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This paper describes an investigation into the accumulation and distribution of Cu and Pb in Helichrysum splendidum, to assess the potential use of the plant in the phytoremediation of Cu- and Pb-contaminated soils. Plants of the same age were exposed to soils artificially contaminated with 10, 50, 100, 500 and 1000 mg/L Cu(II) and Pb(II). Although plant growth was not affected by the presence of Cu, high concentrations of Pb (> 500 mg/L) stunted the plants. Chlorophyll production was slightly reduced when plants were exposed to high concentrations of Cu (> 500 mg/L), whereas all the plants growing in Pb-contaminated soils indicated low levels of chlorophyll. Significant amounts of Cu and Pb were accumulated in the roots, with corresponding bioconcentration factors of > 1. Translocation of Cu to the shoots was restricted for plants growing in > 500 mg/L Cu-treated soils, whereas Pb was translocated to the shoots for all treatments, with translocation factors > 1. The Cu and Pb tolerance mechanism was ascribed to chelation and free radical scavenging, through the production of phenolic compounds. Helichrysum splendidum is able to adapt to and ameliorate soils contaminated with low to high levels of Cu(II) and Pb(II) and has potential for application in phytoextraction and phytostabilization of contaminated soils.

Helichrysum petiolare and Helichrysum odoratissimum, collectively known as ‘Imphepho’, are popular medicinal herbs that are indigenous to South Africa. The species are used interchangeably to relieve gastrointestinal and respiratory conditions, fever and urinary tract infections, implying that they produce antibacterial compounds. Although these herbs are sold in informal markets, they have not been commercialized. The aim of this study was to compare the chemical profiles and the antimicrobial activities of solvent extracts of the two species. The profiles of the smoke condensates were also compared, since the smoke from both species is inhaled during cultural rituals to induce central nervous system effects. After optimizing the extractant, the chemical profiles of aqueous methanol extracts of the aerial parts of twelve H. odoratissimum and fifteen H. petiolare samples, collected from several different localities, were obtained using ultra performance liquid chromatography-mass spectrometry (UPLC-MS). Three isomeric compounds were found to be present in both species. The construction of chemometric models from the data indicated a large degree of inter-population variation within H. odoratissimum, with the samples loosely clustering into two main groups. In contrast, the chemistry of H. petiolare was highly conserved. Discriminant analysis identified four distinguishing marker compounds for each species. One of the isomers common to both species, was isolated from H. odoratissimum and identified as 4,5-dicaffeoylquinic acid using nuclear magnetic resonance (NMR) spectroscopy, and is herein reported for the first time from the two species. Following validation of a developed UPLC-photodiode array (PDA) detection method, it was determined that the concentrations of the compound ranged from 3.89 - 31.1 µg/g in H. odoratissimum and from 2.3 to 13.4 µg/g in H. petiolare. High performance thin layer chromatography (HPTLC) fingerprints also indicated a greater degree of chemical variation within H. odoratissimum than in H. petiolare. This was confirmed through multivariate analysis of densitograms obtained from images of the fingerprints through the application of rTLC software, effectively mirroring the UPLC-MS results. The antibacterial activity of extracts of all the samples was determined using the minimum inhibitory concentration (MIC) serial dilution assay against selected Gram-positive (Staphylococcus saprophyticus, Enterococcus faecalis, Streptococcus agalactiae) and Gram-negative (Proteus mirabilis, Escherichia coli, Moraxella catarrhalis) pathogens that affect the urinary, respiratory and gastro-intestinal tracts. The MICs obtained for H. odoratissimum extracts indicated that these were more effective towards a larger range of bacteria than extracts of H. petiolare. The methanol extracts yielded average MICs denoting noteworthy or moderate activity towards three pathogens (E. coli, M. catarrhalis and S. agalactiae), while the chloroform:methanol extracts displayed noteworthy or moderate activity towards the same three pathogens, but also towards E. faecalis. These results justify to some degree the traditional use of H. odoratissimum to treat conditions of the gastro-intestinal, respiratory and urinary tracts. The UPLC-PDA profiles of smoke condensates of the two species, obtained under controlled conditions, revealed five major compounds that were common to both species. Although the similarity of the smoke profiles rationalises their interchangeable use for inhalation, the solvent extracts displayed little congruence regarding their chemical profiles or antibacterial activities.

