Development of new and validated HPTLC methods for the qualitative and quantitative analysis of hyperforin, hypericin and hyperoside contents in Hypericum species

Abstract

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.Methods: 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.Results: 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.

Chemical Composition of Essential Oils of Different Hypericum Species Growing Wild in Shouf Biosphere Reserve, Lebanon

“…Typical constituents of Hypericum species essential oils include the monoterpenes α-and β-pinene (Figure 1), limonene and myrcene, the sesquiterpenes β-caryophyllene (Figure 1), and caryophyllene oxide, and hydrocarbons such as n-decane, C 16 -and C 29 alkanes and C 24 , C 26 and C 28 alkanols [3,4]. More specifically, flavonols (catechins), naphthodianthrons (hypericin, pseudohypericin), xanthones, coumarins, lycosides, anthraquinones, phloroglucinols (hyperforin, adhyperforin), flavonoids (rutin, hyperoside, quercitrin), flavonol glycosides, lactones, pyrones, lipids, triterpenes, and tannins represent the main constituents in H. perforatum [21]. Whereas, α-pinene, β-pinene, (E)-β-ocimene, β-caryophyllene and germacrene-D dominate constituents of H. helianthemoides, H. scabrum and H. perforatum from Iran [3].…”

Hyperoside as a Potential Natural Product Targeting Oxidative Stress in Liver Diseases

“…A recent study reported that Hyp contents obtained from Hypericum species can be analyzed quantitatively and qualitatively using high-performance thin-layer chromatography (HPTLC) methods [ 26 ]. However, the intact detection of Hyp using high-performance liquid chromatography-ultraviolet (HPLC-UV) or liquid chromatography-tandem mass spectrometry (LC-MS-MS) methods is nearly impossible because of the galactose sugar moiety of Hyp.…”

Bioactivity Profiling and Quantification of Gastrodin in Gastrodia elata Cultivated in the Field versus Facility via Hyphenated High-Performance Thin-Layer Chromatography

“…For complex sample analysis, hyphenated high-performance thin-layer chromatography (HPTLC) is a versatile tool that can be used for simultaneous sample separation, allowing authenticity screening [ 27 , 28 , 29 , 30 , 31 , 32 ], non-target bioprofiling [ 33 , 34 , 35 , 36 ], as well as quantification of chemical marker compounds [ 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 ] from a wide variety of samples, including foods, plants, and other natural products. The combination of multiple analytical steps and identification of bioactivity by the same workflow makes hyphenated HPTLC a very sustainable technique and allows complex mixtures, such as plant extracts, to be studied in parallel in a much faster and straightforward way [ 45 , 46 ].…”