Exploring the phytochemical variation of non-volatile metabolites within three South African Salvia species using UPLC-MS fingerprinting and chemometric analysis.

The South African Salvia species, Salvia africana-lutea, S. lanceolata and S. chamelaeagnea, are widely used to treat fever and inflammation associated with skin and lung infections. The aim of this study was to explore the non-volatile secondary metabolites and the phytochemical variation within these lesser known species, to support product development and commercialisation. Chemical profiles of the methanol extracts of 81 wild-harvested samples were obtained using ultra performance-quadrupole-Time-of-Flight-mass spectrometry (UPLC-qToF-MS). Forty-one compounds, including caffeic acid, rosmarinic acid, carnosol, carnosic acid and ursolic acid, were detected and confirmed across the three species. Nineteen compounds were tentatively identified of which 14 have not been reported in these species. Principal component analysis revealed distinct clusters corresponding to the three species, confirming chemical differences. Marker compounds for each species were revealed using orthogonal projection to latent structures-discriminant analysis. Further chemometric analysis reflected a degree of intraspecies variation, although the chemistry within populations was mostly conserved. Potential chemotypes for each species were identified through unique compounds associated with each group. The concentrations of medicinally important metabolites, namely, rosmarinic acid, carnosol, carnosic acid and ursolic acid, were determined, using validated UPLC-PDA methods. Ursolic acid was present at levels up to 38.2 mg/g, confirming that these species are a rich source of this compound. No similar studies combining liquid chromatography with chemometric analysis, and utilising a large sample size from various habitats, have been reported for these three Salvia species. The results will guide selection of cultivars with the best attributes for the intended therapeutic application, thereby protecting wild populations from over-exploitation.

A chemometric assessment of essential oil variation of three Salvia species indigenous to South Africa.

Indigenous Salvia species from southern Africa are popular traditional medicines for the treatment of a variety of conditions. They produce fragrant volatiles that can be isolated as essential oils. Some of these volatile organic compounds may play a role in the biological activities of the extracts. Three indigenous Salvia species, Salvia africana-lutea, S. lanceolata and S. chamelaeagnea, were selected for this study as they are commonly used in traditional medicine in South Africa, and the essential oils from these species have potential for commercialisation. Although some studies have described the essential oil compositions and some biological activities, only single composite samples were used. The aim of this study was to investigate the intra- and interspecies variation of the essential oils, sampled over a wide geographical area and using a representative sample size, to encourage commercialisation of the essential oil. Essential oils were isolated from individual plants using conventional hydrodistillation of the aerial parts, harvested from several localities. Gas chromatography coupled simultaneously to mass spectrometry/flame ionisation detection (GC-MS/FID) was used to identify and quantify the volatile constituents. The essential oils of S. africana-lutea consisted mainly of terpinene-4-ol + ß-caryophyllene (1.4 - 29.0%), T-cadinol (1.2 - 20.0%), α-eudesmol (trace - 23.0%) and ß-eudesmol (trace - 26.0%), those of S. lanceolata comprised mainly terpinene-4-ol + ß-caryophyllene (4.3 - 31.0%), α-humulene (2.3 - 15.0%), bicyclogermacrene (trace - 37.0%) and spathulenol (trace - 25.0%), while the essential oils of S. chamelaeagnea were characterised by δ-3-carene (trace - 18.0%), limonene (1.6 - 36.0%), viridiflorol (9.8 - 61.0%) and 1,8-cineole (not detected - 11.0%). The compounds identified in the essential oils of the three selected Salvia species have been identified in other Salvia essential oils. To add to the novelty of this study, the superior resolving power of two-dimensional gas chromatography was demonstrated through analysis of selected essential oils. Many additional compounds were identified, and previously co-eluting compounds were clearly separated. Chemometric modelling of the GC-MS data using SIMCA P+ 14 software allowed distinct clustering patterns to be discerned. The unsupervised principal component analysis model revealed separate clusters for the three species, confirming substantial chemical differences between their essential oils. Quantitative, rather than qualitative differences were evident when individual essential oil samples representing the same species, were compared. For each species, two chemically distinct groups were observed and unique marker compounds could be identified. This study has contributed detailed information on the major and minor volatile compounds present in the essential oils of the three Salvia species investigated.