The knowledge of the levels of polyphenols present in green coffee beans from different geographical origins is important to identify beans that can provide better coffee quality. The aim of this study was to determine the concentrations of both soluble and cell wall-bound polyphenols in green coffee beans (Coffea arbica L.) from different cultivation regions of Ethiopia. A total of 100 samples were collected from four geographical regions (East, West, South, Northwest) and comprised eight commercial types of coffee (Harar, Jimma, Kaffa, Wollega, Sidama, Yirgacheffe, Finoteselam and Benishangul). The total concentrations of soluble and cell wall-bound polyphenols and flavonoids in the sample extracts were determined using the Folin-Ciocalteu and aluminum chloride reagent methods, respectively. Qualitative analysis was also performed by using high performance thin layer chromatography (HPTLC). Results revealed that chlorogenic acids as the predominant constituents of the phenolic fraction of the green coffee beans. The total soluble polyphenol, cell wall-bound polyphenol and flavonoid contents of the green coffee beans were in the range of 21.8–43.6 milligrams of gallic acid equivalents per gram (mg GAE g−1), 8.6–15.3 mg GAE g−1 and 3.3–6.2 milligrams of (+)-catechin equivalents per gram (mg CE g−1) of dry mass, respectively. Cell wall-bound polyphenols were found to contribute significantly (21.0–32.5%) to the total polyphenol contents (TPC) of the green coffee beans. The polyphenol contents of green coffee beans have been found to vary with geographical origins. Harar coffee contained significantly (p

Chromones are naturally occurring phenolic compounds that are universally present in a healthy human diet. To date, thousands of chromone derivatives, including chromone glycosides have been discovered, both from natural and synthetic origin. Many plant genera, including Aloe, Aquilaria, Cassia, Hypericum and Polygonum, are rich sources of chromones. Several genera of fungi, such as Aspergillus, Mycoleptodiscus, Orbiocrella, and Penicillium produce bioactive chromones. This class of compounds is mainly associated with anti-oxidant, antimicrobial, anticancer and anti-inflammatory activities. Various in vitro, in vivo and clinical studies have established the role of chromones in alleviating allergies, reducing oxidative damage, inhibiting cancer, infections and inflammation, and for treating neurological and psychiatric disorders. This review focuses mainly on natural chromones, and elaborates on their nutritional benefits, pharmacological activities, and mechanisms of action. Various scientific databases, including PubMed, Scopus and Google Scholar, were used to find relevant literature on chromones covering all available literature up to, and including the year 2019. This critical analysis of completed research regarding the nutraceutical properties of various chromone derivatives may provide new insight to scientists working in related fields.

Acacetin (5,7-dihydroxy-4-methoxyflavone) is a naturally occurring flavonoid, known to possess numerous pharmacological properties, including neuroprotective, cardioprotective, anticancer, anti-inflammatory, antidiabetic, and antimicrobial activities. The molecule is widespread throughout the Plant Kingdom, but is particularly abundant in the Asteraceae, Euphorbiaceae, Passifloraceae, Lamiaceae and Malvaceae families. It has been shown to exhibit strong inhibitory effects against glutathione reductase, cyclo-oxygenase, acetylcholinesterase, aldose reductase and xanthine oxidase enzymes. A comprehensive overview of the potential therapeutic applications of acacetin was prepared after collating selected peer-reviewed papers, accessed from online scientific databases, specifically PubMed, Web of Science and Scopus. The mechanism of action, dose forms and structure-activity relationships of the molecule are some of the aspects discussed in the review, to facilitate further research and use of this molecule.

The National Research Foundation, the Department of Science and Technology of South Africa (through the SARChI initiative) as well as the South African Medical Research Council.

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.

Terminalia sericea is used throughout Africa for the treatment of a variety of conditions and has been identified as a potential commercial plant. The study was aimed at establishing a high-performance thin layer chromatography (HPTLC) chemical fingerprint for T. sericea root bark as a reference for quality control and exploring chemical variation within the species using HPTLC metabo3lomics. Forty-two root bark samples were collected from ten populations in South Africa and extracted with dichloromethane: methanol (1:1). An HPTLC method was optimized to resolve the major compounds from other sample components. Dichloromethane: ethyl acetate: methanol: formic acid (90:10:30:1) was used as the developing solvent and the plates were visualized using 10% sulfuric acid in methanol as derivatizing agent. The concentrations of three major bioactive compounds, sericic acid, sericoside and resveratrol-3-O-&beta, rutinoside, in the extracts were determined using a validated ultra-performance liquid chromatography-photodiode array (UPLC-PDA) detection method. The rTLC software (written in the R-programming language) was used to select the most informative retardation factor (Rf) ranges from the images of the analysed sample extracts. Further chemometric models, including principal component analysis (PCA) and hierarchical cluster analysis (HCA), were constructed using the web-based high throughput metabolomic software. The rTLC chemometric models were compared with the models previously obtained from ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS). A characteristic fingerprint containing clear bands for the three bioactive compounds was established. All three bioactive compounds were present in all the samples, although their corresponding band intensities varied. The intensities correlated with the UPLC-PDA results, in that samples containing a high concentration of a particular compound, displayed a more intense band. Chemometric analysis using HCA revealed two chemotypes, and the subsequent construction of a loadings plot indicated that sericic acid and sericoside were responsible for the chemotypic variation, with sericoside concentrated in Chemotype 1, while sericic acid was more abundant in Chemotype 2. A characteristic chemical fingerprint with clearly distinguishable features was established for T. sericea root bark that can be used for species authentication, and to select samples with high concentrations of a particular marker compound(s). Different chemotypes, potentially differing in their therapeutic potency towards a particular target, could be distinguished. The models revealed the three analytes as biomarkers, corresponding to results reported for UPLC-MS profiling and thereby indicating that HPTLC is a suitable technique for the quality control of T. sericea root bark.

