Phenolic Content Analysis of Two Species Belonging to the Lamiaceae Family: Antioxidant, Anticholinergic, and Antibacterial Activities.

The Lamiaceae family are utilized as ornamental, medicinal, and food supplements throughout the world. The current study focuses on a comparative analysis of the phenolic compositions and bioactivities (including antioxidant, anticholinergic, and antibacterial activities) of ethanolic extracts derived from the aerial parts of the two species (Lavandula stoechas L. and Thymus sipyleus Boiss). The presence of phenolic compounds and phytochemicals in the plant extracts was identified using the LC-MS/MS technique. The LC-MS/MS analysis revealed that vanillic acid (125,596.66 µg/L) was the most abundant phytochemical in L. stoechas. Kaempferol (8550.52 µg/L) was the most abundant substance in Thymus sipyleus. The assessment of the antioxidant efficacy of the species extracts was conducted using the DPPH (2.2-diphenyl-1-picryl-hydrazyl-hydrate), ABTS (2.2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)), Fe3+-Fe2+ reducing, and CUPRAC (Cu2+-Cu+ reducing) assays. The anticholinergic activity of the samples was determined using the acetylcholinesterase (AChE) inhibition assay. The results of antioxidant activity were higher in the T. sipyleus than in the L. stoechas ethanol extracts. The extracts of L. stoechas exhibited radical scavenging activity ranging from 15 to 18%, while T. sipyleus had activity effects ranging from 34% to 38%. The AChE inhibition potential for L. stoechas and T. sipyleus extracts as IC50 values were 0.221 ± 0.01 mg/mL and 0.067 ± 0.02 mg/mL, respectively. The antibacterial effects of the ethanolic extracts of these species against pathogenic bacteria isolates were determined using the MIC (minimal inhibitory concentration) method. These findings indicated that the extracts from L. stoechas and T. sipyleus possess the potential to be natural antioxidants in the realm of food preservation. Additionally, their antioxidant, anticholinergic, and antimicrobial properties suggest potential therapeutic utility in the management of certain diseases.

A General Review of Methodologies Used in the Determination of Cholesterol (C27H46O) Levels in Foods.

Cholesterol (C27H46O) is a lipid-derived substance found in lipoproteins and cell membranes. It is also one of the main sources for the production of bile acids, vitamin D, and steroid hormones. Today, foods are evaluated by consumers not only according to their taste and nutritional content but also according to their effects on consumer health. For example, many consumers choose foods according to their cholesterol level. The cholesterol in the food can directly affect the blood cholesterol level when consumed, which can lead to cardiovascular diseases. High levels of cholesterol can lead to diet-related human diseases such as cardiac arrest, paralysis, type II diabetes, and cerebral hemorrhage. In societies with high living standards, interest in and consumption of foods that lower or have low cholesterol levels have increased recently. Accordingly, efforts to increase the variety of foods with reduced cholesterol levels are on the rise. This has indirectly led to the accurate measurement of cholesterol levels in blood and food being of great importance. Classical chemical, enzymatic, colorimetric, polarographic, chromatographic, and spectrophotometric methods; enzymatic, nonenzymatic, and electrochemical sensors; and biosensors are used for the determination of cholesterol in foods. The purpose of this review is to reveal and explore current and future trends in cholesterol detection methods in foods. This review will summarize the most appropriate and standard methods for measuring cholesterol in biological components and foods.

Heavy Metal Pollution in the Environment and Its Impact on Health: Exploring Green Technology for Remediation.

