Box–Behnken factorial design to obtain a phenolic-rich extract from the aerial parts of Chelidonium majus L.

• First systematic HPLC–DAD–ESI/MSn characterization of phenolics from C. majus.
• Validation of HPLC–DAD method for phenolics quantification.
• Optimization of phenolics extraction by a 4-factor Box–Behnken design.
• Good fitting of the model to the experimental results.
• % MeOH affects both extraction yield and phenolics content.

A Box–Behnken design (BBD) was developed to study the influence of four parameters (X1: % methanol; X2: extraction time; X3: extraction temperature; X4: solid/solvent ratio) on two responses, namely extraction yield and phenolics content of the aerial parts of Chelidonium majus L. The model presented a good fit to the experimental results for the extraction yield, being significantly influenced by X1 and X4. On the other hand a parameter reduction was necessary to run the model for phenolics content, showing that only X1 and X2 had great influence on the response. Two best extraction conditions were defined: X1=76.8% MeOH, X2=150.0 min, X3=60.0 °C and X4=1:100 and X1=69.2%, X2=150 min, X3=42.5 °C and X4=1:100.Moreover, the HPLC–DAD–ESI/MSn analysis conducted with the center point sample revealed the presence of 15 alkaloids and 15 phenolic compounds, from which the 9 flavonoids and 3 hydroxycinnamic acids are described for the first time. Only phenolic compounds were quantified by a validated HPLC–DAD method, the pair quercetin-3-O-rutinoside+quercetin-3-O-glucoside dominating all the 29 extracts. This study is of great importance for future works that seek to apply the phenolic profile to the quality control of C. majus samples.

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