Artemisia afra (African wormwood) is a popular medicinal plant of southern Africa and is an excellent candidate for commercialisation. This current study was aimed at exploring the phytochemistry and chemical variation of non-volatile compounds within wild populations of A. afra, and developing chromatographic quality control protocols for raw materials based on the identification of marker compounds. Chromatographic data, from samples representing 12 distinct populations, were obtained using liquid chromatography-mass spectrometry. An untargeted chemometric approach revealed three clusters. Marker compounds for each cluster, revealed through discriminant analysis, were isolated and identified using NMR spectroscopy, as acacetin (1) (Group 1), chrysoeriol (2) (Group 2), and 3,5-di-O-caffeoylquinic acid (3) and scopoletin (4) (Group 3). In addition, (3) and rutin (5), (both reported for the first time from A. afra), and (1), (2), (4) and 4-caffeoylquinic acid (6) were established as reliable markers for species identification, since they were abundant in most samples. Quantitative analysis using a validated method established (4) as the dominant compound in the samples (1080–19,600 μg/g dry weight (d.w.)), followed by (5) (49.5–2490 μg/g d.w.). A high performance thin layer chromatography (HPTLC) method was developed. The Rf values and colours of the bands corresponding to the marker compounds were recorded so that these compounds could be easily identified for quality control purposes. Multivariate analysis of the data using the rTLC online application confirmed the presence of different chemical groupings within the samples. It was deduced that quantitative, rather than qualitative differences, characterised the samples. Future research should focus on comparing the efficacy of the various chemical clusters in multi-target biological assays aligned to the traditional use of the plant.

Terminalia sericea Burch. ex. DC. (Combretaceae) is a popular remedy for the treatment of infectious diseases. It is widely prescribed by traditional healers and sold at informal markets and may be a good candidate for commercialisation. For this to be realised, a thorough phytochemical and bioactivity profile is required to identify constituents that may be associated with the antibacterial activity and hence the quality of raw materials and consumer products. The aim of this study was to explore the phytochemistry and identify the antibacterial constituents of T. sericea root bark, using a metabolomic approach. The chemical profiles and antibacterial activities of 42 root bark samples collected from three districts in the Limpopo Province, South Africa, were evaluated. Dichloromethane:methanol (1:1) extracts were analysed using ultraperformance liquid chromatography (UPLC)-mass spectrometry (MS), and chemometric models were constructed from the aligned data. The extracts were tested against Bacillus cereus (ATCC 11778), Staphylococcus epidermidis (ATCC 12223), Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 8739), Klebsiella pneumoniae (ATCC 13883), Pseudomonas aeruginosa (ATCC 27853), Shigella sonnei (ATCC 9292) and Salmonella typhimurium (ATCC 14028), using the minimum inhibition microdilution assay. Nine compounds, sericic acid, sericoside, resveratrol-3-O-&beta, rutinoside, ellagic acid, flavogallonic acid dilactone, methyl-flavogallonate, quercetin-3-(2&prime, &prime, galloylrhamnoside), resveratrol-3-(6&prime, galloyl)-O-&beta, d-glucopyranoside and arjunetin, were isolated from the root bark. All the compounds, with the exception of sericic acid, sericoside and resveratrol-3-O-&beta, rutinoside, were isolated for the first time from the root bark of T. sericea. Chemometric analysis revealed clustering that was not population specific, and the presence of three groupings within the samples, characterised by sericic acid, sericoside and an unidentified compound (m/z 682/4.66 min), respectively. The crude extracts from different populations displayed varied antibacterial activities against S. typhimurium (minimum inhibitory concentrations (MICs) 0.25&ndash, 1.0 mg/mL), but similar activity towards Bacillus cereus (1.0 mg/mL). Several compounds present in the root bark were highly active towards all or most of the pathogens tested, but this activity was not reflected by the chemical profiles of extracts prepared from the individual samples. Among the pure compounds tested, only flavogallonic acid dilactone and methyl-flavogallonate exhibited broad-spectrum activity. A biochemometric analysis indicated that there was no consistent association between the levels of phytochemicals and the activity of the active or non-active extracts. Although it was deduced that the major constituents of T. sericea root bark contributed to the chemotypic variation, further investigation of the interactions of compounds present in the root bark may provide antibacterial efficacies not evident when examining compounds singularly. The data reported herein will provide information that is fundamentally important for the development of quality control protocols.

