Panara Anthi, Biliraki Dimitra, Nussbaumer Markus, Filiou Michaela D, Thomaidis Nikolaos S, Kostakis Ioannis K, Gikas Evagelos
Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771 Athens, Greece.
Laboratory of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771 Athens, Greece.
Molecules. 2025 Apr 24;30(9):1900. doi: 10.3390/molecules30091900.
Anxiety and stress-related disorders affect all ages in all geographical areas. As high anxiety and chronic stress result in the modulation of mitochondrial pathways, intensive research is being carried out on pharmaceutical interventions that alleviate pertinent symptomatology. Therefore, innovative approaches being currently pursued include substances that target mitochondria bearing an antioxidant moiety. In this study, a newly synthesized antioxidant consisting of triphenylphosphine (TPP), a six-carbon alkyl spacer, and hydroxytyrosol (HT) was administered orally to mice via drinking water. Cerebellum and liver samples were collected and analyzed using ultra-high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (UHPLC-MS/MS) to assess the levels of TPP-HT in the respective tissues to evaluate in vivo administration efficacy. Sample preparation included extraction with appropriate solvents and a preconcentration step to achieve the required sensitivity. Both methods were validated in terms of selectivity, linearity, accuracy, and limits of detection and quantification. Additionally, a workflow for evaluating and statistically summarizing multiple fortified calibration curves was devised. TPP-HT penetrates the blood-brain barrier (BBB), with a level of 11.5 ng g quantified in the cerebellum, whereas a level of 4.8 ng g was detected in the liver, highlighting the plausibility of orally administering TPP-HT to achieve mitochondrial targeting.
焦虑及应激相关障碍影响着所有地理区域的所有年龄段人群。由于高度焦虑和慢性应激会导致线粒体通路的调节，因此正在对缓解相关症状的药物干预措施展开深入研究。所以，目前正在探索的创新方法包括针对带有抗氧化部分的线粒体的物质。在本研究中，一种由三苯基膦（TPP）、一个六碳烷基间隔基和羟基酪醇（HT）组成的新合成抗氧化剂通过饮用水口服给予小鼠。收集小脑和肝脏样本，并使用超高效液相色谱 - 串联三重四极杆质谱（UHPLC - MS/MS）进行分析，以评估相应组织中TPP - HT的水平，从而评价体内给药效果。样品制备包括用适当溶剂萃取和一个预浓缩步骤，以达到所需的灵敏度。两种方法在选择性、线性、准确性以及检测和定量限方面均经过验证。此外，还设计了一个用于评估和统计汇总多个强化校准曲线的工作流程。TPP - HT能够穿透血脑屏障（BBB），在小脑中测得的含量为11.5 ng/g，而在肝脏中检测到的含量为4.8 ng/g，这突出了口服TPP - HT实现线粒体靶向的可行性。