The in vitro antimicrobial evaluation of commercially essential oils and their combinations against acne.

OBJECTIVE: The study investigated the efficacy of commercial essential oil combinations against the two pathogens responsible for acne with the aim to identify synergy and favourable oils to possibly use in a blend. MATERIALS AND METHODS: Antimicrobial activity was assessed using the minimum inhibitory concentration (MIC) assay against Staphylococcus epidermidis (ATCC 2223) and Propionibacterium acnes (ATCC 11827), and the fractional inhibitory concentration index (ΣFIC) was calculated. Combinations displaying synergistic interactions were further investigated at varied ratios and the results plotted on isobolograms. RESULTS: From the 408 combinations investigated, 167 combinations were identified as displaying noteworthy antimicrobial activity (MIC value ≤1.00 mg mL-1 ). Thirteen synergistic interactions were observed against S. epidermidis, and three synergistic combinations were observed against P. acnes. It was found that not one of the synergistic interactions identified were based on the combinations recommended in the layman's aroma-therapeutic literature. Synergy was evident rather from leads based on antimicrobial activity from previous studies, thus emphasizing the importance of scientific validation. Leptospermum scoparium J.R.Forst. and G.Forst (manuka) was the essential oil mostly involved in synergistic interactions (four) against S. epidermidis. Cananga odorata (Lam.) Hook.f. and Thomson (ylang ylang) essential oil was also frequently involved in synergy where synergistic interactions could be observed against both pathogens. The combination with the lowest MIC value against both acne pathogens was Vetiveria zizanioides Stapf (vetiver) with Cinnamomum verum J.Presl (cinnamon bark) (MIC values 0.19-0.25 mg mL-1 ). Pogostemon patchouli Benth. (patchouli), V. zizanioides, C. verum and Santalum spp. (sandalwood) could be identified as the oils that contributed the most noteworthy antimicrobial activity towards the combinations. The different chemotypes of the essential oils used in the combinations predominantly resulted in similar antimicrobial activity. CONCLUSIONS: The investigated essential oil combinations resulted in at least 50% of the combinations displaying noteworthy antimicrobial activity. Most of the synergistic interactions do not necessarily correspond to the recommended layman's aroma-therapeutic literature, which highlights a need for scientific validation of essential oil antimicrobial activity. No antagonism was observed.

Antimicrobial activity and chemometric modelling of South African propolis.

AIMS: This study reports on the inhibitory and bactericidal properties of 39 South African (SA) propolis samples and three propolis samples from Brazil. METHODS AND RESULTS: Ethanolic extracts of propolis (EEP) were prepared and their antimicrobial activities tested using the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Some samples displayed substantial antimicrobial activity with MIC and MBC values as low as 6 µg ml(-1) against Staphylococcus aureus. The correlation between liquid chromatography-mass spectrometry (LC-MS) chemical data and the antimicrobial activity of propolis extracts was investigated using multivariate data analysis tools. Orthogonal projections to latent structures (OPLS) models were created for the two Gram-positive bacteria (Enterococcus faecalis and S. aureus) and Candida albicans. Using the S-plot function, it was possible to identify the bioactive constituents in propolis as chrysin, pinocembrin, galangin and pinobanksin-3-O-acetate. CONCLUSION: The SA propolis samples tested displayed noteworthy antimicrobial activity, favourably comparable to that of the Brazilian comparator and 'gold standard'. The observed antimicrobial activity of SA propolis can possibly be attributed to its flavonoid content. SIGNIFICANCE AND IMPACT OF THE STUDY: Based on the good antimicrobial activity observed for SA propolis, this natural resource shows promise and should be considered for development which may contribute to growing the bio-economy in the region.

The impact of plant volatiles on bacterial quorum sensing.

