Identification of Major Constituents ofHypericum perforatumL. Extracts in Syria by Development of a Rapid, Simple, and Reproducible HPLC-ESI-Q-TOF MS Analysis and Their Antioxidant Activities.

Hypericum perforatum linn (st.John's wort) is a popular and widespread medicine in syria, which is used for a wide range of conditions, including gastrointestinal diseases, heart disease, skin diseases, and psychological disorders.This widespread use prompted us to identify the main compounds of this plant from syria that are responsible for its medicinal properties, especially since its components differ between countries according to the nature of the soil, climate, and altitude.This is, to the best of our knowledge, the first report in which st.John's wort, a plant native to syria, is extracted using different solvents and its most important compounds are identified.In this study, the dried above-ground parts, i.e., leaves, stem, petals, and flowers, were extracted using different solvents (water, ethanol, methanol, and acetone) and extraction protocols.By increasing the polarity of the solvent, higher yields were obtained, indicating that mainly hydrophobic compounds were extracted.Therefore, we conclude that extraction using the tea method or using a mixture of water and organic solvents resulted in higher yields compared with pure organic solvents or continuous boiling with water for long periods.The obtained extracts were analyzed using high-performance liquid chromatography equipped with a diode array detector (hplc-dad), coupled with uv-visible spectrophotometry at a full spectrum (200-800 nm).The hplc spectra of the extracts were almost identical at three wavelengths (260 nm for phloroglucinols (hyperforin and derivates), 590 nm for naphthodianthrones (hypericins), and 350 nm for other flavonols, flavones, and caffeoylquinic acids), with differences observed only in the intensity of the peaks.This indicates that the same compounds were obtained using different solvents, but in different amounts.Five standards (chlorogenic acid, quercetin, quercitrin hydrate, hyperoside, and hypericin) were used, and a comparison with retention times and ultraviolet (uv) spectra reported in the literature was performed to identify 10 compounds in these extracts: hyperforin, adhyperforin, hypericin, rutin, quercetin, quercitrin, quercitrin hydrate, hyperoside, biapigenin, and chlorogenic acid.Although the european pharmacopoeia still describes ultraviolet spectroscopy as a method for determining the quantity of hyperici herba, interference from other metabolites can occur.Combined hplc-dad and electrospray ionization-mass spectrometry (lc-esi-ms) in the positive mode have therefore also been used to confirm the presence of these compounds in the extracts by correlating known masses with the identified masses or through characteristic fragmentation patterns.Total phenolic contents of the extracts were determined by the folin-ciocalteu assay, and antioxidant activity was evaluated as free radical scavenging capacity using 2,2-diphenyl-1-picrylhydrazyl (dpph) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (abts) assays.The results indicate that the aqueous extracts prepared by the tea method gave the highest total phenols, while the pure organic solvents gave very low phenols.Also, the extracts that contain the largest amount of phenols gave lower ic50values or higher antioxidant activity than that of others.

Journal: ACS omega (2022)
Publication types: Journal Article