2010 - Fernanda Giacomini Bueno
MSc. Thesis
Author: Fernanda Giacomini Bueno
Postgraduate Program in Pharmaceutical Sciences - State University of Maringá
Date of Defense: October 01, 2010
Advisor: Prof. Dr. João Carlos Palazzo de Mello
Examination Board: Prof. Dra. Elaine Rosely Lepri
Prof. Dra. Gisely Cristiny Lopes
Title: Quality control, development of analytical methodology and evaluation of the healing activity of Caesalpinia peltophoroides BENTH
Abstract: Caesalpinia peltophoroides, known as sibipiruna, is a widely used plant in Brazilian cities for afforestation. This work conducted a pharmacognosy study, antioxidant capacity and tannin content of the crude extract (CE) and semipurified fractions (EAF, AF and F1-F5), the analytical methodology development using a spectrophotometer UV/VIS, beyond evaluating the healing activity of the bark of crude extract of C. peltophoroides in Wistar rats skin wounds. The quality control was performed by loss on drying (LD), ash content (AC), extractives content (EC) and dry content with alcohol 100% (DR1), alcohol 70% (DR2) and alcohol 50% (DR3). The total tannins (TT) content was used the reaction with Folin- Ciocalteu. The antioxidant capacity was determined by DPPH assay, determining the IC50 (μg/mL). The analytical methodology development followed the requirements demanded by RE nº 899/2003-ANVISA. The wound healing activity was analyzed at 4, 7, 10, and 14 days of treatment with gel containing 1% of the CE, through the length and thickness of reepithelialized area, photoacoustic spectroscopy permeation analysis, number of cells in\ metaphase and collagen fibers (type I and III). The bark of the vegetal drugs showed values of LD=8,25%, AC=8,66%, EC=7,59%, DR1=10,90%, DR2=9,83% and DR3=14,0%. The TT content was determined in the samples: CE=16,80%, EAF=18,62%, AF=10,27%, F1=28,23%, F2=20,02%, F3=30,40%, F4=40,92% and F5=23,05%. The samples antioxidant capacity tested were: CE=7,40, FAE=4,90, FA=9,57, F1=2,95, F2=3,95, F3=4,10, F4=2,38, F5=4,42, and standards Trolox=3,97 and Vitamin C=4,36. F1 and F4 demonstrated a IC50 lower than Trolox and F1, F2, F3, and F4 lower than Vitamin C (P<0,05). In the analytical methodology development, all the specifications of RE nº 899/2003 were applied. The wound healing activity evaluation showed the formation of a bigger re-epithelialized area in 4 days, and thicker in 10 and 14 days, as well as the stimulation of maturation of collagen fibers (type I) in 10 days and metaphases cell proliferation in 4 and 7 days. The quality control tests were in according with the pharmacopoeic specifications. The fractions (F1-F5) showed a higher amount of TT and therefore a higher antioxidant capacity than the CE and EAF. The analytical methodology development it has been proved that the method for determining the total polyphenol content in CE is valid and reliable, and pyrogalol may be used as standard. In the wound healing activity evaluation, the permeation of the CE was proved in all analyzed times and the gel containing CE showed action on the tissue repair, providing conditions that favor a faster and resistant healing in14 days of treatment, compared to the control.
Keywords: Caesalpinia peltophoroides, Polyphenols, Antioxidant capacity, analytical Methodology development using a spectrophotometer UV/VIS, Wound healing.
Dissertation PDF: Fernanda Giacomini Bueno
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