Abstract
Azadirachta indica A. Juss. (Meliaceae) is a plant whose leaves present several pharmacological activities, among them: antioxidant, antimicrobial and hypoglycemic. The present work aims to determine and validate a spectrophotometric analytical method for quantification of total flavonoids, expressed as rutin, in A. indica leaves and to optimize the extraction conditions of these flavonoids. For the selection of the spectrophotometric method, four analytical methods were investigated. The method selected as the most efficient, simple, fast and sustainable has been validated and optimized. The validation was performed according to criteria of national and international regulatory agencies. Box Behnken design (33) and the response surface methodology (RSM) were used to estimate the best extraction conditions. The analytical method that met the study criteria consisted of ultrasound-assisted extraction (UAE) of the plant material in hydroethanolic solution, at a temperature of 60 ºC, for 30 minutes, followed by filtration, dilution of the extract in ethanol/0.02 M acetic acid solution (99:1) and absorbance reading in a spectrophotometer at 364nm. In the validation stages, this method was shown to be linear (linear interval - 20 to 80 μg/mL), precise (intraday and interday), accurate (mean recovery of 120.8%), robust and selective (absence of matrix effect). The best conditions for the UAE process were: Ethanol content 30% (p/p), plant material/solvent ratio 0.2 g/mL and extraction time of 33 minutes. The spectrophotometric method and the UAE conditions determined in this study are suitable for use in the quality control of plant drugs and ethanol extracts from the leaves of Azadirachta indica.
Highlights
Indica leaves For the determination of total flavonoids in A. indica leaves, four spectrophotometric analytical methods were investigated
We considered the practicality of the flavonoid extraction process, the lower energy consumption of the method and the low toxicity of the reagents used in the execution of the steps, as well as the extraction of higher flavonoid contents from A. indica leaves
Response Surface Methodology and Box-Behnken design were used to optimize the extraction process, and the results suggested that homogenate and ultrasonic cavitation system (HUCS) is a green and efficient method for the extraction of flavonoids from C. camphora, as well as other natural plant products
Summary
In the leaves of this species were identified compounds such as: limonoids, flavonoids, tannins, alkaloids and coumarins (Silva Neto et al, 2020), with possible correlation with their biological activities described in the literature. Phenolic compounds, such as flavonoids, can play an important role in antioxidant and hypoglycemic activities. Flavonoids have the ability to nullify reactive oxygen species, such as the superoxide anion (O2-) (Simões et al, 2016) This action may play a role in the treatment of diabetes mellitus, since the onset of insulin resistance is associated with oxidative stress (Sarian et al, 2017)
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