**1. Introduction**

Mycotoxins are hazardous chemicals produced by *Aspergillus*, *Fusarium Penicillium,* and *Claviceps* genus. Mycotoxins can contaminate foods and feeds and agricultural products [1]. To date, there are more than four hundred mycotoxins with different toxicity, which have been identified in cereals, fruits, vegetables, and other agricultural commodities, resulting in potential adverse effects on human and animal health, and economic losses [2–4]. Moreover, mycotoxins are persistent in food and feeds and not completely eliminated during processing operations [3]. Recently, mycotoxins were a major category

Klangkaew, N.; Zhang, Z.; Giorgi, M.; Logrieco, A.F.; Poapolathep, A. Simultaneous Determination of Ergot Alkaloids in Swine and Dairy Feeds Using Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry. *Toxins* **2021**, *13*, 724. https://doi.org/10.3390/ toxins13100724

**Citation:** Poapolathep, S.;

Received: 13 September 2021 Accepted: 9 October 2021 Published: 13 October 2021

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**Copyright:** © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).

in border rejection in the European Union (EU) according to the annual report of the Rapid Alert System for Food and Feed (RASFF) [3]. The Food and Agriculture Organization (FAO) suggested one-fourth of global food crops is contaminated by mycotoxins [5]. Because of their pathogenicity and lethality, worldwide authorities including the World Health Organization (WHO) have called to monitor mycotoxins in foodstuff and feeds and set up strict maximum levels and legislation, in order to provide an early warning about mycotoxin contamination and reduce the national and international losses. In addition, the impact of climate change on *Calviceps* spp. infection of crops could result in a potential to increase the higher food safety risks for humans and animals due to mycotoxin contamination in the end products [6].

Ergot alkaloids (EAs) are toxic secondary metabolites produced by fungi of the *Claviceps* genus, mainly by the parasitic fungus *Claviceps purpurea,* which parasitize the seed heads of living plants at the time of flowering [7]. EAs are known to cause adverse health effects in humans and animals and have been found in cereals, cereal products, barley, oats, and both rye- and wheat-containing foods [8–11]. Outbreaks of ergotism in livestock do still occur, and EAs can induce abortion by its toxicity [12]. Pigs and cattle have shown symptoms after being infected with EAs, causing financial problems to both breeders and the meat industry [12,13]. Animals, including pigs exposed to EAs from grains, can cause liver and intestinal alterations [14]. In Directive 2002/32/EC on undesirable substances in animal feed and its amendment, the maximum content of rye ergot (*Claviceps purpurea*) in feed containing unground cereals has been established at 1000 mg/kg. EAs have been reported in cereals in European countries, Canada, the United States, and China [15–17]. There have also been some reports on the presence of EAs in feed from other countries, with 86–100% of EAs detected in feed samples from Germany [18] and 83% of compound feeds containing EAs with an average concentration of 89 μg/kg and a maximum concentration of 1231 μg/kg in the Netherlands [19]. The main ergot alkaloids produced by *Claviceps* species are ergometrine, ergotamine, ergosine, ergocristine, ergokryptine, and ergocornine, and the group of agroclavines [20]. Ergotamine and ergosine are heat stable whereas ergocristine, ergokryptine, ergocornine, and ergometrine are decreased by heating [21]. The conversion of ergopeptines to ergopeptinines was accelerated either by acidic or alkaline solutions. However, ergopeptinines can also be transformed to ergopeptines in organic solvents [7,22].

Studies have developed reliable analytical methods of EAs in agricultural commodities [12,15,17,19,22–26], mainly using HPLC-MS/MS. However, the challenge remains in the UHPLC-MS/MS method of optimizing the sample preparation procedure. However, signal suppression and enhancement usually occur due to the interferences in the matrix (matrix effect), leading to unreliable results [25]. To compensate for the matrix effect, some methods developed for the analysis of EAs in agricultural commodities have used a MycoSep® multifunctional column [2].

To the best of the authors' knowledge, to date, there have been a few reports on contaminations of EAs in any kinds of foodstuffs and feeds in Thailand. The current study investigated the occurrence of 11 EAs in swine and dairy feeds using a validated UHPLC-MS/MS with a multifunctional SPE column procedure. We used an SPE column for sample cleanup. Under optimization, the limit of detection, limit of quantification and linearity were studied. Accuracy and precision were evaluated as well. This work provides a promising manner to monitor EAs in feed samples.
