## 1 Introduction
Biobased solvents, sourced from renewable materials like plant biomass, encompass a range of options, including hydrophilic solvents such as glycerol, ethyl acetate (EtOAc), and ethyl lactate, as well as more hydrophobic alternatives like D-limonene, 2-methyltetrahydrofuran (2-MTHF), and cyclopentyl methyl ether (CPME), among others [15; 16; 17]. These solvents are widely utilized for extracting phenolic compounds and terpenoids from various matrices [15; 16; 18]. In some cases, they have been paired with advanced extraction methods, including ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), and pressurized liquid extraction (PLE), to enhance the recovery of phenolic compounds and terpenoids from natural sources [15; 16; 19]. Particularly, ethyl lactate has been used in combination with PLE for the recovery of phenolic compounds like rosmarinic acid and caffeic acid from _Ocimum basilicum_ L., while D-limonene has been used for the PLE extraction of terpenoids from _Ruta graveloens_ L. [15; 20]. Despite their potential to obtain higher extraction yields than conventional solvents, biobased solvents have not been widely investigated in the extraction of phenolic compounds from natural plants [15; 16; 19]. In fact, there are no studies on the extraction of phenolic compounds by PLE with biobased solvents from burdock roots, even using conventional solvents. In addition, little attention has been paid to the non-polar compounds, like terpenoids, that can be co-extracted from burdock roots when hydrophobic solvents such as 2-MTHF or CPME are used. On the other hand, the presence of secondary metabolites in burdock roots depends on the growing conditions or the plant treatment at the place where the plant is collected. It has been reported that phenolic compounds and terpenoids are synthesized by plants in response to stress, playing key roles in the defense against biotic and abiotic factors [21; 22; 23]. They are involved in protecting against herbivores, pathogens, UV radiation, drought, and extreme temperatures. Particularly, different researchers have observed that weeding treatments or altitude can influence the content of bioactive compounds in plants [24; 25; 26]. However, to our knowledge, there are no studies that determine the influence of factors such as altitude and weeding of burdock on the content of bioactive compounds in its roots. Their study can help understanding how these factors affect burdock cultivation, enhance the quality of its derived products, and promote agricultural sustainability. Considering the above, this study addresses the issue of the limited research on obtaining neuroprotective compounds from burdock roots, which has primarily focused on their phenolic composition. Little attention has been given to the extraction of non-polar compounds, such as terpen, and most studies have relied on conventional extraction methods using solvents like ethanol and methanol with water. Additionally, no previous studies have explored how growth conditions, such as altitude and weeding, influence the composition and bioactivity of burdock roots. Taking this into account, the hypothesis of this study is that PLE with biobased solvents will provide higher yields of antioxidant and neuroprotective compounds compared to conventional extraction and that growth at higher altitudes without weeding will enhance the production of bioactive compounds. Therefore, this work proposes, for the first time, the development of a sustainable extraction methodology for the extraction of antioxidant and anticholinergic non-polar and polar compounds from burdock roots based on the combination of biobased solvents with PLE. A simplex-centroid experimental design was used to determine the optimal mixture proportion of biobased solvents by using CPME, 2-MTHF, and EtOAc to obtain the richest extracts in phenolic compounds according to their total peak areas determined by HPLC-DAD.