Introduction Ergot alkaloids (EAs) are toxic substances naturally produced by Claviceps fungi. These fungi infest a wide range of cereals and grasses. When domestic animals are exposed to EAs through contaminated feeds, it is detrimental to them and leads to significant economic losses. For that reason, it is important to monitor feed for the presence of EAs, especially with methods enabling their determination in processed materials. Material and Methods Ergot alkaloids were extracted with acetonitrile, and dispersive solid phase extraction (d-SPE) was used for clean-up of the extracts. After evaporation, the extracts were reconstituted in ammonium carbonate and acetonitrile and subjected to instrumental analysis using high-performance liquid chromatography with fluorescence detection. The developed method was validated in terms of linearity, selectivity, repeatability, reproducibility, robustness, matrix effect, limits of quantification and detection and uncertainty. The EA content of 40 compound feeds was determined. Results All the assessed validation parameters fulfilled the requirements of Regulation (EU) 2021/808. At least one of the monitored alkaloids was determined in 40% of the samples. The EAs with the highest incidence rate were ergocryptine, ergometrinine and ergocornine. The total concentrations of EAs ranged from under the limit of quantification to 62.3 μg kg−1. Conclusion The results demonstrated that the developed method was suitable for simultaneously determining twelve EAs in compound feed and could be used for routine analysis.

中文翻译：

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使用高效液相色谱荧光检测 (HPLC-FLD) 方法的开发、内部验证和应用，用于定量配合饲料中的 12 种麦角生物碱

简介 麦角生物碱 (EA) 是天然产生的有毒物质麦角头鱼菌类。这些真菌侵染多种谷物和草类。当家畜通过受污染的饲料接触EA时，对其有害并导致重大经济损失。因此，监测饲料中是否存在 EA 非常重要，特别是采用能够测定加工材料中 EA 的方法。材料与方法用乙腈提取麦角生物碱，并使用分散固相萃取（d-SPE）对提取物进行净化。蒸发后，将提取物在碳酸铵和乙腈中复溶，并使用带有荧光检测的高效液相色谱进行仪器分析。所开发的方法在线性、选择性、重复性、再现性、稳健性、基质效应、定量和检测极限以及不确定性方面得到了验证。测定了 40 种配合饲料的 EA 含量。结果 所有评估的验证参数均满足法规 (EU) 2021/808 的要求。40% 的样品中至少检测到一种受监测的生物碱。发病率最高的 EA 是麦角隐亭、麦角新宁和麦角康宁。EA 的总浓度范围为定量限以下至 62.3 μg kg−1。结论该方法适用于配合饲料中12种EA的同时测定，可用于常规分析。