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HPLC Analysis of Supercritical Carbon Dioxide and Compressed Propane Extracts from Piper amalago L. with Antileishmanial Activity
Departamento de Farmácia, Universidade Estadual de Maringá, 87020-900, Maringá, Paraná, Brazil
Departamento de Engenharia Química, Universidade Estadual de Maringá, 87020-900, Maringá, Paraná, Brazil
Laboratório de Bioquímica de Tripanossomatídeos, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, 21040-360, Brazil
* Author to whom correspondence should be addressed.
Received: 2 October 2011; in revised form: 5 December 2011 / Accepted: 14 December 2011 / Published: 22 December 2011
Abstract: Piper amalago L. leaves were extracted with supercritical carbon dioxide and compressed propane under different conditions, and with chloroform by the conventional maceration method. These methods were compared for the pyrrolidine alkaloid content. Supercritical carbon dioxide (SFE-CO2) at 313 K and 12.55 MPa showed the highest selectivity for the main compound (600.53 mg/g of extract). A gradient high-performance liquid chromatography (HPLC) method was developed and validated to quantify the alkaloid N-[7-(3′,4′-methylenedioxyphenyl)-2(Z),4(Z)-heptadienoyl]pyrrolidine (1) in the extracts. The HPLC method showed linearity, precision and accuracy, allowing the quantitative analysis of the alkaloid in all the samples. All the extracts were tested against the promastigote and intracellular amastigote forms of Leishmania amazonensis. The antileishmanial activity was evaluated in terms of inhibitory concentration for 50% of protozoa (IC50). The cytotoxicity was also evaluated against J774A1 macrophages, and the cytotoxic concentrations for 50% of macrophages were obtained (CC50). The SFE-CO2 (313 K; 12.55 MPa) extract showed the highest antileishmanial activity with the following IC50 values of 16 and 7 µg/mL against the promastigotes and intracellular amastigotes forms, respectively. The extract showed low cytotoxicity with a CC50 value of 93 µg/mL.
Keywords: Piper amalago L.; pyrrolidine alkaloid; supercritical carbon dioxide; HPLC validation; antileishmanial activity
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Carrara, V.S.; Serra, L.Z.; Cardozo-Filho, L.; Cunha-Júnior, E.F.; Torres-Santos, E.C.; Cortez, D.A.G. HPLC Analysis of Supercritical Carbon Dioxide and Compressed Propane Extracts from Piper amalago L. with Antileishmanial Activity. Molecules 2012, 17, 15-33.
Carrara VS, Serra LZ, Cardozo-Filho L, Cunha-Júnior EF, Torres-Santos EC, Cortez DAG. HPLC Analysis of Supercritical Carbon Dioxide and Compressed Propane Extracts from Piper amalago L. with Antileishmanial Activity. Molecules. 2012; 17(1):15-33.
Carrara, Vanessa da Silva; Serra, Lara Zampar; Cardozo-Filho, Lúcio; Cunha-Júnior, Edézio F.; Torres-Santos, Eduardo C.; Cortez, Diógenes Aparício Garcia. 2012. "HPLC Analysis of Supercritical Carbon Dioxide and Compressed Propane Extracts from Piper amalago L. with Antileishmanial Activity." Molecules 17, no. 1: 15-33.