Simultaneous detection of 12 mycotoxins in cereals using RP-HPLC-PDA-FLD with PHRED and a post-column derivatization system

Abstract

A new method for the simultaneous quantification of 12 mycotoxins was developed and optimized using reverse phase high performance liquid chromatography (RP-HPLC) with a photodiode array (PDA) and fluorescence detector (FLD), a photochemical reactor for enhanced detection (PHRED) and post-column derivatization. The mycotoxins included aflatoxins (AFB(1), AFB(2), AFG(1), and AFG(2)), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisins (FB(1), FB(2), and FB(3)), T-2 and HT-2 toxins. A double sample extraction with a phosphate-buffered saline solution (PBS) and methanol was used for co-extraction of mycotoxins, and a multifunctional immunoaffinity column was used for cleanup. Optimum conditions for separation of the mycotoxins were obtained to separate 12 mycotoxins in FLD and PDA chromatograms with a high resolution. The method gave recoveries in the range 72-111% when applied to spiked corn samples. The limits of detection (LOD) were 0.025 ng/g for AFB(1) and AFG(1), 0.012 ng/g for AFB(2) and AFG(2), 0.2 ng/g for OTA, 1.5 ng/g for ZEA, 6.2 ng/g for FB(1), FB(3) and HT-2 toxin, 9.4 ng/g for FB(2) and T-2 toxin, and 18.7 ng/g for DON. In addition, the limits of quantification (LOQ) ranged from 0.04 ng/g for AFB(2) and AFG(2) to 62 ng/g for DON. The method was successfully applied to the determination of these mycotoxins in 45 cereal samples obtained from the Malaysian market. The results indicated that the method can be applied for the multi-mycotoxin determination of cereals.

Advanced LC–MS-based methods to study the co-occurrence and metabolization of multiple mycotoxins in cereals and cereal-based food

“…Finally, after washing, analytes are eluted from the magnetic material by a proper solvent mixture. So far, only one study on the use of dispersive magnetic SPE in mycotoxin determination (AFs, ZEN, and OTA) in cereals has been published [96].…”

“…The manufacturer claims that highly accurate results are achieved by use of 13 Clabeled standards [66]. Dispersive magnetic SPE might be an attractive technique as an alternative to classic SPE in the future [96]. However, compared with a classic on-column SPE, dispersive magnetic SPE requires the same time for a single extraction.…”

Screening of aflatoxin M1, a metabolite of aflatoxin B1 in human urine samples in Malaysia: A preliminary study

“…The occurrence of aflatoxin in the foodstuff is widely reported in the literatures. In Malaysia, aflatoxin is detected in cereals and grains (Reddy & Salleh, 2010;Soleimany, Jinap, Rahmani, & Khatib, 2011), nuts (Leong, Ismail, Latif, & Ahmad, 2010) and spices (Reddy et al, 2011). In fact, a study has shown that some of the contaminated foods had level above the permissible level adopted by the Malaysia Food Act 1983 and Malaysia Food Regulation 1985 (Sulaiman, Yee, Hamid, & Yatim, 2007).…”

The relationship between ergosterol and mycotoxin contamination in maize from various countries

“…Due to the chemical diversity of mycotoxins and their varying concentrations between samples, simultaneous analysis of multiple mycotoxins is a great challenge for analytical chemists (Ibáñez-Vea et al 2011). A few reports have described simultaneous assessment of AFs, OTA and ZEA by HPLC-FLD after immunoaffinity column (IAC) purification in cereals (maize, wheat and rice) from Malaysia (Rahmani et al 2010;Soleimany et al 2011), breakfast cereals and barley from Spain (Ibáñez-Vea et al 2011, 2012 and breakfast cereals from Pakistan (Iqbal et al 2014).…”