Response Surface Methodology IV-Optimal design applied to the performance improvement of an RP-HPLC-UV method for the quantification of phenolic acids in Cecropia glaziovii products

Chlorogenic and caffeic acids are bioactive phenolic compounds present in Cecropia glaziovii Snethl., Urticaceae, products that have been used as analytical markers. This paper reports a chemometric study aimed at improving chromatographic performance for quantification of these markers by RP-HPLC. The organic to aqueous content ratio, the acid content of the mobile phase, and the elution method were analyzed using a Response Surface Methodology IV-Optimal design. The resolution between peaks, retention time, tailing and retention factors, number of theoretical plates and peak widths were evaluated. The optimized conditions were mathematically determined as (A) trifluoroacetic acid 0.05% (v/v), (B) 12% (v/v) acetonitrile and (C) increasing gradient. The method was considered specific, fast, precise, reliable and linear in the ranges of 1.0–200.0 and 2.5–100.0 μg/ml for the chlorogenic and caffeic acids, respectively. The adequate conditions to separate and quantify both phenolic acids in C. glaziovii products were demonstrated. Satisfactory resolution was achieved when compared to a previously published chromatographic method which is unable to separate the chlorogenic acid and an interfering compound presented under certain extractive conditions, demonstrating the importance of systematic studies, specifically when analyzing complex plant matrices.

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