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Iriflophenone-3-C-glucoside from Cyclopia genistoides: Isolation and quantitative comparison of antioxidant capacity with mangiferin and isomangiferin using on-line HPLC antioxidant assays
... Isomangiferin and mangiferin has a regioisomeric relationship due to their sugar moieties are substituted at the 2 or 4 positions [15] . Also, the antioxidant potential of these compounds cannot be assumed to be the same. ...
... Also, the antioxidant potential of these compounds cannot be assumed to be the same. The contrast on flavone C-glycosides, vitexin and isovitexin, was clearly demonstrated at sugar moiety position and can affect their radical scavenging ability [15,16] . The current demonstration of benzophenones, a putative precursor of xanthones [10,12,17,18] , found in C. genistoides [19] showed the antioxidant capacity of iriflophenone-3-C-glucoside when compared to mangiferin and isomangiferin [15,20] . ...
... The contrast on flavone C-glycosides, vitexin and isovitexin, was clearly demonstrated at sugar moiety position and can affect their radical scavenging ability [15,16] . The current demonstration of benzophenones, a putative precursor of xanthones [10,12,17,18] , found in C. genistoides [19] showed the antioxidant capacity of iriflophenone-3-C-glucoside when compared to mangiferin and isomangiferin [15,20] . In previous works, isomangiferin showed slightly lower antioxidant capacity than mangiferin in diphenyl-1-picrylhydrazyl (DPPH)evaluation, nevertheless it displayed higher capacity against ABTS and peroxyl radical assays [12] . ...
The electronic performance between mangiferin and isomangiferin as antioxidants was achieved using electron and hydrogen atom transfer mechanisms at DFT/B3LYP/6-31+G(d,p) level. According to theoretical properties of these molecules a structural and electronic symmetry among 3-hydroxyl of xanthone, 2’-hydroxyl of sugar, and ether moiety of sugar were stablished. The sugar moiety changes on 2 and 4 positions showed that isomangiferin is more potent than mangiferin by electron transfer and mangiferin by hydrogen transfer. Hydrogen bonds between sugar and resorcinol rings can be involved to their electronic behavior and favored conformations of low energies. Sugar moiety in both compounds is responsible for the antioxidant capacity increase in xanthone ring.
... Malherbe et al. determined the antioxidant activity of isomangiferin (70) from Cyclopia genistoides, using on-line high-performance liquid chromatography (HPLC) antioxidant assays, namely HPLC-oxygen radical absorbance capacity (HPLC-ORAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals on-line assays. Isomangiferin (70) revealed high capacity to scavenge DPPH, ABTS and peroxyl radicals in these online assays [47]. ...
... Furthermore, xanthone 178 showed antioxidant activity, namely oxygen radical scavenging property (ORAC value of 33.6 U ml -1 at 25 μM) and a good oxidative protective capacity in a hydrogen peroxide-impaired pancreatic β cell line at 0.3 and 0.5 μM [49,50]. Mangiferin (179), a natural glucosylxanthone, has been described as having many biological activities, including antioxidant, AChE and MAO inhibitory activities [43,47,68,[79][80][81][82][83][84][85][86][87][88][89][90][91]. The antiacetylcholinesterase [37,68,[79][80] and antioxidant [47,68,[83][84][85][86][87][88][89][90][91] activities of mangiferin (179) are recognized by various authors. ...
... Mangiferin (179), a natural glucosylxanthone, has been described as having many biological activities, including antioxidant, AChE and MAO inhibitory activities [43,47,68,[79][80][81][82][83][84][85][86][87][88][89][90][91]. The antiacetylcholinesterase [37,68,[79][80] and antioxidant [47,68,[83][84][85][86][87][88][89][90][91] activities of mangiferin (179) are recognized by various authors. Besides these biological properties, MAO inhibitory activity of mangiferin (179) was investigated [81,82]. ...
To date, the current therapy for Alzheimer's disease (AD) based on acetylcholinesterase inhibitors is only symptomatic, being its efficacy limited. Hence, the recent research has been focused in the development of different pharmacological approaches. Here we discuss the potential of xanthone derivatives as new anti-Alzheimer agents. The interference of xanthone derivatives with acetylcholinesterase and other molecular targets and cellular mechanisms associated with AD have been recently systematically reported. Therefore, we report xanthones with anticholinesterase, monoamine oxidase and amyloid β aggregation inhibitory activities as well as antioxidant properties, emphasizing xanthone derivatives with dual/multitarget activity as potential agents to treat AD. We also propose the structural features for these activities that may guide the design of new, more effective xanthone derivatives.
[Formula: see text]
... Authentic reference standards with purity >95 % were obtained from Sigma-Aldrich (hesperidin), Chemos GmbH (Regenstauf, Germany; isomangiferin), Phytolab (Vestenbergsreuth, Germany; vicenin-2) and Extrasynthese (Genay, France; mangiferin, eriocitrin, luteolin). Iriflophenone-3-C-β-D-glucoside (97 %; [19]), iriflophenone-3-C-β-D-glucoside-4-O-β-D-glucoside (99 %; [17]) and maclurin-3-C-β-D-glucoside (95 %; [17]) were isolated from Cyclopia genistoides. Phloretin-3′,5′-C-β-D-glucoside (94 % by LC-MS) was from the compound library of the ARC. ...
... Semi-preparative HPLC separation of the crude HPCCC fraction enriched in scolymoside was achieved on a Gemini C18 column (150 ×10 mm; 5 μm; 110 Å; Phenomenex, Torrance, CA, USA) protected by a guard column of the same stationary phase (10×10 mm), using equipment described by Malherbe et al. [19]. For isolation of scolymoside, an isocratic run comprising of 16 % acetonitrile and 84 % of 0.1 % aqueous formic acid at 4.8 mL/min and 35°C, followed by column flushing and re-equilibration, was employed. ...
... The identity of the isolated compound was confirmed by HPLC with high-resolution electrospray ionisation mass spectrometric detection (HR-ESI-MS) performed using a Waters Acquity UPLC coupled to a Synapt G2 Q-TOF system (Waters, Milford, MA, USA) and 1 H NMR (Varian Unity Inova 600) at 600 MHz frequency. HPLC-HR-ESI-MS parameters was as described by De Beer et al. [15], whilst NMR was performed as described by Malherbe et al. [19]. ...
Similarity analysis of the phenolic fingerprints of a large number of aqueous extracts of Cyclopia subternata, obtained by high-performance liquid chromatography (HPLC), was evaluated as a potential tool to screen extracts for relative bioactivity. The assessment was based on the (dis)similarity of their fingerprints to that of a reference active extract of C. subternata, proven to enhance glucose uptake in vitro and in vivo. In vitro testing of extracts, selected as being most similar (n = 5; r ≥ 0.962) and most dissimilar (n = 5; r ≤ 0.688) to the reference active extract, showed that no clear pattern in terms of relative glucose uptake efficacy in C2C12 myocytes emerged, irrespective of the dose. Some of the most dissimilar extracts had higher glucose-lowering activity than the reference active extract. Principal component analysis revealed the major compounds responsible for the most variation within the chromatographic fingerprints, as mangiferin, isomangiferin, iriflophenone-3-C-β-d-glucoside-4-O-β-d-glucoside, iriflophenone-3-C-β-d-glucoside, scolymoside, and phloretin-3′,5′-di-C-β-d-glucoside. Quantitative analysis of the selected extracts showed that the most dissimilar extracts contained the highest mangiferin and isomangiferin levels, whilst the most similar extracts had the highest scolymoside content. These compounds demonstrated similar glucose uptake efficacy in C2C12 myocytes. It can be concluded that (dis)similarity of chromatographic fingerprints of extracts of unknown activity to that of a proven bioactive extract does not necessarily translate to lower or higher bioactivity.
Graphical Abstract
(Dis)similarity of HPLC fingerprints is not predictive of relative in vitro glucos uptake efficacy of C. subternata extracts
... Based on in vitro studies conducted on the mono 3-C-glucoside and 3,5-di-C-glucoside derivatives of iriflophenone, possible biological activities of the iriflophenone derivatives could include α-glucosidase inhibition [20] and triglyceride accumulation inhibition [8], as well as pro-apoptotic activity [7]. In accordance with the antioxidant activity demonstrated for iriflophenone-3-C-glucoside using on-line HPLC antioxidant assays [21], these compounds could potentially also exhibit antioxidant activity and thus contribute to the antioxidant activity of C. genistoides extracts [10]. The UV-Vis, ESI-MS and -MS/MS characteristics of compound c were in agreement with literature reports on maclurin-3-C-glucoside [7,8]. ...
... The UV-Vis, ESI-MS and -MS/MS characteristics of compound c were in agreement with literature reports on maclurin-3-C-glucoside [7,8]. This compound has been identified previously in C. genistoides [7,21] and has also been tentatively identified in C. maculata hot water extracts [13]. A compound with similar characteristics has likewise been observed in hot water extracts of C. subternata (unidentified compound 3) [12]. ...
... The presence of these flavanone derivatives in extracts of C. genistoides exemplifies the high degree of structural variation in natural plant extracts. Such glycosylated derivatives of eriodictyol and naringenin, other than the two most common compounds eriocitrin and narirutin, have been detected in extracts of C. genistoides (unidentified compound 7 [21]) and C. subternata (eriodictyol-di-C-hexoside, eriodictyol-O-glucoside, naringenin-di-C-hexoside and naringenin-O-dihexoside [12]). The flavanone aglycones, eriodictyol and naringenin, as well as their rutinoside derivatives, have phytoestrogenic activity, indicating the potential of these glycosylated flavanone derivatives to contribute to the phytoestrogenic potential of C. genistoides extracts (as reviewed by [33]). ...
A high-performance liquid chromatographic (HPLC) method coupled with diode-array detection (DAD) was optimized for the qualitative analysis of aqueous extracts of Cyclopia genistoides. Comprehensive insight into the phenolic profile of unfermented and fermented sample extracts was achieved with the identification of ten compounds based on comparison with authentic reference standards and the tentative identification of 30 additional compounds by means of electrospray ionization mass spectrometry (ESI-MS) and tandem MS detection. Three iriflophenone-di-O,C-hexoside isomers, three xanthone-dihydrochalcone derivatives and one dihydrochalcone are herein tentatively identified for the first time in C. genistoides. Of special interest is one iriflophenone-di-O,C-hexoside present in large amounts. New compounds (tentatively) identified for the first time in this species, and also in the genus Cyclopia, include two aromatic amino acids, one flavone, an iriflophenone-di-C-hexoside, a maclurin-di-O,C-hexoside, two tetrahydroxyxanthone-C-hexoside isomers, a tetrahydroxyxanthone-di-O,C-hexoside, two symmetric tetrahydroxyxanthone-C-hexoside dimers, nine glycosylated flavanone derivatives and five glycosylated phenolic acid derivatives. The presence of new compound subclasses in Cyclopia, namely aromatic amino acids and glycosylated phenolic acids, was demonstrated. The HPLC-DAD method was successfully validated and applied to the quantitative analysis of the paired sample extracts. In-depth analysis of the chemical composition of C. genistoides hot water extracts gave a better understanding of the chemistry of this species that will guide further research into its medicinal properties and potential uses.
... Mango (Mangifera indica) is a tropical fruit widely consumed worldwide and is one of the fruits which has shown antioxidant activity (11). This activity has been attributed to its content of ascorbic acid, carotenoids, and polyphenols, among them mangiferin, one of its most studied components (12)(13). It is a glucosylxanthone (2-b-D-glucopyranosyl-1,3,6,7-tetrahydroxy-9Hxanthen-9-one), which is more abundant in the leaves and bark of Mangifera indica L (Anacardiaceae) and it can also be found in about sixteen plant families (14)(15). ...
