van Coillie, Geertrui Vlaemynck & Els Daeseleire

  2. Faculty of Bioscience Engineering, Department of Food Safety and Food Quality, Laboratory of Food Microbiology and Food Preservation, Member of Food2Know, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Frank Devlieghere

  3. Faculty of Bioscience Engineering, Department of Food Safety and Food Quality, Laboratory of Food Chemistry and Human Nutrition, Member of Food2Know, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Bruno De Meulenaer

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  1. Nikki De Clercq
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  2. Els Van Pamel
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  4. Geertrui Vlaemynck
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Correspondence to Nikki De Clercq.

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De Clercq, N., Van Pamel, E., Van Coillie, E. et al. Optimization and Validation of a Method Without Alkaline Clean-Up for Patulin Analysis on Apple Puree Agar Medium (APAM) and Apple Products. Food Anal. Methods 9, 370–377 (2016). https://doi.org/10.1007/s12161-015-0190-y

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