The aim of this study was to screen the antioxidant, anti-hyaluronidase, and anti-elastase activity of the lyophilized juice of Cucumis sativus fruit (CSLJ). The CSLJ was subjected to DPPH and superoxide radical scavenging assay in reference to butylated hydroxytoluene. The hyaluronidase and elastase inhibitory assay was performed in reference to oleanolic acid. Furthermore, the activities have been rationalized with HPLC analysis of the CSLJ with standard reference compound of ascorbic acid. The CSLJ exhibited DPPH-free radical and superoxide radical scavenging activity, IC(50) at a concentration of 14.73 ± 1.42 and 35.29 ± 1.30 μg/mL, respectively. The CSLJ also showed strong anti-hyaluronidase ((c) P < 0.001) and anti-elastase ((c) P < 0.001) activity, IC(50) at a concentration of 20.98 ± 1.78 and 6.14 ± 1.74 μg/mL, respectively. The HPLC 'chromatogram' of standard and CSLJ showed specific peak at retention time 2.905 and 3.066 min, respectively. Content of ascorbic acid was calculated with respect to the standard compound and it was found to be 3.5 ± 0.23% w/w. CSLJ is the rich source of ascorbic acid and this study thereby rationalizes the use of C. sativus as potential anti-wrinkle agent in cosmetic products.