Along with expanding urbanization and industrialization, environmental pollution which negatively affects the surroundings, has been rising quickly. As a result, induces heavy metal contamination which poses a serious threat to living organisms of aquatic and soil ecosystems. Therefore, they are a need to ameliorate the effects cost by cost pollution on the environment. In this review, we explore methods employed to mitigate the effects caused by heavy metals on the environment. Many techniques employed to manage environmental pollution are tedious and very costly, necessitating the use of alternative management strategies to resolve this challenge. In this concept, bioremediation is viewed as a future technique, due to its environmental friendliness and cost-effective measures aligned with sustainable or climate-smart agriculture to manage contaminants in the environment. The technique involves the use of living entities such as bacteria, fungi, and plants to deteriorate toxic substances from the rhizosphere. Currently, bioremediation is thought to be the most practical, dependable, environmentally benign, and long-lasting solution. Although bioremediation involves different techniques, they are still a need to find the most efficient method for removing toxic substances from the environment. This review focuses on the origins of heavy metal pollution, delves into cost-effective and green technological approaches for eliminating heavy metal pollutants from the environment, and discusses the impact of these pollutants on human health.

Unveiling the Dual Nature of Heavy Metals: Stressors and Promoters of Phenolic Compound Biosynthesis in Basilicum polystachyon (L.) Moench In Vitro.

The global industrial revolution has led to a substantial rise in heavy metal levels in the environment, posing a serious threat to nature. Plants synthesize phenolic compounds under stressful conditions, which serve as protective agents against oxidative stress. Basilicum polystachyon (L.) Moench is an herbaceous plant of the Lamiaceae family. Some species within this family are recognized for their capacity to remediate sites contaminated with heavy metals. In this study, the effects of mercury (II) chloride and lead (II) nitrate on the in vitro propagation of B. polystachyon were investigated. Shoot tips from in vitro plantlets were cultured in Murashige and Skoog's (MS) media with heavy metals ranging from 1 to 200 µM to induce abiotic stress and enhance the accumulation of phenolic compounds. After three weeks, MS medium with 1 µM of lead (II) supported the highest shoot multiplication, and the maximum number of roots per explant was found in 100 µM of lead (II), whereas a higher concentration of heavy metals inhibited shoot multiplication and root development. The plantlets were hardened in a greenhouse with a 96% field survival rate. Flame atomic absorption spectroscopy (FAAS) was used to detect heavy metal contents in plant biomass. At both 200 µM and 50 µM concentrations, the greatest accumulation of mercury (II) was observed in the roots (16.94 ± 0.44 µg/g) and shoots (17.71 ± 0.66 µg/g), respectively. Similarly, lead (II) showed the highest accumulation in roots (17.10 ± 0.54 µg/g) and shoots (7.78 ± 0.26 µg/g) at 200 µM and 50 µM exposures, respectively. Reverse-phase high-performance liquid chromatography (RP-HPLC) identified and quantified various phenolic compounds in B. polystachyon leaves, including gallic acid, caffeic acid, vanillic acid, p-coumaric acid, ellagic acid, rosmarinic acid, and trans-cinnamic acid. These compounds were found in different forms, such as free, esterified, and glycosylated. Mercury (II)-exposed plants exhibited elevated levels of vanillic acid (1959.1 ± 3.66 µg/g DW), ellagic acid (213.55 ± 2.11 µg/g DW), and rosmarinic acid (187.72 ± 1.22 µg/g DW). Conversely, lead (II)-exposed plants accumulated higher levels of caffeic acid (42.53±0.61 µg/g DW) and p-coumaric acid (8.04 ± 0.31 µg/g DW). Trans-cinnamic acid was the predominant phenolic compound in control plants, with a concentration of 207.74 ± 1.45 µg/g DW. These results suggest that sublethal doses of heavy metals can act as abiotic elicitors, enhancing the production of phenolic compounds in B. polystachyon. The present work has the potential to open up new commercial opportunities in the pharmaceutical industry.

Nutritional Supplementation of Yogurt with Jerusalem Artichoke Tubers: Organic Acid Profiles and Quality Parameters.