In this study, the spatial and temporal variations in the levels of C8-C40 n-alkanes and 18 polycyclic aromatic hydrocarbons (PAHs) in water and sediment from Loskop Dam (Mpumalanga Province South Africa), were investigated between 2015 and 2017. In addition, their sources, which have not been well defined, were also studied over the period. This water body is sourced from a historically contaminated water body, the Olifants River, which flows through areas where a range of industrial and agricultural activities take place. Mass crocodile and fish mortalities have been recorded in this aquatic system, and contamination by organic pollutants were highlighted as a contributing factor. The total average n-alkane concentrations in water and sediments ranged from 0.574±00811 to 18.8±1.39 µg/L and 4760±243 to 30700±906 µg/kg, respectively. Similarly, PAHs were detected at total average concentrations of between 0.150±00494 and 49.8±6.86 µg/L in water and 61.6±5.95 to 2618±300 µg/kg. n-Alkane and PAH diagnostic ratios indicated a mixture of sources of these compounds, attributed to terrestrial, submerged and floating plant material, as well as petrogenic and pyrogenic combustion. Inlet, middle and upper segment site clustering was observed with non-metric multidimensional scaling (NMDS) and hierarchical cluster analysis (HCA), mainly driven by the prevalence of PAHs at the inlet sites and n-alkanes in the upper reaches. By using indicator compounds, the sources of contamination could be predicted. The strategy described here can be applied to any water body for continuous long-term monitoring of pollutant levels and to identify sources attributing to water pollution.

Sceletium tortuosum (Aizoaceae) is widely recognised for the treatment of stress, anxiety and depression, as well as for obsessive compulsive disorders. A comprehensive intraspecies chemotypic variation study, using samples harvested from two distinct regions of South Africa, was done using both proton nuclear magnetic resonance (1H-NMR) spectroscopy of methanol extracts (N = 145) and ultra performance liquid chromatography-mass spectrometry (UPLC-MS) of acid/base extracts (N = 289). Chemometric analysis of the 1H-NMR data indicated two main clusters that were region-specific (Northern Cape and Western Cape provinces). Two dimensional (2D) NMR was used to identify analytes that contributed to the clustering as revealed by the S-plot. The sceletium alkaloids, pinitol and two alkylamines, herein reported for the first time from S. tortuosum, were identified as markers that distinguished the two groups. Relative quantification of the marker analytes conducted by qNMR indicated that samples from the Northern Cape generally contained higher concentrations of all the markers than samples from the Western Cape. Quantitative analysis of the four mesembrine alkaloids using a validated UPLC-photo diode array (PDA) detection method confirmed the results of qNMR with regard to the total alkaloid concentrations. Samples from the Northern Cape Province were found to contain, on average, very high total alkaloids, ranging from 4938.0 to 9376.8 mg/kg dry w. Regarding the Western Cape samples, the total yields of the four mesembrine alkaloids were substantially lower (averages 16.4–4143.2 mg/kg). Hierarchical cluster analysis of the UPLC-MS data, based on the alkaloid chemistry, revealed three branches, with one branch comprising samples primarily from the Northern Cape that seemed somewhat chemically conserved, while the other two branches represented mainly samples from the Western Cape. The construction of an orthogonal projections to latent structures-discriminant analysis model and subsequent loadings plot, allowed alkaloid markers to be identified for each cluster. The diverse sceletium alkaloid chemistry of samples from the three clusters may facilitate the recognition of alkaloid profiles, rather than individual compounds, that exert targeted effects on various brain receptors involved in stress, anxiety or depression.

Athrixia phylicoides, known as “bush tea”, grows abundantly in South Africa. An infusion of the leaves is used as a beverage and to treat a multitude of health conditions. The aim of this study was to investigate the chemical variation within A. phylicoides and to identify characteristic compounds for quality control. Samples from 12 locations in South Africa were analysed using ultra-performance liquid chromatography. Hierarchical cluster analysis of the aligned ultra-performance liquid chromatography-time-of-flight mass spectrometry data indicated two groups on the resulting dendrogram, representing 48 samples. Five marker compounds, identified through visual inspection and the construction of a discriminant analysis model, were evident on the ultra-performance liquid chromatography-MS profiles. Four of these compounds were isolated and identified, three as hydroxy methoxyflavones and the fourth as a coumarate, using nuclear magnetic resonance spectroscopy. An ultra-performance liquid chromatography-photodiode array method was developed and validated for the determination of the marker compounds using the isolates as standards. The limits of detection for the four compounds ranged from 0.92 – 2.50 µg/mL. Their recoveries at three concentration levels (1.00, 10.0, and 100 µg/mL) were between 97.0 and 101%, while acceptable intra- and inter-day precision was obtained as reflected by percentage relative standard deviation values below 2.24%. The concentrations of all the marker compounds were found to be higher in samples corresponding to Group 1 of the dendrogram than in those from Group 2. This may be attributable to differences in altitude, climate, and some edaphic factors. Identification of these marker compounds will make a valuable contribution towards the quality control and sustainable commercialisation of bush tea.

