Development of an analytical method by HPLC for quantification of galloylquinic compounds from Copaifera langsdorffii leaves

Copaifera langsdorffii Desf. (Fabaceae, Caesalpinioideae), popularly known as “copaiba”, is a large tree widely distributed in Brazil and used in folk medicine. Its leaves are composed mainly by phenolic compounds, such as galloylquinic acid derivatives (GQA) and flavonoids. The water soluble fraction of the crude hydroalcoholic extract was submitted to Sephadex LH-20 using a gradient of methanol-water (1:9 to 9:1) and purified by preparative RP-HPLC. Sixteen GQA were isolated and used as chromatographic standards. HPLC-PDA analyses were carried out on a Synergi Polar-RP (100 × 3.0 mm, 2.5 µm) column. The mobile phase was made up of formic acid-water (0.1:99.9, solvent A), and isopropanol-methanol-acetonitrile (0.5:4:6, solvent B). The elution gradient was A:B (90:10 to 85:15) in 8.0 minutes, followed by A:B (85:15 to 64:36) up to 30.0 minutes using a flow rate of 0.7 mL/min. The HPLC-PDA developed method was suitable to quantify nine GQA and two flavonoids, quercitrin and afzelin.

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