Image-Assisted Quantification Using Arc GIS and LC-Ms Based Characterization of Pigments in Pycnoporus sanguineus

Pycnoporus sanguineus is a medicinally important macrofungus recognized for its ability to produce diverse bioactive secondary metabolites, including extracellular pigments with potential biotechnological and therapeutic potential. The present study evaluated the growth characteristics, biomass production under solid-state and submerged culture conditions, pigment formation, and bioactivity of P. sanguineus. Among the submerged conditions, agitated cultivation in potato dextrose broth (PDB) produced the highest mycelial biomass (1.16 g dry weight) after 25 days of incubation. Comparative evaluation on six solid media revealed maximum radial growth on potato dextrose agar (PDA) with a growth rate of 74.66 mm within 120 h and a specific growth rate of 0.09048 h^-1. Pigment production was further assessed using modified PDA culture formulations and quantified by a novel ArcGIS-based digital image analysis approach. Pigment from all five formulations was extracted using accelerated solvent extraction (ASE) with ethanol and subjected to UV-Visible spectrophotometric analysis for comparative assessment of relative pigment accumulation. Spectrophotometric analysis at 416 nm showed a positive correlation (R² = 0.8005) with the ArcGIS-derived pigmented area, validating image-based assessment of relative pigment accumulation. LC-MS analysis identified pigment-associated metabolites, including cinnabarinic acid, tramesanguin, lambertellin, and coibanoles A and B. The pigment extracts exhibited significant antioxidant and concentration-dependent antibacterial activities against Escherichia coli and Enterococcus faecalis, with MIC values supporting their antibacterial potential. Collectively, the results demonstrated that integrating Arc-GIS-based image analysis, UV-visible spectrophotometric, and LC-MS profiling provides a robust approach for the characterization and biological evaluation of P. sanguineus pigments, highlighting their potential for further application.