Abstract

Picroside-I, andrographolide and silybin exhibit manifold biological activities, present as bioactive components in Picrorhiza kurroa (roots), Andrographis paniculata (aerial parts) and Silybum marianum (seeds), respectively. A validated HPTLC method was developed for the simultaneous determination of picroside-I, andrographolide and silybin in the combined hydroalcoholic extract of the plants as per ICH guidelines. On silica gel F₂₅₄ HPTLC plates, the combined hydroalcoholic extract of plants and standard solutions were applied. The plates were run in twin chamber consisting n-butanol: glacial acetic acid: water (6:1:3, v/v/v) as a mobile phase and documented at 254 and 366 nm using Camag TLC scanner III with WinCATS software. The analysis revealed that a good linear relationship was established between response (peak area) and amount in the range of 60–600 ng/spot. The LOD (ng/spot) and LOQ (ng/spot) for estimation of picroside-I, andrographolide and silybin were found to be 15.089, 21.969, 25.702 and 45.725, 66.573, 77.885, respectively. Recovery experiments showed 97.75–103.74%, 99.75–101.12% and 98.96–101.69% for picroside-I, andrographolide and silybin, respectively. The HPTLC method showed good linearity, recovery and high precision of all the three bioactive components, which can be applied for the routine analysis of polyherbal formulations containing these plants as their ingredients.

摘要

Picroside-I、穿心莲内酯和水飞蓟宾表现出多种生物活性，分别作为Picrorhiza kurroa（根）、穿心莲（地上部分）和水飞蓟（种子）中的生物活性成分存在。根据 ICH 指南，开发了一种经过验证的 HPTLC 方法，用于同时测定植物混合水醇提取物中的苦杏仁苷-I、穿心莲内酯和水飞蓟宾。在硅胶 F ₂₅₄ HPTLC 板上，应用组合的植物水醇提取物和标准溶液。平板在由正丁醇：冰醋酸：水（6:1:3，v/v/v) 作为流动相，并使用带有 WinCATS 软件的 Camag TLC 扫描仪 III 在 254 和 366 nm 处记录。分析表明，响应（峰面积）与 60–600 ng/点范围内的量之间建立了良好的线性关系。发现用于估计苦味苷-I、穿心莲内酯和水飞蓟宾的 LOD (ng/spot) 和 LOQ (ng/spot) 分别为 15.089、21.969、25.702 和 45.725、66.573、77.885。回收率实验表明苦味苷-I、穿心莲内酯和水飞蓟宾的回收率分别为 97.75–103.74%、99.75–101.12% 和 98.96–101.69%。HPTLC 方法显示三种生物活性成分均具有良好的线性、回收率和高精度，可用于以这些植物为成分的多草药制剂的常规分析。