Jerusalem artichoke (Helianthus tuberosus L.), also called wild sunflower, belongs to the Asteraceae family and is cultivated widely across the temperate zone for its nutritious tuber, which is used as a root vegetable. In this study, the Jerusalem artichoke (JA) was used as a supplementary additive for producing a functional yogurt, with enhanced health benefits and improving the microbiological, rheological, and sensorial quality characteristics of the product. The effects of the three different concentrations (1%, 2%, and 3%, w/w) of JA on the physicochemical properties, bacterial counts, sensorial properties, and organic acid profiles of yogurts were determined after 1, 7, 14, and 21 days of storage at ±4 °C. The results obtained revealed that with the addition of different concentrations of JA the overall parameters were statistically significant (p < 0.01 and p < 0.05) except for apparent viscosity, Streptococcus thermophilus, yeast and mold count, pyruvic ratios, and scores of flavor. Similarly, some parameters (fat ratio, yeast and mold count, concentrations of propionic, acetic, pyruvic, orotic, and lactic acids, and scores of appearance, consistency, and odor) changed depending on the storage time, while some did not show any changes regarding storage time. There was a relationship found between the concentration of JA and organic acid ratio (except for pyruvic acid) concentration in the yogurt samples. In conclusion, the research revealed the effect of JA in yogurt production as a thickener, flavor enhancer, prebiotic agent, and source of organic acids and bioactive compounds. The results indicate that JA has a good potential for enhancing the nutritional and physicochemical properties of the studied yogurt.

Ethnobotany and Toxicity Status of Medicinal Plants with Cosmeceutical Relevance from Eastern Cape, South Africa.

The indigenous people of the Eastern Cape residing within the richest plant biodiversity in the world, including Africa's floral 'gold mine', have a long history of plant use for skincare. However, such rich flora comes with numerous plants that have the potential to cause harm to humans through their usage. Therefore, the study was aimed at documenting the toxicity status of important medicinal plants used by the indigenous people from the Eastern Cape for skincare and supported by literature for cosmeceutical relevance. A list of plants used for skincare was produced following an ethnobotanical survey. In addition, data on the level of toxicity and cosmeceutical relevance of plants listed from the survey were collected from literature resources. The study listed a total of 38 plants from 25 plant families, the majority being represented by the Asphodelaceae and Asteraceae, both at 13.2%. The most preferred plant parts were the leaves (60.4%) indicating sustainable harvesting practices by the community. The literature reports validated 70% of the medicinal plants surveyed for skincare were nontoxic. Most of the plants can be incorporated in the formulation of products intended for skincare due to their low toxicity and high cosmeceutical relevance.

Exploring the anti-HIV-1 reverse transcriptase, anti-inflammatory, anti-cancer activities and cytotoxicity of two fermented commercial herbal concoctions sold in Limpopo Province of South Africa.

BACKGROUND AND OBJECTIVES: The use of herbal concoctions is very popular in South Africa, including Limpopo Province. The herbal concoctions are claimed to be capable of treating numerous illnesses such as ulcers, cancer, HIV/AIDS, diabetes, certain STDs, blood cleansing to mention but a few. The focus of this study was to evaluate the anti-HIV 1 reverse transcriptase, anti-inflammatory and anti-cancerous activities as well as cytotoxic effects of 2 fermented herbal concoctions used for the treatment of the related ailments in Limpopo province of South Africa. METHOD: Two fermented herbal concoctions obtained from a herbalist in Polokwane were extracted with 80% acetone. The anti-HIV activity of the herbal concoctions was determined using the anti-HIV reverse transcriptase assay. The anti-cancer and cytotoxic effects of the herbal concoctions were evaluated using cancerous Human Colon (HT-29) cells and the normal human Hepatoma cells (C3A) respectively. RESULTS: Notable anti-HIV reverse transcriptase activity was observed from the 80% acetone fraction of herbal concoction 1 (IC50 38.031 µg/mL) which exhibited better activity than the positive control Lamivudine (IC50 40.90 µg/mL). There was variation in the anti-inflammation activity as determined by the sPL2, 15-LOX and COX enzyme assays. The only concerning matter was the high COX-1 activity in some of the extracts, which is not desirable due to the mucosal protection action of COX-1 enzyme. The herbal concoctions did not exhibit cytotoxic effects on normal human cells, however, toxicity against cancerous cells was observed. CONCLUSION: The herbal concoctions displayed some considerable pharmacological effects against various ailments as claimed by the herbalist. More work to ascertain the toxicity of both concoctions against cancerous cells need to be followed as this could lead to the discovery of anticancer drugs.