UNLABELLED: Studies describing the use of essential oil constituents as antimicrobial agents have steadily increased; however, some phyto-constituents are often overlooked due to unfavourable minimum inhibitory concentration (MIC) values. Virulence depends on transcriptional factors which are regulated by cell-to-cell communication called quorum sensing (QS). This study was undertaken to evaluate the antimicrobial and anti-QS properties of 29 compounds commonly found in essential oils using two bioreporter strains. QS-inhibitory activity was assessed qualitatively by agar diffusion and quantitatively by spectrophotometric assays. MICs of all the tested compounds ranged from 0·032 to >5 mg ml(-1). Twenty-two compounds displayed varying levels of QS inhibitory activity with zones of violacein inhibition ranging from 9 to 16 mm. Majority of tested molecules inhibited violacein and pyocyanin production in Chromobacterium violaceum and Pseudomonas aeruginosa, while seven compounds increased violacein and pyocyanin production. Interestingly, it was observed that the (+)-enantiomers of carvone, limonene and borneol increased violacein and pyocyanin production, while their levorotary analogues inhibited this production. α-Terpineol and cis-3-nonen-1-ol exhibited >90% violacein inhibition, suggesting their potential as QS inhibitors. This preliminary study indicates that plant volatiles have the potential to impede or promote bacterial communication and further studies need to be undertaken to explore the contribution of structural analogues and stereochemistry of molecules in this process. SIGNIFICANCE AND IMPACT OF THE STUDY: Antimicrobial and antiquorum sensing (QS) properties of 29 common essential oil compounds were evaluated. Interruption of QS may lead to the development of therapeutic, antivirulence agents to control disease-causing pathogens which are preferable over antimicrobial agents as the latter drives selection pressure on microbial communities to acquire resistance. Twenty-two compounds inhibited QS, while seven promoted the QS to a variable extent in Chromobacterium violaceum and Pseudomonas aeruginosa. Preliminary results suggest that QS-inhibitory compounds of natural origin may inspire the formulation of new generation of antimicrobial agents to control infectious pathogens.

The additive and synergistic antimicrobial effects of select frankincense and myrrh oils--a combination from the pharaonic pharmacopoeia.

AIMS: The in vitro antimicrobial activity of three essential oil samples of frankincense (Boswellia rivae, Boswellia neglecta and Boswellia papyrifera) and two essential oil samples of myrrh and sweet myrrh (Commiphora guidotti and Commiphora myrrha), collected from different regions of Ethiopia, was investigated independently and in combination to determine their anti-infective properties. METHODS AND RESULTS: The microdilution minimum inhibitory concentration (MIC) assay was performed, whereby it was noted that generally Cryptococcus neoformans (MIC values in the range of 0·8-1·4 mg ml(-1)) and Pseudomonas aeruginosa (MIC values in the range of 0·5-1·3 mg ml(-1)) often appeared to be the most susceptible micro-organisms against oils of both Boswellia and Commiphora spp. When assayed in various combinations, the frankincense and myrrh oils displayed synergistic, additive and noninteractive properties, with no antagonism noted. When investigating different ratio combinations against Bacillus cereus, the most favourable combination was between B. papyrifera and C. myrrha. The composition of the oils was determined by gas chromatography coupled to mass spectrometry (GC-MS) to document the specific chemotypes used in the study, and the chemical profiles were found to be congruent with previously reported data. CONCLUSIONS: The majority of interactions identified synergistic and additive effects, with strong synergism noted between B. papyrifera and C. myrrha. SIGNIFICANCE AND IMPACT OF THE STUDY: Frankincense and myrrh essential oils have been used in combination since 1500 bc; however, no antimicrobial investigations have been undertaken to confirm their effect in combination. This study validates the enhanced efficacy when used in combination against a selection of pathogens.

In vitro transport of the steroidal glycoside P57 from Hoodia gordonii across excised porcine intestinal and buccal tissue.

