Quantitative Thin Layer Chromatography Analysis, Antioxidant, and Anti-Inflammatory Activities of Polyherbal Formulation (Ammarit-Osot) Extracts

Thaniya Wunnakup; Thanapat  Songsak; Nirun  Vipunngeun; Lukman  Sueree

doi:10.59796/jcst.V14N3.2024.65

Authors

Thaniya Wunnakup Drug and Herbal Product Research and Development Center, College of Pharmacy, Rangsit University, Pathum Thani 12000, Thailand
Thanapat Songsak Department of Pharmacognosy, College of Pharmacy, Rangsit University, Pathum Thani 12000, Thailand
Nirun Vipunngeun Department of Pharmacognosy, College of Pharmacy, Rangsit University, Pathum Thani 12000, Thailand
Lukman Sueree Faculty of Science and Technology, Hatyai University, Hatyai, Songkhla 90110, Thailand

DOI:

https://doi.org/10.59796/jcst.V14N3.2024.65

Keywords:

Ammarit-Osot, TLC, Piperine, Antioxidant, Anti-inflammatory activity

Abstract

This study was carried out to develop a TLC method for quantifying one of bioactive, piperine, from polyherbal formulation (Ammarit-Osot) extracts. The extracts were prepared by different solvents such as methanol (ME) and water (WE). The mobile phase of toluene:acetone:formic acid (7:2:1, v/v/v) was used for separation. The calibration curve showed a good linearity (r² = 0.9972) in the range of 50 – 500 ng/spot. This assay was assessed by intra-and interday precision (RSD 0.25 – 2.49%), and accuracy (101.05 – 101.56 %). The piperine content was found to be 23.06 ± 0.06 and 0.44 ± 0.03% w/w for ME and WE, respectively. Both ME and WE demonstrated significant activity in DPPH and FRAP assays, indicating their antioxidant potential. The TPC of ME and WE were 53.40 ± 2.21 and 101.18 ± 0.46 (mg GAE/g extract), respectively. The TFC of ME and WE were 43.35 ± 3.94 and 25.31 ± 1.74 (mg catechin/g extract), respectively. In addition, the cytotoxicity of extracts on RAW264.7 was investigated by MTT assay. The ME and piperine showed IC₅₀ with 162.40 ± 7.84 and 115.68 ± 11.96 µg/mL, respectively. While the WE exhibited low cytotoxicity (IC₅₀ > 1,000 µg/mL), the anti-inflammatory activity was assessed by LPS-stimulated NO overproduction. The ME and piperine displayed the ability to inhibit NO production, but WE not inhibited. The results indicated that the different solvent extracts of Ammarit-Osot have the potential to be antioxidant, anti-inflammatory and immune-stimulate agent.

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