... Mangiferin has demonstrated antioxidant capacity in different in vitro studies, including a study in which it showed the ability to sequester DPPH and ABTS radicals (13). It has also shown a protective effect against oxidative stress in vivo in Wistar rats subjected to iron overload, where induced an increase in the activity of superoxide dismutase and glutathione peroxidase compared to the control group (16). ...
Background:
Mango (Mangifera indica L.) is one of the world’s most consumed fruit, and it is also a rich source of antioxidants that may prevent oxidative stress.
Objectives:
This study aimed to determine if mango (cv. Azúcar) juice can improve the antioxidant status of healthy individuals with low consumption of vegetables and fruit.
Methods:
This was a cross-over single-blind study carried out with 16 healthy individuals for 73 days. Participants were randomly assigned to either a mango juice period or a placebo period. Total phenolic content, antioxidant capacity, mangiferin, thiobarbituric acid reactive substances (TBARS), total glutathione, and 8-hydroxydeoxyguanosine levels were determined in plasma.
Results:
Plasma antioxidant activity was significantly higher in the juice consumption period than the placebo consumption period; however, total phenolic content, total glutathione, TBARS, and 8-hydroxydeoxyguanosine levels did not show significant differences between juice period and placebo period. Mangiferin was detected in every participant after juice consumption.
Conclusions:
Mango (cv. Azúcar) juice daily consumption improves plasma antioxidant capacity.
... But this result indicated that the extract had lower activities to eliminate ABTS +Å radical, which could be attributed to different free radicals with different mechanisms (Yang, Han, Wan, & Fang, 2011). A particular compound reacted with different radical species showing different activities due to different reaction kinetics (Malherbe et al., 2014) for them, possibly. Therefore, free radical species might be an important reason causing these differences. ...
... The opposite results from DPPH Å and ABTS +Å might be explained by different types of free radicals with different mechanisms (Yang et al., 2011). Because the reaction mechanisms of Å OH radical scavenging and other two methods were different, the same compound displayed different activities in three radical scavenging experiments (Li et al., 2016;Malherbe et al., 2014). No single testing method was sufficient to estimate the antioxidant activity and free radical scavenging potential of a studied sample (Koleva, van Beek, Linssen, de Groot, & Evstatieva, 2002), and thus a combination of several methods based on different free radicals was employed. ...
An off-line high performance liquid chromatography (HPLC) coupled with chemical methods has been developed to evaluate antioxidant activity of 11 standard polyphenol compounds (SPCs) and vitamin C (Vc) in terms of radical scavenging abilities. The structure-activity relationships of each SPC were also discussed. SPCs showed different abilities in scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH·), 2,2'-azinobis-3-ethyl- benzthiazoline-6-sulphonate (ABTS⁺·) and hydroxyl (·OH) free radicals. Among SPCs, quercetin and kaempferol, as typical flavonoids, displayed the greatest radical-scavenging activities and even exhibited higher activity in ·OH radical removal ability than that of Vc. Furthermore, the proposed method was also applied to screening polyphenolic antioxidant components from Cichorium endivia L. (C. endivia) seeds extract. The results indicated that cynarin in the extract was more active compound to scavenge DPPH· and ABTS+· radicals than chlorogenic acid, while chlorogenic acid had stronger capacity in scavenging ·OH free radicals.
... An on-line HPLC-oxygen radical absorbance capacity (ORAC) assay method, which combines HPLC and the microplate ORAC assay with fluorescein as probe, was proposed and compared with HPLC-DPPH/ABTS methods [32]. The results showed that the HPLC-ORAC assay was more sensitive for the detection of radical scavengers. ...
... Screening of enzyme inhibitors has been become an important tool for potential drug discovery, and various off-line and on-line assay methods have been reported to assess enzyme inhibitory activity. The on-line separation Cyclopia genistoides HPLC-ORAC Iriflophenone 3-C-glucoside, mangiferin, isomangiferin λ ex 288 nm, λ em 320 nm [32] Ristretto espresso coffee HPLC-FL-DPPH Caffeine λ ex 278 nm, λ em 366 nm [33] of HPLC with different detection modes, such as UV, fluorescence (FL) and MS, is usually applied for quick detection of enzyme inhibitors in complex extracts [1, 6, 53, 54] (Fig. 3). ...
On-line high-performance liquid chromatography (HPLC) coupled with biochemical detection (BCD) has been developed to screen compounds showing antioxidant action, enzyme inhibition and receptor affinity in complex matrixes. This review summarizes HPLC methods combining different post-column detection methods, such as diode-array detection (DAD), mass spectrometry (MS), chemiluminescence (CL) and nuclear magnetic resonance, for antioxidant screening. The methods based on a single relatively stable reagent such as DPPH• and ABTS•+ were the most popular. Oxygen free radical scavengers mainly depended on post-column CL detection. The on-line hyphenated HPLC–BCD systems based on post-column UV/DAD fluorescence and MS detection were also widely applied to screen enzyme- and receptor-active compounds. These strategies provide a convenient tool for quick identification and quantification of active compounds in complex matrixes.
... The pharmacological activity of mangiferin is well-documented (as reviewed by Vyas et al. 3 ), while only the antioxidant activity of isomangiferin has been assessed to date. 4 The major benzophenones present in Cyclopia extracts include the novel constituent iriflophenone-3-C-glucoside-4-O-glucoside and iriflophenone-3-C-glucoside, while the presence of maclurin-3-C-glucoside in C. genistoides has also been demonstrated. 5,6 These benzophenones are gaining in prominence in light of their α-glucosidase inhibitory and glucose-uptake stimulatory activities. ...
... Mangiferin exhibited slightly higher reactivity toward DPPH • than its C-4 regioisomer, isomangiferin, while the TEAC ORAC value of isomangiferin was slightly higher than that of mangiferin. These observations are in line with results presented by Malherbe et al. 4 using online HPLC-DPPH and -ORAC assays. Similar antioxidant activities have also been demonstrated for positional isomers of chlorogenic acid. ...
Degradation of the major benzophenones, iriflophenone-3-C-glucoside-4-O-glucoside and iriflophenone-3-C-glucoside, and the major xanthones, mangiferin and isomangiferin, of Cyclopia genistoides followed first-order reaction kinetics during high-temperature oxidation of the plant material at 80 and 90 °C. Iriflophenone-3-C-glucoside-4-O-glucoside was shown to be the most thermally stable compound. Isomangiferin was the second most stable compound at 80 °C, while its degradation rate constant was influenced the most by increased temperature. Mangiferin and iriflophenone-3-C-glucoside had comparable degradation rate constants at 80 °C. The thermal degradation kinetics model was subsequently evaluated by subjecting different batches of plant material to oxidative conditions (90 °C/16 h). The model accurately predicted the individual contents of three of the compounds in aqueous extracts prepared from oxidized plant material. The impact of benzophenone and xanthone degradation was reflected in the decreased total antioxidant capacity of the aqueous extracts, as determined using the oxygen radical absorbance capacity and DPPH• scavenging assays.
... The PICO (Patients, Intervention, Comparison, and Outcomes) format was use perform the systematic review, and the flow diagram shows the selection of Antioxidant and antiproliferative [30,31] Antocyanin ...
Mango and its by-products have traditional medicinal uses. They contain diverse bioactive compounds offering numerous health benefits, including cardioprotective and metabolic properties. This study aimed to explore the impact of mango fruit and its by-products on human health, emphasizing its metabolic syndrome components. PUBMED, EMBASE, COCHRANE, and GOOGLE SCHOLAR were searched following PRISMA guidelines, and the COCHRANE handbook was utilized to assess bias risks. In vivo and in vitro studies have shown several benefits of mango and its by-products. For this systematic review, 13 studies met the inclusion criteria. The collective findings indicated that the utilization of mango in various forms—ranging from fresh mango slices and mango puree to mango by-products, mango leaf extract, fruit powder, and mangiferin—yielded many favorable effects. These encompassed enhancements in glycemic control and improvements in plasma lipid profiles. Additionally, mango reduces food intake, elevates mood scores, augments physical performance during exercise, improves endothelial function, and decreases the incidence of respiratory tract infections. Utilizing mango by-products supports the demand for healthier products. This approach also aids in environmental conservation. Furthermore, the development of mango-derived nanomedicines aligns with sustainable goals and offers innovative solutions for healthcare challenges whilst being environmentally conscious.
... Antioxidant Activity: Malherbe et al. 28 Wongwad et al. 17 showed that Aquilaria crassna is a potential source of natural ingredients for preparing anti-aging cosmetics. Numerous chemical constituents are found in A. crassna, like Phenolic acids, steroids, terpenoids, pyranones, quinones, Benzophenones, xanthonoids, flavonoids and nucleosides 20 but the main compounds of interest in that study are Iriflophenone 3,5-C-β-D diglucoside (i), Iriflophenone 3-C-β-D glucoside (ii), mangiferin (iii), genkwanin 5-o-β-primevoside (iv). ...
ABSTRACT: Iriflophenone 3-C-β-D glucoside is a Benzophenone derivative
which can be obtained from various plant sources like Aquilaria crassna, A.
sinensis, A. malaccensis, Cyclopia genistoides, Mangifera indica, Dryopteris
ramosa etc. It can act as an important herbal active constituent as it has various
pharmacological actions, such as Antidiabetic, anti-inflammatory, antioxidant,
and antimicrobial agent. Some scientists demonstrated the antioxidant activity of
Iriflophenone 3-C-β-D glucoside and have seen that this compound has no
radical scavenging ability against DPPH [2,2-diphenylpicrylhydrazyl] but
scavenged ABTS [2,2’-azino-bis(3- ethylbenzothiazoline-6-sulphonic acid)] and
peroxyl radicals. So, it can be used as an anti- oxidant agent. From A. sinensis 8
compounds were isolated among which Iriflophenone 3-C- β-D glucoside was
present. It is proved that all the isolated compounds have α-glucosidase
inhibition activity stronger than acarbose, that is taken as positive drug control.
Aqueous fraction of D. ramosa is used for so long by the inhabitants of the
Galliyat region of Pakistan to treat their GIT ailments caused by bacteria.
Scientists showed that Iriflophenone 3-C-β-D glucoside that is obtained from
Dryopteris ramosa has stronger antibacterial potential against Klebsiella
pneumoniae, Staphylococcus aureus, and Escherichia coli. The anti-
inflammatory activities of this compound are revealed as the aqueous extract of
A. crassna expressed strong IL-1α and IL-8 inhibitions and the 70% Ethanolic
extract showed IL-1α and NO inhibitions. Apart from this there are many other
effects of that compound which are still under research. Such versatile uses
make this compound a highly valuable herbal constituent.
... In contrast, 6-O-(p-hydroxybenzoyl)mangiferin was eluted at a retention time of 27.10 min. Database spectra and a previous article helped the authors to identify the compounds (Malherbe et al., 2014). C-glycosylated xanthone mangiferin was identified in pulp and husks of arabica coffee varieties, although its quantification was omitted by coelution with feruloylquinic acid (Esquivel et al., 2020). ...
The coffee chain generates over two billion tons of solid residues annually. Husk, pulp, silverskin, defective beans, and spent coffee grounds are produced during coffee drying, husking, roasting, storage, coffee beverage preparation, and other steps. These solid materials are rich in fibers, proteins, and bioactive compounds. Therefore, this review demonstrated research trends in using solid coffee residues as a source of valuable compounds. The integral use of coffee through a biorefinery concept has shown the potential of its residues as a source of new food, pharmaceutical, materials, energy, and fertilizer products. Ultrasound, high-pressure, microwave, pulsed electric field, combining technologies, and alternative solvents (deep eutectic solvents and ionic liquids) were proposed as efficient strategies to obtain phenolic compounds, one of the highest-added-value products from solid coffee residues. However, this review verified that more standardization in reporting the studied parameters is necessary to compare the extractive methodologies. Moreover, solid coffee residues are a rich source of phenolic compounds, but researchers have reported their extraction results in total phenolic content (TPC). TPC helps to compare different extraction conditions, but chromatographic methods are much more accurate, directly identifying and quantifying each compound. Furthermore, regulatory and economic aspects regarding the use of products from coffee residues are presented. The trends indicated future studies evaluating the financial features of using emerging technologies combined with alternative solvents and more robust quantification of extracts through chromatographic techniques to obtain phenolic compounds from coffee residues.