Consumption of a refreshing beverage prepared from the dried leaves and twigs of Athrixia phylicoides, commonly referred to as bush tea, is widespread in South Africa. The tea has an illustrious history of use by the indigenous people of southern Africa and has the potential for commercialisation. Several ethnic groups use decoctions and pastes prepared from the plant to treat a multitude of unrelated conditions. This review is aimed at providing a comprehensive overview of the ethnopharmacology, phytochemistry, biological activities, toxicity and commercial aspects of A. phylicoides. Books, containing botanical and ethnopharmacological information were extensively consulted, and electronic databases were searched to acquire relevant articles, with no specific time frame set. Although the majority of the traditional uses indicate that A. phylicoides has strong antimicrobial properties, this is not well supported by scientific data, suggesting that more research is required involving more pathogens and the investigation of a potential alternative mode of action. The secondary metabolites of the plant have been extensively explored and several sesquiterpenes, coumarins, flavonoids and phenol carboxylic acids have been identified. However, there is a paucity of information regarding the chemical variation within the species. A multidisciplinary metabolomic approach involving sophisticated and information-rich technologies, in combination with multivariate analysis, should be undertaken to record chemotypic variation and to identify potential biomarker compounds for quality control. Some of the ethnobotanical uses have been supported through scientific studies, but a window of opportunity exists to contribute data on the pharmacological activities. Despite efforts to facilitate the commercialisation of bush tea in South Africa in an effort to grow the bio-economy of the region, more fundamental research is required to ensure the sustainability and expansion of such an industry. The commercialisation of rooibos and honeybush tea in South Africa is underpinned by basic and applied research and serves as appropriate models for the development of the bush tea industry.

The combustion of coal in thermal power plants may result in high concentrations of elements in the coal fly ash remaining that may be toxic to living organisms or pose a risk to the environment. This study was aimed at determining the concentrations of potentially toxic elements in coal fly ash leachates, in an attempt to simulate natural processes that influence the environment. Powder X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and elemental composition studies were employed to characterize the physicochemical properties of the parent coal fly ash. The effect of various leaching parameters, including the pH of leaching solutions, the volume ratio of leaching solutions to the mass of coal fly ash, leaching time and temperature, were investigated. Moreover, the kinetics of the leaching of toxic elements from coal fly ash was also investigated by considering that the leaching process is governed by dissolution. Zero order, pseudo-first order, pseudo-second order, power function, simple Elovich and parabolic diffusion kinetic models were used to evaluate the leaching process. The experimental results indicate that the pH and leaching time had the most significant effect on the leaching behavior of elements from coal fly ash.

Rivers feeding into the Loskop Dam, South Africa, pass through coal mining and heavily industrialised areas. Previous studies investigating mass mortalities of crocodile and fish in the river system, revealed the presence of organic compounds, including polycyclic aromatic hydrocarbons (PAHs), in their tissues. Samples were collected from nine sites within the dam in winter and summer. Liquid-liquid and microwave-assisted extraction was used for preconcentrating PAHs from water and sediment samples, respectively. Extracts were analysed by gas chromatography-mass spectrometry in the selected ion monitoring mode to determine the presence and levels of the 16 United States Environmental Protection Agency (US EPA)-priority PAHs. Significant levels of PAHs were found in both water and sediment samples. Concentrations were higher in sediments than in water, as well as in samples collected in winter. Levels of PAHs in sediments were generally higher than those reported by many researchers for other water bodies in industrialised areas. Zebrafish embryos were used to assess potential risks associated with the water and sediment, and to determine the effects of PAHs on aquatic life. Embryos, exposed to intact water and sediment samples, as well as to diluted sediment extracts, were monitored for 120 h post fertilisation. Sediment proved more toxic to zebrafish embryos than water, causing delayed embryo development and malformations. These findings are alarming as they indicate that this water system is under stress. These findings can be typical of any water system situated in a coal mining and industrial region.

