Induction and Quantification of Patulin Production in Penicillium Species   

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Original research article

A brief version of this protocol appeared in:
Molecular Plant Microbe Interactions
Jun 2015


Patulin, a worldwide regulated mycotoxin, is primarily produced by Penicillium and Aspergillus species during fruit spoilage. Patulin contamination is a great concern with regard to human health because exposure of the mycotoxin can result in severe acute and chronic toxicity, including neurotoxic, mutagenic, and immunotoxic effects. Penicillium expansum is known as the main producer of patulin. This protocol addresses the cultivation procedure of P. expansum under patulin permissive conditions and describes the method of collection and detection of patulin.

Keywords: Penicillium expansum, Patulin induction, HPLC, Quantification


Patulin is a polyketide lactone mycotoxin and is produced by several species of fungi including Penicillium, Aspergillus and other species. Among them, Penicillium expansum, which is a well-known postharvest pathogen causing decay of pomaceous fruits during storage, is the main producer. Patulin levels in apple products are of great concern because of the severe acute and chronic effects caused by the toxin. Therefore the patulin level in food is limited in many countries around the world. The European Commission (2006) has set maximum permitted levels in apple juices (50 μg/kg), solid apple products (25 μg/kg) and, above all, fruit-derived baby foods (10 μg/kg), as children are major consumers of apple derived products.

Studies on patulin in recent years have focused on environmental factors regulating patulin production, molecular basis of patulin biosynthesis and biodegradation of patulin. The methods of induction and quantification of patulin production are important in these studies. Patulin analysis in fruits usually follows the AOAC method 995.10 (Brause et al., 1996). After treatment with pectinase, patulin is extracted with ethyl acetate from the puree of decayed portion of fruits. Many methods have been developed for measuring patulin such as TLC, mass spectrometry and gas chromatography/mass spectrometry. Now, high performance liquid chromatography with ultra violet light detection (HPLC-UV) is the most frequently used method (Baert et al., 2007).

In this protocol, we address two methods of patulin induction in vitro and describe the specific parameters appropriate for HPLC-UV analysis of patulin.

Copyright: © 2017 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Chen, Y., Li, B., Zhang, Z. and Tian, S. (2017). Induction and Quantification of Patulin Production in Penicillium Species. Bio-protocol 7(11): e2324. DOI: 10.21769/BioProtoc.2324.

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