Herbal products are preferable to synthetic medicines, and the use of traditional medicines is increasing day-by-day. The current study was designed to evaluate the potentials of bioactive compounds from Citrullus colocynthis by performing FTIR, HPLC, and GC-MS analyses, which explore the good concentration of the secondary metabolites, such as gallic acid (74.854 ppm), vanillic acid (122.616 ppm), and ferulic acid (101.045 ppm) with considerable bioactivities. Antimicrobial protein was estimated by performing SDS-PAGE, ranging from 15 to 70 kDa in all protein fractions. The current study also checked the cytotoxicity of the bioactive compounds in the active fraction of C. colocynthis, and to perform this activity, the groups of rats were arranged with 16 rats randomly divided into four groups (three experimental and one control) by administering various dosage of methanolic fractions in dose-dependent manner. Histopathology was conducted on the livers of the rats after 15 days of sacrifice under deep anesthesia. In liver cell slides examined at the maximum dose of 600 mg/kg, minimal morphological changes, such as slight ballooning, nuclear variation, vacuolar degeneration, and hydropic degeneration, were observed. Furthermore, the in silico analysis identified bioactive compounds as potential drug candidates.

Medicinal plants have been extensively exploited for their immense pharmacological and immune-supporting potential. Fruit of Citrullus colocynthis has several active secondary metabolites such as phenolics, flavonoids, and essential oils that are used in traditional medicines as antidiabetic, anti-inflammatory, antioxidant, and antimicrobial agents. In this study, phytoconstituents in organic fractions ( n-hexane, chloroform, and ethyl acetate) of the methanolic extract of C. colocynthis were analyzed and identified by FT-IR, HPLC, and GC-MS analysis. Ethyl acetate fraction showed the highest antioxidant scavenging (76 ± .769%) and anti-inflammatory (40 ± .473%) activities at the concentration of 3 mg/mL. Similarly, antidiabetic effect was measured by inhibition of α-amylase where, ethyl acetate fraction (77 ± .844%) exhibited the highest antidiabetic activity. Among all organic fractions, ethyl acetate exhibited strong antimicrobial potential followed by n-hexane and chloroform fractions against selected pathogenic bacteria. Various concentrations of the ethyl acetate extract were tested in-vivo for cytotoxicity and results indicated minor morphological changes in liver cells including ballooning, fatty droplets, and slight accumulation of extracellular matrix even at concentrations of 400 mg/kg. In-silico study showed that stigmasta-7,16-dien-3-ol had a strong interaction with COX-1 and COX-2 to reduce inflammation. The abovementioned results indicate the pharmacological strengths of C. colocynthis to fight several diseases.