Strong antioxidant phenolics from Acacia nilotica: profiling by ESI-MS and qualitative-quantitative determination by LC-ESI-MS

Acacia nilotica (L.) Del. syn is a species rich in polyphenolic constituents, in which catechins are hypothesized to possess antioxidant properties and to play a role in the anti-inflammatory activity of several plants. Due to the complexity of catechin derivatives, the investigation of this class of natural compounds has been limited by difficulties in their separation. In this paper, rationalization of the phenolics occurring in the 80% EtOH extract of Acacia nilotica pods, on the basis of ESI-MS and ESI-MS/MS profiles, has been proposed. Additionally, an LC-ESI-MS qualitative study has been performed by using a C18 polar endcapped stationary phase. The fragmentation pattern obtained evidenced the presence in A. nilotica pods of galloylated catechin- and gallocatechin derivatives along with galloylated glucose derivatives. The structures were confirmed by NMR, after isolation of the pure compounds.In addition, the radical scavenging activities of extracts and pure compounds were investigated, by using the TEAC assay. Furthermore quantitative analyses were performed by LC-ESI-MS/MS, confirming the interest of this species as a rich source of very strong antioxidant principles.     

雷公藤属醋酸乙酯部位化学成分及药理作用研究进展

雷公藤属植物化学成分复杂,具有广泛的药理作用。现在已经从雷公藤属醋酸乙酯提取部位分离得到多种化学成分,主要为二萜类、三萜类、倍半萜类、生物碱类及其他化合物;药理研究表明其具有抗炎、免疫抑制、抗血小板聚集作用和杀虫活性。对雷公藤属醋酸乙酯部位化学成分及药理作用研究进展进行综述,为后续研究提供理论依据。     

Comparative analysis of the antioxidant and DNA protection capacities of Anadenanthera colubrina, Libidibia ferrea and Pityrocarpa moniliformis fruits

This study aimed to explore the antioxidant and DNA protection abilities of hydroalcoholic extracts from fruits of Anadenanthera colubrina (ACHE), Libidibia ferrea (LFHE) and Pityrocarpa moniliformis (PMHE). These extracts were tested by five antioxidant methods (phosphomolibdenium and reducing power assays; superoxide, hydrogen peroxide and nitric oxide scavenging) and DNA protection capacity. Total phenolic content was measured by Folin-Ciocalteu method. ACHE exhibited the highest phenolic content (578 mg/g GAE), followed by LFHE (460 mg/g GAE) and PMHE (448 mg/g GAE). In phosphomolibdenium assay, ACHE showed 24.81% of activity in relation to ascorbic acid, whereas LFHE and PMHE had 21.08% and 18.05%, respectively. These plants showed high ability to inhibit reactive species tested with IC50 values ranged from 10.66 to 14.37 μg/mL for superoxide radical; 26.05 to 45.43 μg/mL for hydrogen peroxide; 178.42 to 182.98 μg/mL for reducing power; and 199.2 to 283 μg/mL for nitric oxide. Furthermore, these extracts had capacity to break the DNA damage induced by hydroxyl radicals. The antioxidant activity of these plants is related with their higher phenolic content and show that they may be used as source of bioactive compounds, relevant to the maintenance of oxidative stability of the food matrix, cosmetics and/or pharmaceutical preparations.     

Antidiabetic activity of 50% ethanolic extract of Ricinus communis and its purified fractions

We investigated the antidiabetic activity of 50% ethanolic extract of roots of Ricinus communis (RCRE) along with its bioassay-guided purification. Five-hundred milligram per kilogram body weight appeared to be the effective dose as it caused the maximum lowering of the fasting blood glucose, both in normal as well as type 1 diabetic animals. The maximum hypoglycemic effect was always observed at the 8th h up to which the study has been conducted.     

Potential analgesic,anti-inflammatory and antioxidant activities of hydroalcoholic extract of Areca catechu L. nut

The hydroalcoholic extract of Areca catechu L. (ANE) nut was screened for its analgesic, anti-inflammatory and in vitro antioxidant potential. Three doses of ANE (250, 500 and 1000 mg/kg orally) were tested for analgesic and anti-inflammatory activities. Evaluation of analgesic activity of ANE was performed using hot plate and formalin test in mice. ANE showed maximum increase in hot plate reaction time (56.27%, p < 0.01), while reduced the duration of licking/biting behaviors in first (39.45%, p < 0.05) and second (92.71%, p < 0.01) phases of the formalin test indicating significant analgesic activity. ANE reduced the paw edema considerably (86.79% inhibition after 24 h, p < 0.01) in dose-dependent manner compared to carrageenan-induced rat. In addition, in vitro antioxidant activity of ANE was investigated by total phenolic content (TPC) and hydrogen peroxide assay. The IC₅₀ observed in hydrogen peroxide assay was 83.14 μg/ml and TPC 120.56 ± 21.09 mg QE/g. Altogether, these results suggest that the hydroalcoholic extract of Areca catechu could be considered as a potential analgesic, anti-inflammatory and antioxidant agent.     

