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| Author(s): |
Juhan Augusto Scardelato Pereira
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Araraquara. 2017-05-03. |
| Institution: | Universidade Estadual Paulista (Unesp). Faculdade de Ciências Farmacêuticas. Araraquara |
| Defense date: | 2017-03-31 |
| Advisor: | André Gonzaga dos Santos |
| Abstract | |
Cordia verbenacea DC. belongs to the Boraginaceae family which contains about 100 genera with more than 2,000 species. C. verbenacea is popularly named erva-baleeira and occurs throughout the Brazilian coast including the Atlantic Rainforest. It is traditionally used to treat various inflammatory processes and was employed by Aché® laboratory for the development of Acheflan®, an anti-inflammatory topical medicine containing the essential oil of C. verbenacea. Considering the relevance of chemical variability studies for medicinal plants we propose to develop a study of the metabolism dynamics focused on seasonal, circadian and population variations. The leaves of C. verbenacea were collected, dried, crushed and macerated with ethanol (1:15 m/v). The solid phase extraction with ethanolic extract yielded 10 fractions which were analyzed by TLC and HPLC. Fraction 4.1 was selected for column chromatography yielding 120 subfractions. Four compounds were isolated using preparative HPLC-UV and three compounds presented high purity (≤95 %; HPLC-UV) and were analyzed by spectrophotometric techniques (UV, IR, NMR). Compounds were identified as the triterpenes cordialin A and cis-cordialin A and the flavonoides 4',5-dihydroxy-2',3,5',6,7-pentametoxiflavone or 2',5-dihydroxy-4',3,5',6,7-pentametoxiflavone and artemetin. An HPLC-DAD method was developed for the analysis of the ethanolic extract. The ethanolic extracts of seasonal, circadian and population studies were analyzed by HPLC-DAD. All these studies showed quantitative differences in relation to the majority compounds, being the substance dihydroxypentamethoxyflavone found in a larger quantity in specimen 13II4 of the population study, already in the circadian study the time of 04:30 h had a higher yield and in the seasonal study a greater trend of production of this flavonoid in the period from january to march of 2015/2016, while for the triterpenes the specimen 3IV1 presented higher yield in the population study and the specimens 2II5 and 3II5 greater yield in the circadian and seasonal studies, being the periods of march to may of 2015 and january to march of 2016 the ones of greater tendency of production of these triterpenes. The statistical analysis using ANOVA showed that the yield of the extract and the content of triterpenes and flavonoid were affected by the periods of the year. Essential oils were obtained from the 27 specimens from the population study and the seven specimens from the circadian and seasonal studies. The essential oil content in the plant drug and its color (light yellow, yellow, green and blue) differed among the specimens. The TLC analysis also showed significant differences among the oils. Data obtained by TLC analyzes of the 27 specimens of the population study were used in PCA and HCA analyzes that showed two groups of oils - blue / green oil specimens and yellow color oil specimens. These clusters were also verified in PCA and HCA obtained with GC-MS data, evidencing that the sample of the 13I5 of the specimen has a greater trend of production of trans-caryophyllene and α-humulene. The blue/green oils showed the presence of 3 specific compounds: elemene, camazulene and spatulenol. The statistical analysis using ANOVA showed that the yield of the essential oil and the content of its components were affected by the periods of the year, but not by circadian rythm. (AU) | |
| FAPESP's process: | 13/24298-9 - Evaluation of secondary metabolism variability in Cordia verbenacea DC |
| Grantee: | Juhan Augusto Scardelato Pereira |
| Support Opportunities: | Scholarships in Brazil - Doctorate |