The volatile constituents of nine liverworts species ( Asterella marginata , Dumortiera hirsuta , Fossombronia swaziensis , Marchantia pappeana , Marchantia polymorpha subsp. ruderalis , Pallavicinia lyellii , Plagiochasma rupestre , Riccia albolimbata and Symphyogyna podophylla ) from South Africa were determined by gas chromatography. Where possible the volatile constituents identified were compared to those reported for the same species from other countries. Among the analysed liverworts, the chemical compositions of A. marginata , F. swaziensis , M. pappeana , and R. albolimbata are reported for the first time. Each of the analysed liverwort species produced its own characteristic components. Thujopsanes, chamigranes and cuparanes were found to be the most characteristic components of M. polymorpha subsp. ruderalis . The presence of dumortane-type sesquiterpenoids in D. hirsuta indicated that this species is similar in chemistry to an Argentinean sample. This is one of only a handful of reports on the presence of this sesquiterpene-type in liverworts. Simple thallose liverworts, S. podophylla and P. lyellii , were characterized by the presence of labdane-type diterpenoids. In addition, the antimicrobial activities of chloroform:methanol (1:1) extracts of the liverworts were evaluated against several important human pathogens using the serial dilution assay. Four of the liverwort species were active against Pseudomonas aeruginosa with MIC values ranging from 0.50 to 1.0 mg/mL. Some activity (MIC value of 1.0 mg/mL) was also recorded for the crude extracts of P. lyellii and M. pappeana against Escherichia coli. The extract of S. podophylla displayed the best activity towards the yeast Cryptococcus neoformans (MIC = 1.0 mg/mL). Although approximately 300 liverwort species occur in southern Africa, a Scopus search confirmed that this is the first report of the volatile profiles and biological properties of species from the region.

Mango blossom malformation, caused by infection of mango panicles and young shoots by Fusarium species, leads to significant reductions in fruit yield. Previously, chemometric models were established that allowed biomarkers associated with resistance in tolerant cultivars to be identified. High concentrations of these biomarkers, mangiferin, maclurin and maclurin O-galloyl-glucoside, are inherent genetic traits of some cultivars and have been linked to their ability to confine Fusarium infection. In this study, phenolic profiles of mature leaf extracts from cultivars exhibiting different levels of resistance to Fusarium infection were obtained by UPLC-Q-ToF x MS, five tolerant and seven susceptible cultivars. A robust prediction model, was developed that could be used throughout the season to predict the likelihood of new cultivars being susceptible or tolerant towards mango malformation disease. The levels of biomarkers revealed by the models in tolerant and susceptible cultivars were compared and significant differences were observed. These models can serve as an important tool to investigate appropriate cultivars, prior to their introduction to areas prone to the disease.

Forty-nine green coffee (Coffea arabica L.) bean samples, representing the Harar, Jimma, Kaffa, Wollega, Sidama, and Yirgachefe varieties from east, west, and south Ethiopia were analyzed for Mg, P, S, Ca, Mn, Fe, Cu, Ba, Si, and K using inductively coupled plasma-optical emission spectroscopy. Principal component analysis was used to group the samples based on their elemental compositions. Application of linear discriminant analysis provided models with an accuracy of 92% for region of production and 79% for variety. The elements P, Mn, S, Cu, and Fe were the most discriminating elements. These results show that initial screening may be performed to characterize possible fraudulently labelled products with verification by the existing paper-based traceability system and sensory evaluation.

A total of 100 samples of green coffee (Coffea arabica L.) beans from the major producing regions of Ethiopia were studied using ultra performance liquid chromatography–mass spectroscopy to determine if the phenolic content could be linked to their geographical origins for authentication purposes. Principal component analysis allowed the most discriminating compounds to be identified. Based on their concentrations, 3-O-caffeoylquinic and 4,5-O-dicaffeoylquinic acids were found to be characteristic markers for Northwest and East (Harar) region coffees, respectively. Sub-regional coffee types from West, except Jimma B, could be distinguished by their 3,5-O-dicaffeoylquinic to 4,5-O-dicaffeoylquinic acid concentration ratios, while Yirgachefe coffees from South could be distinguished by their 4,5-O-dicaffeoylquinic to 3,4-O-dicaffeoylquinic acid concentration ratios. Linear discriminant analysis provided a classification model with recognition and prediction abilities of 91% and 90%, respectively, at regional level, and 89% and 86%, respectively, at sub-regional level. This is important for the detection of fraud, including the selling of inferior Ethiopian coffees under the label of the more expensive Harar coffees.

Contamination of maize by fumonisin mycotoxins is of major concern to consumer health. This study was designed to improve our understanding of the binding of fumonisin B (FB) to maize and its major macrocomponents, during the preparation of porridge. None of the processing treatments (pH 4, 7 and 10) or temperatures (30 and 90 °C) used resulted in a significant decrease in the recovery of FB1 from spiked starch, zein or maize meal. These results confirm the stability of FB1 in maize porridge, and the absence of modified forms of FB. The bioaccessibility of fumonisins in the gastrointestinal tract, following consumption of contaminated stiff maize porridge, was determined using the TNO Gastrointestinal Model (tiny-TIM). The porridge was digested in the tiny-TIM using parameters simulating human digestion. The concentrations of fumonisins were determined in the dialysates at fixed time intervals using ultra performance liquid chromatography-quadrupole Time-of-Flight high definition mass spectrometry (UPLC-QToF-HDMS). The results proved that fumonisins B1, B2 and B3 are released rapidly from stiff porridge prepared from contaminated maize meal. This confirms that the low bioavailability of fumonisins cannot be attributed to strong association of the toxin with the intestinal content. This study presents new insights into the stability of fumonisins in maize porridge and the kinetics of fumonisin-release from porridge through simulated human digestion, suggesting that consumers using maize as a staple may be at risk.

