Analytical determination, antioxidant and anti-inflammatory activities of Bhamrung-Lohit a traditional Thai medicine

Background and purpose: Bhamrung-Lohit (BRL) remedy is a traditional Thai medicine (TTM). There are few reports of biological activity, the activity of its constituent plants, or quantitative analytical methods for the content of phytochemicals. In this study, we investigated antioxidant, anti-inflammatory activity, and total phenolic and flavonoid content and validated a new analytical method for BRL. Experimental approach: Antioxidant activity was evaluated by a 2,2-diphenyl-1-picrylhydrazyl (DPPH and 2,2 -azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging. The cellular antioxidant activity was evaluated by inhibition of the superoxide anion (O2●-) production from HL-60 cells and antiinflammatory activity by inhibition of nitric oxide production in RAW264.7 cells. The total phenolic and flavonoid contents were analyzed using the Folin-Ciocalteu method and an aluminum chloride colorimetric assay, respectively. Validated analytical procedures were conducted according to International Conference on Harmonization (ICH) guidelines. Findings/Results: An ethanolic extract of BRL exerted potent DPPH radical scavenging activity and moderate antioxidant and anti-inflammatory activity. Caesalpinia sappan exerted the greatest effect and the highest content of total phenolics and flavonoids. The HPLC method validated parameters that complied with ICH requirements. Each peak showed selectivity with a baseline resolution of 2.0 and precision was less than 2.0% CV. The linearity of all compounds was 0.999 and the recovery % was within 98.0%-102.0%. The validated results demonstrated specificity/selectivity, linearity, precision, and accuracy with appropriate LOD and LOQ. Conclusion and implication: BRL remedy, a TTM demonstrated antioxidant and anti-inflammatory properties. This study is the first report on the biological activity and the validation of an HPLC method for BRL remedy.