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 ~- ~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
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