Chlorophytum borivilianum is an important medicinal plant and its tuberous roots are used for various health treatments. The plant regeneration protocol in in-vitro conditions have been developed for this medicinal plant. Best callus was induced when flower buds were cultured on MS medium supplemented with 1.0 mg/L BAP + 1.0 mg/L NAA. The above callus was subcultured on MS medium + BAP 1.0 mg/L + NAA 0.5 mg/L for root differentiation and on MS medium + BAP 1.0 mg/L for shoot differentiation. Metabolic changes occurring during callus differentiation were studied to quantify metabolites. Metabolites like total soluble carbohydrates, free amino acids increase during root and shoot differentiation from calli while reducing sugar decreases.