Cognitive enhancing and antioxidant activity of ethyl acetate soluble fraction of the methanol extract of Hibiscus rosa sinensis in scopolamine-induced amnesia

Objective:

The objective of the present study was to evaluate the cognitive enhancing and antioxidant activity of Hibiscus rosa sinensis.

Materials and Methods:

The learning and memory was impaired by administration of scopolamine (1 mg/kg, i.p.) in mice which is associated with altered brain oxidative status. The object recognition test (ORT) and passive avoidance test (PAT) were used to assess cognitive enhancing activity. Animals were treated with an ethyl acetate soluble fraction of the methanol extract of H. sinensis (25, 50 and 100 mg/kg, p.o).

Results:

The ethyl acetate soluble fraction of the methanol extract of H. sinensis (EASF) attenuated amnesia induced by scopolamine and aging. The discrimination index (DI) was significantly decreased in the aged and scopolamine group in ORT. Pretreatment with EASF significantly increased the DI. In PAT, scopolamine-treated mice exhibited significantly shorter step-down latencies (SDL). EASF treatment showed a significant increase in SDL in young, aged as well as in scopolamine-treated animals. The biochemical analysis of brain revealed that scopolamine treatment increased lipid peroxidation and decreased levels of superoxide dismutase (SOD) and glutathione reductase (GSH). Administration of extract significantly reduced LPO and reversed the decrease in brain SOD and GSH levels. The administration of H. sinensis improved memory in amnesic mice and prevented the oxidative stress associated with scopolamine. The mechanism of such protection of H. sinensis may be due to augmentation of cellular antioxidants.

Conclusion:

The results of the present study suggested that H. sinensis had a protective role against age and scopolamine-induced amnesia, indicating its utility in management of cognitive disorders.     

In vitro and in vivo antioxidant properties of different fractions of Moringa oleifera leaves

The antioxidant potency of different fractions of Moringa oleifera leaves were investigated by employing various established in vitro systems, such as β-Carotene bleaching, reducing power, DPPH/superoxide/hydroxyl radical scavenging, ferrous ion chelation and lipid peroxidation. On the basis of in vitro antioxidant properties polyphenolic fraction of M. oleifera leaves (MOEF) was chosen as the potent fraction and used for the DNA nicking and in vivo antioxidant properties. MOEF shows concentration dependent protection of oxidative DNA damage induced by HO and also found to inhibit the toxicity produced by CCl₄ administration as seen from the decreased lipid peroxides (LPO) and increased glutathione (GSH) levels. Among the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) levels were restored to almost normal levels compared to CCl₄ intoxicated rats. The HPLC analysis indicated the presence of phenolic acids (gallic, chlorogenic, ellagic and ferulic acid) and flavonoids (kaempferol, quercetin and rutin). Thus, it may be concluded that the MOEF possess high phenolic content and potent antioxidant properties, which may be mediated through direct trapping of the free radicals and also through metal chelation.     

Antioxidant potential of aqueous extract of Phyllanthus amarus in rats

Objective:

Increased levels of oxidative stress may be implicated in the etiology of many pathological conditions. Protective antioxidant action imparted by many plant extracts and plant products make them promising therapeutic drugs for free radical induced pathologies. In this study we assessed the antioxidant potential of Phyllanthus amarus (Euphorbiaceae).

Materials and Methods:

Experimental rats were divided into two groups: Control and Phyllanthus amarus (P. amarus) treated. Treated rats received P. amarus aqueous extract (PAAEt) at a dose of 200 mg/kg body wt/day for 8 weeks. After the treatment period of 8 weeks lipid peroxidation (LPO), vitamin C, uric acid and reduced glutathione (GSH) were estimated in plasma and antioxidant enzymes: Glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD) were also assayed. Genotoxicity of PAAEt was assessed by single cell gel electrophoresis (SCGE) of lymphocytes under both in vitro and in vivo conditions. The protective role of PAAEt against hydrogen peroxide (H₂O₂), streptozotocin (STZ) and nitric oxide generating system induced lymphocyte DNA damage was also assessed by SCGE.