The Potential Therapeutic Value of Medicinal Plants in the Management of Metabolic Disorders.

Metabolic syndrome (MetS) is a prevalent, multifactorial and complex disease that is associated with an increased risk of developing diabetes and other major cardiovascular complications. The rise in the global prevalence of MetS has been attributed to genetic, epigenetic, and environmental factors. The adoption of sedentary lifestyles that are characterized by low physical activity and the consumption of high-energy diets contributes to MetS development. Current management criteria for MetS risk factors involve changes in lifestyle and the use of pharmacological agents that target specific biochemical pathways involved in the metabolism of nutrients. Pharmaceutical drugs are usually expensive and are associated with several undesirable side effects. Alternative management strategies of MetS risk factors involve the use of medicinal plants that are considered to have multiple therapeutic targets and are easily accessible. Medicinal plants contain several different biologically active compounds that provide health benefits. The impact of phytochemicals present in local medicinal plants on sustainable health and well-being of individuals has been studied for many years and found to involve a plethora of complex biochemical, metabolic, and physiological mechanisms. While some of these phytochemicals are the basis of mainstream prescribed drugs (e.g., metformin, reserpine, quinine, and salicin), there is a need to identify more medicinal plants that can be used for the management of components of MetS and to describe their possible mechanisms of action. In this review, we assess the potential health benefits of South African ethnomedicinal plants in protecting against the development of health outcomes associated with MetS. We aim to provide the state of the current knowledge on the use of medicinal plants and their therapeutically important phytochemicals by discussing the current trends, with critical examples from recent primary references of how medicinal plants are being used in South African rural and urban communities.

Effect of crude Moringa oleifera Lam. seed extract on the blood markers of metabolic syndrome in high-fructose diet-fed growing Sprague-Dawley rats.

Background Moringa oleifera seed has anti-diabetic and anti-obesogenic properties. This study interrogated the effect of crude hydroethanolic M. oleifera seed extract on the blood markers of metabolic syndrome (MetS) in high-fructose diet fed growing Sprague-Dawley rats. Methods Sixty 21-day old female and male Sprague-Dawley rat pups were randomly allocated to and administered one of the following treatment regimens daily for twelve weeks: group I - plain drinking water (PW)+plain gelatine cube (PC), group II - 20% (w/v) fructose solution (FS)+PC, group III - FS+100 mg/kg body mass fenofibrate in gelatine cube (FN), group IV - FS+low dose (50 mg/kg body mass) of M. oleifera in gelatine cube (LMol) and group V - FS+high dose (500 mg/kg body mass) of M. oleifera in gelatine cube (HMol). The rats in each treatment regimen had ad libitum access to a standard rat chow. After the 12-week trial, the rats were subjected to an oral glucose tolerance test and then euthanised 48 h later. Blood was collected. Plasma triglyceride, cholesterol and insulin concentration were determined. HOMA-IR was then computed. Results The high-fructose diet increased (p<0.05) plasma insulin concentration and HOMA-IR in female rats only. It increased plasma triglyceride concentration in both female and male rats and plasma cholesterol concentration in male rats only. The crude hydroethanolic M. oleifera seed extract prevented the high-fructose diet-induced metabolic derangements in male and female rats. Conclusion Crude hydroethanolic M. oleifera seed extract can potentially be used as a prophylactic intervention for diet-induced MetS in children.