... Antioxidant activity of mangiferin has also been demonstrated using the 2,2 0 -azinobis-(3-ethylbenzothiazolin-6 -sulfonic acid (ABTS) and DPPH assays (Tang et al., 2004). Mangiferin showed in vitro values of 0.61, 1.67, and 3.69 mmol of Trolox, using DPPH, ABTS, and Oxygen Radical Absorbance Capacity (ORAC) assays, respectively (Malherbe et al., 2014). Mangiferin has also been shown to protect against oxidative stress in Wistar rats and in healthy older adults (Pardo-Andreu et al., 2008). ...
The mango fruit is of great economic value worldwide and of great nutritional importance. Its functional effects are due to numerous important nutrients and bioactive compounds. In this review, the benefits of mango fruit (pulp, peel, and seed) for human nutrition and health are detailed. The first part of the review presents the nutrient and phytochemical content of the mango fruit, and the second part addresses its health effects. The very diverse phytochemical components in mango fruit can be classified into several groups including macronutrients such as carbohydrates (sugars, pectin, and cellulose), proteins, lipids (Ω-3 and Ω-6 fatty acids); micronutrients such as vitamins A and C, minerals, pigments (chlorophylls, carotenoids, and anthocyanins, depending on the cultivar), phenolic compounds (phenolic acids and flavonoids), and volatile compounds. The beneficial effects of mango fruit and its components have been studied on different non-communicable diseases such as obesity, type 2 diabetes mellitus, hypertension, and cancer. A great diversity of research approaches have been used to study mango fruit health benefits, and those can be grouped into three main categories: a) those that present the associations or direct effects of the edible portion of the fruit on health through observational epidemiological studies and clinical trials; b) in vivo and in vitro experimental approaches that address either the physiological effects or mechanisms of several kinds of extracts or purified components of mango fruit (pulp, peel and seed), and c) preclinical and clinical studies that explore the possible pharmacological uses of mango fruit. Current scientific understanding of mango health benefits suggest that consumption of mango fruit and its byproducts containing bioactive components can be useful as part of a healthy diet in order to reduce the incidence of health problems.
... Fraction 4 enriched in hesperidin was less effective than the extract in scavenging O •− 2 . Orallo et al. [26] reported that [27,28]. Their weak activity could be attributed to the replacement of the C-7 OH-group with a glucopyranosyl moiety, which sterically prevents the remaining free OH-groups from accessing the radical site [29]. ...
Anti-allergic activity was previously demonstrated for extracts of Cyclopia subternata Vogel plant material, containing substantial amounts of xanthones, benzophenones, dihydrochalcones, flavanones and flavones. Fractionation of a hot water extract on macroporous resin was performed aiming to increase its potency. Operating conditions for scaled-up fractionation of the extract were determined, using small-scale static and dynamic sorption/desorption experiments. The anti-allergic potential of the fractions was assessed based on inhibition of β-hexosaminidase release from IgE-sensitized RBL-2H3 cells. Given the role of oxidative stress in allergic reactions, the extract and fractions were also tested for their ability to scavenge the superoxide anion radical and inhibit xanthine oxidase (XO), an enzyme involved in its generation. The routine DPPH and ORAC assays were used for determination of the antioxidant capacity of the fractions. 3-β-D-Glucopyranosyl-4-O-β-D-glucopyranosyliriflophenone (IDG) had the lowest affinity for the resin, dictating selection of the optimal separation conditions. The extract was separated into four fractions on XAD1180N, using step-wise gradient elution with EtOH-water solutions. The major phenolic compounds present in the fractions were IDG and 3-β-D-glucopyranosyliriflophenone (fraction 1), mangiferin, isomangiferin, 3′,5′-di-β-D-glucopyranosyl-3-hydroxyphloretin and vicenin-2 (fraction 2), 3′,5′-di-β-D-glucopyranosylphloretin, eriocitrin and scolymoside (fraction 3) and hesperidin and p-coumaric acid (fraction 4). Fractionation was only partially effective in increasing activity compared to the extract, i.e., fractions 2, 3 and 4 in the DPPH• and XO assays, fractions 1 and 2 in the ORAC assay and fraction 1 in the β-hexosaminidase release assay. In vivo testing will be required to determine whether the increased activity of fractions is worth the effort and expense of fractionation.
... In vitro and in vivo studies reported that mangiferin exhibits analgesic, anti-ageing, anti-allergic, anticancer, antidiabetic, antimicrobial, antioxidant, antipyretic, cardioprotective, hypocholesterolaemic, immunomodulatory and neuroprotective effects (Du et al., 2018). Isomangiferin has been shown to exert anti-breast cancer effects via functional inhibition of vascular endothelial growth factor , α-glucosidase inhibitory activity , anti-arthritic activity (Kokotkiewicz et al., 2013) and antioxidant activity (Malherbe et al., 2014). The routine generation of chemical fingerprints of raw materials destined for tea production can ensure that sufficient mangiferin/isomangiferin is present to offer health benefits. ...
High performance thin layer chromatography (HPTLC) has been widely implemented for the rapid chemical fingerprinting of medicinal plant extracts for identification and authentication purposes. Despite herbal teas of rooibos (Aspalathus linearis) and honeybush (Cyclopia spp.) being important local and export products of South Africa, characteristic and well-defined HPTLC chemical profiles for the species that could assist in identification and authentication, have not been established and reported. ‘Fermentation’, an oxidation processing step for flavour and colour development, is known to alter the chemical composition of these plant materials, with consequences for their therapeutic properties. In this study, HPTLC fingerprints were established that characterise these herbal teas in terms of phenolic constituents linked to their health benefits. Selected flavonoids present in rooibos were clearly distinguished from the extract matrix, and the marked depletion of aspalathin in the fermented samples was demonstrated. The analytical method developed for honeybush achieved partial separation of the major xanthone constituents, mangiferin and isomangiferin. Lower relative concentrations of the regio-isomers were evident in the fermented samples of C. genistoides compared to their unfermented counterparts. The chemical profile of fermented C. subternata was markedly different to those of C. genistoides and C. intermedia in that mangiferin/isomangiferin was detected in only one of the five samples of C. subternata. Cost-effective HPTLC methods are herein described that can be used for the rapid analysis of multiple samples in industrial environments, as a tool for the quality control, in particular in identification of these herbal teas of South African origin.
... Benzophenones are a class of natural compounds abundant in plants such as the Clusiaceae family [230], but recently, the C-glucosyl benzophenones, 3-β-D-glucopyranosyliriflophenone, 3-β-D-glucopyranosyl-4-β-D-glucopyranosyloxyiriflophenone and 3-β-D-glucopyranosylmaclurin have been detected in several Cyclopia extracts [83,120,124,125,[231][232][233][234]. In vitro studies reporting on the anti-obesity potential of Cyclopia benzophenones are limited (Table 2), while there is currently no in vivo animal and human studies that have reported on the anti-obesity potential of these benzophenones. ...
Obesity is a significant contributor to increased morbidity and premature mortality due to increasing the risk of many chronic metabolic diseases such as type 2 diabetes, cardiovascular disease and certain types of cancer. Lifestyle modifications such as energy restriction and increased physical activity are highly effective first-line treatment strategies used in the management of obesity. However, adherence to these behavioral changes is poor, with an increased reliance on synthetic drugs, which unfortunately are plagued by adverse effects. The identification of new and safer anti-obesity agents is thus of significant interest. In recent years, plants and their phenolic constituents have attracted increased attention due to their health-promoting properties. Amongst these, Cyclopia, an endemic South African plant commonly consumed as a herbal tea (honeybush), has been shown to possess modulating properties against oxidative stress, hyperglycemia, and obesity. Likewise, several studies have reported that some of the major phenolic compounds present in Cyclopia spp. exhibit anti-obesity effects, particularly by targeting adipose tissue. These phenolic compounds belong to the xanthone, flavonoid and benzophenone classes. The aim of this review is to assess the potential of Cyclopia extracts as an anti-obesity nutraceutical as underpinned by in vitro and in vivo studies and the underlying cellular mechanisms and biological pathways regulated by their phenolic compounds.
... Mangiferin (MA) is a poorly water-soluble polyphenolhydroxyl diphenyl and pyranoid ketone carbon glycoside, with a solubility reported as 0.111 mg/mL in water. 28 Modern pharmacological studies have shown that MA has a variety of pharmacological effects, such as anti-inflammatory effects, 29 antioxidation, 30,31 immune regulation, 1,20 and other effects. 32,33 However, its poor solubility in water limits the development of its preparation, and severely affects the bioavailability of its solid formulations. ...
Objective
This study aimed to investigate the interaction between the ion-complementary self-assembling peptide RADA16-I and the hydrophobic drug mangiferin (MA), and the potential of the self-assembling peptide to be exploited as a drug carrier of MA.
Methods
The RADA16-I–MA suspension was prepared by magnetic stirring, followed by fluorescence spectrophotometry, particle size determination, rheological properties analysis, and in vitro release assay to characterize the interaction between RADA16-I and MA. Then, the effects of in situ MA-loaded hydrogel on the proliferation of KYSE 30 and DLD-1 tumor cells and the toxic effect of the hydrogel on 293T renal epithelial cells were studied by the Cell Counting Kit 8 method.
Results
The RADA16-I–MA suspension was formed in water under magnetic stirring; the in situ hydrogel was formed when the suspension was added to PBS. The particle size in the RADA16-I–MA suspension was around 300–600 nm with an average size of 492 nm. Within 24 h, the cumulative release of MA from the RADA16-I–MA hydrogel was about 80%. The release rate of MA from the hydrogel was dependent on the concentration of RADA16-I and the release can be fitted with a first-order kinetic equation. The results suggested that the self-assembling peptide can stabilize MA in water to form a relatively stable suspension; the results also indicated that controlled release of MA from the RADA16-I–MA in situ hydrogel formed from the RADA16-I–MA suspension can be achieved by adjusting the concentration of the peptide in suspension. The cell viability studies showed that the RADA16-I–MA in situ hydrogel not only can maintain or enhance the intrinsic proliferation inhibition effects of MA on tumor cells, but also can reduce the toxicity of MA to normal cells.
Conclusion
The self-assembling peptide RADA16-I can be a potential candidate for constructing a delivery system of the hydrophobic drug MA.
... Bhuvaneshwari et al. (2014) reported on similar studies focusing on the activities of tender and mature leaves. Similarly, Malherbe et al. (2014) reported on the anti-oxidant activities of mangiferin from Cyclopia genistoides. There are many such reports published on the anti-oxidant capacity of mangiferin or mango leaves but these researchers did not compare the anti-oxidant capabilities of mangiferin and mango leaves. ...
