The plant Nothapodytes nimmoniana belongs to the Icacinaceae family, and is a significant source of camptothecin (CPT), an effective quinoline alkaloid with an array of pharmacological benefits including anti-cancer, anti-HIV, antibacterial, and antioxidant etc. The elicitation method is the most effective strategy to increase and maintain the synthesis of particular compounds in vitro culture. In this study, to understand the impact of various abiotic elicitors on CPT production by using established N. nimmoniana cell suspension culture obtained from friable leaf callus grown in Murashige & Skoog basal media with sucrose and agar augmented with 2 mg/L of 2,4-dichloro phenoxy acetic acid (2,4-D). A total of six abiotic elicitors were used such as cadmium chloride, ethanol, salicylic acid, tryptophan, copper chloride, and calcium chloride. The impact of these abiotic elicitors on the CPT yield at three different concentrations by cell suspension was evaluated for up to 15 days. High-performance liquid chromatography (HPLC) was used to quantify the CPT production. A maximum of 9.43 and 6.26-fold increment was recorded in CPT production in suspension culture elicitated with 2 mM of tryptophan and 1 mM of tryptophan on the 12th day of inoculation followed by ethanol 7.5% treated cell suspension which enhanced the CPT production up to 1.44-fold increase while compared to control. The cell viability of the suspension culture treated with calcium chloride and cadmium chloride was observed in decreased biomass as per the increment in the concentration. The findings of the current study indicate the potential use of abiotic elicitation as another possible option for boosting up the synthesis of CPT in N. nimmoniana cell suspension cultures.