2.3. Laboratory analyses
To evaluate the concentration of TSPCs and AOA, the samples were extracted according to the modified methodology of Iqbal et al., 2005 and Pérez-Jiménez and Saura-Calixto, 2005. A one-gram sample was homogenized with 20 mL of 80% methanol in a 50 mL Falcon tube and agitated for 1 h at room temperature. After this period, the sample was centrifuged for 10 min at 3000 rpm, and the supernatant was separated. To the remaining residue, 20 mL of 80% methanol (pH 2.0) was added, and the same procedure of agitation, centrifugation and separation of the supernatant was completed. To this residue, 20 mL of 70% acetone was added, and the same procedure of agitation, centrifugation and separation of the supernatant was completed. The three supernatants were mixed and used for the analysis of TSPCs and AOA. The extractions were performed in triplicate.
2.3. Laboratory analysesTo evaluate the concentration of TSPCs and AOA, the samples were extracted according to the modified methodology of Iqbal et al., 2005 and Pérez-Jiménez and Saura-Calixto, 2005. A one-gram sample was homogenized with 20 mL of 80% methanol in a 50 mL Falcon tube and agitated for 1 h at room temperature. After this period, the sample was centrifuged for 10 min at 3000 rpm, and the supernatant was separated. To the remaining residue, 20 mL of 80% methanol (pH 2.0) was added, and the same procedure of agitation, centrifugation and separation of the supernatant was completed. To this residue, 20 mL of 70% acetone was added, and the same procedure of agitation, centrifugation and separation of the supernatant was completed. The three supernatants were mixed and used for the analysis of TSPCs and AOA. The extractions were performed in triplicate.
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