89 ISSN 0326-2383 Acta Farm. Bonaerense 24 (1): 89-90 (2005) Recibido el 17 de agosto de 2004 Aceptado el 19 de diciembre de 2004 Comunicaciones breves KEY WORDS: HPLC, Hypericin, H. styphelioides, H. tetrapetalum, Hypericum nitidum, TLC. PALABRAS CLAVE: Hipericina, HPLC, H. styphelioides, H. tetrapetalum, Hypericum nitidum, TLC. * Autora a quien dirigir la correspondencia. E-mail: sylvia.prieto@infomed.sld.cu Determination of Hypericin in Hypericum Species Grown in Cuba Lauro NUEVAS-PAZ 1 , Jorge MOLINA-TORRES 2 and Sylvia PRIETO-GONZÁLEZ 1 * 1 Centro de Química Farmacéutica; Calle 200 y 21, Atabey, Playa; P.O.Box 16042, La Habana, Cuba; C.P. 11600; Tel. (537)2713994, Fax (537) 33746. 2 Departamento de Biotecnología y Bioquímica, CINVESTAV-IPN, Unidad Irapuato, México. SUMMARY . Three species of Hypericum grown in Cuba were collected in different seasons and years, dried and powered. The plant material was extracted with methanol and analyzed by HPTLC and reverse phase HPLC for hypericin content. In the species of Hypericum analyzed hypericin was not found. RESUMEN. “Determinación de Hipericina en Especies de Hypericum crecidas en Cuba”. Tres especies de Hype- ricum crecidas en Cuba se colectaron, secaron y molieron en estaciones y años diferentes. El material vegetal se extrajo con metanol y se realizó el análisis para determinar el contenido de hipericina mediante HPTLC y HPLC de fase reversa. No se encontró hipericina en las especies de Hypericum analizadas. INTRODUCTION The genus Hypericum consists of more than 370 species. This genus is widely used in folk medicine since antiquity. Several pharmacologi- cal uses of Hypericum are known worldwide. The most popular plant of this genus is the Hy- pericum perfotatum L. (Saint John’s Wort) main- ly due to its use as a healing agent and by its anti-depressive and anti-viral applications 1 . St John’s Wort extract contains at least 10 con- stituents or groups of components that could contribute to its pharmacological effects. These components include naphthodianthrones (hy- pericin and pseudohypericin), flavonoids (rutin, hyperoside, isoquercitrin, quercitrin and quer- cetin), phloroglucinols (hyperforin and adhyper- forin), tannins (cathechin and epicatechin), proanthocyanidins (procyanidin B2) and bi- flavonoids (biapigenin and amentoflavone) 2 . Hypericin and pseudohypericin have been con- sidered effective against several viruses such as the influenza virus and the Herpes simplex virus 1 . These two compounds also have been found to inhibit retroviral infections such as the HIV 3 . For the determination and identification of Hypericin several methods by HPTLC 4 , HPLC 5 and HPLC-MS 6,7,8 have been reported. Among the Hypericum species that grow in Cuba are found Hypericum styphelioides Rich, H. nitidum Lam., and H. tetrapetalum Lam. In the literature consulted there are not reports re- lated to phytochemicals studies on these species. The purpose of this work was the determina- tion of hypericin content in these Hypericum species to promote their use as antiviral agents in Cuban folk medicine. MATERIAL AND METHODS Sample Collection All the Hypericum species were collected from various locations in the region of Pinar del Río, Cuba. Specimen samples were all collected at blossoming time with most of the flowers opened from April to November in 2000 and 2001. The samples were dried at room tempera- ture for seven days and powdered. Voucher specimens of H. tetrapetalum (HPPR 9192), H. nitidum (HPPR 9212 and 9278) and H. styphe- lioides (HPPR 9190, 9120-2 and 9277) were de- posited at the Herbarium of the Pinar del Río Botanical Gardens, Cuba. Sample preparation Powered samples (0.5 g ) were extracted with 10 ml of methanol using an ultrasonic bath for 10 min. For TLC and HPLC analysis the ex- tracts were first filtered. Standard preparation Commercial hypericin formulation tablets (Nutricia Manufacturing, Greenville, USA. Lot