Establishing the safety of impurities in drug substances or products is crucial. The assessment of genotoxicity for these impurities and determining the acceptable limits pose considerable challenges, as recognized in recent guidelines. While the genotoxicity profile of vildagliptin—an oral hypoglycemic drug—is well established, there is limited knowledge about the genotoxic potential of its impurities. In this study, vildagliptin cyclic amidine, vildagliptin diketopiperazine, and vildagliptin amide were assessed in silico and in vitro for mutagenic and clastogenic/aneugenic potential using Ames and micronucleus tests. None of the investigated impurities showed mutagenic or clastogenic potential, thus, are considered non-mutagenic and non-clastogenic/aneugenic in vitro. These findings are consistent with negative in silico predictions for mutagenicity and clastogenicity/aneugenicity in vitro, indicating a good correlation between in silico and in vitro data. In conclusion, this study provides valuable information for the safety assessment of vildagliptin, confirming that its impurities are neither clastogenic/aneugenic nor mutagenic.

Tea is a widely consumed beverage that may contain both essential and toxic metals, raising nutritional and health concerns. This study evaluated the transfer of selected essential and toxic metals from black tea leaves to their infusions and assessed the associated health risks and nutritional contributions. Black tea leaves and their infusions from eight commercially available brands sold in Türkiye were analyzed. Toxic elements [arsenic (As), aluminium, cadmium (Cd), lead (Pb)] were quantified using inductively coupled plasma mass spectrometry, while essential elements [copper, iron (Fe), magnesium (Mg), zinc] were quantified using flame atomic absorption spectrometry. Health risks were evaluated using the Estimated Daily Intake, the Target Hazard Quotient (THQ), the Hazard Index (HI), and the Incremental Lifetime Cancer Risk (ILCR). In dry tea leaves, aluminum was the predominant toxic element (12.6-25.3 mg/g), followed by Pb (21.0-39.0 μg/g); As and Cd were present at lower levels. Mg (523.0-1421.0 μg/g) and Fe (263.1-445.5 μg/g) were the most abundant essential elements. Metal concentrations in tea infusions were significantly lower than those in dry leaves (p < 0.05), indicating limited transfer during brewing. In infusions, aluminum (0.5-1.0 mg/g) and Mg (88.5-413.4 μg/g) were the dominant toxic and essential elements, respectively. All THQ values and the combined HI were below 1, indicating no significant non-carcinogenic risk. ILCR values for Pb were below 10^-6, whereas those for As ranged from 10^-6 to 10^-4, indicating a moderate carcinogenic risk for As. The transfer of metals from tea leaves to infusions is limited, likely due to chelation and complexation during brewing. Under typical consumption conditions, black tea infusions were within established safety limits, with risk indices below thresholds of concern. While As warrants continued monitoring, essential elements contribute only a small fraction of the recommended daily intakes. The small sample size is a limitation.

Objective: This study aimed to evaluate the genotoxic potential of verbascoside using a standard battery of assays across bacterial and mammalian cell systems. Materials and Methods: The genotoxic potential of verbascoside was evaluated using a standard battery of assays, including the bacterial reverse mutation (Ames) test, cytokinesis-block micronucleus (CBMN) assay, and alkaline Comet assay. The Ames test was performed on Salmonella typhimurium TA98 and TA100 strains, with and without S9 metabolic activation, at concentrations ranging from 1 to 1000 µg/plate. Chinese hamster ovary (CHO-K1) cells were used for the CBMN and Comet assays at concentrations between 25 and 200 µg/mL. Results: The mutagenic index remained below 2.0 across all tested concentrations, showing no significant variation with increasing dose in the Ames test. No significant differences were observed in micronucleus frequency between the negative control and any concentration of verbascoside. The Comet assay results revealed no significant difference in DNA tail percentage between the negative control and verbascoside-treated groups. Conclusion: Under the tested conditions, verbascoside showed no mutagenic or genotoxic effect in bacterial or mammalian cell models, supporting a favorable genotoxicity safety profile and warranting further pharmacological development. Keywords: Micronucleus assay, Comet assay, Ames test, verbascoside, genotoxicity

Tanacetum parthenium (L.) Sch.Bip., a well-known member of the Asteraceae family, has long been used in traditional medicine against different ailments. Although the biological activities of its extracts and constituents such as parthenolide have been extensively studied, investigations on its essential oil remain limited. This study aimed to investigate the chemical composition and in vitro biological activities of essential oil obtained from T. parthenium wild-growing in Türkiye. The essential oil was obtained via steam distillation and analyzed by GC-FID and GC-MS. Its anti-inflammatory potential was evaluated by measuring nitric oxide (NO) inhibition in LPS-stimulated RAW 264.7 macrophages using the Griess assay. In addition, the cytotoxic effects of the essential oil were assessed on two human breast cancer cell lines (MCF-7 and MDA-MB-231) using the MTT assay. The results showed that the essential oil reduced NO production by 27% and 11% with 0.5 and 1 mg/mL, and exhibited weak cytotoxicity with IC₅₀ values of 333 ± 2.5 µg/mL and 214 ± 13.8 µg/mL against MCF-7 and MDA-MB-231, respectively. These findings highlight the potential of T. parthenium essential oil as a natural source of bioactive compounds with anti-inflammatory and anticancer properties, warranting further pharmacological and phytochemical investigations.

Advanced wound dressings should actively promote tissue repair while preventing microbial infection and excessive inflammation. In this study, multifunctional electrospun nanofibers composed of poly(vinyl alcohol)/chitosan/gelatin were developed by incorporating green-synthesized mesoporous silica nanoparticles (MSNs), Acacia gum, and Hypericum perforatum extract (HPE). Biological activity and safety were evaluated using disk-diffusion antimicrobial testing, MTT cell viability assays, nitric oxide measurement in LPS-stimulated RAW 264.7 macrophages, an L929 fibroblast scratch assay, and Ames and comet assays. The optimized nanofibers exhibited a uniform morphology with an average fiber diameter of approximately 400 nm and high encapsulation efficiencies (86–96%). MSN incorporation significantly prolonged hypericin release, producing a biphasic release profile with approximately 1.4-fold slower release than nanofibers without MSNs, while increasing Young’s modulus from 2.1 to 2.5 MPa. The hybrid nanofibers demonstrated broad-spectrum antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria, and Candida albicans, achieving approximately 70% of the efficacy of standard antimicrobial agents. In lipopolysaccharide-stimulated RAW 264.7 macrophages, the optimized formulation reduced nitric oxide production by approximately 51%, comparable to indomethacin and L-NAME. Although fibroblast migration was not significantly enhanced, the nanofibers exhibited acceptable cytocompatibility and showed no mutagenic or genotoxic effects in Ames and comet assays. These findings demonstrate that green-synthesized MSN-enhanced herbal nanofibers represent a safe and promising platform for sustained drug delivery and advanced wound-healing applications.