A new cytotoxic flabelliferin from palmyrah ( Borassus flabellifer L . ) flour

The cytotoxic compound 1 was isolated from palmyrah flour methanol: water (1:1) extracts, during the course of isolation of the mosquito larvicidal compound4 using MPLC in petroleum ether: methylene chloride: ethyl acetate: methanol gradient at a flow rate of 18 mL/ min against gravity. Further purification of compound 1 was conducted using chromatotron with methanol: water (1: 1) isocratic system. Compound 1 was re-crystallized using water, MeOH and EtOAc solvent mixtures and dissolved in DMSO at a concentration of 100 μg/mL. The resulting solution was introduced into micro-wells containing melanoma cancer cells obtained from the Institute of Cancer Research (ICR), USA. Doxorubicin 20 μg/mL solution was used as a positive control and the growth was measured microscopically after 24 h according to a well established screening method10.

Palmyrah dried seed shoots were purchased from the Palmyrah Development Board.These seed shoots have been collected from home gardens in the Kalpitiya area during the year 2004.They were stored in a freezer (< 0 o C) until use.
The cytotoxic compound 1 was isolated from palmyrah flour methanol: water (1:1) extracts, during the course of isolation of the mosquito larvicidal compound 4 using MPLC in petroleum ether: methylene chloride: ethyl acetate: methanol gradient at a flow rate of 18 mL/ min against gravity.Further purification of compound 1 was conducted using chromatotron with methanol: water (1: 1) isocratic system.Compound 1 was re-crystallized using water, MeOH and EtOAc solvent mixtures and dissolved in DMSO at a concentration of 100 µg/mL.The resulting solution was introduced into micro-wells containing melanoma cancer cells obtained from the Institute of Cancer Research (ICR), USA.Doxorubicin 20 μg/mL solution was used as a positive control and the growth was measured microscopically after 24 h according to a well established screening method 10 .
Compound 1 obtained as white coloured crystals (~ 10 mg, 0.001%) had a retention factor (Rf) value of 0.49 on thin layer chromatography (TLC) in butanol: ethanol: ammonia; (7: 3: 4) solvent system.The compound 1 was shown to have the molecular formula C 51 H 82 O 20 by high resolution fast atom bombardment mass spectrometry (HRFABMS) (negative mode) at m/z = (calculated: 1014.5399)found 1013.5318 for [M -H] -.This shows the possibility of the presence of three-ramnosyl and a glucosyl residues, leaving room for IntroductIon Palmyrah (Borassus flabellifer L.) flour (Odiyal) is known to have a number of toxins.These toxins possess mutagenic 1 , clastogenic 2 , immunosuppressive 3 , dengue mosquito larvicidal 4 and neurotoxic 5 effects.Odiyal contains a number of saponins including a hyperhaemolytic factor 6 , a variety of steroidal saponins (flabelliferins) and an antimicrobial flabelliferin F B
During medium pressure liquid chromatography (MPLC) separation for isolating the dengue mosquito larvicide, a white amorphous solid was obtained.The objectives of this study were purification, identification and determination of bioactivity of this compound.

RESEARCH NOTE
A -87% of growth was observed at a 100µg/mL concentration on the melanoma cell line, where Doxorubicin resulted a -37% at a concentration of 20 µg/mL.Thus, this experiment indicated cytotoxic effect on differentiating cancer cells.Logistic reasons prevented determining the anti cancer activity of the compound.