Antinociceptive and Anti‐Inflammatory Effects of Alchemilla daghestanica and Alchemilla minusculiflora: Quantitative Phytochemical Profiling and In Vivo Evidence

Abstract

Inflammatory pain mediated by pro‐inflammatory factors remains a major clinical challenge, highlighting the need for safer alternatives to conventional anti‐inflammatory drugs. Here, the antinociceptive effects of methanolic extracts from the aerial parts of Alchemilla daghestanica (AD) and Alchemilla minusculiflora (AM) were evaluated in mice using the formalin test, and anti‐inflammatory activity was assessed by measuring formalin‐induced paw edema. Phytochemical characterization was performed by LC–MS/MS, and potential activities of major constituents were predicted using PASS Online. LC–MS/MS identified miquelianin as the predominant compound in both extracts. AD (100–200 mg/kg) and AM (200 mg/kg) significantly inhibited the inflammatory phase of the formalin test (p < 0.001 and p < 0.01, respectively) without affecting the neurogenic phase, and AM extract reduced paw edema (p < 0.05). PASS predicted high probabilities (Pa > 0.7) for anti‐inflammatory activity of miquelianin, isoquercitrin, and tannic acid. These findings suggest that these extracts may represent valuable candidates for further investigation in inflammatory pain management.