Gamma radiation treatment activates glucomoringin synthesis in Moringa oleifera

Abstract Plants are a very rich source of pharmacologically relevant metabolites. However, the relative concentrations of these compounds are subject to the genetic make-up, the physiological state of the plant as well as environmental effects. Recently, metabolic perturbations through the use of abiotic stressors have proven to be a valuable strategy for increasing the levels of these compounds. Oxidative stress-associated stressors, including ionizing radiation, have also been reported to induce metabolites with various biological activities in plants. Hence, the aim of the current study was to investigate the effect of gamma radiation on the induction of purported anti-cancerous metabolites, glucomoringin and its derivatives, in Moringa oleifera Lam., Moringaceae. Here, an UHPLC-qTOF-MS-based targeted metabolic fingerprinting approach was used to evaluate the effect of gamma radiation treatment on the afore-mentioned health-beneficial secondary metabolites of M. oleifera. Following radiation, an increase in glucomoringin and three acylated derivatives was noted. As such, these molecules can be regarded as components of the inducible defense mechanism of M. oleifera as opposed to being constitutive components as it has previously been assumed. This might be an indication of a possible, yet unexplored role of moringin against the effects of oxidative stress in M. oleifera plants. The results also suggest that plants undergoing photo-oxidative stress could accumulate higher amounts of glucomoringin and related molecules.

Wound Healing Properties of Selected Plants Used in Ethnoveterinary Medicine.

Plants have arrays of phytoconstituents that have wide ranging biological effects like antioxidant, anti-inflammatory and antimicrobial properties key in wound management. In vivo wound healing properties of ointments made of crude methanolic extracts (10% extract w/w in white soft paraffin) of three plant species, Cissus quadrangularis L. (whole aerial plant parts), Adenium multiflorum Klotzsch (whole aerial plant parts) and Erythrina abyssinica Lam. Ex DC. (leaves and bark) used in ethnoveterinary medicine were evaluated on BALB/c female mice based on wound area changes, regular observations, healing skin's percentage crude protein content and histological examinations. White soft paraffin and 3% oxytetracycline ointment were used as negative and positive controls, respectively. Wound area changes over a 15 day period for mice treated with C. quadrangularis and A. multiflorum extract ointments were comparable to those of the positive control (oxytetracycline ointment). Wounds managed with the same extract ointments exhibited high crude protein contents, similar to what was observed on animals treated with the positive control. Histological evaluations revealed that C. quadrangularis had superior wound healing properties with the wound area completely returning to normal skin structure by day 15 of the experiment. E. abyssinica leaf and bark extract ointments exhibited lower wound healing properties though the leaf extract exhibited some modest healing properties.

Antimicrobial, Anthelmintic Activities and Characterisation of Functional Phenolic Acids of Achyranthes aspera Linn.: A Medicinal Plant Used for the Treatment of Wounds and Ringworm in East Africa.

Achyranthes aspera Linn. (Amaranthaceae) commonly known as Prickly Chaff flower (English) is traditionally used for treating a number of ailments. Different parts of the plant are used in treating wounds and ringworm in East Africa and elsewhere for a number of ailments. In this study, leaf extracts of A. aspera collected from two different geographical locations (Ciaat, Eritrea and Ukulinga, South Africa) were evaluated for antibacterial, antifungal, anthelmintic activities and the plant characterized for functional phenolic acids as well as protein binding capacity. The pathogens used in the tests were, two Gram-negative (Escherichia coli and Klebsiella pneumoniae), two Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), a filamentus yeast-like fungus (Candida albicans) and a free-living nematode (Caenorhabditis elegans). The water and acetone extracts of the samples collected from Ciaat exhibited good antibacterial, antifungal and anthelmintic activity (MIC < 1 mg/ml) except the water extract against E. coli which showed moderate activity. In contrast, the extracts collected from Ukulinga exhibited moderate to weak activities except for the acetone (aq.) extracts which had good activity against some of the tested organisms. UHPLC-MS/MS revealed variation in the levels of some functional phenolic compounds, with rutin, chlorogenic acid and genistein not being detected in the extracts from Ukulinga. The variation was also observed in the protein binding capacity, which could offer a predictive wound healing model. All extracts from plant samples collected at Ciaat expressed significant dominant potency compared to similar extracts from Ukulinga.

Antioxidant, antimicrobial and phytochemical variations in thirteen Moringa oleifera Lam. cultivars.