Hoodia gordonii was traditionally used by the Khoisan people as a thirst and appetite suppressant and is currently commercially available as a popular weight-loss supplement. The perceived active ingredient isolated from this plant is a steroidal glycoside named P57. This study aimed at investigating the in vitro transport of P57 across excised porcine intestinal and buccal mucosa in a Sweetana-Grass diffusion apparatus. For both the intestinal and buccal experiments, the transport of pure P57 was compared to that obtained from a crude plant extract. Bi-directional transport experiments were conducted across the intestinal tissue in two different media namely Krebs-Ringer bicarbonate buffer and simulated intestinal fluid. Apical-to-basolateral transport experiments were conducted across the buccal tissue in two different media namely Krebs-Ringer bicarbonate buffer and artificial saliva. Apparent permeability coefficient (P(app)) and flux values were calculated and analysed by means of a one-way repeated analysis of variance (ANOVA) to determine if differences were significant (p≤0.05). The transport of pure P57 across intestinal tissue was significantly higher in the secretory direction than in the absorptive direction indicating efflux by membrane transporters. Much higher intestinal transport was obtained for P57 in both directions when applied in the form of a crude extract, possibly due to inhibition of efflux as indicated by lower secretory transport compared to absorptive transport. For the buccal tissue, no transport was obtained for the pure P57, while relatively high transport was obtained when applied in the form of a crude extract. Furthermore, the intestinal transport of P57 was significantly decreased when the crude extract was prepared in simulated intestinal fluid compared to when it was prepared in buffer. On the other hand, buccal transport was higher in artificial saliva than in buffer. It is therefore evident that the transport of P57 across mucosal tissues is significantly affected on exposure to conditions simulating the in vivo situation.

The in vitro antibiofilm activity of selected culinary herbs and medicinal plants against Listeria monocytogenes.

AIMS: The antibiofilm activity of extracts obtained from selected herbs, spices, beverages and commercially important medicinal plants was investigated on Listeria monocytogenes. METHODS AND RESULTS: The growth and development of the biofilm was assessed using the crystal violet (CV) assay. The respiratory activity was assessed using the 2, 3-bis [2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT) reduction assay. The majority of extracts tested prevented cell adhesion to the polyvinyl chloride (PVC) surface. Seven of the 15 extracts reduced biofilm adhesion of both the clinical and the type strains by at least 50%. In contrast, inhibition of a preformed biofilm was more difficult to achieve, with only three extracts (Rosmarinus officinalis, Mentha piperita and Melaleuca alternifolia) inhibiting the growth of both strains by at least 50%. CONCLUSIONS: Although most extracts were able to reduce initial cell attachment, inhibition of growth in a preformed biofilm was more difficult to achieve. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability to reduce biofilm biomass as shown by several plant extracts warrants further investigation to explore the use of natural products in antibiofilm adhesion.

The antimicrobial activity of four commercial essential oils in combination with conventional antimicrobials.

AIMS: Due to the emergence of multi-drug resistance, alternatives to conventional antimicrobial therapy are needed. This study aims to investigate the in vitro pharmacological interactions between essential oils (considered valuable as natural therapeutic treatments) and conventional antimicrobials (ciprofloxacin/amphotericin B) when used in combination. METHODS AND RESULTS: Interactions of the essential oils (Melaleuca alternifolia, Thymus vulgaris, Mentha piperita and Rosmarinus officinalis) when combined with ciprofloxacin against Staphylococcus aureus indicate mainly antagonistic profiles. When tested against Klebsiella pneumoniae the isobolograms show antagonistic, synergistic and additive interactions depending on the combined ratio. The R. officinalis/ciprofloxacin combination against K. pneumoniae displayed the most favourable synergistic pattern. The interactions of M. alternifolia (tea tree), T. vulgaris (thyme), M. piperita (peppermint) and R. officinalis (rosemary) essential oils with amphotericin B indicate mainly antagonistic profiles when tested against Candida albicans. CONCLUSION: While a number of interactions show complete antagonism, others show varied (synergistic, additive and/or antagonistic) interactions, thus the efficacy is dependent on the ratio in which the two components co-exist. SIGNIFICANCE AND IMPACT OF THE STUDY: The predominant antagonistic interactions noted here, suggests that some natural therapies containing essential oils should be used with caution when combined with antibiotics.