Results:

PAAEt treated rats showed a significant decrease in plasma LPO and a significant increase in plasma vitamin C, uric acid, GSH levels and GPx, CAT and SOD activities. SCGE experiment reveals that PAAEt was devoid of genotoxicity and had a significant protective effect against H₂O₂, STZ and nitric oxide (NO) induced lymphocyte DNA damage.

Conclusion:

The results suggest the non-toxic nature of PAAEt and consumption of PAAEt can be linked to improved antioxidant status and reduction in the risk of oxidative stress.     

In vivo assessment of genotoxic effects of Annona squamosa seed extract in rats

Widespread use of pesticides represents a potential risk to human and environmental health. Hence, biopesticides from plants are some of the future strategies for plant protection. In this regard, a seed extract of Annona squamosa was prepared and found to be a promising pesticide. In order to establish the inherent toxicity and non-target safety required for registration and marketing of pesticides, toxicological studies are conducted. The genotoxicity potential was evaluated in rats with 75, 150 and 300 mg/kg Annona squamosa by the comet assay in leucocytes, micronucleus and chromosomal aberration tests in bone marrow. We also studied the effects of 300 mg/kg of extract on lipid peroxidation, reduced glutathione level and glutathione S transferase activity in liver, lungs, brain, kidneys, heart and spleen of treated rats. The comet assay showed a statistically significant dose related increase in DNA migration. The micronucleus and chromosomal aberration tests revealed a significant induction in frequency of micronuclei and chromosomal aberrations at 150 and 300 mg/kg. Annona squamosa treatment significantly enhanced lipid peroxidation, decreased glutathione and glutathione S transferase levels revealing the oxidative stress condition. Our results warrant careful use of Annona squamosa seed extract as a biopesticide till more tests are carried out.     

蛴螬脂溶性成分的研究

目的 研究蛴螬提取物中的脂溶性成分。 方法 采用乙酸乙酯回流提取蛴螬中的脂溶性成分,对其进行气相色谱-质谱法(GC-MS)分析,并对主要脂溶性成分进行定性分析。 结果 从蛴螬脂溶性成分中鉴定出33个化合物,十五酸、Z-11-十六烯酸、十六酸、十七酸、十八烯酸和十八酸为其主要的脂溶性成分。 结论 本研究为进一步研究动物药和确认蛴螬的有效成分奠定了基础。     

Evaluation of metal concentration and antioxidant activity of three edible mushrooms from Mugla,Turkey

This study is designed for the determination of metal concentrations, antioxidant activity potentials and total phenolics of Amanita caesarea, Clitocybe geotropa and Leucoagaricus pudicus. Concentrations of four heavy metals (Pb, Cd, Cr, Ni) and five minor elements (Zn, Fe, Mn, Cu, Co) are determined. In the case of A. caesarea, Cr and Ni concentrations are found in a high level. Concentrations of the metals are found to be within safe limits for C. geotropa. In β-carotene/linoleic acid test, L. pudicus showed the highest activity potential. In DPPH system, A. caesarea showed 79.4% scavenging ability. Additionally, reducing power and chelating capacity of the mushrooms increased with concentration. The strongest super-oxide anion scavenger was A. caesarea. In the case of total phenolics, L. pudicus found to have the highest content.     

Acetylcholinesterase inhibitory and antioxidant properties of Cyclotrichium niveum, Thymus praecox subsp. caucasicus var. caucasicus, Echinacea purpurea and E. pallida

The dichloromethane, ethyl acetate, ethanol, and aqueous extracts of Cyclotrichium niveum (CN) and Thymus praecox subsp. caucasicus var. caucasicus (TP), Echinacea purpurea (EPU), and E. pallida (EPA) along with the essential oils of CN and TP were assessed for their anti-acetylcholinesterase (AChE) and antioxidant activities. AChE inhibition was estimated using spectrophotometric method of Ellman. Antioxidant activity was evaluated by 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and ferrous ion-chelating power tests. Ferric-reducing antioxidant power (FRAP) of CN and TP were also tested. CN essential oil was found to contain isomenthone (56.21%) and pulegone (19.76%). The ethyl acetate (83.11–87.98%) and dichloromethane (73.45–84.02%) extracts of CN showed the highest AChE inhibition. The ethyl acetate and ethanol extracts of TP exerted significant DPPH scavenger effect. The water extracts of CN and TP and the chloroform extract of the aerial parts of EPU displayed the highest ferrous ion-chelating effect. The leaf and flower essential oils of TP had the best FRAP.     

