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In this study we aimed to compare the chemical composition and biological activity between <i>Morus alba</i> L. leaf extract obtained with 95% <i>v</i>/<i>v</i> ethanol using a pulsed electric field (PEF) and the conventional maceration method. Extracts of <i>M. alba</i> leaves collected from Chiang Mai (CM), Sakon Nakon (SK), and Buriram (BR), Thailand, were investigated for 1-deoxynojirimycin content by high-performance liquid chromatography and for total phenolic content by the Folin–Ciocalteu method. Antioxidant activity was investigated by 2,2′-diphenyl-1-picrylhydrazyl (DPPH), 2,2’-azinobis-3-ethylbenzothiazoline-6-sulphonate (ABTS), and ferric reducing antioxidant power (FRAP) assay. Anti-tyrosinase and anti-hyaluronidase activity was investigated by in vitro spectrophotometry. The results show that this is the first study to indicate PEF as a novel method for enhancing the phenolic content and antioxidant, anti-tyrosinase, and anti-hyaluronidase activity of <i>M. alba</i> leaf extract (<i>P</i> < 0.05). PEF extract of <i>M. alba</i> leaves collected from BR had comparable ABTS^•+ scavenging activity to <span style="font-variant: small-caps;">l</span>-ascorbic acid and comparable anti-tyrosinase activity to kojic acid (<i>P</i> > 0.05). On the other hand, PEF extract of <i>M. alba</i> leaves collected from SK exhibited significantly high anti-hyaluronidase activity, comparable to that of oleanolic acid (<i>P</i> > 0.05). Therefore, PEF is suggested for further <i>M. alba</i> leaf extraction in the production of natural whitening and anti-aging cosmetic ingredients.