Sceletium--a review update.

It is probable that plants of the genus Sceletium (Mesembryanthemaceae) have been used as masticatories and for the relief of thirst and hunger, to combat fatigue, as medicines, and for social and spiritual purposes by San hunter-gatherers (historically referred to as Bushmen) and Khoi pastoralists (historically referred to as Hottentots) for millennia before the earliest written reports of the uses of these plants by European explorers and settlers. The oral-tradition knowledge of the uses of Sceletium by indigenous peoples has largely been eroded over the last three centuries due to conflicts with settlers, genocidal raids against the San, loss of land, the ravages of introduced diseases, and acculturation. Wild resources of Sceletium have also been severely diminished by over-harvesting, poor veld-management, and possibly also by plant diseases. Sceletium was reviewed almost a decade ago and new results have emerged substantiating some of the traditional uses of one of South Africa's most coveted botanical assets, and suggesting dietary supplement, phytomedicine and new drug applications. This review aims to collate the fragmented information on past and present uses, the alkaloid chemistry and pharmacological evidence generated on Sceletium.

Validation of smoke inhalation therapy to treat microbial infections.

AIM OF THE STUDY: In traditional healing, the burning of selected indigenous medicinal plants and the inhalation of the liberated smoke are widely accepted and a practiced route of administration. This study elucidated the rationale behind this commonly practiced treatment by examining the antimicrobial activity for five indigenous South African medicinal plants commonly administered through inhalation (Artemisia afra, Heteropyxis natalensis, Myrothamnus flabellifolius, Pellaea calomelanos and Tarchonanthus camphoratus). MATERIAL AND METHODS: An apparatus was designed to simulate the burning process that occurs in a traditional setting and the smoke fraction was captured for analysis and bioassay. Methanol and acetone extracts as well as the essential oil (for the aromatic species) were prepared and assayed in parallel with the smoke fraction. RESULTS: Antimicrobial data revealed that in most cases, the 'smoke-extract' obtained after burning had lower minimum inhibitory concentration (MIC) values than the corresponding solvent extracts and essential oils. The combustion, acetone and methanol extracts produced different chromatographic profiles as demonstrated for Pellaea calomelanos where several compounds noted in the smoke fraction were not present in the other extracts. CONCLUSION: These results suggest that the combustion process produces an 'extract' with superior antimicrobial activity and provides in vitro evidence for inhalation of medicinal smoke as an efficient mode of administration in traditional healing.

'Buchu' -Agathosma betulina and Agathosma crenulata (Rutaceae): a review.

South Africa has offered the world two indigenous aromatic plants from which commercially important natural products have been developed: Pelargonium graveolens (and its hybrids) the source of geranium oil and Agathosma betulina, from which 'Buchu' oil is produced. Despite the historical use of 'Buchu' and the commercial interest developed around this coveted indigenous resource the (limited) research has not been coherently assembled. This overview aims to unite aspects on the botany, traditional and modern day uses, chemistry and pharmacological data on 'Buchu' which is undeniably one of South Africa's most renowned botanical assets.

Phytochemistry and in vitro pharmacological activities of South African Vitex (Verbenaceae) species.

