Andrographis paniculata (Family: Acanthaceae) locally known as Kiriyattu or Kalmegh, finds its use in various Ayurvedic formulations. Andrographolide is the major constituent extracted from the leaves of the plant which is a bicyclic diterpenoid lactone. Isolation and structural elucidation by spectral analysis of any plant active constituent from ethanolic extract of Andrographis paniculata with the aid of DPPH radical scavenging assay by column chromatography. Ethanolic extract was subjected to activity guided column chromatographic fractionation using methanol as eluent. 200 fractions of 5 ml each were collected and were screened for free radical scavenging activity using DPPH (2, 2 diphenyl 1-picryl hydrazyl) radical scavenging assay. The consecutive fractions showing highest similar activity were pooled to obtain pooled fractions (Pf), concentrated and named as Pf1-Pf10. Pfs were subjected to TLC (Thin-layer chromatography) and among all of these, Pf 10 gave a single spot. UV (Ultraviolet)-Visible spectra of Pf 10 showed a single peak at 324 nm. The FT-IR (Fourier transform Infrared) and NMR (Nuclear Magnetic Resonance as HNMR) Spectra of Pf 10 were carried out and the structure was elucidated using www.nmrdb.org software. UV-Visible �» max (H2O) at 324 nm and IR (KBr) cm-1 exhibited bands at 3242, 2920, 1734 and HNMR (CDCl3) �´ : 0.896, 0.904, 1.247, 6.425, 7.226, 7.229. By combining the spectral results and with the help of the software from www.nmrdb.org, which helps in revealing HNMR results from the structure, the structure was established as 19-dehydroxy 9,11- didehydro 3,14- dihydroxy methyl andrographolide.
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