Successive extracts guided isolation of plumbagin from Plumbago zeylanica L. in the course of development of marker compound

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BACKGROUND: Certain phytochemical compounds that act as therapeutic or chemical markers play a crucial role in the identification of raw drugs or formulations in the course of standardization and quality control. The isolation and development of marker compounds are important. In this study, the isolation of plumbagin was aimed at the development of a phytochemical marker compound from Plumbago zeylanica L. METHODS: The P. zeylanica L. root was purchased from the crude drug market from Chennai, authenticated at Captain Srinivasa Murthy Central Ayurveda Research Institute (CSMCARI), Chennai, Tamil Nadu, India. Successive extraction of Soxhlet was carried out, and high-performance thin layer chromatography (HPTLC) profiles and thin layer chromatography (TLC) array were developed and documented at UV 254 nm. The bulk sample was extracted through the cold maceration method. The crude extracts of n-hexane and chloroform were combinedly loaded on the silica-gel column for separation and isolation of the marker compound. Various spectral data were recorded using instruments such as UV, IR, 1H and 13C nuclear magnetic resonance (NMR), electrospray ionization-mass, HPTLC, and high-performance liquid chromatography for further confirmation of the isolated compound. RESULTS: Isolation was carried out through the separation of components of combined crude extracts of n-hexane and chloroform through chromatography over column loaded with silica gel using a mixture of n-hexane and chloroform as a mobile phase in a gradient manner. Finally, the yellow orange color solid compound (0.09% yield W.R.T. raw material) was eluted in a mobile phase of n-hexane: chloroform (6: 4, v/v). The eluted compound was cross-verified with the R f of plumbagin on co-TLC and further confirmed by spectral data such as UV, IR, 1H and 13C NMR, and EI-Mass and are found according to the reported data. CONCLUSION: The developed TLC and HPTLC profiles of successive extracts are used for identification of P. zeylanica L. in the course of standardization. The developed profiles and observed data for all spectra are compared with the data reported in the literature and found to be correlated. Thus the isolated compound is confirmed as Plumbagin.