AIM OF THE STUDY: The in vitro phytochemical and pharmacological investigation of the non-volatile extracts of five South African Vitex species (Verbenaceae); V. obovata ssp. obovata, V. obovata ssp. wilmsii, V. pooara, V. rehmannii and V. zeyheri were investigated in order to validate their traditional use to treat a wide range of ailments such as malaria, wounds, skin diseases and body pains. MATERIAL AND METHODS: The antimicrobial activity was assessed using the minimum inhibitory concentration assay. Through bioactivity-guided fractionation, the fraction responsible for the antimicrobial activity was determined. The toxicity profile, anti-oxidant and anti-inflammatory activity was evaluated using the tetrazolium cellular viability, 2,2-diphenyl-1-picrylhydrazyl and 5-lipoxygenase assays respectively. The antimalarial activity of the extracts and isolated compound from V. rehmannii was also investigated on the chloroquine-resistant Gambian FCR-3 strain of Plasmodium falciparum using the tritiated hypoxanthine incorporation assay. RESULTS: Mostly good antimicrobial inhibition was evident against Gram-positive bacteria (0.02-8.00 mg/ml) and lower activity against the Gram-negative bacteria and the yeast (0.50-8.00 mg/ml). The fraction responsible for antimicrobial activity of V. rehmannii was purified to give a labdane diterpene as an inseparable epimeric mixture of 12S,16S/R-dihydroxy-ent-labda-7,13-dien-15,16-olide. Cirsimaritin was also isolated and identified from V. rehmannii. All the species, apart from V. zeyheri, exhibited scavenging activity (IC50: 22.14+/-1.74 to 33.06+/-1.68 microg/ml) in the anti-oxidant assay. None of the species displayed any anti-inflammatory activity at 100 microg/ml. All the extracts and the labdane diterpene exhibited good antimalarial activity, with the labdane diterpene being the most active (IC50: 2.39+/-0.64 microg/ml). The test extracts were shown to be highly toxic, displaying safety index values ranging from 0.53 to 2.59. CONCLUSION: Of all the pharmacological investigations, the antimalarial and antimicrobial activity exhibited greatest activity and may provide a scientific basis for the ethnomedical use of Vitex species.

South African Helichrysum species: a review of the traditional uses, biological activity and phytochemistry.

AIMS OF THE STUDY: In South Africa, the genus Helichrysum is widely used in traditional medicine. The uses are well documented although renaming of species and the resulting confusing taxonomic nomenclature may cause uncertainty as to which specific species was referred to in some reports. The aim of this paper is to present a collated and coherent overview of the documented traditional uses of Helichrysum species and to update the botanical identity of previously studied species. MATERIALS AND METHODS: Databases (Scifinder, ISI Web of Knowledge) and several books were used to collect in information on South African Helichrysum species. RESULTS: The traditional uses, chemistry and biological activity of Helichrysum species have been summarized. It was attempted to give clarity as to exactly which species is refer to in the ethnobotanical literature. CONCLUSIONS: Although a large number of ethnopharmacological uses have been documented and the chemistry of the genus has been studied extensively, only a few South African species have been investigated for their biological activity.

In vitro evidence of phyto-synergy for plant part combinations of Croton gratissimus (Euphorbiaceae) used in African traditional healing.

AIM OF THE STUDY: Despite the extensive traditional use of Croton gratissimus Burch. var. gratissimus for medicinal purposes, scientific studies validating the therapeutic properties of this indigenous plant are lacking. As the bark, roots and leaves of C. gratissimus are used separately as well as in combination, this study focused on determining antimicrobial efficacies of the plant parts independently and in combination to assess possible pharmacological interactions (e.g. synergy, antagonism). MATERIAL AND METHODS: The hydro-distilled leaf essential oil and extracts of bark, root and leaf were comparatively assessed for antimicrobial activity by means of microdilution minimum inhibitory concentration (MIC). The fractional inhibitory concentrations (FIC) were determined for the leaf and root (1:1), bark and root (1:1), leaf and bark (1:1) combination. Isobolograms were plotted to demonstrate interactions between various ratios of the roots and leaves. RESULTS: The MIC and FIC results indicated variable efficacies for the various plant part combinations, the greatest of which was noted for Cryptococcus neoformans in the root and leaf combination (MIC 0.4 mg/ml and FIC of 0.4). Isobolograms indicated the greatest synergy for Bacillus cereus, Candida albicans and Cryptococcus neoformans. CONCLUSION: The observed synergistic interactions clearly indicate that the reductionist approach may often be short-sighted and that biological activity may be improved through combination therapy, where different complex metabolic pools collectively contribute to the enhanced effect.

The in vitro biological activity of selected South African Commiphora species.

