Walnut (Juglans regia L.) leaves: phenolic compounds, antibacterial activity and antioxidant potential of different cultivars.

Different cultivars of walnut (Juglans regia L.) leaves (Cv. Lara, Franquette, Mayette, Marbot, Mellanaise and Parisienne) grown in Portugal, were investigated in what concerns phenolic compounds and antimicrobial and antioxidant properties. Phenolics analysis was performed by reversed-phase HPLC/DAD and 10 compounds were identified and quantified: 3- and 5-caffeoylquinic acids, 3- and 4-p-coumaroylquinic acids, p-coumaric acid, quercetin 3-galactoside, quercetin 3-pentoside derivative, quercetin 3-arabinoside, quercetin 3-xyloside and quercetin 3-rhamnoside. The antimicrobial capacity was screened against Gram positive (Bacillus cereus, B. subtilis, Staphylococcus aureus) and Gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae) and fungi (Candida albicans, Cryptococcus neoformans). Walnut leaves selectively inhibited the growth of Gram positive bacteria, being B. cereus the most susceptible one (MIC 0.1mg/mL). Gram negative bacteria and fungi were resistant to the extracts at 100mg/mL. Lara walnut leaves were also submitted to antibacterial assays using 18 clinical isolates of Staphylococcus sp. Antioxidant activity was accessed by the reducing power assay, the scavenging effect on DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and beta-carotene linoleate model system. In a general way, all of the studied walnut leaves cultivars presented high antioxidant activity (EC(50) values lower than 1mg/mL), being Cv. Lara the most effective one.     

Total phenols, antioxidant potential and antimicrobial activity of walnut (Juglans regia L.) green husks

The total phenols content and antioxidant and antimicrobial activities were studied in walnut (Juglans regia L.) green husks aqueous extracts of five different cultivars (Franquette, Mayette, Marbot, Mellanaise and Parisienne). Total phenols content was determined by colorimetric assay and their amount ranged from 32.61 mg/g of GAE (cv. Mellanaise) to 74.08 mg/g of GAE t (cv. Franquette). The antioxidant capacity of aqueous extracts was assessed through reducing power assay, scavenging effects on DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and beta-carotene linoleate model system. A concentration-dependent antioxidative capacity was verified in reducing power and DPPH assays, with EC50 values lower than 1 mg/mL for all the tested extracts. The antimicrobial capacity was screened against Gram positive and Gram negative bacteria, and fungi. All the extracts inhibited the growth of Gram positive bacteria, being Staphylococcus aureus the most susceptible one with MIC of 0.1 mg/mL for all the extracts. The results obtained indicate that walnut green husks may become important in the obtainment of a noticeable source of compounds with health protective potential and antimicrobial activity.     

Chemical composition, and antioxidant and antimicrobial activities of three hazelnut (Corylus avellana L.) cultivars.

Hazelnut (Corylus avellana L.) is a very popular dry fruit in the world being consumed in different form and presentations. In the present work, three hazelnut cultivars (cv. Daviana, Fertille de Coutard and M. Bollwiller) produced in Portugal, were characterized in respect to their chemical composition, antioxidant potential and antimicrobial activity. The samples were analysed for proximate constituents (moisture, fat, crude protein, ash), nutritional value and fatty acids profile by GC/FID. Antioxidant potential was accessed by the reducing power assay, the scavenging effect on DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and beta-carotene linoleate model system. Their antimicrobial capacity was also checked against Gram positive (Bacillus cereus, B. subtilis, Staphylococcus aureus) and Gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae) and fungi (Candida albicans, Cryptococcus neoformans). Results showed that the main constituent of fruits was fat ranging from 56% to 61%, being the nutritional value around 650 kcal per 100 g of fruits. Oleic was the major fatty acid varying between 80.67% in cv. F. Coutard and 82.63% in cv. Daviana, followed by linoleic, palmitic, and stearic acids. Aqueous hazelnut extract presented antioxidant activity in a concentration-dependent way, in general with similar behaviour for all cultivars. Hazelnut extracts revealed a high antimicrobial activity against Gram positive bacteria (MIC 0.1 mg/mL) showing a good bioactivity of these fruits.     

Larvicidal action of ethanolic extracts from fruit endocarps of Melia azedarach and Azadirachta indica against the dengue mosquito Aedes aegypti

Ethanolic extracts from the kernels of ripe fruits from the Indian Lilac Melia azedarach and from the well-known Neem tree, Azadirachta indica were assayed against larvae of Aedes aegypti, the mosquito vector of dengue fever. The lethality bioassays were carried out according to the recommendations of the World Health Organization. Extracts were tested at doses ranging from 0.0033 to 0.05 g% in an aqueous medium for 24 and 48 h, at 25 or 30 °C, with or without feeding of the larvae. LC₅₀, LC₉₅ and LC₉₉ were determined. Both seed extracts proved lethal for third to fourth instar larvae. Non-fed A. aegypti larvae were more susceptible to Azadirachta extracts at both temperatures. Under a more realistic environmental situation, namely with fed larvae at 25 °C, the death rates caused by the Melia extract were higher, although at 30 °C the extract of Azadirachta had an even higher lethality. Inter allia, the LC₅₀ values for the crude extracts of these two members of the Meliaceae ranged from 0.017 to 0.034 g% while the LC₉₉ values ranged from 0.133 to 0.189 g%. Since no downstream processing was undertaken to purify the active agents in the extracts, our findings seem very promising, suggesting that it may be possible to increase the larvicidal activity further by improving the extraction and the fractionation of the crude limonoids, for instance removing the co-extracted natural fats.     

