Polcaro_2026_Fitoterapia__107324

Andrographis paniculata Burm. F. (Acanthaceae) is receiving significant interest from the scientific and medical communities due to its immunological, anti-inflammatory, antibacterial, and neuroprotective properties. This study aimed at comparing green extraction methods for A. paniculata in order to identify an environmentally friendly process capable of yielding a product rich in bioactive compounds. Andrographis paniculata aerial parts were submitted to extraction methods (maceration, Ultrasound Assisted Extraction UAE, and Solid Liquid Dynamic Extraction SLDE) using different solvent mixtures (100% EtOH, 75% EtOH/H(2)O, and 50% EtOH/H(2)O). Chemical profiling of the extracts was performed using an integrated approach based on high-resolution mass spectrometry (HRMS) via liquid chromatography-tandem mass spectrometry (LC-MS) and Nuclear Magnetic Resonance (NMR) spectroscopy, facilitating the identification of metabolites. A comparative analysis, conducted using multivariate statistical techniques, revealed that the extracts obtained by SLDE were the richest in metabolites. A detailed phytochemical investigation of the 75% EtOH/H(2)O extract obtained by SLDE was carried out to isolate and unambiguously characterize compounds putatively identified by LC-MS analysis. Among diterpenes, 7-hydroxy-14-deoxyandrographolide (15) is here reported for the first time from a natural source. Furthermore, the ability of extracts and isolated compounds to inhibit tyrosinase and acetylcholinesterase (AChE) enzymes was tested by spectrophotometric assays. Flavonoids (IC(50) = 54.8-97.0 microM) inhibited tyrosinase enzyme slightly better than diterpenoids (IC(50) = 98.8-173.8 microM), while the diterpenes 7-hydroxy-14-deoxyandrographolide (IC(50) = 87.2 microM) and andrographolide (IC(50) = 97.1 microM) showed the highest AChE inhibitory activity.