Comprehensive evaluation of Pulicaria dysenterica subsp. dysenterica decoction: Phenolic profile by UHPLC-HESI-Orbitrap®-HRMS2, In vitro gastrointestinal stability of biological activities, acute oral toxicity, and antihemorrhoidal potential.

ETHNOPHARMACOLOGICAL RELEVANCE: Pulicaria dysenterica Gaertn. subsp. dysenterica has been traditionally used for gastrointestinal and inflammatory disorders; however, its phenolic stability during digestion and its anti-inflammatory efficacy have not been comprehensively investigated.

AIM OF THE STUDY: This study aimed to evaluate the phenolic profile, antioxidant capacity, gastrointestinal stability, acute oral toxicity, and therapeutic efficacy of P. dysenterica subsp. dysenterica decoction extract.

MATERIALS AND

METHODS

Total phenolic content (TPC) and antioxidant activity (DPPH and CUPRAC assays) were assessed before digestion and after simulated gastric and intestinal digestion. Phenolic compounds were characterized using a validated UHPLC-HESI-Orbitrap®-HRMS2 method. Acute oral toxicity was evaluated according to OECD guideline 423. The therapeutic effects of the extract at two different doses, low dose (PDE-L, 3.2 mg/kg) and high dose (PDE-H, 6.4 mg/kg), were investigated in an experimentally induced hemorrhoid model by assessing rectal-anal coefficient (RAC), inflammatory index, and TNF-α levels in serum and rectal tissues.

RESULTS

The undigested extract exhibited the highest TPC (1541.03 ± 17.34 μg GAE/mL) and DPPH activity (616.22 ± 0.39 μg TE/mL), whereas gastric digestion showed superior CUPRAC activity (1255.36 ± 16.14 μg TE/mL). Among 75 screened phenolics, 28 were identified, with chlorogenic acid (8543.88 ± 38.04 μg/g, gastric phase) and caffeic acid (6412.00 ± 96.50 μg/g, intestinal phase) being predominant. No acute oral toxicity was observed up to 2000 mg/kg. PDE-H significantly reduced RAC, inflammatory index, and TNF-α a levels in rats with croton oil-induced hemorrhoids, while PDE-L exhibited limited effects. TNF-α levels correlated positively with inflammation severity.

CONCLUSION

P. dysenterica subsp. dysenterica is a safe, phenolic-rich plant with strong antioxidant and anti-inflammatory properties, supporting its potential as a natural therapeutic candidate for pharmaceutical and nutraceutical applications.