Hepatoprotective potential of polyphenol rich extract of Murraya koenigii L.: an in vivo study

The present study investigates hepatoprotective effects of polyphenol rich Murraya koenigii L. (MK) hydro-ethanolic leaf extract in CCl(4) treated hepatotoxic rats. Plasma markers of hepatic damage, lipid peroxidation levels, enzymatic, and non-enzymatic antioxidants in liver and histopathological changes were investigated in control and treated rats. MK pretreated rats with different doses (200, 400 and 600mg/kg body weight) showed significant decrement in activity levels of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total protein, and bilirubin. Also, MK treated rats recorded a dose dependent increment in hepatic super oxide dismutase, catalase, reduced glutathione and ascorbic acid and, a decrement in lipid peroxidation. Microscopic evaluations of liver revealed CCl(4)-induced lesions and related toxic manifestations that were minimal in liver of rats pretreated with MK extract. These results demonstrate that hydro-ethanolic leaf extract of MK possesses hepatoprotective potentials.     

Systematic comparison of nutraceuticals and antioxidant potential of cultivated, in vitro cultured and commercial Melissa officinalis samples

Melissa officinalis (lemon balm) infusions are used worldwide for digestive, analgesic and other pharmaceutical applications. Herein, the nutraceuticals production and antioxidant potential in garden cultivated, in vitro cultured and two commercial samples (bags and granulated) of lemon balm was compared. The profile of in vitro cultured lemon balm is closer of garden cultivated sample than of both commercial samples (bag or granulate). It presented the highest levels of proteins and ash, and the lowest energetic value. The most favorable n6/n3 ration, as also the highest PUFA (mostly α-linolenic acid), tocopherols (including α-, γ- and δ-isoforms) and ascorbic acid contents were also observed in this sample. Nevertheless, it was the commercial bag lemon balm that gave the highest antioxidant activity and the highest levels of phenolics and flavonoids. As far as we kwon, this is the first comparison of nutraceuticals and antioxidant potential of cultivated, in vitro cultured and commercial lemon balm samples. Moreover, it proved that in vitro culture might be used to stimulate vitamins production.     

Antioxidant activity and protective effect of Turnera ulmifolia Linn. var. elegans against carbon tetrachloride-induced oxidative damage in rats

The present study aimed to determine whether the leaves of Turnera ulmifolia Linn. var. elegans extract exert significant antioxidant activity. The antioxidant activity of its hydroethanolic extract (HEETU) was evaluated by assessing (a) its radical scavenging ability in vitro, and (b) its in vivo effect on lipid peroxidation and antioxidant enzyme activities. The in vitro antioxidant assay (DPPH) clearly supported HEETU free radical scavenging potential. Moreover, glutathione content and antioxidant enzyme activities (glutathione peroxidase, superoxide dismutase and catalase) were significantly enhanced in CCl₄-treated rats due to oral HEETU-treatment (500 mg/kg b.w.) over 7 and 21 days. In addition, an improvement was observed in lipid peroxidation and serum biochemical parameters (aspartate aminotransferase and alanine aminotransferase), indicating a protective effect against CCl₄-induced liver injuries, confirmed by histopathological studies. The HEETU effect was comparable to the standard drug Legalon® (50 mg/kg b.w.) under the same experimental condition. Quantitative analysis of the HPLC extract revealed the presence of flavonoids, wich mediate the effects of antioxidant and oxidative stress. In conclusion, extract components exhibit antioxidant and hepatoprotective activities in vitro and in vivo.     

Anti-inflammatory, antinociceptive and antioxidant activities of the endemic Soqotraen Boswellia elongata Balf. f. and Jatropha unicostata Balf. f. in different experimental models

In the present study, the two endemic Soqotraen plants Boswellia elongata and Jatropha unicostata were investigated for their anti-inflammatory, antinociceptive and antioxidant potential. To assess the anti-inflammatory and antinociceptive activities, two concentrations of each extract (200 and 400 mg/kg, p.o.) were tested in carrageenan-induced rat paw edema, cotton pellet granuloma in rats, acetic acid-induced abdominal writhing and hot-plate test model in mice. Moreover, the antioxidant activity was determined in vitro, using scavenging activity of DPPH radical and β-carotene-linoleic acid assays. Both plants produced significant (P < 0.05-0.01) anti-inflammatory and antinociceptive effects; however the results suggest that B. elongata possesses the highest activities. B. elongata and J. unicostata at (400 mg/kg) reduced the paw edema considerably (82% and 53%) and the weight of cotton pellet granuloma (51% and 32%), respectively. Furthermore, they diminished the abdominal constriction induced by acetic acid with a 67% and 41% inhibition respectively, and prolonged significantly the reaction time of animal with relatively extended duration of stimulation. In addition, both plants showed considerable antioxidant activity in both assays. These results clearly confirmed the traditional anti-inflammatory indication of B. elongata and suggest that B. elongata could be a potential source for anti-inflammatory, antinociceptive and antioxidant agents.     

