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Background: Seasonal climatic variations significantly influence the biosynthesis and accumulation of secondary metabolites in medicinal plants. Bauhinia variegata Linn. bark is an important traditional medicinal resource, but the optimal harvesting season for maximum phytochemical yield remains unclear. Objective: To investigate the effect of seasonal variation on the physicochemical properties, phytochemical composition, antioxidant, and anti-inflammatory activities of B. variegata stem bark. Methods: Bark samples were collected during monsoon (S1), winter (S2), and summer (S3). Physicochemical parameters, total phenolic, flavonoid, and tannin contents were evaluated. Phytochemical profiling was performed using TLC and HPTLC. Antioxidant activity was assessed by DPPH and FRAP assays, while anti inflammatory activity was determined using HRBC membrane stabilization and BSA protein denaturation assays. Results: Significant seasonal variations were observed. The winter sample (S2) showed the highest alcohol soluble extractive value (7.5%), total phenolics (15.6 µg/mL), flavonoids (25.6 µg/mL), and tannins (5.40%). HPTLC profiling demonstrated distinct seasonal chemical variations. The S2 extract also exhibited the strongest antioxidant and anti-inflammatory activities, with 75.45% HRBC membrane stabilization and 37.03% inhibition of BSA denaturation at 400 µg/mL. Conclusion: Seasonal variation markedly affects the phytochemical composition and biological activity of B. variegata bark. Winter is the optimal harvesting season, providing maximum bioactive constituents and superior antioxidant and anti-inflammatory potential, thereby supporting its standardization for phytopharmaceutical applications. Keywords: Bauhinia variegata Linn.; Kanchanar; seasonal variation; phytochemical standardization; HPTLC fingerprinting; antioxidant activity; anti-inflammatory activity.