A study was undertaken to assess variation in antioxidant, antimicrobial and phytochemical properties of thirteen Moringa oleifera cultivars obtained from different locations across the globe. Standard antioxidant methods including the DPPH scavenging, ferric reducing power (FRAP) and ß-carotene-linoleic acid model were used to evaluate the activity. Variation in the antioxidant activity was observed, with TOT4951 from Thailand being the most active, with activity five times higher than that of ascorbic acid (reference compound). A different trend was observed for the activity in the FRAP and ß-carotene-linoleic acid assays. Antimicrobial activity was tested against Gram-positive (Staphylococcus aureus) and Gram-negative (Klebsiella pneumoniae) strains using the microdilution method. Acetone extracts of all cultivars exhibited good antibacterial activity against K. pneumoniae (MIC values of 0.78 mg/mL). The remaining extracts exhibited weak activity against the two microorganisms. For the antifungal activity, all the extracts exhibited low activity. Variations were observed in the total phenolic and flavonoid contents. Cultivars TOT5169 (Thailand) and SH (South Africa) exhibited highest amounts of total phenolic compounds while TOT5028 (Thailand) exhibited the lowest amounts of five times lower than the highest. The information offer an understanding on variations between cultivars from different geographical locations and is important in the search for antioxidant supplementation and anti-ageing products.

Hypoxis (Hypoxidaceae) in African traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Hypoxis species (Hypoxidaceae) are popular medicinal plants used in the African traditional medicine to treat numerous ailments and are reported to have a wide spectrum of pharmacological properties. In this paper we reviewed and evaluated the traditional uses, pharmacological, phytochemistry and toxicity aspects of the genus Hypoxis. Potential medicinal prospects and possible knowledge gaps and pitfalls are discussed. MATERIALS AND METHODS: A comprehensive and systematic review of literature on Hypoxis species was done using numerous resources such as books and scientific databases that include Pubmed, Scopus, Scirus, Google scholar, Web of Science and others. Information on the various pharmacological and chemical properties of Hypoxis extracts was critically analysed and discussed under the various topics. RESULTS: The literature indicated a broad range of uses, pharmacological and toxicological properties of different Hypoxis species extracts and their relevance to African healthcare systems. Several compounds, mostly glucosides, sterols and sterolins, have been isolated, identified and tested in various in vitro and in vivo models as well as in a couple of clinical trials with a lot of promising prospects reported in some studies. CONCLUSION: A critical analysis of the available literature and studies identifies positive potential for the future use of Hypoxis species in both traditional and modern medicine and concurrently so with possible pitfalls and research gaps in current knowledge. An integrated and holistic approach to addressing research issues, particularly toxicology aspects could be more effectively applied and incorporate conservation strategies for the species.

Antimicrobial and selected in vitro enzyme inhibitory effects of leaf extracts, flavonols and indole alkaloids isolated from Croton menyharthii.

Croton species are used in folk medicine in the management of infections, inflammation and oxidative stress-related diseases. In order to isolate, characterize and evaluate the bioactive constituents of Croton menyharthii Pax leaf extracts, repeated column fractionation of the ethyl acetate fraction from a 20% aqueous methanol crude extract afforded three flavonols identified by NMR (1D and 2D) spectroscopic methods as myricetrin-3-O-rhamnoside (myricetrin, 1), quercetin-3-O-rhamnoside (2) and quercetin (3) along with an indole alkaloid, (E)-N-(4-hydroxycinnamoyl)-5-hydroxytryptamine, [trans-N-(p-coumaroyl) serotonin, 4]. All the compounds are reported from the leaf extract of this plant for the first time. The crude extracts, four solvent fractions (hexane, DCM, ethyl acetate and butanol) and isolated compounds obtained from the leaves were evaluated for their inhibitory effects on selected bacteria, a fungus (Candida albicans), cyclooxygenase (COX-2), α-glucosidase and acetylcholinesterase (AChE). Amongst the compounds, quercetin (3) was the most active against Bacillus subtilis and Candida albicans while myricetrin-3-O-rhamnoside (1) and trans-N-(p-coumaroyl) serotonin (4) were the most active compounds against Escherichia coli, Klebsiella pneumonia and Staphylococcus aureus. The inhibitory activity of myricetrin-3-O-rhamnoside (1) against COX-2 was insignificant while that of the other three compounds 2-4 was low. The AChE inhibitory activity of the alkaloid, trans-N-(p-coumaroyl) serotonin was high, with a percentage inhibitory activity of 72.6% and an IC50 value of 15.0 µg/mL. The rest of the compounds only had moderate activity. Croton menyharthii leaf extracts and isolated compounds inhibit α-glucosidase at very low IC50 values compared to the synthetic drug acarbose. Structure activity relationship of the isolated flavonols 1-3 is briefly outlined. Compounds 1-4 and the leaf extracts exhibited a broad spectrum of activities. This validates the ethnomedicinal use of the plant in folk medicine.

