Comparative HPLC Analysis of Bioactive Compounds from Stem, Root and Callus of Caralluma stalagmifera C.E.C. Fisch

Objectives: This study aims to conduct HPLC profiling of the stem, root and callus extracts of C. stalagmifera in order to identify, evaluate and compare their bioactive phytoconstituents, particularly pregnane glycosides, across both natural and in vitro derived plant callus. Method: In this study, the stem, root and callus tissues of C. stalagmifera were collected, dried, powdered, and extracted using suitable solvents. The extracts were then analyzed using High Performance Liquid Chromatography (HPLC). A standard reference compound (epoxy- polyoxypregnane glycosides) was used for comparison. The chromatographic profiles obtained from the stem, root and callus extracts were examined and compared based on their retention times and peak intensities to identify common and unique bioactive phytoconstituents and to evaluate similarities between natural and in vitro samples. Findings: The HPLC analysis revealed distinct chromatographic profiles for the stem, root and callus extracts of C. stalagmifera, confirming the presence of bioactive pregnane glycosides across all samples. This was evidenced by a prominent peak in the standard compound at a retention time of 3.521 minutes, which closely matched the major peaks observed in the stem (3.562 min) and root (3.540 min) extracts. Furthermore, variations in peak intensity and the presence of minor peaks indicated differential metabolite accumulation influenced by tissue type and physiological state. These results confirm that tissue culture techniques have potential for biotechnological applications and providing a sustainable alternative to wild plant harvesting. Novelty: This is one of the first comparative HPLC studies on C. stalagmifera, simultaneously analyzing stem, root and callus extracts. It demonstrates that in vitro callus cultures can biosynthesize bioactive pregnane glycosides, similar to those found in natural tissues. The study provides a unique phytochemical fingerprint of Caralluma stalagmifera which is useful for species authentication and standardization. Keywords: Caralluma stalagmifera, Callus, HPLC analysis, Bioactive Compounds, Pregnane glycosides