There has been an alarming increase in the prevalence of diabetes in India and worldwide. As an alternative medicine to treat diabetes mellitus, many herbal extracts and bioactives are being assessed all over the world. One of the strategic approaches for diabetes management is to control gastrointestinal glucose production and its absorption into the blood stream. The purpose of the study was to evaluate the anti-diabetic potential of aqueous extract of Mangifera indica leaves (mango leaves) and to compare it with its main biomolecule (mangiferin). An in vitro method using carbohydrate metabolism (α-amylase and α-glucosidase inhibition) was performed. The concentrations required for 90% inhibition of enzymes activity were determined to investigate the relative potency of the extract and mangiferin. The results showed that mangiferin strongly inhibited both α-amylase and α-glucosidase activity, with much higher potency than Acarbose and mango leaves extract, while mango leaves extract showed higher anti-oxidant activity compared to mangiferin. The results also showed that mango leaves extract and mangiferin have a bright future in the therapy of diabetes mellitus. Such a comparison helps to evaluate whether the active biomolecules are playing a role or if the whole extract is required for the management of a disease. This would be beneficial in designing a proper process for utilization of natural medicines.
... Conversely, it has been reported that M. indica kernels mostly contain gallotannin derivatives, with penta-O-galloylglucoside as the major one. Of interest, this compound has been previously shown to only exert moderate radical scavenging activities when submitted to ORAC evaluation [30] while mangiferin and iriflophenone 3-C-毬 -D-glucoside are both known to give excellent results with that assay [31]. Taken together, these data tend to explain why, in the present study, kernel extracts possess the lowest ORAC values despite being the richest source of phenolic compounds. ...
Objective: To evaluate the total phenolic contents, antioxidant and antiglycation activities of leaves, barks, roots and kernels from two cultivars of Mangifera indica (Anacardiaceae). Methods: Total phenolic contents were determined by using Folin-Ciocalteu's method. The antioxidant activities were assessed by three different protocols including DPPH, oxygen radical absorbance capacity and iron (II) chelation assays. In addition, in vitro bovine serum albumin/D-ribose assay was chosen to evaluate the antiglycation properties of the extracts. Results: All the investigated extracts were found to contain high level of total phenols as well as potent antioxidant activities. Kernel extracts showed the highest total phenol contents and DPPH radical scavenging activities whereas higher oxygen radical absorbance capacity values were observed for leave, root and bark extracts. Besides, extracts from leaves, roots and barks from both cultivars exhibited potent inhibitory effects against the formation of advanced glycation end products, with IC50 values lower than the standard positive control aminoguanidine. Conclusions: The potent antiglycation and antioxidative activities of these two Mangifera indica cultivars suggest a possible role in targeting aging, diabetic complications and oxidative stress related diseases.
... The plant material, extract preparation, high performance counter-current chromatography (HPCCC) separation and HPLC equipment were previously described (Malherbe et al., 2014). Briefly: Dried, green shoots of C. genistoides plant material (150 g), defatted with dichloromethane, were extracted using methanol (600 mL). ...
Extraction of xanthones and benzophenones from Cyclopia genistoides was optimised in terms of solvent composition (ethanol concentration) and extraction temperature, using a central composite design, response surface methodology and multi-response desirability profiling. Compounds of interest were the xanthones, mangiferin and isomangiferin, and the benzophenones, 3-beta-D-glucopyranosyl-4-beta-D- glucopyranosyloxyiriflophenone and 3-b-D-glucopyranosyliriflophenone, present in high levels in C. genistoides. Optimum extraction conditions (40% ethanol; 70 �C; 10:1 solvent:solid ratio; 30 min) applied to different batches of plant material (n = 10), delivered mean extract, xanthone and benzophenone yields of 37.8, 4.12 and 1.99 g/100 g plant material, respectively, with a mean alpha-glucosidase inhibitory activity of 39.5% at 100 lg/mL. Similar a-glucosidase inhibitory activity was demonstrated for mangiferin and isomangiferin, indicating that the position of the sugar moiety on the dibenzo-gamma-pirone structure did not affect activity. Superior C. genistoides genotypes in terms of xanthone and benzophenone content were identified.
... The scientific interest shown in this herbal tea is lacking, evident by the few experimental studies especially with regards to its antimicrobial and antioxidant bioactivities. A handful of studies have evaluated the antioxidant (Hubbe and Joubert, 2000;Garcia et al., 2003;Leiro et al., 2003;Joubert et al., 2008;Van der Merwe et al., 2012;Malherbe et al., 2014) and antimicrobial (Coetzee et al., 2008) activity of some Cyclopia spp. Coetzee and colleagues focused mainly on the anti-fungal aspect of the green honeybush herbal tea, with their main interest in the ethanolic solvent extracts. ...
Ethnopharmacological relevance
Cyclopia intermedia is indigenous to South Africa and used to prepare honeybush herbal tea. This aromatic herbal tea has been associated with numerous health benefits, mostly based on anecdotal evidence and with very few studies reporting on the antimicrobial activities.
Aim
The inhibitory effect on the growth of important nosocomial microorganisms and possible association with the antioxidant capacity/content of various solvent extracts of green/unfermented and fermented honeybush plant material were determined in the current study.
Materials and methods
The agar disk diffusion assay was used as a screening assay for the antimicrobial activity of the various honeybush extracts, while the minimal inhibitory concentration (MIC) values, using the broth micro-dilution method, were determined against Streptococcus pyogenes, Staphylococcus aureus and Candida albicans. Active antimicrobial compounds were then shown using thin layer chromatography bioautography. Total antioxidant capacities and -content were also determined for each extract, while the main phenolic compounds were quantified using HPLC.
Results
Six of the eight solvent extracts of honeybush showed antimicrobial activity, with the fermented and green methanol extracts being most effective against S. aureus and C. albicans respectively, whilst the green chloroform extract was most potent against S. pyogenes. Thin layer chromatography-bioautography acknowledged the existence of active antimicrobial fractions within these different solvent honeybush extracts. Regardless of the assay, the green honeybush extracts generally exhibited the highest antioxidant capacity when compared to the fermented honeybush. The total polyphenols were also observed to be highest in green extracts when using water and methanol as solvents. In general the mangiferin and hesperidin contents were higher in the green than the fermented extracts of honeybush plant material.
Conclusions
The inhibitory activity of the various extracts against specific microorganisms was observed to be linearly proportional to the extract concentration. Although the different solvent extracts can only be considered weak antimicrobial agents, three compounds showed specific activity and should be further elucidated in future. It appears the antioxidant capacity of the various solvent herbal extracts did not relate to the antimicrobial activities; however, further work will be required to ascertain this observation. The current data also suggested that the various extracts of honeybush can be considered a good source of a unique blend of natural phytochemical antioxidants and antimicrobial compounds and should be further elucidated.
... Quercetin-3-O-robinobioside was quantified as rutin equivalents due to unavailability of an authentic standard. Extracts of Cyclopia species (honeybush) were analysed by HPLC as described by de Beer and Joubert and Malherbe et al. [19,33] Eriodictyol glucoside, phloretin-3 0 ,5 0 -di-C-glucoside, iriflophenone-3-C-glucoside and scolymoside were quantified as eriocitrin, phloretin-3 0 -C-glucoside (nothofagin), hesperidin and luteolin equivalents, respectively. The two xanthones and the flavone, scolymoside, were quantified at 320 nm, while remaining compounds were quantified at 288 nm. ...
Objectives:
The relationship between polyphenol constituents, antioxidant properties of aqueous and methanol extracts of green tea (Camellia sinensis), the herbal teas, rooibos (Aspalathus linearis) and honeybush (Cyclopia spp.), against skin cell viability was investigated in vitro.
Methods:
The effect of extracts, characterised in terms of polyphenol content and antioxidant properties, on cell viability of premalignant, normal and malignant skin cells was determined.
Key findings:
Phenolic composition, particularly high levels of potent antioxidants, of rooibos and green tea methanol extracts was associated with a strong reduction in cell viability specifically targeting premalignant cells. In contrast, the aqueous extracts of Cyclopia spp. were more effective in reducing cell viability. This correlated with a relatively high flavanol/proanthocyanidin content and ABTS radical cation scavenging capacity. The major green tea flavanol (epigallocatechin gallate) and rooibos dihydrochalcone (aspalathin) exhibited differential effects against cell viability, while the major honeybush xanthone (mangiferin) and flavanone (hesperidin) lacked any effect presumably due to a cytoprotective effect. The underlying mechanisms against skin cell viability are likely to involve mitochondrial dysfunction resulting from polyphenol-iron interactions.
Conclusions:
The polyphenol constituents and antioxidant parameters of herbal tea extracts are useful tools to predict their activity against skin cell survival in vitro and potential chemopreventive effects in vivo.
... Since then, many studies have shown the in vitro antioxidant activity of MF at doses up to 100 lg/mL through chemical assays such as the already reported DPPH and lipid peroxida-tion assays, and other assays as oxygen radical absorbance capacity (ORAC) at doses between 1.04 and 3.55 lmol, Trolox Equivalent, Antioxidant Capacity at doses between 10 and 100 lg/mL, and 2,2 0 -azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS 1 ) at doses between 20 and 220 lg/mL [46][47][48][49][50][51][52]. Besides the determination of the antioxidant activity with chemical assays, this property has been evaluated using cell cultures. ...
Mangiferin (1,3,6,7-tetrahydroxyxanthone-C2-β-D-glucoside) is a natural bioactive xanthonoid that can be found in many plant species, among which the mango tree (Mangifera indica L), a plant widely used in the traditional medicinal, is one of its primary sources. The use of mangiferin for cancer treatment has attracted the attention of research groups around the World. Single administration of mangiferin or in combination with known anticancer chemicals has shown the potential benefits of this molecule in lung, brain, breast, cervix, and prostate cancers, and leukemia. Mangiferin mechanisms of action against cancer cells through in vitro, ex vivo, or in vivo models are discussed besides its antioxidant and anti-inflammatory properties. Nevertheless, pharmaceutical development and, therefore, clinical trials on cancer targets are still lacking. © 2016 BioFactors, 2016.
... Two other important C. genistoides constituents, isomangiferin (IM) and iriflophenone-3-C-b-glucoside (IG) constituting ca. 2 and 1 % DW, respectively (Kokotkiewicz et al. 2012(Kokotkiewicz et al. , 2013bJoubert et al. 2014), have so far not been extensively studied with respect to their biological effects. However, both compounds were recently shown to be potent antioxidants (Malherbe et al. 2014) and also demonstrated strong pro-apoptotic activity on rheumatoid arthritis synoviocytes (Kokotkiewicz et al. 2013b). Moreover, IG and IGcontaining Cyclopia extracts were shown to inhibit adipogenesis in 3T3-L1 cell line (Dudhia et al. 2013). ...
In vitro shoots of the commercially important South-African legume Cyclopia genistoides were grown in different liquid culture systems [membrane rafts (MR) and a temporary immersion bioreactor (TIB)] and evaluated for the accumulation of phenolic secondary metabolites. The major constituents of the investigated cultures were medicinally relevant xanthones [mangiferin (M) and isomangiferin (IM)] and benzophenone derivatives [iriflophenone 3-C-β-glucoside (IG)]. The highest concentrations of M, IM and IG in MR-grown shoots were 1,843.59, 712.02 and 594.29 mg 100 g−1 dry wt, respectively. Bioreactor cultivation provided higher peak concentrations of M (2,622.70 mg 100 g−1 dry wt), IM (757.40 mg 100 g−1 dry wt) and IG (648.30 mg 100 g−1 dry wt) which corresponded to the respective productivities of 5.48, 1.58 and 3.04 mg l−1 d−1. The results indicate that TIB cultures of C. genistoides may be utilized as an alternative source of the above constituents, particularly IM and IG, which are relatively expensive and so far hardly available from commercial sources.
... [14]. A recent study compared the peroxyl radical activity of mangiferin with that of two other major polyphenols found in the unfermented extract of C. maculata used in this study i.e. isomangiferin and the benzophenone, iriflophenone-3-C-glucoside, showing the relative order of activity to be isomangiferin > mangiferin > iriflophenone-3-C-glucoside [15]. The unfermented extract used in this study has been characterised by Dudhia et al. ...
