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Chemical characterization of the bulbs of <i>Drimia pancration</i> was conducted to isolate four steroidal saponins (<b>1</b>–<b>4</b>). Earlier, we focused on the structural elucidation of compounds <b>1</b>–<b>3</b>. Herein, by means of ¹H-NMR, ¹³C-NMR, Nuclear Overhauser Effects (NOE), and 2D-NMR spectra, the full stereochemical structure of <b>4</b> is reported, and all the ¹H and ¹³C signals are assigned. Compounds <b>1</b>–<b>4</b> were tested for their acaricidal properties against the two-spotted spider mite <i>Tetranychus urticae</i>. Our results showed excellent activity of compound <b>1,</b> with an LD₅₀ (µg/cm²) of 0.29 and a LD₉₀ (µg/cm²) of 0.96, whereas compounds <b>2</b>, <b>3</b>, and <b>4</b> showed moderate activity. Furthermore, the acaricidal and cytotoxic properties of the crude extract were also investigated. Of note, after 96 h of exposure, the acaricidal activity of compound 1 was higher than that of the positive control, hexythiazox. Indeed, for compound <b>1</b>, LD₅₀ and LD₉₀ were 0.29 and 0.96 µg/cm², respectively, while hexythiazox LD₅₀₍₉₀₎ was 18.7 (132.5) µg/cm². Additionally, <i>D. pancration</i> extract, after 72 h, induced a high cytotoxic effect in HaCaT and THP-1 cell lines, with an IC₅₀ of 7.37 ± 0.5 µg/mL and 3.50 ± 0.15 µg/mL, respectively. Overall, <i>D. pancration</i> can be considered as a green source of novel acaricides effective against mites of agricultural importance, such as <i>T. urticae</i>, pending proper field validation and the assessment of non-target effects on other invertebrate species.