Department of Pharmacognosy, Faculty of Pharmacy, Yeditepe University, Istanbul, Turkey.
Phytochem Anal. 2022 Apr;33(3):355-364. doi: 10.1002/pca.3093. Epub 2021 Nov 4.
Hypericum perforatum L. (St. John's wort) is a medicinally important member of Hypericaceae. Many pharmacological activities have been mostly attributed to its hyperforin, hypericin and/or hyperoside contents. Therefore, qualitative and quantitative determinations of these ingredients are essential to justify the beneficial effects of St. John's wort on health. In the European Pharmacopoeia, the TLC and HPLC methods were given for this purpose. High performance thin layer chromatography (HPTLC) has recently become increasingly used as a suitable technique for analysing herbal drugs. This study aims to develop new and validated HPTLC methods to analyse these active components in different Hypericum spp. to find other suitable species to replace the official plant.
Three different mobile phases were developed: n-hexane-ethyl acetate (8:2) for hyperforin analysis, toluene-chloroform-ethyl acetate-formic acid (8:5:3.5:0.6) for hypericin analysis and ethyl acetate-formic acid-acetic acid-water (15:2:2:1) for hyperoside analysis. These newly developed and validated HPTLC systems were further applied to determine their concentrations in different Hypericum species.
Hyperforin concentration was found between 6.40 to 26.40 mg/g only in H. triquetrifolium, H. scabrum and two H. perforatum samples; hypericin was detected between 0.81 and 1.41 mg/g only in H. bithynicum, H. perfoliatum, H. triquetrifolium and two H. perforatum samples; and hyperoside was identified in all tested specimens ranging from 1.01 to 9.73 mg/g. The new HPTLC methods developed and validated in the present study may ensure reliable results for the qualification and quantification of hyperforin, hypericin and hyperoside contents in Hypericum species.
贯叶金丝桃（贯叶连翘）是金丝桃科的一种药用植物。其许多药理学活性主要归因于其贯叶金丝桃素、金丝桃素和/或金丝桃苷的含量。因此，对这些成分进行定性和定量测定对于证明贯叶连翘对健康的有益作用是必要的。在《欧洲药典》中，为此提供了 TLC 和 HPLC 方法。高效薄层色谱（HPTLC）最近已越来越多地用作分析草药的合适技术。本研究旨在开发新的和经过验证的 HPTLC 方法来分析不同贯叶连翘属物种中的这些活性成分，以找到其他合适的物种来替代官方植物。
开发了三种不同的流动相：正己烷-乙酸乙酯（8:2）用于贯叶金丝桃素分析，甲苯-氯仿-乙酸乙酯-甲酸（8:5:3.5:0.6）用于金丝桃素分析，乙酸乙酯-甲酸-乙酸-水（15:2:2:1）用于金丝桃苷分析。这些新开发和验证的 HPTLC 系统进一步应用于确定它们在不同贯叶连翘属物种中的浓度。
仅在 H. triquetrifolium、H. scabrum 和两个 H. perforatum 样品中发现贯叶金丝桃素浓度在 6.40 至 26.40mg/g 之间；仅在 H. bithynicum、H. perfoliatum、H. triquetrifolium 和两个 H. perforatum 样品中检测到金丝桃素浓度在 0.81 至 1.41mg/g 之间；而在所有测试样本中均鉴定出金丝桃苷，浓度范围为 1.01 至 9.73mg/g。本研究中开发和验证的新 HPTLC 方法可确保贯叶连翘属物种中贯叶金丝桃素、金丝桃素和金丝桃苷含量的定性和定量的可靠结果。