New strategies, which include β-cell protection, are required in the treatment of T2D, as current drugs demonstrate little or no capacity to directly protect the vulnerable β-cell against diabetes-induced cytotoxicity. In this study we investigated the ameliorative effect of pre-treatment with an aqueous extract of unfermented Cyclopia maculata (honeybush) on STZ-induced diabetes and pancreatic β-cell cytotoxicity in Wistar rats after demonstrating a protective effect in vitro in RIN-5F cells. The amelioration of STZ-induced diabetes was seen in the reduction of the area under the curve, determined by the oral glucose tolerance test, as well as fasting glucose levels in extract-treated rats. Pre-treatment with extract also improved serum triglyceride levels and the glucose-to-insulin ratio. Pre-treatment with the extract or the drug, metformin, increased the β-cell area in islets, with a concomitant increase in β-cell proliferation at the higher extract dose (300 mg/kg/d), but not the lower dose (30 mg/kg/d). Subsequently, the in vitro tritiated thymidine incorporation assay showed that the extract was not mitogenic in RIN-5F cells. STZ-induced elevation of plasma nitrite levels was reduced in extract-treated rats, but no changes were observed in their serum catalase, serum glutathione, liver lipid peroxidation and liver nitrotyrosine levels. Pre-treating the rats with extract ameliorated the diabetic effect of STZ in Wistar rats, with evidence of pancreatic β-cells protection, attributed to the presence of high levels of antioxidants such as the xanthones, mangiferin and isomangiferin.
Naturally occurring benzophenones represent a relatively small group of plant metabolites with narrow distribution, mainly in members of Clusiaceae, Gentianaceae, Hypericaceae, Polygalaceae, Myrtaceae, etc.; however, there were reports of several compounds derived from microorganisms belonging to the Aspergillaceae and Valsaceae families and propolis. Benzophenones exhibit many biological activities, such as antioxidant, anti-inflammatory, cytotoxic, antimicrobial, etc. Few reviews on benzophenones that have appeared in the literature were focused on their prenylated derivatives. Summarized information on structural diversity, distribution, and biological activities of simple oxygenated naturally occurring benzophenones and their glycosides has not been found in the literature. Until 2000, only benzophenone C-glycosides were known to occur in nature. Since then, many O-glycosides have been isolated, structurally, and biologically characterized. This review covers the years from 1850 to 2023 and was compiled using databases such as Chemical Abstracts, Scopus, Google Scholar, PubMed, and ResearchGate. Based on their degree of oxidation, 210 chemical structures of benzophenone derivatives and glycosides were grouped into six categories. In addition, in one group of 40 miscellaneous benzophenones, where one or several protons are replaced by a methyl, alcohol, carboxyl, or acyl group, glycosidic forms with such an aglycone and dimeric compounds with xanthone was included. Simple oxygenated benzophenones and their glycosides were found in 77 plant genera belonging to 44 families. The allergy-associated bezophenone-1, benzophenone-2 and benzophenone-3 have limited distribution across natural sources. A wide range of biological activities (antioxidant, anti-inflammatory, cytotoxic, antitumor, cytoprotective, antimicrobial, MAO-A, antiarthritic, anticholinesterase, anti-atherosclerotic, laxative, etc.) of simple oxygenated benzophenones and their glycosides that appeared in the literature were discussed.
An online high-performance liquid-chromatography-diode-array detector coupled with detection of antioxidant compounds using oxygen radical absorbance capacity (ORAC) assay and electrospray ionization-high-resolution mass spectrometer (HPLC-DAD-antioxidant assay (ORAC)/ESI-HRMS) was developed for the identification of antioxidant compounds in complex mixtures. The method was validated using quercetin and a mixture of antioxidant compounds with different antioxidant activities (resveratrol, dihydroxymethoxy-dihydrochalcone, ferulic acid, baicalein and luteolin). Accuracy of the system was established by comparing the results from the developed system with those from ORAC microplate assay determination and reveals the ability of the system to determine the respective contribution of antioxidant compounds to the whole activity of complex mixtures. Application of the system to the identification of antioxidants in a commercial Yerba Mate extract (Ilex paraguariensis St. Hil.) reveals the occurrence of seven actives, which were characterized as chlorogenic acids isomers (3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid and 5-O-caffeoylquinic acid), dicaffeoylquinic acid isomers (3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid and 4,5-di-O-caffeoylquinic acid) and rutin based on UV/Vis spectra, HRMS and MS/MS data. This on-line system is able to generate HPLC-DAD fingerprints, UV/Vis spectra, ORAC activity profile and high-resolution mass spectrometric data.
Coffea arabica L. leaves represent a viable alternative to the canonical matrices used for preparation of beverages, such as tea leaves and grounded coffee beans. Coffee leaves infusions are rich in antioxidant phenolic compounds and have a lower concentration of caffeine. Due to increasing interest in this field, a complete study of the bioactive compounds as chlorogenic acids, xanthones and alkaloids is noteworthy. C. arabica leaves were subjected to ultrasound-assisted extraction, and the extracts were studied via nuclear magnetic resonance spectroscopy (NMR) and chromatographic techniques coupled with mass spectrometry (HPLC-MSn) to identify and quantify the secondary metabolites profile through an untargeted data dependent approach. A quantitative analysis was performed for the major components—chlorogenic acids, mangiferin, caffeine and trigonelline—via HPLC-MS in Single Ion Monitoring (SIM) mode. In total, 39 compounds were identified. The presence of these bioactive compounds proved the strong potential of C. arabica leaves as functional food and as an alternative to classic infused beverages.
In this study, we aimed to identify the utility of pruned mango (Mangifera indica ‘Irwin’) leaves as a resource for ingredients with antioxidant activity. Firstly, we examined the antioxidant activity of extracts obtained from the pericarps, flesh, flowers, barks, seeds, young dark reddish brown leaves (YDL-ext), young yellow leaves (YYL-ext), and pruned old dark green leaves (OML-ext) obtained from ‘Irwin’ mango. Among them, methanolic extract of flower and OML-ext showed the most potent 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and superoxide dismutase (SOD)-like activity. The flesh extract showed weak DPPH radical scavenging activity, but did not show SOD-like activity. Secondly, we investigated the relationship between the maturation of leaves and their antioxidant activity by considering the contents of their two active polyphenolic components, 3-C-β-D-glucosyl-2,4,4’,6-tetrahydroxybenzophenone (1) and mangiferin (2), in addition to chlorophyll (3) and anthocyanins represented by cyanidin-3-O-glucoside (4). The DPPH radical scavenging activity of YDL-ext, YYL-ext and OML-ext were mainly attributable to 1, 2 and 3, whereas their SOD-like activity was partly attributable to 2. The DPPH radical scavenging and SOD-like activities of YDL-ext and YYL-ext were attributable to 1 and 2. These activities were also due to anthocyanins whose content is highest in YDL-ext. Considering the amounts of leaves obtained from pruning, old dark green leaves may be a reasonable natural resource for preparing cosmetics and/or supplemental ingredients with health-enhancing properties, antioxidant activity and inhibitory effect on AGEs formation and pancreatic lipase.
In this work, we have fabricated a novel amperometric platform for the sensing of mangiferin (MAG) based on application of an interesting modification strategy to a bare glassy carbon electrode (GCE). The first layer of the modification process consisted of loading of multiwalled carbon nanotubes-ionic liquid (MWCNTs-IL) onto the GCE surface which was then, modified with reduced fullerene C60 (C60). The C60/MWCNTs-IL/GCE was further modified with chitosan-IL (CS-IL) which was an excellent platform for electrodeposition of the Au and Pt nanoparticles (AuPt NPs). Our modification procedure allowed us to obtain AuPt NPs/CS-IL/C60/MWCNTs-IL/GCE as the sensing platform for determination of MAG in real samples. After characterization and confirmation of the modification process, the sensor response to MAG was calibrated by standard solutions which showed us that the sensor response was linear in the concentration range of 2–40 nM with a detection limit of 0.61 nM. After that, the sensor was applied to the determination of MAG in spiked human plasma samples which confirmed the capability of the sensor for determination of MAG in real samples. Our results confirmed that the sensor response was repeatable, reproducible, selective, sensitive and stable towards determination of MAG.
Extracts of honeybush (Cyclopia genistoides) containing glucosylated xanthones and benzophenones inhibit α-glucosidases, a group of key glycolytic enzymes and drug targets in the treatment of type 2 diabetes mellitus. Ultrafiltered C. genistoides extract was used as the starting material for the development of an optimized protocol to produce xanthone- and benzophenone-enriched fractions (XEFs and BEFs, respectively) by macroporous adsorbent resin chromatography. Static sorption experiments were performed to determine resin loading capacity. Adsorption isotherm data were fitted to the Langmuir and Freundlich models. A fractionation protocol using step-wise gradient elution with 0–30% EtOH-water mixtures was developed, based on the results of dynamic sorption experiments. All XEFs contained 3-β-D-glucopyranosyliriflophenone due to co-elution with the xanthones, mangiferin and isomangiferin, whereas BEFs contained zero or trace amounts of xanthones. Phenolic variation between different batches of the raw material (n = 10) manifested as variation in the composition and degree of enrichment of BEFs and XEFs. This scalable, eco-friendly enrichment strategy should find application for the preparation of enriched fractions from other herbal plants containing the target compound.
Increasing demand for honeybush tea (Cyclopia spp.) and the need for industry expansion have created interest in non-utilised species such as Cyclopia pubescens Eckl. & Zeyh. Very limited information is available on the phenolic composition of this species. A reversed phase core-shell biphenyl column was used to develop and validate a quantitative HPLC diode-array detection method for separation of the major phenolic compounds in C. pubescens. Eight phenolic compounds were identified and a further six tentatively identified by comparison of retention time, UV–vis and high resolution mass spectrometric characteristics with those of authentic reference standards and literature, respectively. Genotypic variation in the phenolic composition of C. pubescens was determined by analysing the leaves and stems of seedlings (n = 17) in a field gene bank. The xanthone and benzophenone present in the highest levels in the leaves were mangiferin and 3-β-D-glucopyranosyl-4-β-D-glucopyranosyloxyiriflophenone, respectively. The leaves contained higher quantities of all compounds, except hesperidin, the major compound in the stems, and a second hesperetin glycoside. Statistical analysis included hierarchical clustering to determine the degree of dissimilarity between genotypes, which provided valuable information for future breeding programs. Comparison of the dendrograms for leaves and stems indicated different clustering patterns for genotypes.
A novel Au-Ag nanoparticles (NPs) on multi-walled carbon nanotubes/sulfonated graphene sheets modified glassy carbon electrode (Au-AgNPs/MWNTs-SGSs/GCE) was prepared to determine mangiferin (MIN) and icariin (IIN) by electrochemical methods Scanning electron microscopy and energy-dispersive spectroscopy showed that Au-AgNPs were electrodeposited on drape and tubular structure of MWNTs-SGSs/GCE successfully. The modified electrode had high electroactivities against both MIN and IIN. The anodic peak currents of MIN and IIN on the GCE modified by Au-AgNPs/MWNTs-SGSs were much higher than those on the bare GCE. Differential pulse voltammetry showed a wide detection range of 0.05. ∼. 500.0. μM for MIN and 0.05. ∼. 100.0. μM for IIN, with low detection limits (0.017. μM both) as well as high sensitivity, selectivity and repeatability. Au-AgNPs/MWNTs-SGSs/GCE was successfully applied to simultaneously detect MIN and IIN in Rhizoma anemarrhenae and to independently determine MIN in Artemisia capillaris Herba and IIN in the leaves of Epimedium macranthum.
