Purification of Lepidilines A, B, C, and D from Lepidium meyenii Walpers by Centrifugal Partition Chromatography Followed by Semi-Preparative HPLC and Preliminary Evaluation of Anticancer Activity Against Neuroblastoma Cell Lines

<i>Lepidium meyenii</i> Walpers (syn. <i>Lepidium peruvianum</i> Chacon) has been cultivated for centuries in the Peruvian Andes as both a vegetable and a traditional medicine resource. Maca is classified as a superfood and is widely used as a dietary supplement, particularly noted for its potential to enhance endurance, fertility, and endocrine balance. In recent years, there has been a growing interest in the cytotoxic effects of lepidilines and their derivatives; however, these compounds have been less extensively studied due to challenges associated with their isolation. This study aims to establish optimal extraction conditions to enrich lepidiline content in the extracts and to propose an efficient isolation method for four lepidilines using a green purification technique known as Centrifugal Partition Chromatography (CPC). The isolated compounds will be evaluated for their anticancer potential utilizing the MTT assay on SK-N-SH (ATCC^® HTB-11™) and SK-N-AS (ATCC^® CRL-2137™) neuroblastoma cell lines. The findings indicate that Soxhlet extraction with dichloromethane resulted in the highest recovery of lepidilines, with a content of 10.24% expressed as lepidiline A. The optimal biphasic solvent mixture suitable for CPC chromatographic applications was identified as a combination of chloroform, methanol, and water (4:3:2 <i>v</i>/<i>v</i>/<i>v</i>) containing 60 mM HCl. When utilized in conjunction with semi-preparative high-performance liquid chromatography (HPLC), this method successfully isolated lepidilines A-D, achieving a purity exceeding 95%. Notably, lepidiline B exhibited the highest cytotoxic potential, with an IC50 value of 14.85 µg/mL in SK-N-AS cells.