An HPLC-MS/MS Method Using a Multitoxin Clean up Column for Analysis of Seven Mycotoxins in Aquafeeds

Abstract

Background In Guangdong Province of China, the climate here is very wet, so there are many different fungus living in the aquatic feeds, which produce mycotoxins. These compounds contaminate agriculture products world-wide and represent a great threat to human health. It is necessary to determine their contamination level in aquatic feeds. Objective A high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method was developed for the quantitative analysis of aflatoxin B1, aflatoxin M1, T-2 toxin, HT-2 toxin, deoxynivalenol, ochratoxin, and zearalenone in the fish and shrimp feed. Methods Samples were extracted with acetonitrile-water (V: V = 3:1), and degreased with acetonitrile-saturated hexane. Such obtained extract was cleaned up with a multitoxin column. The target compounds were separated on a C18 chromatographic column and analyzed simultaneously by electrospray ionization mass spectrometry in both positive or negative ion mode. Detected compounds were quantified by using the matrix-matched external standard method. Results Under the optimized conditions, good linearities for the analytes in corresponding concentration range were obtained with correlation coefficients (r2) higher than 0.9948. LOD ranged from 1.83 to 12.63 μg/kg, and LOQ ranged from 5.49 to 37.89 μg/kg. Average recoveries for the target mycotoxins at three spiked levels ranged from 80.5% to 116.5% with RSD ranging from 2.4% to 10.4%. 23 real aquafeed samples were determined by this method, and 7 kinds of toxins were all detected. Conclusions Obtained results showed that developed method could be successfully applied for the simultaneous determination of mycotoxins in aquatic feeds.

Recent Advances in Mycotoxin Determination in Fish Feed Ingredients

“…A limited number of multi-mycotoxin techniques, particularly for finished fish feeds and shrimp feeds, has been reported. Aquatic feeds are complex matrices consisting of minerals, vitamins, fatty acids, and proteins in high concentrations that are challenging to remove [ 88 ]. As a result, choosing an appropriate clean-up step that reduces matrix effects (MEs) and interferences during chromatographic analysis is essential [ 89 ].…”

“…The samples were extracted with a mixture of acetonitrile and water followed by a defatted step by hexane and a clean-up step with a multi-toxin purification column. LODs ranged between 1.83 and 12.63 μg/kg and the method was successfully applied in several fish feeds in China [ 88 ] The LODs of the method ranged between 0.15 and 61 μg/kg [ 59 ]. Another method for the simultaneous determination of ZEN and DON in fish feed was reported based on an HPLC-DAD system and an SLE extraction step followed by clean-up with an IAC column [ 100 ].…”

Risks for animal health related to the presence of ochratoxin A (OTA) in feed

“…In the last few years much of the literature has focused on multi‐mycotoxin methods especially using QuEChERS and LC–MS/MS in different feed materials (e.g. Nakhjavan et al., 2020 ; Nualkaw et al., 2020 ; Jo et al., 2021 ; Gonzales‐Jartin et al., 2021 ; Konak et al., 2021 ; Seo et al., 2021 ; Bi et al., 2022 ; Mackay et al., 2022 ; Nochetto and Li, 2022 ). However, there has also been focus on other and often faster methods including those with biosensors that often involve specific antibodies, aptamers or molecular imprinting polymers in the analytical work (e.g.…”

Essential oils: An eco‐friendly alternative for controlling toxigenic fungi in cereal grains

“…Around 400 types of mycotoxins have been identified simultaneously in food, resulting in synergistic effects on health (Bi et al., 2021). Aflatoxins, ochratoxin A (OTA), zearalenone, deoxynivalenol, patulin, citrinin, fumonisins, and T‐2 and HT‐2 toxins represent major concerns for the health of vertebrates (Rahmani et al., 2009; Zinedine et al., 2007).…”