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 + β-caryophyllene (1.4 – 29.0%), T-cadinol (1.2 – 20.0%), α-eudesmol (trace – 23.0%) and β-eudesmol (trace – 26.0%), those of S. lanceolata comprised mainly terpinene-4-ol + β-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.

Piper methysticum Forst., commonly referred to as kava, has been used medicinally and recreationally by inhabitants of the South Pacific Islands for centuries. Kavalactones present in roots and aerial parts are regarded as the bioactive compounds responsible for the relaxant effects, and for mitigating anxiety and stress-related conditions. The development of pharmaceutical products containing root extracts led to a boom in kava sales in Europe in 1998. However, reported cases of toxicity led to the subsequent banning of kava products in several countries. This study was initiated to develop rapid, robust and alternative spectroscopic methods for quality control that can be implemented at the point of export, to authenticate the use of kava roots as legislated by the Vanuatu Amended Kava Act no. 6 (2015). Roots, peeled stems, and stump peelings samples (n = 47) were sourced from Fiji, Hawaii, Samoa, the Solomon Islands and Tonga. The sample extracts were analysed using ultra performance liquid chromatography coupled to a photodiode array detector and mass spectrometer (UHPLC-PDA/MS), while powdered material was analysed using spectroscopic techniques. These included single-point (near-infrared (NIR) and mid-infrared (MIR) spectroscopy), as well as an imaging (hyperspectral imaging). Principal component analysis of both the raw UPLC-MS and the quantitative UPLC-PDA data revealed chemical differences between the root and non-root samples. Kavain, methysticin and yangonin were identified as the compounds largely responsible for the chemical differences between the plant parts. Discriminant analysis models (OPLS-DA and PLS-DA) were developed for all the techniques, to reliably discriminate kava roots from non-roots. All the discriminant models indicated a good prediction ability (Q2XCum ≥ 60 %) and were successfully used to accurately identify external roots and non-root samples. However, hyperspectral imaging yielded superior results, with a prediction ability above 90 %. This technique can be automated and is capable of continuously scanning multiple samples, making it ideal for quality control.

A molecularly imprinted polymer (MIP) was prepared using (-)-norephedrine as the template, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linker and chloroform as the porogen. The MIP was used as a selective sorbent in the molecularly imprinted solid-phase extraction (MIP-SPE) of the psychoactive phenylpropylamino alkaloids, norephedrine and its analogs, cathinone and cathine, from Khat (Catha edulis Vahl. Endl.) leaf extracts prior to HPLC-DAD analysis. The MIP was able to selectively extract the alkaloids from the aqueous extracts of Khat. Loading, washing and elution of the alkaloids bound to the MIP were evaluated under different conditions. The clean baseline of the Khat extract obtained after MIP-SPE confirmed that a selective and efficient sample clean-up was achieved. Good recoveries (90.0-107%) and precision (RSDs 2.3-3.2%) were obtained in the validation of the MIP-SPE-HPLC procedure. The content of the three alkaloids in Khat samples determined after treatment with MIP-SPE and a commercial Isolute C18 (EC) SPE cartridge were in good agreement. These findings indicate that MIP-SPE is a reliable method that can be used for sample pre-treatment for the determination of Khat alkaloids in plant extracts or similar matrices and could be applicable in pharmaceutical, forensic and biomedical laboratories. Copyright © 2015 John Wiley & Sons, Ltd.

The alkaloid compositions of 99 green coffee (Coffea arabica L.) bean samples comprising eight varieties (Harar, Jimma, Kaffa, Wollega, Sidama, Yirgachefe, Benishangul and Finoteselam) from the major production regions of Ethiopia were investigated. High performance liquid chromatography was applied for the simultaneous determination of four coffee alkaloids in the aqueous extracts of the beans. The limits of detection for the method were established as 13 mg kg−1 for trigonelline, 7 mg kg−1 for theobromine, 8.5 mg kg−1 for caffeine and 4 mg kg−1 for theophylline in the dry coffee beans. Theophylline was not detected in any of the samples. The determined concentrations (% w/w dry coffee beans) ranged from 0.98 to 1.32 % for trigonelline, 0.0048 to 0.0094 % for theobromine and 0.87 to 1.38 % for caffeine. The concentrations of the alkaloids varied significantly, depending on the geographical origin of the beans. Theobromine was not detected in coffee beans from the East (Harar coffees), and its absence in samples can be used to ascertain whether the coffee originates from this region. Coffee beans from the Northwest were characterized by higher concentrations of caffeine. Application of linear discriminant analysis provided 75 % correct classification of samples into the respective production regions, with a 74 % prediction success rate. The moderate classification efficiency obtained when using alkaloid data demonstrates the potential of using this class of compounds in discriminant models for determination of the geographical origin of green coffee beans from Ethiopia.