Natural anti-oxidative compounds have important disease prevention and food preservation properties, in addition to anti-bacterial, anti-inflammation, anti-cancer, and skin whitening effects. High-performance liquid chromatography (HPLC), with an ultra vilolet (UV) detector coupled to a reverse phase C18 column and an online measurement system for 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radicals, was used to search for potent antioxidative compounds in crude extracts. The online HPLC-DPPH assay was then applied to confirm antioxidative compounds in water extracts from Radix of Pueraria lobata, Rhizoma of Zingiber officinale, Fructus of Chaenomeles sinensis, Cortex of Ulmus pumila, and Radix of Astragalus membranaceus. To determine the yields of the extracts, the Brix% of each extract solution was measured using a refractometer. When the relative DPPH radical scavenging ability values of the water extracts were compared with those of a positive control (ascorbic acid), the water extracts of P. lobata, C. sinensis, and U. pumila were 7.77%, 4.71%, and 4.19%, respectively. The results suggest that this method provides a useful assay for rapid measurement of DPPH radical scavenging abilities and conformation of antioxidative compounds in natural products. Moreover, it can reduce the time spent on the separation of active compounds from natural materials, such as medicinal plants, in addition to the use of reagents for separation.
A sensitive and specific liquid chromatography with tandem mass spectrometry method was firstly developed for the measurement of isomangiferin in rat plasma. Chloramphenicol was selected as the internal standard. Sample preparation was carried out through a simple one-step protein precipitation procedure with methanol. Negative electrospray ionization was performed using multiple reaction monitoring mode with transitions of m/z 421.1/301.1 for isomangiferin, and 321.1/151.9 for chloramphenicol. The calibration curve was linear over the range of 0.1-600 ng/mL, with a lower limit of quantification at 0.1 ng/mL. The intra- and inter-day precisions (relative standard deviation) were no more than 8.2% and accuracies (relative error) were within the range of -8.4 to 2.2%. The recovery, matrix effect and stability under different conditions were all proved acceptable. The validated method has been successfully applied to a preclinical pharmacokinetic study of isomangiferin in rats for the first time. This article is protected by copyright. All rights reserved.
BACKGROUND: Bone marrow mesenchymal stem cells have particularly applied prospects in tissue engineering. However, the death of transplanted cells limits the tissue regeneration. To search a new drug of anti-free radicals and protecting bone marrow mesenchymal stem cells is of great significance. OBJECTIVE: To investigate the protective effects of mangiferin on bone marrow mesenchymal stem cells against hypoxia. METHODS: Rat bone marrow mesenchymal stem cells were cultured in vitro and hypoxia cell model was established by cobalt chloride. Cells were divided into normal control group, hypoxia group (treated with cobalt chloride), and mangiferin groups (cobalt chloride+20, 40, 80, 160 μmol/L mangiferin). After 12 and 24 hours of hypoxia, superoxide dismutase, malondialdehyde, and catalase levels in the cell supernatant were determined. After 3, 6, 12, 24 hours of hypoxia, reactive oxygen species change was detected in each group. RESULTS AND CONCLUSION: Mangiferin significantly improved the survival rate of bone marrow mesenchymal stem cells exposed to hypoxia, increased the intracellular superoxide dismutase and catalase activities, decreased intracellular malondialdehyde and reactive oxygen species levels, thereby effectively protecting bone marrow mesenchymal stem cells against hypoxia. These findings indicate that mangiferin has effective protection against hypoxia and strong antioxidant ability, and can significantly reduce oxidative damage. © 2014, Chinese Journal of Tissue Engineering Research. All Rights Reserved.
Cyclopia genistoides Vent is a commercially important species of the genus Cyclopia, endemic to the Cape Floristic region of South Africa. In recent years, the utilization of Cyclopia spp. has progressed from their traditional use as herbal tea (honeybush) to the production of hot water extracts destined for the food and nutraceutical industries. The functional ingredient status of Cyclopia extracts is supported by their biological properties [1], which may partially be attributed to their phenolic composition [2]. Comprehensive data regarding the nature of these phenolic constituents, as well as their individual content values, are therefore required. High-performance liquid chromatographic (HPLC) methods currently employed in the quantitative analysis of C. genistoides extracts are, however, hampered in terms of limited resolution and the number of quantifiable analytes [3 – 4]. In light of the above, a validated HPLC method was developed to quantify the major polyphenols present in hot water extracts of C. genistoides. The phenolic constituents were identified with electrospray-ionisation mass spectrometry (ESI-MS) and characterized on the basis of their unique fragmentation pattern (ESI-MS/MS). The optimized method enabled accurate quantification of twenty compounds, with the xanthones (mangiferin, isomangiferin) and benzophenones (iriflophenone-3-C-β-D-glucoside, iriflophenone-3-C-β-D-glucoside-4-O-β-D-glucoside) representing the major phenolic constituents. This is the first report of the presence of iriflophenone-3-C-β-D-glucoside-4-O-β-D-glucoside in C. genistoides extracts, unambiguously identified by appropriate procedures including nuclear magnetic resonance after isolation. C. genistoides was ultimately shown to be a rich source of bio-active xanthones and benzophenones and is, to our knowledge, the only known natural source of iriflophenone-3-C-β-D-glucoside-4-O-β-D-glucoside. Keywords: HPLC, ESI-MS, mangiferin, isomangiferin, iriflophenone-3-C-β-D-glucoside, iriflophenone-3-C-β-D-glucoside-4-O-β-D-glucoside, quantification, isolation References: [1] Joubert E, Joubert ME, Bester C, de Beer D, De Lange JH. Honeybush (Cyclopia spp.): From local cottage industry to global markets – The catalytic and supporting role of research. S Afr J Bot 2011; 77: 887 – 907. [2] Joubert E, Gelderblom WCA, Louw A, de Beer D. South African herbal teas: Aspalathus linearis, Cyclopia spp. and Athrixia phylicoides – A review. J Ethnopharmacol 2008; 119: 376 – 412. [3] Joubert E, Otto F, Gruner S, Weinreich B. Reversed-phase HPLC determination of mangiferin, isomangiferin and hesperidin in Cyclopia and the effect of harvesting date on the phenolic composition of C. genistoides. Eur Food Res Technol 2003; 216: 270 – 273. [4] de Beer D, Joubert E. Development of HPLC method for Cyclopia subternata phenolic compound analysis and application to other Cyclopia spp. J Food Compos Anal 2010; 23: 289 – 297.
Mangiferin (MgFn) is a naturally occurring bioactive glucosylxanthone having numerous pharmacological activities. A feasible microwave assisted extraction (MAE) technique for extraction of MgFn from Curcuma amada (mango ginger) was developed in our laboratory. To prove the efficacy of MAE technique, it was compared with other methods like maceration extraction (ME) and ultrasonic sound assisted extraction (UAE). A high yield of MgFn was obtained by MAE (1.472 mg/g) compared to 1.017 mg/g yield by UAE and 0.837 mg/g by ME technique in acetone-water (50:50 v/v). Scanning electron microscopy (SEM) photographs of C. amada cells were taken, which were subjected to different extraction forces mentioned above. The efficacy of MgFn as a radical scavenger was studied using 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), nitrate scavenging and ferric reducing power assays. The IC50 value of MgFn extracted with MAE was recorded to be 26 μg/mL; showing it to be an effective radical scavenger more efficient than ascorbic acid.
An enriched fraction of an aqueous extract prepared from the aerial parts of Cyclopia genistoides Vent. yielded a new benzophenone di-C,O-glucoside, 3-C-β-d-glucopyranosyl-4-O-β-d-glucopyranosyliriflophenone (1), together with small quantities of a known benzophenone C-glucoside, 3-C-β-d-glucopyranosylmaclurin (2). The isolated compounds showed α-glucosidase inhibitory activity against an enzyme mixture extracted from rat intestinal acetone powder. Compound 2 exhibited significantly (p < 0.05) higher inhibitory activity (54%) than 1 (43%) at 200 μM. In vitro tests in several cell models showed that 1 and its 3-C-monoglucosylated derivative (3-C-β-d-glucopyranosyliriflophenone) were marginally effective (p ≥ 0.05) in increasing glucose uptake.
Cyclopia species are used for the production of honeybush tea and food ingredient extracts associated with many health benefits. A species-specific high-performance liquid chromatography (HPLC) method for Cyclopia maculata, developed and validated, allowed quantification of the major compounds in extracts from "unfermented" and fermented C. maculata. Two xanthones were tentatively identified for the first time in a Cyclopia species, whereas an additional four compounds were tentatively identified for the first time in C. maculata. "Fermentation" (oxidation) decreased the content of all compounds, with the exception of vicenin-2. Similarity analysis of the chromatographic fingerprints of unfermented C. maculata aqueous extracts showed extremely low variation (r ≥ 0.97) between samples. Some differences between wild-harvested and cultivated seedling plants were, however, demonstrated using principal component analysis. Quantitative data of selected compounds confirmed the low level of variation, making this Cyclopia species ideal for the production of standardized food ingredient extracts.
Models to predict the total antioxidant capacity (TAC) of rooibos tea infusions from their chromatographic fingerprints and peak table data (content of individual phenolic compounds), obtained using HPLC with diode array detection, were developed in order to identify potential antioxidant markers. Peak table data included the content of 12 compounds, namely phenylpyruvic acid-2-O-glucoside, aspalathin, nothofagin, isoorientin, orientin, ferulic acid, quercetin-3-O-robinobioside, vitexin, hyperoside, rutin, isovitexin and isoquercitrin. The TAC values, measured using the oxygen radical absorbance capacity (ORAC) and DPPH radical scavenging assays, could be predicted from the peak table data or the chromatographic fingerprints (prediction errors 9–12%) using partial least squares (PLS) regression. Prediction models created from samples of only two production years could additionally be used to predict the TAC of samples from another production year (prediction errors < 13%) indicating the robustness of the models in a quality control environment. Furthermore, the uninformative variable elimination (UVE)-PLS method was used to identify potential antioxidant markers for rooibos infusions. All individual phenolic compounds that were quantified were selected as informative variables, except vitexin, while UVE-PLS models developed from chromatographic fingerprints indicated additional antioxidant markers, namely (S)-eriodictyol-6-C-glucoside, (R)-eriodictyol-6-C-glucoside, aspalalinin and two unidentified compounds. The potential antioxidant markers should be validated prior to use in quality control of rooibos tea.
Oxidative stress plays an important role in the progression of diabetes complications. The aim of the present study was to investigate the beneficial effect of oral administration of mangiferin in streptozotocin (STZ)-induced diabetic rats by measuring the oxidative indicators in liver and kidney as well as the ameliorative properties. Administration of mangiferin to diabetic rats significantly decreased blood glucose and increased plasma insulin levels. The activities of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) and level of reduced glutathione (GSH) were significantly (P < 0.05) decreased while increases in the levels of lipidperoxidation (LPO) markers were observed in liver and kidney tissues of diabetic control rats as compared to normal control rats. Oral treatment with mangiferin (40 mg/kg b.wt/day) for a period of 30 days showed significant ameliorative effects on all the biochemical and oxidative parameters studied. Diabetic rats treated with mangiferin restored almost normal architecture of liver and kidney tissues, which was confirmed by histopathological examination. These results indicated that mangiferin has potential ameliorative effects in addition to its antidiabetic effect in experimentally induced diabetic rats.
Iterative computations are pervasive among data analysis applications in the cloud, including Web search, online social network analysis, recommendation systems, and so on. These cloud applications typically involve data sets of massive scale. Fast convergence of the iterative computation on the massive data set is essential for these applications. In this paper, we explore the opportunity for accelerating iterative computations and propose a distributed computing framework, PrIter, which enables fast iterative computation by providing the support of prioritized iteration. Instead of performing computations on all data records without discrimination, PrIter prioritizes the computations that help convergence the most, so that the convergence speed of iterative process is significantly improved. We evaluate PrIter on a local cluster of machines as well as on Amazon EC2 Cloud. The results show that PrIter achieves up to 50x speedup over Hadoop for a series of iterative algorithms.
