LC-DAD-MSn and HR-LC-QTOF analysis of Ziziphus budhensis leaves and evaluation of their in vitro and in vivo biological activities.

Ziziphus budhensis is a species described in 2015 in Nepal poorly studied for its phytochemical composition and possible bioactivities. In this study, six extracts were obtained from Z. budhensis leaves using ultrasound assisted extraction with solvents in increasing polarity. The phytochemical contents, along with the antioxidant, antibacterial, and antifungal capabilities, were evaluated.. Chemical composition was studied combining LC-DAD-MSn and LC-HR-MS-QTOF. Fourty-six secondary metabolites belonging to the classes of phenolics, benzyl-isoquinolinic alkaloids, cyclopeptide alkaloids, triterpene aglycone and saponins were identified. Quercetin 3-O-β-neohesperidoside (179.63 mg/g), quercetin-3-O-rutinoside (123.20 mg/g), and quercetin-3-O-rhamnoside (116.62 mg/g) were the most abundant in the methanol extract while quercetin 3-O-β-neohesperidoside (236.06 mg/g), medicagenic acid (192.80 mg/g) were the most abundant in acetone extract and oleanolic acid (163.14 mg/g) was the most abundant in dichloromethane extract. Extracts were tested against gram-positive (Bacillus subtilis and Staphylococcus aureus), gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae), and Candida albicans. Brine Shrimp Lethality Test was used to evaluate the toxicity of the Z. budhensis leaves extracts. In vivo acute oral toxicity tests on mice was performed on extracts and showed no toxic effects. The findings of this study advocate for further in-depth research into the use of Z. budhensis leaves for medicinal purposes.