Phytopharmaceutical Mission Progress Report

(15th may to 31th dec 2018)

•Name of the project fellow:
Deendyal
•Designation:
Project assistant (Level-2)
•Name of the Project Investigator:
Dr. Sandip Bharate
•Date of Joining:
15/05/2018
•Duration of report:
15 may-31 dec 2018

PLANT : Dysoxylum binectaiferum

OBJECTIVE: The objective of this work is “Isolation of four marker
compounds (at least 20 mg of each) from this plant”.
 1 kg dried and crushed leaves of plant was already available in lab.
 The plant material was extracted with methanol (using cold maceration method). The
extraction was repeated 3 times (24 hrs x 3 L). The extract obtained from three cycles was
combined and evaporated on vacuo rotavapor. 98 g of extract was obtained. Column of
extract was started for separation over silica gel with increasing amount of methanol in
ethyl acetate.
EXTRACTIVE VALUE: 9.8%
The TLC was performed on each day to see the extent of extraction
TLC: (15% methanol in DCM)

After Day 1 After Day 2

 Column of extract is started for separation over silica gel with increasing amount of
methanol in ethyl acetate.
 Fraction of non polar compounds were obtained in 5 to 15 % MeOH in EtOAc.
 Column continued by increasing percentage of MeOH and certain mixtures of the
compounds were isolated.
 Mixture of compounds were isolated from first column.
 Second column is started: Column of extract is started for separation over silica gel
with increasing amount of MeOH in EtOAc.
 Non polar compounds were isolated in 5 to 15 % MeOH in EtOAc.
 Column continued by increasing percentage of MeOH and 40 mg of DD 09
(Rohitukine-N-oxide) was isolated.
 Column continued by increasing polarity and mixture of compounds was isolated.
 Further 3rd column was started for sepration of marker compounds over silica gel by
using MeOH and ethyl acetate as solvent.
 From 3rd column two more marker compounds (Rohitukine and Schumaniofoside A)
was isolated.
 Three marker compounds were isolated and submitted to repository (25 mg each).
TLC of final dried extract and Structure of marker compounds

15% methanol in DCM

Rohitukine-N-oxide
Schumaniofoside A
Rohitukine
 2 kg of dried and crushed leaves of plant was taken and extracted with 50 %
aqueous EtOH. The extraction was repeated 3 times (24 hrs x 5 L). The extract
obtained from three cycles was combined and evaporated on vacuo rotavapor. 345 g
of extract was obtained after drying.

 EXTRACTIVE VALUE: 17%

 The TLC of plant extract.
(15% MeOH in DCM)

After charring
• Literature search on Dysoxylum binectaiferum phytochemistry/ pharmacology.
The literature search was performed in order to know the amount of work already
done on this plant, both from phytochemistry and pharmacology point of view.

1. Tetrahedron Lett. 2017; 58; 3974-78.

Reported steroidal compounds from Dysoxylum binectariferum

2. Steroids. 2014; 86; 26-31.

3. Bioorg Med Chem Lett. 2014; 24; 3146-49.
References :-
1. Kumar, V.; Gupta, M.; Gandhi, S. G.; Bharate, S. S.; Kumar, A.;
Vishwakarma, R. A.; Bharate, S. B. Anti-inflammatory
chromone alkaloids and glycoside from Dysoxylum
binectariferum. Tetrahedron Lett. 2017, 58, 3974-3978.
2. Yan, H. J.; Wang, J. S.; Kong, L. Y. Cytotoxic steroids from the
leaves of Dysoxylum binectariferum. Steroids. 2014, 86, 26-31.
3. Jain, S. K.; Meena, S.; Gupta, A. P.; Kushwaha, M.; Uma
Shaanker, R.; Jaglan, S.; Bharate, S. B.; Vishwakarma, R. A.
Dysoxylum binectariferum bark as a new source of anticancer
drug camptothecin: Bioactivity-guided isolation and LCMS-
based quantification. Bioorg.Med. Chem. Lett. 2014, 24, 3146-
3149.
 Spectral data of marker compounds

Spectral Data of Rohitukine

Spectral data of Rohitukine-N-oxide
Spectral data of Schumaniofoside A