Ten South African Commiphora (Burseraceae) species were investigated to validate their use in traditional healing rites. The leaf and stem extracts of each species were analysed for the anti-oxidant (ABTS and DPPH assays), antimicrobial (MIC and death kinetic assays), anti-inflammatory (5-LOX assay), anticancer (SRB assay) properties, as well as the cytotoxic effects (tetrazolium-based assay). The best anti-oxidant activity (ABTS assay) was observed for the stem extracts of Commiphora tenuipetiolata IC(50)=5.10 microg/ml), Commiphora neglecta (IC(50)=7.28 microg/ml) and Commiphora mollis (IC(50)=8.82 microg/ml). Extracts generally exhibited poor anti-oxidant activity in the DPPH assay, with the exception of Commiphora schimperi (stem), Commiphora neglecta (stem), Commiphora tenuipetiolata (stem and leaf), and Commiphora edulis (stem), with IC(50) values ranging between 7.31 and 10.81 microg/ml. The stem extracts exhibited moderate to good 5-LOX inhibitory activity with Commiphora pyracanthoides (stem) displaying the greatest inhibitory effect (IC(50)=27.86+/-4.45 microg/ml). For the antimicrobial (MIC) assay, a greater selectivity was exhibited by the extracts against the Gram-positive bacteria (0.01-8.00 mg/ml) and the yeasts (0.25-8.00 mg/ml) than against the Gram-negative bacteria (1.00-8.00 mg/ml). Using death kinetic studies (time-kill studies), the rate at which Commiphora marlothii (stem) kills Staphylococcus aureus over a 24h period was determined. Mostly, a concentration-dependent antibacterial activity was observed beginning after ca. 30 min. All concentrations exhibited antibacterial activity, with complete bactericidal effect achieved by the 24(th) hour. The most active Commiphora species against the HT-29 cells (SRB anticancer assay) were Commiphora glandulosa (leaf and stem) and Commiphora marlothii (leaf). The MCF-7 cells (SRB anticancer assay) exhibited the highest sensitivity to indigenous Commiphora species, with Commiphora edulis (leaf and stem), Commiphora glandulosa (leaf and stem), Commiphora marlothii (leaf), Commiphora pyracanthoides (leaf and stem), Commiphora schimperi (stem), and Commiphora viminea (stem) all possessing a percentage inhibition greater than 80% at 100 microg/ml. Commiphora glandulosa (leaf and stem) and Commiphora pyracanthoides (leaf and stem) were the two most active species against the SF-268 cells (SRB anticancer assay), with IC(50) values ranging between 68.55+/-2.01 and 71.45+/-1.24 microg/ml. The majority of the Commiphora extracts were largely non-cytotoxic against Graham human kidney epithelial cells when investigated in the MTT assay.

South African Salvia species: a review of biological activities and phytochemistry.

The genus Salvia (sage) belongs to the Lamiaceae and encompasses 900 species worldwide of which ca. 26 indigenous species are found in southern Africa. Salvia is the largest genus in this family and constitutes almost one quarter of the Lamiaceae. In South Africa, the majority of Salvia species are distributed predominantly in the Cape region. Salvia species are used in many parts of the world to treat various conditions. Many sages, if not all, form an integral part of traditional healing in South Africa particularly in regions where they occur in abundance. Several species are used to treat microbial infections, cancer, malaria, inflammation, loss of memory and to disinfect homes after sickness. Despite the extensive traditional use and the general interest in phytoconstituents of Salvia it remains ironic that research on the South African counterparts has until recently been neglected. The review aims to collate recent research results on the phytochemistry and pharmacological properties of indigenous species. Bio-active compounds with antiplasmodial and antibacterial activities have been isolated and structurally elucidated from Salvia chamelaegnea, Salvia radula and Salvia verbenaca. The essential oil composition of Salvia showed the dominance of monoterpene hydrocarbons, oxygen-containing monoterpenes and oxygen- containing sesquiterpenes. Salvia runcinata is identified as an alternative source of natural alpha-bisabolol. Many pharmacological activities are summarised (anti-oxidant, antimicrobial, antiplasmodial, analgesic, antipyretic, anticancer, anti-inflammatory and antinociceptive) as a first attempt to provide scientific support for past and present local traditional uses.