Phenolic compounds are abundant secondary metabolites in plums, with potential health benefits believed to be due to their antioxidant activity, amongst others. Phenolic characterisation of South African Prunus salicina Lindl. plums is necessary to fully evaluate their potential health benefits. An HPLC method using diode-array detection (DAD) for quantification of phenolic compounds was improved and fluorescence detection (FLD) was added for quantification of flavan-3-ols. Validation of the HPLC-DAD-FLD method showed its suitability for quantification of 18 phenolic compounds, including flavan-3-ols using FLD, and phenolic acids, anthocyanins and flavonols using DAD. The method was suitable for characterisation of the phenolic composition of 11 South African plum cultivars and selections, including various types with yellow and red skin and flesh. The method was used in conjunction with mass spectrometry (MS) to identify 24 phenolic compounds. Neochlorogenic acid and cyanidin-3-O-glucoside were the major compounds in most of the plums, while cyanidin-3-O-glucoside was absent in Sun Breeze plums with yellow skin and flesh. Post-column on-line coupling of the ABTS•+ scavenging assay with HPLC-DAD enabled qualitative evaluation of the relative contribution of individual phenolic compounds to the antioxidant activity. The flavan-3-ols, neochlorogenic acid and cyanidin-3-O-glucoside displayed the largest antioxidant response peaks.
Cyclopia subternata plants are traditionally used for the production of the South African herbal tea, honeybush, and recently as aqueous extracts for the food industry. A C. subternata aqueous extract and mangiferin (a major constituent) are known to have anti-diabetic properties. Variation in phenolic composition and antioxidant capacity is expected due to cultivation largely from seedlings, having implications for extract standardization and quality control. Aqueous extracts from 64 seedlings of the same age, cultivated under the same environmental conditions, were analyzed for individual compound content, total polyphenol (TP) content and total antioxidant capacity (TAC) in a number of assays. An HPLC method was developed and validated to allow quantification of xanthones (mangiferin, isomangiferin), flavanones (hesperidin, eriocitrin), a flavone (scolymoside), a benzophenone (iriflophenone-3-C-β-glucoside) and dihydrochalcones (phloretin-3',5'-di-C-β-glucoside, 3-hydroxyphloretin-3',5'-di-C-hexoside). Additional compounds were tentatively identified using mass spectrometric detection, with the presence of the 3-hydroxyphloretin-glycoside, an iriflophenone-di-O,C-hexoside, an eriodictyol-di-C-hexoside and vicenin-2 being demonstrated for the first time. Variability of the individual phenolic compound contents was generally higher than that of the TP content and TAC values. Among the phenolic compounds, scolymoside, hesperidin and iriflophenone-3-C-β-glucoside contents were the most variable. A combination of the measured parameters could be useful in product standardization by providing a basis for specifying minimum levels.
This review details the vernacular names, origin, distribution, taxonomy and variety of Mangifera indica L. (Anacardiaceae), a medicinal plant traditionally used in tropical regions. Mangiferin, a major C-glucosylxanthone from M. indica stem bark, leaves, heartwood, roots and fruits occurs widely among different angiosperm families and ferns. The reported pharmacological activities of mangiferin include antioxidant, radioprotective, antitumor, immunomodulatory, anti-allergic, anti-inflammatory, antidiabetic, lipolytic, antibone resorption, monoamine oxidase inhibiting, antiviral, antifungal antibacterial and antiparasitic properties, which may support the numerous traditional uses of the plant.
A reversed-phase HPLC method for separation of polyphenols in honeybush tea (Cyclopia spp.) is presented. Separation of eriodictyol, luteolin, medicagol, formononetin, mangiferin, isomangiferin, hesperetin and hesperidin was investigated. A C12 stationary phase was required to separate mangiferin and isomangiferin. The method was used to quantify the three major polyphenols (mangiferin, isomangiferin and hesperidin) in C. genistoides, C. intermedia, C. maculata and C. sessiliflora and to study the effect of harvesting date on these compounds in two types of C. genistoides. The highest levels of the xanthones, mangiferin (3.61 g/100 g) and isomangiferin (0.54 g/100 g), and the flavanone, hesperidin (1.74 g/100 g), were found for C. genistoides (both xanthones) and C. intermedia, respectively. Cyclopia sessiliflora contained the lowest levels of mangiferin (1.04 g/100 g) and hesperidin (0.29 g/100 g). The mangiferin content of both the Overberg and West Coast types decreased with harvesting date (P <0.05). The Overberg type contained more mangiferin, but hesperidin was more prominent in the West Coast type.
Biochemical detection (BCD) methods are commonly used to screen plant extracts for specific biological activities in batch assays. Traditionally, bioactives in the most active extracts were identified through time-consuming bio-assay guided fractionation until single active compounds could be isolated. Not only are isolation procedures often tedious, but they could also lead to artifact formation. On-line coupling of BCD assays to high performance liquid chromatography (HPLC) is gaining ground as a high resolution screening technique to overcome problems associated with pre-isolation by measuring the effects of compounds post-column directly after separation. To date, several on-line HPLC-BCD assays, applied to whole plant extracts and mixtures, have been published. In this review the focus will fall on enzyme-based, receptor-based and antioxidant assays.
The role of oxidative stress in streptozotocin (STZ)-induced toxicity and its prevention by a xanthone glucoside, mangiferin was investigated. To induce diabetes mellitus, adult male Wistar rats were injected STZ intravenously at 55 mg/kg body weight. The effect of mangiferin (10 and 20 mg/kg, i.p., 28 days) was investigated in STZ-induced diabetic male rats. Insulin-treated rats (6 U/kg, i.p., 28 days) served as positive control. Diabetic rats given normal saline served as negative control. Normal rats that neither received STZ nor drugs served as normal control. On day 28, the diabetic rats showed significant increase in serum creatine phosphokinase (CPK) and total glycosylated haemoglobin. Kidney revealed tubular degeneration and decreased levels of superoxide dismutase (SOD) and catalase (CAT) with an elevation of malonaldehyde (MDA). Cardiac SOD, CAT and lipid peroxidation were significantly increased. Histopathological findings revealed cardiac hypertrophy with haemorrhages. Analysis of erythrocyte revealed significantly elevated levels of MDA with insignificant decrease in CAT and SOD. Repeated intraperitoneal injections of mangiferin (10 and 20 mg/kg) and insulin (6 U/kg) controlled STZ-induced lipid peroxidation and significantly protected the animals against cardiac as well as renal damage. From the study, it may be concluded that oxidative stress appears to play a major role in STZ-induced cardiac and renal toxicity as is evident from significant inhibition of antioxidant defence mechanism in renal tissue or a compensatory increase in antioxidant defence mechanism in cardiac tissue. Intraperitoneal administration of mangiferin exhibited significant decrease in glycosylated haemoglobin and CPK levels along with the amelioration of oxidative stress that was comparable to insulin treatment.
From the heartwood of Garcinia pedunculata,2,4,6,3′,5′-pentahydroxybenzophenone and 1,3,5,7-tetrahydroxyxanthone have been isolated. Their occu
A fast and efficient method for the isolation of the C-glucosidated xanthones mangiferin and isomangiferin from the South-African plant Cyclopia genistoides was developed for the first time. The procedure involved extraction, liquid-liquid partitioning with ethyl acetate and subsequent precipitation of mangiferin and isomangiferin from methanol and acetonitrile-water fractions, respectively. Additionally, two benzophenone derivatives: 3-C-β-glucosides of maclurin and iriflophenone, were isolated from C. genistoides extracts using semi-preparative HPLC. Apart from the above, the isolation procedure also yielded hesperidin and small amounts of luteolin. The structures of the compounds were determined by 1D and 2D NMR experiments and/or LC-DAD-ESI-MS. The selected Cyclopia constituents were screened for pro-apoptotic activity on TNF-α-stimulated synovial cells isolated from rheumatoid arthritis patients. The strongest effect, measured as percent of apoptotic cells, was recorded for isomangiferin (75 %), followed by iriflophenone 3-C-β-glucoside (71 %), hesperidin (67 %) and mangiferin (65 %). The results are encouraging for further studies on the use of the above compounds in the treatment of rheumatoid arthritis.
In the presented work, an insight was made into the polyphenolic composition of intact plant material and in vitro cultures of indigenous South African plant Cyclopia subternata Vogel (honeybush). Ethyl acetate fractions of methanol extracts were separated by means of gravity column chromatography and/or semipreparative HPLC on two serially connected monolithic RP-18 columns. The structures of the isolated compounds were determined by means of 1D and 2D NMR techniques and additionally confirmed by LC-DAD-ESI-MS. Apart from the previously described honeybush components, that is mangiferin (1), scolymoside (2), hesperidin (3) and narirutin (4), three additional compounds: iriflophenone 3-C-beta-glucoside (benzophenone) (5), phloretin 3',S'-di-C-beta-glucoside (dihydrochalcone) (6), and isorhoifolin (flavone) (7) were identified for the first time in the herb of C. subternata. Additionally, three isoflavone glucosides, namely calycosin 7-O-beta-glucoside (8), rothindin (9) and ononin (10), which had not been previously reported in Cyclopia plants, were identified in the callus of the above species. As far as the authors are concerned, this is the first report on the presence of benzophenone and dihydrochalcone derivatives in Cyclopia genus.
70% ethanol-water extract from the leaves of Mangifera indica L. (Anacardiaceae) was found to show inhibitory effect on triglyceride (TG) accumulation in 3T3-L1 cells. From the active fraction, six new benzophenone C-glucosides, foliamangiferosides A3 (1), A4 (2), C4 (3), C5 (4), C6 (5), and C7 (6) together with 11 known benzophenone C-glucosides (7-17) were obtained. In this paper, the isolation, structure elucidation (1-6) and the MS fragment cleavage pathways of all the 17 isolates were studied. 1-6 showed inhibitory effects on TG and free fatty acid accumulation in 3T3-L1 cells at 10 μM.
Honeybush tea (Cyclopia spp.), one of the traditional South African herbal teas with a long history of regional use, remained a cottage industry until the mid-1990s when researchers were instrumental in the development of a formal agricultural and agro-processing industry. It is one of the
few indigenous South African plants that made the transition from the wild to a commercial product during the past 100 years. Research activities during the past 20 years included propagation, production, genetic improvement, processing, composition and the potential for value-adding. The present review provides an up-to-date and comprehensive record of the development of the South African honeybush industry, against the background of the historical highlights in the making of an industry. It provides a blueprint of the processes and actions involved in the development of a new agricultural and agro-processing industry from an herbal plant. Insight into challenges faced by the industry and future research needed to keep it competitive are provided.
Lippia multiflora, a perennial, aromatic shrub commonly known as bush tea has recently been identified as an African plant with high commercial potential due to its medicinal properties. The plant material was subjected to steam pasteurisation to improve its microbial quality. The major compounds of L. multiflora herbal infusion, i.e. the phenylethanoid glycosides (PhGs), verbascoside, isoverbascoside, nuomioside A and isonuomioside A, and the flavone, luteolin-7-O-glucuronide were quantified by HPLC. Verbascoside was the most abundant PhG. The PhGs are of interest due to their pharmacological properties. Liquid chromatography tandem-mass spectrometry (LC-MS(2)) was used to tentatively identify the compounds. The on-line DPPH() (2,2'-diphenyl-1-picrylhydrazyl radical) scavenging assay (reaction time=0.45s) applied to the infusion in "quantitative" mode, showed the relative order of activity: isonuomioside A>isoverbascoside>verbascoside>nuomioside A. In the microplate assay (reaction time=2h), isoverbascoside and verbascoside had similar activity. Both compounds were less active in the latter assay than the well-known flavan-3-ol antioxidant, (-)-epigallocatechin gallate, but more active than caffeic acid and an ester, rosmarinic acid. Steam pasteurisation of L. multiflora leaves at maximum exposure (150s at ca 99°C) for improved microbial quality did not decrease the soluble solids content, phenolic content and antioxidant activities of the infusion compared to the untreated control (p<0.05). The PhG content of the water soluble solids was as high as 15%, underscoring the potential of L. multiflora extract as functional ingredient.
