Insights into UPLC- QqQ- MS Analysis of Ammi visnaga Fruits and Potential Hepatic Cytotoxicity of Nano-khellin and Nano-visnagin: Comparative Evaluation and Mechanistic Study

Hepatocellular carcinoma (HCC) is the most prevalent type of liver cancer, necessitating innovative therapeutic approaches due to high mortality rates. Nano-encapsulated compounds from plant sources could offer a promising avenue for hepatic cancer management. Khellin and visnagin are natural furanochromones from Ammi visnaga fruits known many years ago, but their cytotoxic activity recently gained attention and approval.This study aimed to explore the phytoconstituents of A. visnaga fruits and evaluate the role of nanotechnology in improving the therapeutic efficacy of khellin and visnagin nanoparticles against HepG2 and Huh7 cell lines.The phytochemical constituents of A. visnaga fruit were determined by UPLC- QqQ- MS. Khellin and visnagin were isolated, identified and then formulated into nanostructured lipid carriers for improved delivery. Cytotoxic activity of nano-khellin and nano-visnagin was studied using Doxorubucin as a positive control.The chromatogram of UPLC- QqQ- MS identified 17 phytochemical constituents, including furanochromones and flavonoids, from Ammi visnaga fruits. Cytotoxicity assessments revealed that nano-khellin and nano-visnagin exhibited significant activity against HepG2 and Huh7 cell lines, with IC50 values of 12.99 ± 0.587 µM and 9.561 ± 0.305 µM for khellin, and 10.843 ± 0.431 µM and 8.671 ± 0.398 µM for visnagin, respectively. Moreover, treatment with these compounds resulted in decreased levels of miRNA-181a, increased caspase-3 activity, and decreased levels of AKT and p38 MAPK.These findings support the therapeutic potential of nano-khellin and nano-visnagin, highlighting their biochemical mechanisms in HCC management.Keywords: Ammi visnaga fruit, UPLC-QqQ-MS profiling, nano-khellin, nano-visnagin, hepatic cytotoxicity.