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Abstract

ABSTRACT:The non-infected bark of Alstonia scholaris R. Br. was carefully peeled off, shade dried, and coarsely powdered with the help of a hand club. Powdered dried bark was extracted in 95% ethanol at room temperature and evaporated in vacuo (40°C). The ethanolic extract was suspended in distilled water and subsequently extracted in hexane, chloroform, ethyl acetate and n-butanol. The solvent of each fraction was removed in a rotary vacuum evaporator under reduced pressure. The crude chloroform extract was mixed with 3% HCl and ethanol. The aqueous layer was collected and the pH was adjusted to 10 by adding NaOH, thereafter it was further extracted with chloroform to get a crude alkaloidal fraction. The alkaloidal fraction was subjected to column chromatography over silica gel and eluted with chloroform: methanol (50:50). The elutes between fractions 40-100 were collected and subjected to preparative HPTLC in the solvent system of ethyl acetate: benzene (1:1). The HPTLC showed the presence of two alkaloids, which were evaluated for their cytotoxic effects in neoplastic cell lines by MTT assay. The compound which showed greater cytotoxicity was subjected to UV, NMR, Mass, and IR spectra and identified as echitamidine-N-oxide-19--β-D-glucopyranoside (EOG). The presence of EOG in the chloroform fraction was further confirmed by semipreparative RPHPLC (H 2 O/CH 3 CN-0%-80%; CH 3 CN in 80 min, flow rate 3 mL/min, UV detector - 250 nm) and was also identified based on the literature.

Chromosome-level Alstonia scholaris genome unveils evolutionary insights into biosynthesis of monoterpenoid indole alkaloids

“…Akuammiline alkaloids are a class of MIAs in A. scholaris , and more than 300 compounds with various pharmacological activities have been identified in A. scholaris . 23 , 24 For example, echitamines exhibit both in vitro and in vivo cytotoxicity, 25 while strictamines 26 inhibit the transcription factor nuclear factor κB (NF-κB). 27 Additionally, the renal cortex protein SGLT2 is inhibited by the derivatives of picraline, 28 , 29 , 30 whereas aspidophylline A reverses drug resistance in cancerous cell lines.…”