Biofunctional activity and phytochemical composition by LC-QToF-MS of Mitragyna speciosa Korth leaves extracted in Northern Thailand

Mitragyna speciosa Korth, a traditional Thai herb, is known for its medicinal properties, including enhancing physical strength and promoting relaxation. However, its misuse can lead to addiction. This study evaluated the antioxidant capacity, anti-inflammatory activity, and chemical composition of M. speciosa using liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QToF-MS). Leaves were extracted with ethanol, methanol, and water. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay showed that ethanol extraction had the best IC50 value (0.23 ± 0.01 mg/mL). Methanol extraction excelled in the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay (2.24 ± 0.33 mg/mL), while water extraction displayed the highest antioxidant capacity in the fluorescence recovery after photobleaching (FRAP) assay (231.87 ± 11.51 mg ascorbic acid equivalent [AAE]/100 g). The water extract exhibited the highest total phenolic content (153.41 ± 0.06 mg gallic acid equivalent [GAE]/g) and total flavonoid content (6.24 ± 0.83 mg QE/g). It also provided the maximum anti-inflammatory protection, with a protection rate of 96.35% and a hemolysis of 3.65%. LC-QToF-MS analysis identified key bioactive compounds, such as stigmatellin Y (99.17%) and mitragynine (98.58%), along with other compounds with antioxidant and anti-inflammatory activities. These findings highlight M. speciosa’s potential of being a therapeutic agent. Future research should focus on elucidating the molecular mechanisms of its bioactive compounds, optimizing extraction techniques, and conducting in vivo and clinical studies to validate its safety and therapeutic efficacy.