Contents lists available at ScienceDirect Food Chemistry journal homepage: www.elsevier.com/locate/foodchem Development, chemometric-assisted optimization and in-house validation of a modified pressurized hot water extraction methodology for multi- mycotoxins in maize Sefater Gbashi a, ⁎ , Patrick Berka Njobeh a, ⁎ , Sarah De Saeger b , Marthe De Boevre b , Ntakadzeni Edwin Madala c a Department of Biotechnology and Food Technology, Faculty of Science, University of Johannesburg, P.O Box 17011, Doornfontein Campus, 2028 Gauteng, South Africa b Centre of Excellence in Mycotoxicology and Public Health, Department of Bioanalysis, Ghent University, 9000 Ghent, Belgium c Department of Biochemistry, School of Mathematical and Natural Sciences, University of Venda, Thohoyandou, South Africa ARTICLE INFO Keywords: Mycotoxins Pressurized hot water extraction Chemometric-assisted optimization Central composite design ABSTRACT Effective management of mycotoxins rely on stringent regulation and routine surveillance of food/feed com- modities via efficient analysis, hence the continuous need for improved methods. The present study developed, optimized and validated a modified pressurized hot water extraction (PHWE) method for the simultaneous extraction of multi-mycotoxins from maize and subsequent quantification on LC-MS/MS. The PHWE system was modified using ethanol (EtOH) as a cosolvent, while a numerical modelling approach, the central composite design (CCD), was adopted for the optimization of the extraction conditions. Using the optimized method, it was possible to effectively extract and quantify 15 different mycotoxins from maize in a single step with satisfactory linearities (0.986–0.999), recoveries (14–124%) and other associated method validation parameters. Further efficacious application of the method to real samples re-affirmed the prospects of PHWE as a suitable, cost- effective and greener alternative to traditional methods of mycotoxin extraction. 1. Introduction Mycotoxins are poisonous secondary metabolites produced by cer- tain fungal species that contaminate various agricultural commodities (González-Jartín et al., 2019). Exposure to mycotoxin-contaminated foods results in various health-related problems, depending on the type of mycotoxin and level of exposure (Misihairabgwi, Ezekiel, Sulyok, Shephard, & Krska, 2017; Wielogorska et al., 2019). Relative to their prevalence and potency, they have been identified as the most sig- nificant nutritional hazard, superior to other natural inherent plant toxins, pesticide residues, food additives and other synthetic con- taminants in foods (Bennett & Klich, 2003). Besides the health-related problems, mycotoxins also exert significant socio-economic effects on societies globally, particularly in the developing nations (Gbashi, Madala, Adebo, Piater et al., 2017). Annual losses in agricultural commodities due to mycotoxin contamination have been estimated at one billion metric tons (Schmale & Munkvold, 2009). All these coupled with the globalization of the food supply chain, has led to more strin- gent regulatory limits and routine surveillance of the levels of these environmental pollutants in food and feed. As such, analysis plays a critical tool in the control and management of mycotoxins. Development of improved methods for mycotoxin analysis, parti- cularly extraction methods, has been an active research field (Arroyo- Manzanares, Huertas-Pérez, García-Campaña, & Gámiz-Gracia, 2014; Dong et al., 2019). The extraction and sample preparation part of an analytical process is often the most critical and difficult, both in terms of difficulty in extracting the desired analyte(s) from the matrix and the time involved, in addition to the fact that each sample matrix has its own unique challenges (Vaghela, Patel, Patel, Vyas, & Patel, 2016). In fact, it has been estimated that up to 70% and perhaps even more of the effort and time that goes into sample analysis comprises the extraction and sample preparation process (Rezaee et al., 2015). Many efforts have been geared towards developing suitable methods to quantitatively extract and detect mycotoxins in agricultural commodities. For any bioanalytical scientist, the goal is to develop methods with improved sensitivity and selectivity, while at the same time maintaining the credibility of the results, as well as reduce cost and time (Augusto, Hantao, Mogollo, & Braga, 2013). A number of studies have aimed at designing methods for myco- toxin analysis that avoid a clean-up step, however, such easy-to-use methods often demonstrate lack of sensitivity or are too expensive (Sulyok, Berthiller, Krska, & Schuhmacher, 2006; Rahmani, Jinap, & https://doi.org/10.1016/j.foodchem.2019.125526 Received 10 May 2019; Received in revised form 16 July 2019; Accepted 12 September 2019 ⁎ Corresponding authors. Food Chemistry 307 (2020) 125526 Available online 30 September 2019 0308-8146/ © 2019 Elsevier Ltd. All rights reserved. T