• Valmir Gomes De Souza Department of Pharmaceutical Sciences, Medicine Development and Assays Unified Laboratories, Federal University of Paraíba, João Pessoa, PB, Brazi
  • FabrÍcio Havy Dantas De Andrade Department of Pharmaceutical Sciences, Federal University of Pernambuco, Recife-PE, Brazil
  • Fabio Santos De Souza Department of Pharmaceutical Sciences, Medicine Development and Assays Unified Laboratories, Federal University of Paraíba, João Pessoa, PB, Brazil
  • Rui Oliveira Macedo Department of Pharmaceutical Sciences, Federal University of Pernambuco, Recife-PE, Brazil


Objective: The Anadenanthera colubrina (Vell.) Brennan var. cebil is a medicinal plant that has been used for the treatment of many diseases in the northeastern region of Brazil. This plant contains secondary metabolites such as quercetin, a flavonoid that is known by its antioxidant and anti-inflammatory effects. The aim of this work is to propose the validation of an analytical method using high-performance liquid chromatography with diode array detector (HPLC-DAD) for the quantification of quercetin and standardization of the hydroalcoholic extract (HAE) of A. colubrina.

Methods: The A. colubrina extracts were prepared by the maceration process with powdered leaves at 20% weight: volume (w/v) and a hydroalcoholic solution at 50% volume: volume (v/v) for 120 h at room temperature. After pretreatment of the hydroalcoholic extract, the quercetin marker was used for quantification and proceeded to the evaluation of validation parameters for the method using HPLC-DAD.

Results: The analytical method proved to be specific. Linear over the range 1.4–26.6 µg/ml, regression analysis showed a good correlation coefficient (R2= 0.999); the limit of detection (LOD) and the limit of quantification (LOQ) were 0.27 and 0.81 μg/ml respectively. The relative standard deviation (RSD) did not exceed 2.5% for precision. The proposed method was validated with an average recovery of 92.5–97.5%.

Conclusion: The method was validated using HPLC-DAD, allowing the quantification of quercetin in the standardisation process of extracts and quality control of the herbal drug containing A. colubrina Phyto complex.

Keywords: Validation, Analytical, Method, Quercetin, Flavonoid, Anadenanthera colubrina


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How to Cite
Souza, V. G. D., F. H. D. D. Andrade, F. S. D. Souza, and R. O. Macedo. “ANALYTICAL METHOD BY HPLC-DAD ALLOWS QUANTIFICATION OF QUERCETIN MARKER IN STANDARDIZED EXTRACT OF ANADENANTHERA COLUBRINA VAR. CEBIL”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 9, no. 8, Aug. 2017, pp. 47-55, doi:10.22159/ijpps.2017v9i8.16468.
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