| dc.description.abstract | Indonesia is rich in medicinal plants, one of which that was believed to be
effective against cancer is “ekor naga” leaf (Rhaphidophora pinnata
(L.f)Schott). The boiled “ekor naga” was traditionally used for the treatment of
east cancer, and from the previorus study by brine shrimp (Artemia salina
Leach) test showed that ethanolic extract of “ekor naga” leaf (EDEN) showed
cytotoxic activity. The objective of this study is to determine the cytotoxic
activity of FEDEN, the fractions and the isolate of ekor naga leaf againt MCF-7
cancer cells. Antiproliferation and apoptosis assay had been conducted for
cytotoxic isolate, followed by the isolation of active compound.
Characterization of crude drug and phytochemical screening were
performed on the dried powdered crude drug and the ethanolic extrat of “ekor
naga” leaf (EEDEN), the phytochemical screening also performed on the n-
hexane, chloroform, ethylacetate, and aqueous fractions. The ekor naga leaf was
extracted. by percolation using ethanol as solvent, followed by fractionation with
Tiguid-liquid extraction using n-hexane, chloroform, and ethylacetate as solvents.
The cytotoxic activity testing was performed for the fractions (n-hexane,
chloroform, ethyl acetate, and aqueous) against MCF-7 cells with MIT assay (3
(4.5-dimethyltiazol-2-e))2,5-diphenyltetrazoliumbromide) The active chloroform
fraction as anticancer was subjected to column chromatography: the resulting
isolate was tested for its cytotoxicity activity. The isolate D and F from
fractionation were tested for its antiproiferative (doubling time) with observation
time 24; 48 and 72 hours using MTT assay. The isolate D was tested for
apoptosis induction by double staining assay using acridine orange-ethidium
bromide and by floweytometry assay using Ancksin V against the MCF - 7 cells.
The result of characterization of dried powdered ekor naga leave showed
brownish colour, odorless, tasteless powder; the water content was 8.99%; water
soluble extract 14.81% ethanol soluble extract 9.76%, total ash content 10.09%,
and acid insoluble ash 0.17%. Phytochemical screening of dried powdered ckor
naga leaves and EEDEN showed the presence of tannin, alkaloid, flavonoid,
saponin, glycoside, and triterpenoid/steroid compounds; the n-hexane fraction
gave 2 positive result for triterpenoid/steroid; the chloroform fraction showed a
positive result for alkaloid, and triterpenoid/steroid; the ethylacetate fraction
showed a positive result for flavonoid, tannin, and glycoside, while the aqueous
fraction contained tannin, saponin, and glycoside. Probit analysis gave ICsp values,
for EEDEN, chloroform fractions and ethyl acetate fractions were 112.240;
59,082; 812.663 ug/mL, respectively while the n-hexane and aqueous fractions
did not inhibit the growth of MCF-7 cells
The result of the cytotoxic activity testing for A through I isolates showed
IC50 values of 190.550; 134.014; 204.474; 87.831; 96.016; 34.161; 9.6215,
758.572; 245.446 uglml. respectively. The doubling time result of D and F
isolates could inhibit cell proliferation depending on the concentration, aliquot
with incubation time the ICs value decreased. at 72 hours observation. Apoptosi
observation for isolate D and isolate F used floweytomerry assay also showed
apoptosis and necrosis.
The separation of fraction F by column chromatography gave six isolates
with FFI and FF2 showed one TLC spot which then characterized by UV, FTIR
and LC-MS methods. The UV spectrum of FFI isolate had maximum
absorption peak at % 238 and 402 nm while FF2 isolate at 2 of 408 nm. The
inftared spectrum showed the presence of OH, -CH aliphatic (CHs and ~CHs) C
C and C = 0 groups. MS data for FF1 isolates showed fragmen peak mie 429
and 319 while FF2 isolate showed fragmen m/e 466.6 that suggested to be
steroidtritepenoid,
Column chromatography separation of isolate A resulted five isolates
with FA: was then purified by column chromatography to give FA2.22 isolate
The PA2.2.2 isolate was characterized by UV spectrophotometry, FT-IR, GC-MS
and NMR. The infrared spectrum showed. the presence of -OH, -CH aliphatic
CH: and -CHh), C= C and C = O groups. The "C-NMR data showed 23 carbon
toms, GC-MS detected seven compounds, with four of them had similarity
indices greater than 90%, | en_US |
