Formulation of Blumea balsamifera, Anredera cordifolia, and Phyllanthus niruri Extracts as Potential Anti-Inflammatory Agents

Inflammation is an immune response to foreign substances, pathogens, and cellular damage, characterized by redness, pain, and swelling. The use of synthetic anti-inflammatory drugs may cause adverse side effects, prompting the need for natural alternatives. This study aimed to evaluate the total flavonoid content and anti-inflammatory activity of single and combined extracts of sembung (Blumea balsamifera) [S], binahong (Anredera cordifolia) [B], and meniran (Phyllanthus niruri) [M] leaves. Extraction was performed using a 70% ethanol and water solution. Total flavonoid content was determined spectrophotometrically using quercetin as the standard. LC-HR-MS/MS results showed the presence of flavonoids, terpenoids, phenolics, and amino acids in each single extract (S, B, and M). Anti-inflammatory activity was assessed through the inhibition of albumin denaturation and the inhibition of cyclooxygenase-2 (COX-2). The highest flavonoid content (35.27 mg QE/g DW) in single extracts was found in B of the ethanolic extract, while the highest flavonoid content (40.24 mg QE/g DW) in all formulations was discovered in a combination of the ethanolic extract S:B:M (1:0:1). Moreover, the ethanolic extract S:B:M (1:0:1) gave the strongest inhibition (76.47%) of the albumin denaturation at 300 µg/mL and the strongest inhibition (98.5%) of the COX-2 inhibition at 100 µg/mL. Sembung extract was highest for reducing the expression of pro-inflammatory cytokines and TNF-α was 97.66% at an extract concentration of 15.625 ppm. As for S, S:B:M (1:0:1) and S:B:M (1:1:1) can reduce the expression of pro-inflammatory cytokines, as IL-6 was 100% in 0 pg/mL at an extract concentration of 15.625 ppm.