HPLC Analysis of Supercritical Carbon Dioxide and Compressed Propane Extracts from Piper amalago L. with Antileishmanial Activity

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.

Supercritical Fluid Extraction of Pyrrolidine Alkaloid from Leaves of Piper amalago L.

“…The isolation of the alkaloid N-[7-(3′,4′-methylenedioxyphenyl)-2(Z),4(Z)-heptadienoyl]pyrrolidine (compound 1 ) from P. amalago L. leaves and its spectral data were described in a previous study [12]. …”

“…The infusions of the roots are used as diuretic and against renal stones [5]. Pharmacological investigations have also shown that the extracts from P. amalago leaves present anti-inflammatory [7], anxiogenic [8], diuretic, natriuretic, antilithiatic [9], healing [10], schistosomicidal [11], and antileishmanial activities [12]. …”

“…The chemical components recently found in leaves of this plant are ( 1 ) N -[7-(3′,4′-methylenedioxyphenyl)-2( Z ),4( Z )-heptadienoyl]pyrrolidine, ( 2 ) N -[7-(3′,4′-methylenedioxyphenyl)-2( E ),4( E )-heptadienoyl]pyrrolidine [12], ( 3 ) N-[3-(6′-methoxy-3′,4′-(methylenedioxyphenyl)-2(Z)-propenoyl]pyrrolidine, ( 4 ) N-[3-(6′-methoxy-3′,4′-methylenedioxyphenyl)-2( E )-propenoyl]pyrrolidine [10], ( 5 ) lupeol, and ( 6 ) vitexin [13]. …”

“…In this sense, supercritical fluid extraction is safer for the human being, due to the reduction of the volume of organic solvent and extraction time [16]. Authors have been extracting alkaloids semipurified by supercritical fluid extraction [17–19], and we have extracted the pyrrolidine alkaloids with better yield than chloroform [12]. Considering the important biological activities of compound 1 , it is indispensable to evaluate the factors which influence its extraction using supercritical carbon dioxide.…”

Comportamento De Fases Do Sistema Binario Dióxido De Carbono + Cumarina a Altas Pressões

“…Além dos significativos rendimentos observados neste tipo de extração Gonçalves et al, 2013;Lemos et al, 2012), uma grande vantagem dessa tecnologia é a produção de um princípio ativo livre de solventes residuais. Na literatura especializada são vários os trabalhos que reportam a utilização de gases (CO 2 , propano e etileno) a altas pressões na extração de óleos e princípios ativos de oleaginosas (Carrara et al, 2011;Gonçalves et al, 2013;Jesus et al, 2013). Diante da possibilidade de utilização de fluidos supercríticos na extração da cumarina de diversas fontes vegetais, torna-se de extrema importância o estudo da solubilidade deste princípio ativo em fluidos supercríticos, visando assim, a determinação das condições ótimas de extração da cumarina.…”