UV-Vis spectrophotometric analysis and in vivo evaluation of safety and neuronal targets of the raw materials <i>Ziziphora clinopodioides</i> (Lam.) and <i>Ferula rigidula</i> (L.)

Background: The analysis of Armenian folk medicine shows that Ferula rigidula and Ziziphora clinopodioides are widely used for their phytotherapeutic and phytonutrient properties. However, these plants should be used with caution in the food and pharmaceutical industries, as prolonged consumption at high concentrations may pose potential toxicity risks. Objectives: The purpose of this investigation was to determine the safety profile of Ziziphora and Ferula through in vivo assessment of hepatic toxicity and to evaluate their potential effects on neuronal activity. Methods: UV-Vis spectrophotometric analysis was used to standardize the raw materials of Ziziphora and Ferula based on their total hydroxycinnamic acid content. Potential hepatotoxic effects were evaluated in Wistar albino rats by measuring serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels following administration of Ziziphora extract at 200 or 400 mg/kg/day and Ferula extract at 200 mg/kg/day for 3 weeks. The 200 mg/kg doses of both extracts were subsequently evaluated in acute electrophysiological experiments using single-neuron spike activity recordings. Results: The results of preliminary standardization showed that the total hydroxycinnamic acid content of the ethanolic extract of hydroponically cultivated Ziziphora was 5.16 ± 0.025%, whereas that of the Ferula extract was 3.13 ± 0.052%. Among the treatment groups, only rats receiving Ziziphora at 400 mg/kg showed a significant difference in body-weight gain compared with the control group (p < 0.05). Serum AST levels did not differ significantly among the experimental groups, whereas ALT was significantly elevated in the Ziziphora 400 mg/kg group. No significant changes in ALT were observed following administration of Ziziphora or Ferula at 200 mg/kg. A single intraperitoneal administration of the 200 mg/kg extracts produced changes in neuronal spike activity within cholinergic projections of the rat brain. Conclusion: Ziziphora and Ferula extracts demonstrated no significant alterations in serum AST or ALT at the tested dose of 200 mg/kg, whereas the higher 400 mg/kg dose of Ziziphora produced a significant increase in ALT, suggesting a dose-dependent safety consideration. Both extracts also influenced neuronal activity within cholinergic brain pathways, indicating potential neuromodulatory activity. Further studies are required to establish their long-term safety, mechanisms of action, and potential relevance to neurodegenerative disorders such as Alzheimer’s disease. Novelty: To our knowledge, this study is among the first to combine the standardization of Ziziphora and Ferula species from Armenian flora based on their hydroxycinnamic acid content with an in vivo assessment of hepatic safety and neuronal responses. Keywords: Ziziphora clinopodioides, Ferula rigidula, UV-Vis spectrophotometric analysis, hydroxycinnamic acids, aminotransferase, neuronal activity