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Abstract
Kalanchoe brasiliensis, known as “saião” and “coirama branca,” is native to Brazil. Although this species is widely used in folk medicine, there is no quantification method of chemical markers to it. To characterize and quantify the chemical markers of hydroethanolic extract from leaves of the K. brasiliensis, the study was divided into two parts: 1) isolation and identification of one chemical marker of the hydroethanolic extract from K. brasiliensis leaves; and 2) development and validation of an analytical methodology to quantify it via HPLC–DAD. The hydroethanolic extract from K. brasiliensis leaves was subjected to column chromatographies using different mobile phases, resulting in the pure compound named Kb1. The Kb1 compound was identified as the flavonoid, patuletin 3-O-α-L-rhamnopyranosyl-7-O-α-L-rhamnopyranoside, through analysis by UV spectrum and MS–MS. The most appropriate chromatographic system by HPLC analysis was as follows: phase A, water:formic acid (99.7:0.3, v/v) and phase B, methanol:formic acid (99.7:0.3, v/v), with an elution gradient of 40% B–58% B in 50 min, a C18 Hichrom (250 × 4.0 mm, 5 µm) column, a flow rate of 0.8 mL/min, and chromatogram recorded at 370 nm. The method proved to be linear, precise, accurate, and reproducible. According to the results, it was observed that patuletin 3-O-α-L-rhamnopyranosyl-7-O-α-L-rhamnopyranoside can be used as a chemical marker of hydroethanolic extract from K. brasiliensis leaves.