Application of Box-Behnken design for optimization of phenolics extraction from Leontodon hispidulus in relation to its antioxidant, anti-inflammatory and cytotoxic activities

A wild plant no one had ever studied just revealed a treasure trove of healing compounds—and scientists used a clever statistical trick to unlock it.

Meet Leontodon hispidulus, a wild member of the daisy family that has been flying under the scientific radar—until now. For the first time, researchers optimized a green extraction method to pull out phenolic compounds from its aerial parts, using a 3-level Box-Behnken design to fine-tune three factors: ethanol/water ratio, material/solvent ratio, and extraction time. The results were striking: the optimized extract packed a total phenolic content of 104.18 µg/mg, achieved with 201 ml of 74.5% ethanol at 72 hours. But the real excitement came from the bioactivity tests. The extract neutralized free radicals with 80% inhibition and showed an antioxidant capacity of 41.89 mg Trolox-equivalent per gram. In rats, a 100 mg/kg dose reduced carrageenan-induced paw edema by 83.5%—edging out the standard drug indomethacin at 81.7%. Against cancer cells, prostate carcinoma lines proved more sensitive (IC50 = 16.5 µg/ml) than cervical ones (23 µg/ml). HPLC analysis identified quercetin, rutin, kaempferol, chlorogenic acid, and ρ-coumaric acid as the key players. This green extraction not only enriches the extract with phenolics but also opens the door to a promising natural source for antioxidant, anti-inflammatory, and anticancer therapies.

Key Points

• First phytochemical study of Leontodon hispidulus.
• Optimized green extraction yielded 104.18 µg/mg phenolics.
• Extract beat indomethacin in anti-inflammatory test.