Contents lists available at ScienceDirect Microbial Pathogenesis journal homepage: www.elsevier.com/locate/micpath Determination of mycotoxins by HPLC, LC-ESI-MS/MS, and MALDI-TOF MS in Fusarium species-infected sugarcane John Martin Jerome Jeyakumar a , Muqing Zhang a , Muthu Thiruvengadam b,* a State Key Lab for Conservation and Utilization of Subtropical Agro-Biological Resources, Guangxi University, Nanning, Guangxi, 530005, China b Department of Applied Bioscience, College of Life and Environmental Sciences, Konkuk University, Seoul, 05029, South Korea ARTICLE INFO Keywords: Fusarium High-performance liquid chromatography Liquid chromatography-electrospray ionization-tandem mass spectrometry Matrix-assisted laser/desorption ionization time of flight mass spectrometry Mycotoxins Sugarcane ABSTRACT Mycotoxins are secondary metabolites of fungi that are damaging to both animals and humans. Extensive contamination of foods and feeds with mycotoxins is an important problem. Fumonisins, trichothecenes, zear- alenone, and aflatoxins are mycotoxins produced by Fusarium species and occur naturally in sugarcane and cereal-based foods, threatening health and food security worldwide. Their distribution in the contaminated material is of great interest for obtaining insight into infection mechanisms and the potential for reducing contamination during food processing. In this study, mycotoxins were evaluated by high-performance liquid chromatography, liquid chromatography-electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS), and matrix-assisted laser/desorption ionization time of flight mass spectrometry (MALDI-TOF MS) of Fusarium species-infected sugarcane materials. A simple, sensitive, and reliable analytical method was developed for ra- pidly detecting eight mycotoxins in Fusarium species: fumonisin B1 and fumonisin B2, B-trichothecene mix (deoxynivalenol, nivalenol, 3-acetyl- deoxynivalenol, 15-acetyl-deoxynivalenol), zearalenone, and aflatoxin G1. Analyses were carried out in multiple reaction monitoring mode using the two primary product ions. The results generated by LC/MS and MALDI-TOF MS/MS revealed various mechanisms regulating mycotoxins production, which may help to clarify the roles of sensitive and selective compounds. The results demonstrate that this procedure is suitable for simultaneous determination of mycotoxins in sugarcane and can be performed in routine analysis in mycotoxin laboratories. 1. Introduction Sugarcane (Saccharum officinarum) is the most important crop worldwide because of its strategic position and extensive uses in the daily life in all countries as well as its industrial uses for producing nutritional and economical foods. Sugar present in the stem of S. offi- cinarum is the primary location of fungal growth. Pokkah boeng is one of the most severe and devastating diseases occurring in sugarcane, causing great damage and yield losses worldwide; this disease is caused by the Fusarium species complex, which includes Fusarium verticillioides, Fusarium proliferatum, and Fusarium sacchari [14]. Fungi produce a large number of metabolites that are not essential for life but may provide the fungus with an ecological advantage in specific environments. Fungal secondary metabolites include plant growth regulators (gibberellins), pharmaceutically useful compounds (penicillin, lovastatin), pigments (carotenoids), and mycotoxins (fumonisins, trichothecenes, zear- alenone, aflatoxins, ochratoxins) [17]. Mycotoxins may accumulate in infected crop plants, and upon ingestion, lead to the development of diseases (mycotoxicoses) in humans and animals [35]. Because of the widespread distribution of fungi in the environment, mycotoxins are considered among the most critical contaminants in foods and feeds. According to the FAO, 25% of the world's crops are contaminated with toxic fungal metabolites [28]. The most significant mycotoxins in terms of food and feed safety are fumonisins (Fs), trichothecenes, zearalenone (ZEA), aflatoxins (AFs), ochratoxin A (OTA), and ergot alkaloids, which have carcinogenic, mutagenic, teratogenic, cytotoxic, neurotoxic, ne- phrotoxic, estrogenic, dermotoxic, and immunotoxic effects [1]. To date, approximately 400 compounds have been identified as myco- toxins including AFs, OTA, Fs, nivalenol (NIV), deoxynivalenol (DON), ZEA, T-2, and HT-2 toxins [4]. https://doi.org/10.1016/j.micpath.2018.06.045 Received 24 April 2018; Received in revised form 16 June 2018; Accepted 28 June 2018 * Corresponding author. E-mail address: muthu@konkuk.ac.kr (M. Thiruvengadam). Abbreviations: 3- AcDON, 3-Acetyl-deoxynivalenol; 15- AcDON, 15-Acetyldeoxynivalenol; AFG1, Aflatoxins G1; CH 3 CN, Acetonitrile; DON, Deoxynivalenol; Fs, Fumonisins; Fv, Fusarium verticillioides; Fp, Fusarium proliferatum; Fo, Fusarium oxysporum; FB1, Fumonisin B1; FB2, Fumonisin B2; HPLC, High-performance liquid chromatography; LC–ESI-MS/MS, Liquid chromatography-electrospray ionization-tandem mass spectrometry; MRM, Multiple reaction monitoring; NIV, Nivalenol; OTA, Ochratoxin A; OPA, Ortho-phthaldialdehyde; TCT, Trichothecenes; TFA, Trifluoroacetic acid; MALDI-TOFMS/MS, Matrix-assisted laser/desorption ionization time of flight MS; ZEA, Zearalenone Microbial Pathogenesis 123 (2018) 98–110 Available online 30 June 2018 0882-4010/ © 2018 Elsevier Ltd. All rights reserved. T