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Phytochemical analysis and antimicrobial activity of Piper capensis L.f.

Dissertation (MSc)--University of Pretoria, 2011.

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Other Authors: Steenkamp, Vanessa
Format: Thesis
Published: University of Pretoria 2013
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access_status_str Open Access
author2 Steenkamp, Vanessa
author_browse Steenkamp, Vanessa
author_facet Steenkamp, Vanessa
collection Thesis
dc_rights_str_mv © 2010, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria.
description Dissertation (MSc)--University of Pretoria, 2011.
format Thesis
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institution University of Pretoria (South Africa)
last_indexed 2026-06-10T12:40:29.643Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository
publishDate 2013
publishDateRange 2013
publishDateSort 2013
publisher University of Pretoria
publisherStr University of Pretoria
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source_str UPSpace — University of Pretoria Institutional Repository
spelling oai:repository.up.ac.za:2263/25752 Phytochemical analysis and antimicrobial activity of Piper capensis L.f. Steenkamp, Vanessa Cromarty, Allan Duncan s22006062@tuks.co.za Thorburn, Anzelle Piper capense l.f. Cytotoxicity Antimicrobial Phytochemical Plumbagin Plant extracts UCTD Dissertation (MSc)--University of Pretoria, 2011. Medicinal plants are the focus of intense study, in particular whether their traditional uses are supported by real pharmacological effects, or merely based on folklore. Piper capense L.f. (Piperaceae) is used traditionally for the treatment of infectious diseases, and has the potential to be a source of novel antimicrobial compound(s). Crude solvent extracts (water, methanol, hexane and acetone) and sequentially extracted subfractions of the root-bark of P. capense were prepared, of which the hexane-soluble subfraction MsAsHs was identified as the most promising antimicrobial subfraction. Phytochemical analyses of the various extracts and subfractions using TLC with numerous mobile phases and compound selective visualising reagents revealed the presence of quinones in all of the crude solvent extracts. Alkaloids, lipids/sterols/steroids, phenolic compounds and amino acids/peptides were detected in select subfractions. Gradient reverse phase HPLC analyses using 0.1% formic acid and methanol indicated three major peaks in MsAsHs. IR spectroscopy indicated that carbonyl and hydroxyl functional groups, and aromatic characteristics were present in the major compound present in MsAsHs. Further analysis using targeted LC-MS Q-TOF and quadrupole LC-MS/MS analyses indicated an empirical formula of C11H8O3. This formula was confirmed for the isolated compound by GC-MS (HP5-MS column) that identified the compound as 5-hydroxy-2-methyl-1,4-naphthoquinone (C11H8O3 MW: 188.18) with 98% certainty using the database. Although 5-hydroxy-2-methyl-1,4-naphthoquinone (also known as plumbagin) is well-known, this is the first time that the presence of this compound is reported in the Piper genus. Antimicrobial activities of P. capense root-bark extracts and the subfractions were determined against Gram-negative and Gram-positive bacteria and a yeast strain using the disk diffusion and broth micro-dilution assays. Antimicrobial activity was observed against Gram-positive bacteria, Gramnegative bacteria as well as a yeast strain, indicating broad spectrum activity. The antimicrobial activities of the crude solvent extracts decreased in the order: acetone > methanol > hexane > water. The MsAsHs subfraction demonstrated the highest antimicrobial activity with an MIC of 29 μg/ml against both Staphylococcus aureus (ATCC 12600) and Candida albicans (ATCC 10231). HPLC eluents of this subfraction that were collected in a drop-wise fashion onto silica TLC plates and assayed by bioautography, indicated that the major compound eluting at 13.6 minutes accounted for most of the antimicrobial activity. Antioxidant activity was observed for the crude water extract, crude methanol extract, crude acetone extract, MsAsAs subfraction as well as the MsAsHs subfraction. Cytotoxicity against mammalian cells in culture was observed for the crude methanol extract, crude acetone extract, crude hexane extract and the MsAsHs subfraction when determined using C2C12 cells as well as resting and PHA stimulated lymphocytes. Stability testing of the MsAsHs subfraction revealed that the antimicrobial compounds found in this subfraction appear to be stable up to 30 days at both 25°C and 40°C when assayed against S. aureus. However, when assayed against C. albicans, there was an increase in antifungal activity from 29 μg/ml to < 7 μg/ml after 30 days at both temperatures tested. This study provides scientific support for the ethnomedical use of the rootbark of P. capense as an antimicrobial. To date, the presence of plumbagin has not been reported in any other plant in the Piper genus. Due to the significant cytotoxic activity against mammalian cells reported in the current study and the mechanism of action of plumbagin, the therapeutic potential of P. capense extracts is very limited due to non-selective cytotoxicity, despite its marked antimicrobial activity. Pharmacology unrestricted 2013-09-06T23:57:47Z 2011-06-23 2013-09-06T23:57:47Z 2011-04-08 2011-06-23 2011-06-22 Dissertation Thorburn, A 2010, Phytochemical analysis and antimicrobial activity of Piper capensis L.f., MSc dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://hdl.handle.net/2263/25752 > E11/298/gm http://hdl.handle.net/2263/25752 http://upetd.up.ac.za/thesis/available/etd-06222011-161306/ © 2010, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria
spellingShingle Piper capense l.f.
Cytotoxicity
Antimicrobial
Phytochemical
Plumbagin
Plant extracts
UCTD
Phytochemical analysis and antimicrobial activity of Piper capensis L.f.
title Phytochemical analysis and antimicrobial activity of Piper capensis L.f.
title_full Phytochemical analysis and antimicrobial activity of Piper capensis L.f.
title_fullStr Phytochemical analysis and antimicrobial activity of Piper capensis L.f.
title_full_unstemmed Phytochemical analysis and antimicrobial activity of Piper capensis L.f.
title_short Phytochemical analysis and antimicrobial activity of Piper capensis L.f.
title_sort phytochemical analysis and antimicrobial activity of piper capensis l f
topic Piper capense l.f.
Cytotoxicity
Antimicrobial
Phytochemical
Plumbagin
Plant extracts
UCTD
url http://hdl.handle.net/2263/25752
http://upetd.up.ac.za/thesis/available/etd-06222011-161306/