Subchronic toxicity of Citrus aurantium L. (Rutaceae) extract and p-synephrine in mice

Extracts of Citrus aurantium L. (Rutaceae) unripe fruits have gained popularity for the treatment of obesity. Due to the wide use of C. aurantium/p-synephrine-containing products, this research was undertaken to evaluate its subchronic toxicity in mice and their actions in oxidative stress biomarkers. Groups of 9–10 mice received for 28 consecutive days a commercial C. aurantium dried extract (containing 7.5% p-synephrine) 400, 2000 or 4000 mg/kg and p-synephrine 30 or 300 mg/kg by oral gavage. There was a reduction in body weight gain of animals treated with both doses of p-synephrine. Organs relative weight, biochemical and hematological parameters were not altered in all treated mice. There was an increase in reduced glutathione (GSH) concentration in groups treated with C. aurantium 4000 mg/kg and p-synephrine 30 and 300 mg/kg. In glutathione peroxidase (GPx), there were an inhibition of the activity in C. aurantium 400 and 2000 mg/kg and p-synephrine 30 and 300 mg/kg treated animals, respectively, and was no alteration in malondialdehyde (MDA) levels. Thus, the results indicate a low subchronic toxicity of the tested materials in mice and a possible alteration in the oxidative metabolism. However, further tests are required to better elucidate the effects of these compounds in the antioxidant system.     

Evaluation of the genotoxic potential and acute oral toxicity of standardized extract of Andrographis paniculata (KalmCold™)

Andrographis paniculata is used in the traditional medicine for cold and influenza remedy. The main endeavor in this study was to assess the genotoxicity of the standardized extract of A. paniculata (KalmCold™) through three different in vitro tests: Ames, chromosome aberration (CA), and micronucleus (MN). Ames test was performed at 5000 μg/ml, 1581 μg/ml, 500 μg/ml, 158 μg/ml, 50 μg/ml, 16 μg/ml, while the clastogenicity tests were performed at 80 μg/ml, 26.6 μg/ml, 8.8 μg/ml for short-term treatment without S9; 345 μg/ml, 115 μg/ml, 38.3 μg/ml for short-term treatment with S9; and 46 μg/ml, 15.3 μg/ml, 5.1 μg/ml for long-term without S9 using DMSO as a vehicle control. Results of Ames test confirmed that KalmCold™ did not induce mutations both in the presence and absence of S9 in Salmonella typhimurium mutant strains TA98 and TAMix. In CA and MN, KalmCold™ did not induce clastogenicity in CHO-K1 cells in vitro. Based on our results, it is evident that KalmCold™ is genotoxically safe.     

Antinociceptive and anti-inflammatory effects of the ethyl acetate stem bark extract of Bridelia scleroneura (Euphorbiaceae)

Bridelia scleroneura is a member of the Euphorbiaceae family. In folk medicine in Cameroon, the stem bark of this plant is used for relieving abdominal pain, contortion, arthritis and inflammation. In this study, the antinociceptive and anti-inflammatory activities of the ethyl acetate stem bark extract have been evaluated. The putative analgesic effect of the plant extract was examined in abdominal constriction, hot plate, formalin and on pain using tail immersion mouse models and in carrageenan-induced paw edema in rats. The extract (150–600 mg/kg) exhibited a dose-dependent analgesic effect (46.27–78.97%) in acetic acid-induced abdominal constriction in mice. B. scleroneura extract increased the pain latency of nociceptive response to thermal stimuli at the higher dose of 600 mg/kg. B. scleroneuna induced significant dose-dependent reduction of the nociception in both early and late phases of the formalin test. The extract at the dose of 300 mg/kg, increased significantly, by 63.70% and 52.01% the tail-immersion latency time, 1 and 2 h post-dosing. In the carrageenan test, B. scleroneura (150–600 mg/kg, p.o) had dose-dependent and significant effects at different time intervals. This behaviour was similar to indometacin (10 mg/kg) used as a standard drug. These results show that the ethyl acetate stem bark extract of B. scleroneura possesses peripheral and central analgesic properties as well as anti-inflammatory activity against acute inflammation processes, in support of the folk medicinal use of the plant.