Toxicology of some important medicinal plants in southern Africa.

Africa is home to two major floral kingdoms: the Paleotropical kingdom of central Africa and the Capensis kingdom of the Western Cape province of South Africa, the latter of which consists of approximately 10,000 species, representing about 20% of Africa's floral 'gold mine', better known as the Cape herbal medicine. Needless to say, such rich flora comes with numerous plants with a potential to cause poisoning to humans. This review document reports important toxic medicinal plants and their toxic ingredients for plant species resident in the southern African region. These include important medicinal uses and pharmacological properties ranging from antimicrobial, antiviral, anticancer, anti-inflammatory as well as those that are used as aphrodisiacs and for maternal health care.

Anti-oxidative and cholinesterase inhibitory effects of leaf extracts and their isolated compounds from two closely related Croton species.

A comparative evaluation of the antioxidant and acetylcholinesterase inhibitory activity of the leaf extracts of Croton gratissimus and Croton zambesicus (subgratissimus) and compounds isolated from the extracts was carried out to determine their potential and suitability or otherwise as a substitute for each other in the management of oxidative and neurodegenerative conditions. Different antioxidant assays (DPPH, FRAP, ß-carotene-linoleic and the lipid peroxidation models) and the microplate assay for acetylcholinesterase (AChE) inhibition were carried out separately to study the activities of the crude leaf extracts and four solvent fractions from each of the two Croton species. Bioassay guided fractionation was used to target antioxidant constituents of the crude extracts and ethyl acetate fractions of 20% aqueous methanol extract of C. gratissimus on silica gel and Sephadex LH-20 columns resulted in the isolation of kaempferol-3-O-ß-6''(p-coumaroyl) glucopyranoside (tiliroside, 2), apigenin-6-C-glucoside (isovitexin, 3) and kampferol (4). The extract of C. zambesicus yielded quercetin-3-O-ß-6''(p-coumaroyl) glucopyranoside-3'-methyl ether (helichrysoside- 3'-methyl ether, 1), kaempferol-3-O-ß-6''(p-coumaroyl) glucopyranoside (tiliroside, 2) and apigenin-6-C-glucoside (isovitexin, 3). Three of the isolated compounds and their different combinations were also included in the bioassays. In all the assays performed, the antioxidant capacity and AChE inhibitory effects of C. zambesicus extracts were weaker than those of C. gratissimus. This suggests that C. gratissimus may not be substituted by C. zambesicus, despite the similarity in some of their constituents. Generally, the combinations made from the isolated compounds showed better activities in most of the assays compared to the individual isolated compounds. This suggests mechanisms such as synergism and/or additive effects to be taking place. This study established low, moderate and high antioxidant activities as well as AChE inhibitory effects by the crude extracts, fractions, compounds and compound combinations. This means some of the extracts, isolated compounds and compound combinations could be useful in the management of neurodegenerative conditions and serve as sources of natural neurodegenerative agents.

