Álvarez Micaela, Rodríguez Alicia, Bermúdez Elena, Roncero Elia, Andrade María J
Food Hygiene and Safety, Meat and Meat Products Research Institute, Faculty of Veterinary Science, University of Extremadura, Avda. de las Ciencias, s/n. 10003 Cáceres, Spain.
Food Quality and Microbiology, University Institute for the Research in Agrifood Resources, School of Agricultural Engineering, University of Extremadura, Avda. Adolfo Suárez, s/n. 06007 Badajoz, Spain.
Foods. 2021 Feb 17;10(2):438. doi: 10.3390/foods10020438.
Antifungal agents are commonly used in the meat industry to prevent the growth of unwanted moulds, such as toxigenic ones, on dry-cured meat products. For enhancing the application of antifungals, their mode of action must be evaluated. Their effect on the mould ergosterol content is one of the most studied ones, since it is the target site of some commercialised antifungals or of those that are in development. The aim of this study was to develop a methodology for determining how the antifungal agents used in the meat industry work. A method for analysing ergosterol was firstly developed using high-performance liquid chromatography with fluorescence detection coupled to a diode array detector (HPLC-FLD/DAD). The chromatographically optimised conditions (gradient and mobile phases) allowed us to reduce the time per analysis with respect to previously published methods up to 22 min. Withing the six checked extraction methods, method 5, showing the best mean recovery values (99.51%), the shortest retention time (15.8 min), and the lowest standard deviation values (9.92) and working temperature (60 °C), was selected. The limit of detection and limit of quantification were 0.03 and 0.1 µg/mL, respectively. All the validation parameters corroborated the method's suitability. Finally, its feasibility for evaluating the effect of a commercial antifungal preparation (AP) and different herbs that are frequently added to meat products on the ergosterol content of several toxigenic moulds was studied. Differences at the strain level were obtained in the presence of AP. Moreover, the addition of herbs significantly reduced the ergosterol content in up to 83.91%. The developed methodology is thus suitable for screening the antifungals' role in altering mould ergosterol biosynthesis before their application in real meat products.
抗真菌剂在肉类工业中常用于防止干腌肉制品上不需要的霉菌生长，如产毒霉菌。为了加强抗真菌剂的应用，必须评估其作用方式。它们对霉菌麦角固醇含量的影响是研究最多的方面之一，因为它是一些商业化抗真菌剂或正在研发的抗真菌剂的作用靶点。本研究的目的是开发一种方法，以确定肉类工业中使用的抗真菌剂的作用方式。首先开发了一种使用高效液相色谱结合荧光检测和二极管阵列检测器（HPLC-FLD/DAD）分析麦角固醇的方法。色谱优化条件（梯度和流动相）使我们能够将每次分析的时间相对于先前发表的方法减少至22分钟。在六种检查的提取方法中，选择了方法5，它显示出最佳的平均回收率（99.51%）、最短的保留时间（15.8分钟）、最低的标准偏差值（9.92）和工作温度（60℃）。检测限和定量限分别为0.03和0.1μg/mL。所有验证参数均证实了该方法的适用性。最后，研究了其在评估市售抗真菌制剂（AP）和肉类产品中常用的不同草药对几种产毒霉菌麦角固醇含量影响方面的可行性。在存在AP的情况下，在菌株水平上获得了差异。此外，添加草药可使麦角固醇含量显著降低达83.91%。因此，所开发的方法适用于在抗真菌剂应用于实际肉类产品之前，筛选其在改变霉菌麦角固醇生物合成中的作用。
