The alkylated cardanol sample was supplied by Grupo de Inovações Tecnológicas e Especialidades Químicas (GRINTEQUI, Brazil) and purified by column chromatography using silica gel as stationary phase, and hexane as eluent (Freitas et al. 2015). AnimalsAlbino mice were used (Mus musculus), Swiss, adults, male lineage, weighing between 25-30 g, 2 months old, from the Central FEDERAL Biologist and accommodated in the Vivarium of LAPNEX the Federal University of Piauí. The animals were packed in plastic cages (35 cm W × 52 cm L × 17 cm H), containing 6 animals per cage, temperature controlled (25 ± 2 °C), with light/dark cycles of 12:00 (06:00 at 18:00), receiving rations type of pellets and water ad libitum. The experimental protocols and procedures were approved by the Committee of Ethics in Animal Experimentation at the Federal University of Piauí, Brazil (Protocol # 417/17). The experiments were conducted in accordance with the Guide for care and use of laboratory of the Department of Health and Human Services of the United States, Washington, DC (1985). Statistical analysisStatistical analysis was performed using unidirectional ANOVA for multiple comparisons and followed by Student-Newman-Keuls as a post-hoc test by GraphPad Prism (version 6.0; GraphPad San Diego, California, USA Copyright © 1994Copyright © -1999. The chi-square test was used to analyse the percentage of animals with seizures and the mortality rate. Differences were considered statistically significant when p < 0.05. Experimental Characterization of alkylated cardanolThe gas-chromatography coupled to mass spectrometry (GC/MS) analysis was performed in a GCMS-QP5050-A spectrometer (Shimadzu, Kyoto, Japan) equipped with an OV-5 column (0.25 mm, 30 m, 0.25 μm film) (Agilent, USA). The initial column temperature of 100 °C was increased to 180 °C at a rate of 40 °C/min and at the sequence was increased to 300 ºC at a rate of 10 °C/min, the column temperature was held at 300 ºC until the end of detection. The injection temperature was set to 250 °C, while interface and ion source temperatures were set to 280 °C. Samples were ionized by an electron impact (EI) ionization method, 70 eV.A high-resolution NMR spectrometer operating at 500 MHz (Bruker Advance DRX-500) was used for the spectra acquisition ( 1 H NMR and 13 C NMR). The samples were prepared dissolving alkylated cardanol in deuterated chloroform (CDCl 3 99.8%, Sigma Aldrich brand). The chemical shifts are expressed in ppm using as internal standard the TMS.
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