Abstract

Background and Aims: Scutellaria barbata D. Don, a perennial herb with a rich history in traditional medicine, is known for its bioactive compounds, including wogonin. It possesses anti-inflammatory, antioxidant, and anticancer properties. Together with other bi-oactive compounds, wogonin contributes to the pharmacological activities associated with S. barbata D. Don, making it a subject of interest in both traditional and modern medicinal re-search. This study aimed to develop and validate a High-Performance Thin-Layer Chroma-tography (HPTLC) method for the precise and accurate quantification of wogonin in the hy-droethanolic extract of S. barbata D. Don.

Methods: The process was developed and underwent rigorous validation following the ICH guidelines. Parameters such as linearity, specificity, instrument precision, precision, accura-cy, limit of detection (LOD), limit of quantification (LOQ), and robustness were systemati-cally evaluated to ensure the method's reliability.

Results: The HPTLC separation was conducted on TLC aluminum plates precoated with sil-ica gel 60 F254, and optimal separation was achieved using Ethyl acetate: toluene: formic ac-id (5: 4.9: 0.1, (v/v/v/v)) as the mobile phase. Densitometric scanning at 254 nm in reflec-tion/absorbance mode revealed compact spots at Rf 0.87, corresponding to wogonin. Linear regression analysis demonstrated a robust linear relationship between peak area and the amount of wogonin in the range of 10-40 (μg/mg). The quantification of wogonin in crude extracts obtained was 3.274 μg/mg of extract.

Conclusion: In summary, the developed HPTLC method emerges as a straightforward, pre-cise, and accurate tool for the rapid screening of active constituents, specifically wogonin, within the petroleum ether extracts of S. barbata D. Don. This method holds promise for routine quality control and analysis of herbal materials and formulations containing S. bar-bata D. Don, contributing to the ongoing exploration of its therapeutic potential.

Keywords: Scutellaria barbata D. Don, Wogonin, HPTLC, Quantitative analysis, Validation of method.