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DOI: 10.1055/s-0031-1282174
Determination of Hypericin in St. John's Wort (Hypericum perforatum L.) extracts using HPLC-ED
The genus Hypericum, which comprises about 450 species of herbs, shrubs and trees, belongs to the family Clusiaceae (Guttiferae), formerly Hypericaceae. One of the topmost and commercially renowned species of the genus Hypericum is Hypericum perforatum L., commonly known as St. John's Wort. Hypericum perforatum, is well known for its profound pharmacological activities as antidepressant, anxiolytic, antiviral, antimicrobial and wound healing. In this study, using HPLC-ED system, quantitative analysis of hypericin was carried out in different water-methanolic extracts of St. John's Wort. Hot of water-methanol extracts of St. John's Wort were prepared. The herba (1g) was powdered and extracted with water-methanol [(1:1) 10 mL]. Afterward 1 mL of that extract was centrifuged, obtaining supernatant which was used for analysis. The standard was hypericin purchased from HWI Analytic GmbH, Germany. Mobile phase: methanol-acetonitrile, water, acetic acid (20+10+70+1), electrochemical detector with range 50 nA, potential + 0.84 V, filter, 0.02Hz, flow rate, 1 mL/min: and temperature 25°C. Injection volume: 20µL. Concentrations of standard were 0.1µg/20µL, 0.2µg/20µL, and 0.4µg/20µL. Retention time of hypericin in the standard solution and herbal extracts were 28.4 minutes. Limit of detection was 100ng of hypericin. Amount of hypericin in extracts of St. John's Wort was 25–26mg/g dry weight. The very high content of hypericin in Bosnian St. John's Wort give more importance to this plant as traditional drug for its use in official medicine.
Keywords: hypericin, Hypericum perforatum, HPLC-ED, determination
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