Aktiviti anti helicobacter pylori ekstrak sepuluh spesies phyllanthus dengan memfokuskan ekstrak kloroform phyllanthus pulcher
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Date
2005
Authors
Wan Ismail, Wan Iryani
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Abstract
Ten spesies of Phyllanthus were screened for their in-vitro anti Helicobacter
· pylori activities using agar diffusion method. The species are P. debilis, P. niruri and P.
urinaria, P. acidus, P. columnaris, P. emblica, P. myrtifolius, P. oxyphyl/us, P. pulcher
and P. ·reticulates. For the first three species as listed above, the whole plants were
used, while only the leaves were used for other species. The plants were sequentially
extracted by hexane, chloroform and methanol using soxhlet. A total of 30 crude
extracts from the plants were screened for their anti bacterial activities and toxicity
using brine shrimp Artemia salina assay. Ten bacterial pathogen i.e. H. pylori,
Enterobacter sp., Escherichia coli, E. coli (ATCC 25922), Pseudomonas stutzeri,
Salmonella sp., Shigella boydii, Sh dysenteriae, Staphylococcus aureus and Vibrio
cholerae were also included in the disc diffusion assay. Chloroform extract of P.
pulcher was selected based on its selectivity, specificity and strong activity (largest
inhibition zone 24.2 ± 1.2 mm) against H. pylori and non-toxicity against A. salina. The
phytochemical screening results showed the occurrence of terpenoid in the chloroform
extract of P. pulcher. The chloroform extract was fractionated into 30 fractions using
the best solvent system i.e. chloroform (8): ethyl acetate (2) by column
chromatography. These fractions were retested on H. pylori and other pathogens using
disc diffusion assay and fraction 18 (F18) was found to be selective, specific and active
against on H. pylori with the largest inhibition zone diameter (30.0 ± 0 mm). Further
testing on A. salina revealed that it was non-toxic LC50 acute = 1.350 ± 0.1 mg/ml and
LCso chronic = 1 .000 ± 0.1 mg/ml. The minimum inhibition concentration (MIG) test of
F18 showed that the MIC50 and MIC90 were 4 J.Jg/ml and 256 J.Jg/ml respectively on 20
clinical isolates of H. pylori. The MIG for H. pylori (SJ 179) was 32 IJQ/ml. Using batch
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~culture of H. pylori in Eugon broth with or without F18 (control), monitoring H. pylori by
colony counting method and direct count, and observing cellular changes by SEM and
TEM, there was evidence that F18 caused direct killing of the culturable spiral cells and
at 16 !Jg/ml caused cell leakage. By LC-MS and referring to the database, F18
tentatively contain 13-sitosterol and some fatty acids (methyl nonadecanoate and
tripalmitin).
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Keywords
Helicobacter pylori , Ekstrak kloroform