Embelin(2,5-dihydroxy-3-undecyl-p-benzoquinone)is known for its potent anthelmintic activity, but also for wound-healing, antidiabetic, anticonvulsant, antitumour, anti-inflammatory, analgesic, hepatoprotective, antioxidant, antibacterial andantispermatogenic activities. A high performance countercurrent chromatography method was developed for the purification of embelin from an extract of the seeds of Embelia schimperi fruit. The optimised solvent system (n-hexane:ethylacetate:ethanol:water (7:3:7:3)) resulted in the isolation of 13.9 mg of embelin, directly from 100 mg of crude extract, in a single step within a short retention time (40 min). Although the compound appeared to be completely pure when analysed by ultra performance liquid chromatography (UPLC) with photo diode array detection (PDA), the purity was established as approximately 90% by UPLC-mass spectrometry (MS). Furthermore, we report the fatty acid composition of the seeds of Embelia schimperi fruit. Nine fatty acids were quantified from the fruit seed extract by gas chromatography-mass spectrometry, with linoleic (46.4%), palmitic (21.5%), and oleic (19.6%) acids making up the largest proportions.

Quorum sensing controls bacterial pathogenesis and virulence, hence, interrupting this system renders pathogenic bacteria non-virulent, and presents a novel treatment for various bacterial infections. In the search for novel anti-quorum sensing (AQS) compounds, 14 common culinary herbs and spices were screened for potential antipathogenicity activity against Chromobacterium violaceum ATCC 12472. Extracts of Glycyrrhiza glabra (liquorice), Apium graveolens (celery), Capsicum annuum (cayenne pepper) and Syzygium anisatum (aniseed) demonstrated good AQS potential, yielding opaque halo zones ranging from 12&ndash, 19 mm diameter at sub-minimum inhibitory concentrations (0.350&ndash, 4.00 mg/mL). For the same species, the percentage reduction in violacein production ranged from 56.4 to 97.3%. Zones with violacein inhibitory effects were evident in a celery extract analysed using high performance thin layer chromatography-bio-autography. The major active compound was isolated from celery using preparative-high performance liquid chromatography-mass spectrometry and identified using gas chromatography-mass spectrometry (GC-MS) as 3-n-butyl-4,5-dihydrophthalide (sedanenolide). Potent opaque zones of inhibition observed on the HPTLC-bio-autography plate seeded with C. violaceum confirmed that sedanenolide was probably largely responsible for the AQS activity of celery. The bacteriocidal properties of many herbs and spices are reported. This study, however, was focussed on AQS activity, and may serve as initial scientific validation for the anti-infective properties ascribed to several culinary herbs and spices.

Henna (Lawsonia inermis) is applied to stain keratin, present in hair, skin and fingernails, a red-orange or rust colour. Producers of temporary tattoos mix the aromatic amine compound, para-phenylenediamine (PPD) into natural henna to create 'black henna' that rapidly stains the skin black. However, PPD may cause severe delayed hypersensitivity reactions following skin contact. This study proposes a rapid direct-analysis method to detect and identify PPD using an atmospheric solids analysis probe (ASAP) coupled to a Q-ToF mass spectrometer (MS). Since laborious, multistep methods of analysis to determine PPD are undesirable, due to the instability of the compound in solution, a screening method involving no sample preparation steps was developed. Experiments were carried out to optimise the corona current, sample cone voltage, source temperature, and desolvation gas temperature to determine ideal ASAP-Q-ToF-MS analysing conditions. Eleven of the 109 henna samples, originating from various countries, tested positive for PPD when henna products were screened using ASAP-MS, without any form of sample preparation other than grinding. Ultra-performance liquid chromatography electrospray ionisation-mass spectrometry (UPLC-Q-ToF-MS) was subsequently used to confirm the results from ASAP and to determine the concentrations of PPD in henna products. The allergen was detected in the same eleven samples, with concentrations ranging from 0.05-4.21% (w/w). It can be concluded that the sensitivity of the ASAP-MS technique is sufficient (limit of detection=0.025% w/w) to allow screening of henna samples for the presence of PPD. This relatively new technique can be applied to commercial products without extraction, sample treatment or chromatographic separation.