Abstract
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.
Highlights
Anadenanthera colubrina (Vell.) Brenan var. cebil (Griseb) Altschul
The quercetin standard (Cas 117-39-5) was purchased from Sigma Alldrich®, Brazil, methanol HPLC grade was purchased from Tedia®, USA, orthophosphoric acid was purchased from Merck®, Germany, ethanol 96% was purchased from Toscano®, Brazil, dichloromethane hplc grade and hexane were purchased from Vetec®, Brazil
The standard methods in pharmacopeias are the most indicated for the evaluation quality control of herbal drugs but not exists any reference related to Anadenanthera colubrina [41,42]
Summary
Anadenanthera colubrina (Vell.) Brenan var. cebil (Griseb) Altschul Hydroalcoholic extracts and decoctions of the leaves and bark of A. colubrina are used in the treatment of gastric and respiratory tract infections and others inflammatory diseases. It is classified among one of ten medicinal plants most frequently used by the communities living in the bioma named Caatinga located in northeastern Brazil [5,6]. Preclinical studies of the healing and antimicrobial activities of A. colubrina extracts endorse their popular usage [7,8,9] These assays demonstrated anti-inflammatory mechanisms, such as the decrease of the production of pro-inflammatory cytokines, inflammatory mediators, increased tyrosine kinase production, and healing action by reducing the synthesis of polymeric collagen and acidic glycoproteins that make healing difficult, and contributing to the development of a phytotherapeutic medicinal products with safety and therapeutic efficacy [10,11,12,13]. Research on the quality of a herbal drug is not based solely on clinical effects and on the standardization of plant extracts through quantitative analyses using active ingredients and secondary metabolites called biomarkers [14,15,16]
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