The phytochemical analysis and biological activities assessment of secondary metabolites in callus cultures of Clitoria ternatea L.

The butterfly pea is a significant source of bioactive secondary metabolites. Callus cultures provide a viable option for the reliable synthesis of important secondary metabolites, overcoming the limitations of inconsistent yields from field-grown plants. The present results indicated that MS medium with 1.0 mg/L BAP and 1.0 mg/L 2,4-D produced a highly significant growth stimulation for callus induction in Clitoria ternatea L. after 40–45 days under darkness. The biomass of the callus augmented with each subsequent subculture, reaching its peak by the sixth subculture, where the biomass increased about 11.8-fold. The micronutrient content, antioxidant assays, flavonoids, phenolics, and HPLC analysis were assessed. Anti-diabetic, anticancer, and anti-inflammatory characteristics were evaluated. The results of total phenolics and total flavonoids were 81.37 ± 0.65 and 4.63 ± 1.03 mg/g DW, respectively, and total antioxidant activities, including DPPH and ABTS, were 15.52 ± 1.07 and 2.32 ± 0.09 mg/g DW, respectively. HPLC analysis revealed that Gallic acid, Chlorogenic acid, and Rosmarinic acid were the highest compounds with concentrations of 346.92, 329.22, and 188.64 µg/g, respectively, which correlated with strong anti-inflammatory and cytotoxic activities in vitro. Also, IC50 for α-amylase and α-glucosidase for C. ternatea L. callus culture was 850.5 ± 0.54 and 198.63 ± 0.67 µg/mL, respectively. Moreover, HYA, Lipoxygenase, and Proteinase Inhibitions of C. ternatea L. callus cultures were 53.41 ± 0.66, 47.35 ± 0.98, and 35.37 ± 0.63, respectively. In conclusion, using the C. ternatea L. callus cultures has shown promise in secondary metabolite synthesis. Moreover, the HPLC analysis of callus cultures of C. ternatea L. comprises a range of flavonoid and phenolic compounds that demonstrated the most anti-diabetic, anti-inflammatory, and cytotoxic effects, in addition to their potential application as a natural source of secondary metabolites.