The in vitro pharmacological activities of 12 South African Hermannia species.

Hermannia species are widely used in traditional medicine in southern Africa, however no extensive study has been conducted on this genus. The acetone extracts of 12 indigenous Hermannia species (flowers, stems and leaves combined) were evaluated for various pharmacological activities. All investigated species displayed promising antimicrobial activity, with Hermannia saccifera being the most active against Staphylococcus aureus (MIC=19.5 microg/ml), Bacillus cereus (MIC=19.5 microg/ml) and Enterococcus faecalis (MIC=125 microg/ml). Time-kill studies on H. saccifera against S. aureus indicated bacteriostatic activity at 1.25, 2.5 and 5.0%, and a concentration of 7.5% achieved complete bactericidal activity after 4h. Ten of the 12 species indicated good free radical scavenging activity, with H. cuneifolia demonstrating the most promising activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH():IC(50)=10.26 microg/ml) and 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS(+)/TEAC: IC(50)=10.32 microg/ml) assays. In addition, all species exhibited moderate anti-inflammatory activity in the 5-lipoxygenase assay with the exception of H. cuneifolia (IC(50)=15.32 microg/ml). Overall, the selected species were low in cytotoxicity, except for H. saccifera and H. trifurca. Several Hermannia species indicated promising in vitro biological activity which relate to their traditional use in treating a number of disease states.

In vitro 5-lipoxygenase and anti-oxidant activities of South African medicinal plants commonly used topically for skin diseases.

An investigation was undertaken to determine the possible mechanisms of action of medicinal plants used for dermatological pathologies. A total of 14 plant species were selected from the readily available ethnobotanical literature. 5-Lipoxygenase and DPPH (2,2-diphenyl-1-picrylhydrazyl) assays were used to determine the anti-inflammatory activity and the anti-oxidant activity of selected medicinal plants, respectively. Both aqueous and methanol extracts were tested. Among the plants screened, four species (Croton sylvaticus, Warburgia salutaris, Pentanisia prunelloides, and Melianthus comosus) displayed promising 5-lipoxygenase inhibitory activity with IC(50) values <61 ppm. A large number of plants exhibited significant anti-oxidant activities with IC(50) values between 5.27 and 83.36 ppm. Aqueous extracts of M. comosus exhibited the most potent anti-inflammatory and anti-oxidant activity.

The in vitro pharmacological activities and a chemical investigation of three South African Salvia species.

Salvia species (sage) are well known in folk medicine throughout the world. In South Africa sage is used against fever and digestive disorders. Three closely related South African species (Salvia stenophylla, Salvia repens and Salvia runcinata) were investigated for their anti-oxidant (DPPH assay); anti-inflammatory (5-lipoxygenase and cyclo-oxygenase assays); antimalarial (tritiated hypoxanthine incorporation assay); antimicrobial (disc diffusion and micro-dilution assays) properties and toxicity profile (tetrazolium-based assay). The solvent extracts exhibited anti-oxidant, antimalarial and antibacterial and poor anti-inflammatory properties. The essential oils exhibited anti-inflammatory and antimalarial properties, but displayed poor anti-oxidant and antimicrobial activity. The extract of Salviastenophylla and the essential oil of Salvia runcinata displayed the highest toxicity profile. Overall, Salvia runcinata displayed the most favorable activity of all three taxa tested with an IC(50) value of 6.09 (anti-oxidant); 29.05 (antimalarial) and 22.82 microg/ml (anti-inflammatory). Analytical procedures (GC-MS and HPLC-UV) were employed to generate chromatographic profiles for the essential oils and solvent extracts respectively. The HPLC analysis revealed the presence of rosmarinic acid in all three taxa while carnosic acid was only present in Salvia repens and Salvia stenophylla. The GC-MS analysis showed that oils were qualitatively and quantitatively variable. beta-Caryophyllene was present in large amounts in all three taxa. Other components present include camphor, alpha-pinene and alpha-bisabolol. The results of the in vitro pharmacological activities provide a scientific basis to validate the use of these Salvia species in traditional medicine in South Africa.