Abstract:
Ochratoxin A (OTA) is a mycotoxin commonly found in various food products, which poses potential health risks to humans and animals. Recently, more attention has been directed towards its potential neurodegenerative effects. However, there are currently no fully validated HPLC analytical methods established for its quantification in mice, the primary animal model in this field, that include pivotal tissues in this area of research, such as the intestine and brain. To address this gap, we developed and validated a highly sensitive, rapid, and simple method using HPLC-FLD for OTA determination in mice tissues (kidney, liver, brain, and intestine) as well as plasma samples. The method was rigorously validated for selectivity, linearity, accuracy, precision, recovery, dilution integrity, carry-over effect, stability, and robustness, meeting the validation criteria outlined by FDA and EMA guidelines. Furthermore, the described method enables the quantification of OTA in each individual sample using minimal tissue mass while maintaining excellent recovery values. The applicability of the method was demonstrated in a repeated low-dose OTA study in Balb/c mice, which, together with the inclusion of relevant and less common tissues in the validation process, underscore its suitability for neurodegeneration-related research.
摘要:
曲霉毒素A(OTA)是一种常见于各种食品中的霉菌毒素,这对人类和动物构成了潜在的健康风险。最近,越来越多的注意力集中在其潜在的神经退行性效应上。然而,目前还没有完全验证的HPLC分析方法在小鼠中建立定量,这个领域的主要动物模型,包括这个研究领域的关键组织,比如肠道和大脑。为了解决这个差距,我们开发并验证了一种高度敏感的,快速,和使用HPLC-FLD测定小鼠组织中OTA的简单方法(肾,肝脏,大脑,和肠)以及血浆样品。该方法的选择性经过严格验证,线性度准确度,精度,recovery,稀释完整性,结转效应,稳定性,和鲁棒性,符合FDA和EMA指南概述的验证标准。此外,所述方法使得能够使用最小的组织质量同时保持优异的恢复值对每个单独样品中的OTA进行定量。在Balb/c小鼠中的重复低剂量OTA研究中证明了该方法的适用性,which,在验证过程中包括相关和较不常见的组织,强调其对神经变性相关研究的适用性。