Chemical Characterization and In Vitro Biological Activity of Edible Desert Truffle Tirmania pinoyi CCASU-TP-03

Tirmania pinoyi is an edible desert truffle valued for its nutritional properties; however, information regarding its biological activity and chemical composition is still limited. The current investigation sought to describe the chemical makeup of T. pinoyi CCASU-TP-03 extract and assess its biological properties. Molecular identification based on 25S rDNA and ITS1–5.8S–ITS2 sequence analysis placed CCASU-TP-03 within the T. pinoyi lineage, supporting the morphological identification. The extract was characterized by its chemical composition, while its antioxidant, antimicrobial, cytotoxic, and anti-inflammatory activities were evaluated using DPPH/ORAC, broth microdilution, SRB, and Griess assays, respectively. Western blotting was used to assess iNOS protein expression. The methanolic extract contained total carbohydrate content expressed as glucose equivalents (354.25 ± 7.71 mg glucose equivalents/g extract), total phenolics (72.57 ± 0.12 mg GAE/g extract), total flavonoids (9.83 ± 0.02 mg RE/g extract), and protein (11.12 ± 0.57 mg/g extract). Targeted qualitative HPLC-PDA profiling detected selected phenolic and flavonoid constituents, including gallic acid, isoferulic acid, ellagic acid, quercetin, and rutin. The T. pinoyi CCASU-TP-03 extract exhibited measurable antioxidant capacity in both DPPH and ORAC assays. Antimicrobial evaluation demonstrated inhibitory activity against several of the tested bacterial and yeast strains, whereas no activity was detected against Aspergillus niger under the tested conditions. Cytotoxicity screening showed the greatest reduction in viability in CaCo-2 cells, whereas PANC-1, HepG2, and HCT-116 cells maintained comparatively high viability over the investigated concentration range. The extract showed measurable inhibition of LPS-induced NO production accompanied by reduced iNOS protein expression. In conclusion, the findings demonstrate measurable antioxidant capacity, inhibitory activity against several tested microorganisms, differential cytotoxic effects among the tested cancer cell lines, and inhibition of NO production accompanied by reduced iNOS protein expression. Further studies are required to isolate the respective pure metabolites followed by preclinical evaluation for potential use in humans.