An on-line HPLC (high performance liquid chromatography) method for the rapid screening of individual antioxidants in mixtures was developed using crocin as a substrate (i.e. oxidation probe) and 2,2'-azobis(2-amidinopropane dihydrochloride (AAPH)) in phosphate buffer (pH 7.5) as a radical generator. The polyene structure of crocin and AAPH-derived peroxyl radicals resemble the lipidic substrates and radicals found in true food more closely than the popular, albeit artificial, DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS(+) (2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate)) do. After separation by a C18 (octadecyl silica) column and UV (ultraviolet) detection, antioxidative analytes react with peroxyl radicals at 90°C and the inhibition of crocin oxidation (i.e. bleaching) is detected as a positive peak by an absorbance detector at 440 nm. The method is simple, uses standard instruments and inexpensive reagents. It can be applied for isocratic HPLC runs using mobile phases containing 10-90% organic solvent in water, weak acids or buffers (pH 3.5-8.5). With baseline correction, gradient runs are also feasible. The radical scavenging activity of several natural antioxidants and a green tea extract was studied. After optimisation of conditions such as reagent concentrations and flows, the limit of detection varied from 0.79 to 7.4 ng, depending on the antioxidant.
Analysis of phenolic compounds in Cyclopia spp. (honeybush) is very important because of commercial interest in the plant material as an herbal tea and in extracts for the nutraceutical and cosmeceutical markets. An existing HPLC-DAD method was adapted and proved reproducible as evidenced by good intra- and inter-day precision. The method allows the quantification of several well-known as well as unidentified phenolic compounds. Its suitability for quantification of the major phenolic compounds present in the water extracts of unfermented and fermented Cyclopia subternata, Cyclopia intermedia, Cyclopia genistoides and Cyclopia sessiliflora was evaluated. Known compounds that were identified and quantified include mangiferin (0.1–11.8 g/100 g), isomangiferin (0.1–3.2 g/100 g), eriocitrin (not detected–0.4 g/100 g), hesperidin (0.2–1.2 g/100 g) and scolymoside (not detected–1.8 g/100 g), while narirutin and luteolin were detected in most samples, but could not be quantified due to low concentrations and co-elution. An unidentified compound (traces–4.8 g hesperidin equivalents/100 g), unidentified hydroxycinnamic acid derivative (not detected–0.3 g mangiferin equivalents/100 g), unidentified flavanone (not detected–2.0 g hesperidin equivalents/100 g) and an eriodictyol-glycoside (not detected–0.5 g hesperidin equivalents/100 g) could also be quantified. Large variation was observed within a sample series of the same extract type due to the nature of the plant material. Qualitative and quantitative differences were observed between species, while “fermentation”, a high-temperature process involving non-enzymatic oxidation, decreased the content of most phenolic compounds. Results using DAD as the only detection method should be handled with care, as the tentative identification of compounds 9 and 10, based on retention time and UV–Vis spectrum, could not be confirmed using liquid chromatography–electrospray ionisation-mass spectrometry.
Aspalathin (2',3,4,4',6'-pentahydroxy-3'-C-beta-d-glucopyranosyldihydrochalcone) is the major flavonoid present in the South African herbal tea rooibos. In vitro metabolism of aspalathin and a structural analogue nothofagin, lacking the A ring catechol group, was investigated by monitoring the formation of glucuronyl and sulfate conjugates using Aroclor 1254 induced and uninduced rat liver microsomal and cytosolic subcellular fractions. Following glucuronidation of both aspalathin and nothofagin, HPLC-DAD, LC-MS, and LC-MS/MS analyses indicated the presence of two metabolites: one major and one minor. Only one aspalathin metabolite was obtained after sulfation, while no metabolites were observed for nothofagin. Two likely sites of conjugation for aspalathin are 4-OH or 3-OH on the A-ring. For nothofagin, the 4-OH (A-ring) and 6'-OH (B-ring) seem to be involved. The glucuronyl conjugates of aspalathin lack any radical scavenging properties in online postcolumn DPPH radical and ABTS radical cation assays. Deconjugation assays utilizing glucuronidase and sulfatase resulted in the disappearance of the metabolites, with the concomitant formation of the unconjugated form in the case of the glucuronidated product. The balance between conjugated and unconjugated forms of aspalathin could have important implications regarding its role in affecting oxidative status in intra- and extracellular environments in vivo.
The antioxidant activity of rooibos flavonoids, including the dihydrochalcones aspalathin and nothofagin and their corresponding flavone glycosides, was evaluated using the ABTS radical cation, metal chelating, and Fe(II)-induced microsomal lipid peroxidation assays. Epigallocatechin gallate (EGCG) and Trolox were used as reference standards. Optimized geometric conformers of aspalathin and nothofagin, in addition to calculated physicochemical properties, were considered to explain interaction with the microsomal membrane structure and thus relative potency of the dihydrochalcones. The most potent radical scavengers were aspalathin (IC50 = 3.33 microM) and EGCG (IC50 = 3.46 microM), followed by quercetin (IC50 = 3.60 microM) and nothofagin (IC50 = 4.04 microM). The least effective radical scavengers were isovitexin (IC50 = 1224 microM) and vitexin (IC50 > 2131 microM). Quercetin (IC50 = 17.5 microM) and EGCG (IC50 = 22.3 microM) were the most effective inhibitors of lipid peroxidation. Aspalathin (IC50 = 50.2 microM) and catechin (IC50 = 53.3 microM) displayed similar potencies. Nothofagin (IC50 = 1388 microM) was almost as ineffective as its flavone glycoside analogues.
Two benzophenones, hypericophenonoside (1) and annulatophenone (2) were isolated from the methanol extract of the herb of Hypericum annulatum. The structures of the benzophenones were established as 2'-O-beta-D-glucopyranosyl-2,4,5',6-tetrahydroxy benzophenone (1) and 2,3',5',6-tetrahydroxy-4-methoxybenzophenone (2) based on spectral and chemical evidence. Hypericophenonside is the second benzophenone O-glycoside found in nature. Acid and enzymatic hydrolysis of (1) led directly to the formation of 1,3,7-trihydroxyxanthone (gentisein). This fact confirmed the hypothesis that some xanthones could be formed in plants by dehydration of 2,2'-dihydroxybenzophenones, and the intermediate precursors appear to be benzophenone O-glycosides ortho to the carbonyl function.
An improved method of oxygen radical absorbance capacity (ORAC) assay has been developed and validated using fluorescein (3',6'-dihydroxyspiro[isobenzofuran-1[3H],9'[9H]-xanthen]-3-one) as the fluorescent probe. Our results demonstrate that fluorescein (FL) is superior to B-phycoerythrin. The oxidized FL products induced by peroxyl radical were identified by LC/MS, and the reaction mechanism was determined to follow a classic hydrogen atom transfer mechanism. In addition, methodological and mechanistic comparison of ORAC(FL) with other widely used methods was discussed. It is concluded that, unlike other popular methods, the improved ORAC(FL) assay provides a direct measure of hydrophilic chain-breaking antioxidant capacity against peroxyl radical.
The oxygen radical absorbance capacity (ORAC) assay has been widely accepted as a standard tool to measure the antioxidant activity in the nutraceutical, pharmaceutical, and food industries. However, the ORAC assay has been criticized for a lack of accessibility due to the unavailability of the COBAS FARA II analyzer, an instrument discontinued by the manufacturer. In addition, the manual sample preparation is time-consuming and labor-intensive. The objective of this study was to develop a high-throughput instrument platform that can fully automate the ORAC assay procedure. The new instrument platform consists of a robotic eight-channel liquid handling system and a microplate fluorescence reader. By using the high-throughput platform, the efficiency of the assay is improved with at least a 10-fold increase in sample throughput over the current procedure. The mean of intra- and interday CVs was <or=15%, and the limit of detection and limit of quantitation were 5 and 6.25 microM, respectively.
Polyphenolics were extracted from peels, pulp and kernels of mango fruits (Mangifera indica L. cv. 'Tommy Atkins') and characterized by high-performance liquid chromatography/electrospray ionization mass spectrometry. In the peel 18 gallotannins and five benzophenone derivatives were detected which were tentatively identified as galloylated maclurin and iriflophenone glucosides. Twenty-one and eight gallotannins were found in the kernels and pulp, respectively, whereas no evidence for the presence of benzophenone derivatives was obtained. Gallotannins quantified by the rhodanine assay amounted to 1.4 mg/g dm in the peels (expressed as gallic acid), while only small amounts (0.2 mg/g dm) were found in the pulp. In contrast, mango kernels contained 15.5 mg/g dm and thus proved to be a rich source of gallotannins.
Cyclopia genistoides, normally used for the preparation of an herbal tea, honeybush, is a good source of the bio-active compounds mangiferin and hesperidin and is in demand for the preparation of xanthone-enriched extracts. Near-infrared spectroscopy (NIRS) was used to develop calibration models to predict the mangiferin and hesperidin contents of the dried green plant material. NIRS measurements of plant material and pure compounds were performed in diffuse reflectance mode. The calibration sets for mangiferin and hesperidin contents ranged from 0.7 to 7.21 and 0.64-4.80 g/100 g, respectively. Using independent validation, it was shown that the NIRS calibration models for the prediction of mangiferin (SEP=0.46 g/100 g; R2=0.74; and RPD=1.96) and hesperidin (SEP=0.38 g/100 g; R2=0.72; and RDP=1.90) contents of the dried plant material are adequate for screening purposes, based on RPD values.
An analytical separation was performed on an analytical J-type counter-current chromatography (CCC) instrument using a 5.4 ml column, with a 1 ml/min mobile phase flow rate. This separation had a resolution of 0.69 and was achieved in 10 min. The same separation was performed using two 2300 ml columns connected in series at a flow rate of 850 ml/min using a production scale J-type centrifuge. This production scale separation was also obtained in 10 min with a resolution of 0.71. This represents an 850 times increase in productivity. This paper presents these separations and the underlying scale up theory.
The contents of secondary plant substances in solvent extracts of various byproducts (barks, kernels, peels, and old and young leaves) in a range of Brazilian mango cultivars were identified and quantitated. The results show that the profiles of secondary plant substances such as xanthone C-glycosides, gallotannins, and benzophenones in different byproducts vary greatly but are fairly consistent across cultivars. The free radical scavenging activity of the solvent extracts was evaluated using a high-performance liquid chromatography-based hypoxanthine/xanthine oxidase assay and revealed dose-dependent antioxidant capacity in all extracts. Four (mangiferin, penta- O-galloyl-glucoside gallic acid, and methyl gallate) of the major phenolic compounds detected were also evaluated in additional in vitro bioassay systems such as oxygen radical absorbance capacity, 2,2-diphenyl-1-picrylhydrazyl, and ferric reducing ability of plasma. Mangiferin in particular, detected at high concentrations in young leaves (Coite = 172 g/kg), in bark (Momika = 107 g/kg), and in old leaves (Itamaraka = 94 g/kg), shows an exceptionally strong antioxidant capacity.
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