Antioxidant activity, acetylcholinesterase inhibition, iridoid content and mutagenic evaluation of Leucosidea sericea.

Leucosidea sericea is an important medicinal plant widely used in traditional medicine in southern Africa. Leaf and stem petroleum ether (PE), dichloromethane (DCM) and 50% aqueous methanol (MeOH) extracts were investigated for antioxidant and acetylcholinesterase inhibitory activities. The safety of the extracts was evaluated using the Ames test. In addition, the iridoid content of L. sericea stems and leaves were quantified. For DPPH radical-scavenging activity, the stem MeOH extract (EC(50) value: 1.6 µg/ml) was more potent than ascorbic acid (EC(50) value: 1.7 µg/ml). In the ß-carotene-linoleic acid model system, antioxidant activity of the leaf DCM extract (89.8%) was not significantly different to that of butylated hydroxytoluene (BHT) (98.9%). All extracts showed a dose-dependent acetylcholinesterase inhibition; in terms of the IC(50) value, the leaf DCM extract (0.14 mg/ml) was the most potent sample. Total iridoid content was 35% higher in the stem extract than in the leaf extract. Based on the Ames test, L. sericea extracts were not mutagenic, either with or without S9 metabolic activation. These findings suggest the safety as well as the potential of L. sericea as a possible source of novel/alternative antioxidant and acetylcholinesterase inhibitory compounds.

In vitro antioxidant properties, HIV-1 reverse transcriptase and acetylcholinesterase inhibitory effects of traditional herbal preparations sold in South Africa.

The antioxidant potentials for fourteen multipurpose traditional herbal preparations sold in South Africa were determined using the DPPH radical scavenging, ferric reducing power and ß-carotene-linoleic acid model system, the anti-HIV-1 reverse transcriptase (RT) enzyme inhibitory effects using an ELISA kit and acetylcholinesterase (AChE) enzyme inhibition using the microtitre plate assay. Nine of the herbal mixtures (Umzimba omubi, Umuthi wekukhwehlela ne zilonda, Mvusa ukunzi, Umpatisa inkosi, Imbiza ephuzwato, Vusa umzimba, Supreme one hundred, Sejeso herbal mixture Ingwe® and Ingwe® special muti) exhibited higher antioxidant potentials, while only four (Imbiza ephuzwato, Ingwe® muthi mixture, Sejeso herbal mixture Ingwe® and African potato extract™ showed potent activity against the RT enzyme. Nine mixtures (Imbiza ephuzwato, Umpatisa inkosi, African potato extract™, Sejeso herbal mixture Ingwe®, Vusa umzimba; Ingwe® muthi mixture, Ibhubezi™, Lion izifozonke Ingwe® and Ingwe® special muti) showed AChE enzyme inhibitory activity greater than 50%. The observed activity exhibited by some of the herbal mixtures gives some credence to the manufacturers' claims and goes part of the way towards validating their use against certain conditions such as oxidative stress, HIV/AIDS proliferation and some mental conditions. It is however, desirable to carry out further studies to determine the effects of mixing plant species/parts in one mixture on the antioxidant potency as well as isolating active constituents from the herbal mixtures.

Natural antioxidants: fascinating or mythical biomolecules?

Research on the use, properties, characteristics and sources of antioxidants especially phenolic compounds, flavonoids, vitamins, synthetic chemicals and some micronutrients began in the late 18(th) century. Since then antioxidant research has received considerable attention and over a hundred thousand papers have been published on the subject. This has led to a rampant use of antioxidants in order to try to obtain and preserve optimal health. A number of nutraceuticals and food supplements are frequently fortified with synthetic or natural antioxidants. However, some research outcomes have led to the belief that antioxidants exist as mythical biomolecules. This review provides a critical evaluation of some common in vitro antioxidant capacity methods, and a discussion on the role and controversies surrounding non-enzymatic biomolecules, in particular phenolic compounds and non-phenolic compounds, in oxidative processes in an attempt of stemming the tidal wave that is threatening to swamp the concept of natural antioxidants.