Brazilin content and potential biological properties of Caesalpinia sappan L. heartwood extracts from different extraction methods

Caesalpinia sappan L. has generally been used in Asian folk medicines for disease treatment. C. sappan heartwood is commonly utilized as a dyeing agent in food items, beverages, cosmetics, and clothing. The effects of three extraction techniques (maceration, reflux, and ultrasonic-assisted extraction) of C. sappan heartwood extract (CSE) on the brazilin content and potential of biological activities were investigated. The brazilin content was determined using high-performance liquid chromatography (HPLC). Quantifications of the antioxidant properties (DPPH and ABTS), the total phenolic contents, and the antimicrobial activity were also examined. Additionally, the effect of CSE on pro-inflammatory cytokine and mediator production was investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. The results showed that the CSE from the maceration and reflux methods produced high levels of brazilin and total phenolic contents, as well as having excellent antioxidant and antimicrobial activities. The macerated extract showed significant antioxidant activities with IC50 values of 2.88 ± 0.01 and 0.32 ± 0.01 mg/mL from DPPH and ABTS assays, respectively. Furthermore, the CSE from the maceration method which was rich in brazilin had an anti-inflammatory response by decreasing the levels of nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)-α on LPS-stimulated RAW 264.7 macrophage cells. These results confirmed CSE as a potential source of biological components. Antioxidant and anti-inflammatory assays proved maceration was the best extraction method.