2011 - Lia Akina Ito
MSc. Thesis
Author: Lia Akina Ito
Postgraduate Program in Pharmaceutical Sciences - State University of Maringá
Date of Defense: August 26, 2011
Advisor: Prof. Dr. João Carlos Palazzo de Mello
Examination Board: Prof. Dr. Claudio Celestino de Oliveira
Prof. Dra. Juliana Rodrigues
Title: Development of analytical method by capillary electrophoresis for identification and quantification of cafein and falvan-3-ol in Paullinia cupana var. sorbilis (Mart.) Ducke – Sapindaceae extract
Abstract: Paullinia cupana var. sorbilis (Mart.) Ducke, known as Guarana, is a plant native from Brazilian Amazon. Its seeds are popularly used as a stimulator of brain functions, aphrodisiac, tonic, diuretic, febrifuge and sedative. Guarana has a high content of methylxanthines and considerable amounts of tannins, which are related to its medicinal effects. Already been mentioned a wide range of biological activity, such as antioxidant, antibacterial, antidepressant, antigenotoxic, anti-platelet aggregating and disintegrating, improved cognitive and physical performance, and stimulating. In order to obtain a rapid and accurate methodology to identify the substances responsible for this biological activity, in this study was developed a methodology using capillary electrophoresis for separation, identification and quantification of substances in the semipurified fraction (EPA) of Paullinia cupana var. sorbilis (Mart.) Ducke. Two techniques were employed for separation, the capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC). For CZE was used borate buffer 50 mmol l-1 at pH 8,5 with 1,0 mmol l-1 of β-cyclodextrin, applied voltage 20 kV, detection at 214 nm, sample injection for 3 s at 0,3 psi, capillary temperature of 25 ºC, resulting in separation of the peaks of caffeine, ent-catechin, catechin and epicatechin, however, the electropherogram did not show symmetrical peaks. The methodology used by MEKC was phosphate buffer 40 mmol l-1 at pH 7,0 supplemented with 30 mmol l-1 SDS, applied voltage of 20 kV, detection at 214 nm, sample injection time of 5 s at 0,5 psi and capillary temperature at 25 ° C, showing an electropherogram with good resolution, allowing the identification of the peaks of caffeine, catechin, epicatechin, procyanidin B1, procyanidin B2 and procyanidin B4. The analytical method by MEKC proved to be selective, sensitive, simple and high-frequency, allowing separate analytical reference substances caffeine, catechin and epicatechin in 20 min with migration times of 8,94 ± 0,22, 9,27 ± 0,24 and 18,29 ± 0,77, and limits of detection and quantification of 0,638, 1,465 and 0,965 mg/ml and 2,126, 4,883 and 3,217 mg/ml, respectively. However, the method did not provide robustness to small variations in voltage.
Keywords: Paullinia cupana, guaraná, methylxanthines, tannins, capillary electrophoresis, validation.
Dissertation PDF: Lia Akina Ito
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