Aloe vera leaf skin (AVLS) is a major byproduct of plant gel treatment and a promising source of bioactive compounds. The present study aims at intensifying the extraction of AVLS using an ultrasound-assisted technique. Central composite design (CCD) protocol as a subset of Response Surface Methodology (RSM) was employed to investigate the interaction of process variables on the amount of aloin A and aloin B extraction from AVLS. The optimum extraction conditions of 30 min sonication time, EtOH 80%, and 65% sonication power were obtained corresponding to maximize extraction of aloin A (4.33 mg.g−1), and aloin B (4.46 mg.g−1), and the extraction yield was 9.43%. It was revealed that the amounts of aloin A and aloin B increased as the extraction yield decreased. To evaluate the cytotoxic effect of the extract on cancer cells MTT assay was performed on A549 cells (a human lung cancer cell line). Our preliminary results indicated that the AVLS extract could inhibit the proliferation of A549 cells (IC50 =166 µg.mL−1). Moreover, compared with conventional methods, ultrasound radiation made effective cracks in the surface structures of the leaf skin. The results of the present study may have contributed to the development of extraction methods with probe-sonic applicable to extract of aloin A and B.