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Abstract
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inefficient extraction limit pharmaceutical development. In this study, we introduce, for the first time, a lowvoltage
electric field (LV-EF) method for extracting bioactive M¨obius cyclotides from Viola ignobilis. The key
variables tested were voltage (20, 40, 60 V) and electrode gap (1.0–2.5 cm). Optimal conditions (20 V, 1.5 cm;
field strength 13.33 V/cm) significantly improved cyclotide yield compared to conventional maceration and
ultrasound-assisted extraction. Molecular dynamics simulations revealed that the electric field strengthens
hydrogen-bonding between cyclotide Vigno 5 and the acetonitrile/water/formic acid solvent system, providing a
mechanistic basis for enhanced extractability. Scanning electron microscopy confirmed greater cellular disruption
under LV-EF. The optimized extract showed potent and selective cytotoxicity against PC-3 prostate cancer
cells (IC₅₀ = 47.61 μg mL 1) via apoptosis, with minimal toxicity to normal L929 cells (IC₅₀ ≥ 300 μg mL 1).
Practical implications include a scalable, low-cost, and green extraction platform for peptide-based therapeutics.
Future work should focus on continuous-flow LV-EF systems and isolation of individual cyclotides for structureactivity
studies. This establishes LV-EF as a superior technology for producing high-quality bioactive cyclotide
extracts.
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