In vitro toxicity of cereulide on porcine pancreatic Langerhans islets

Cereulide is a K⁺ ionophore cytotoxic and mitochondriotoxic to primary cells and cell lines of human and other mammalian origins. It is a heat-stable, highly lipophilic (log K_ow 5.96) peptide (1152 g mol⁻¹) produced by certain strains of Bacillus cereus, a bacterium connected to emetic food poisonings. In this study the pancreatic toxicity of purified cereulide, and cereulide-containing bacterial extracts, was studied using fetal porcine Langerhans islets in culture. Exposure to 1 ng ml⁻¹ of purified cereulide caused necrotic cell death of the islet cells impairing their insulin content within 2 days. Cell extracts of cereulide-positive B. cereus strains connected to food poisoning or isolated from foodstuffs were toxic, corresponding to their measured cereulide content. Extracts of B. cereus strains producing or not producing the B. cereus diarrheal toxin, but no cereulide, were tolerated by the porcine islet cultures up to concentrations 1000-fold higher compared to extracts from strains containing cereulide, and up to exposure times of 7 d. Cereulide thus was identified as the B. cereus-produced substance toxic towards porcine fetal Langerhans islets and beta cells.     

夹竹桃花脂溶性成分提取工艺优化及其生物活性分析

目的 研究夹竹桃花脂溶性成分的提取工艺、组成成分及生物活性。方法 利用Box-Behnken响应面法优化提取工艺,GC-MS分析组成成分,滤纸片扩散法和二倍连续梯度稀释法分析抑菌活性,并测定抗氧化活性。结果 最佳提取工艺为索氏提取4 h,连续提取4次,料液比1∶ 15,得率9.16%,比优化前提高了13.09%。脂溶性成分含有烷烃类、醛类和酯类等30种化合物,具有还原能力和清除DPPH自由基、羟自由基、超氧自由基的能力,对枯草芽孢杆菌、金黄色葡萄球菌、大肠埃希菌和铜绿假单胞菌的最小抑菌浓度分别为1.22,1.72,0.75,1.96 g·L^–1,最小杀菌浓度分别为2.34,2.84,1.88,3.32 g·L^–1。结论 脂溶性成分最佳提取工艺得率为9.16%,以烷烃类、醛类、酯类为主,具有一定的抗氧化和抑菌活性,可为夹竹桃资源的开发利用提供参考。     

Subcritical water extraction of nutraceuticals with antioxidant activity from oregano. Chemical and functional characterization

In the present work, oregano leaves (Origanum vulgare L.) are explored as natural source of nutraceuticals with antioxidant activity. To do this, subcritical water extraction (SWE), a new environmentally friendly technique, is employed as extraction procedure and HPLC coupled to DAD is used for the chemical characterization of the extracts. Moreover, the radical scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) method and the determination of the total phenolic content (measured with the Folin test) are applied to evaluate the antioxidant activity of the extracts. The extraction of antioxidants from oregano leaves by SWE is studied considering different temperatures (25, 50, 100, 150 and 200 °C) to investigate the selectivity of the process. The highest antioxidant activity is observed for the extract obtained at the highest temperature, 200 °C (EC₅₀ equal to 10 μg/ml). Moreover, the extraction yield was also the highest (54% dry weight) at these extraction conditions. The total phenolic content showed no differences among the different extracts, concluding that the amount of phenolic compounds extracted was similar but the type and structure of the phenolics was different, providing in this way different antioxidant activity. Some compounds could be tentatively identified, proposing some probable chemical structures for some of them, such as flavanones, dihydroflavonols, favonols and flavones.     

In vitro assays of the antibacterial and antioxidant activity of aqueous leaf extracts from different Prunus salicina Lindl. cultivars

The growing interest in the substitution of synthetic food antioxidants and antimicrobial additives by natural ones has fostered research on vegetable sources and on the screening of raw materials, for identifying new antioxidants and antimicrobial natural agents. The aim of the present study was to assess total phenolic contents and determine polyphenolic composition, related antioxidative and antimicrobial properties of plum leaves extracts from six cultivars. It was observed that the content of total phenolic compounds was cultivar dependent. High antioxidant capacity has been observed and related to the relative amounts of polyphenolic compounds with good antioxidant properties. The antimicrobial activity was confirmed against Listeria innocua and Escherichia coli, and it has found to be related with the high phenolic contents. Our results suggest that the use of plum leaf extracts is a feasible alternative as antibacterial and antioxidant agents to prevent the deterioration of stored foods by bacteria and oxidation.     

Characterization of phenolic content,in vitro biological activity,and pesticide loads of extracts from white grape skins from organic and conventional cultivars

Grape skin extracts of Riesling Vitis vinifera L. grapes from conventionally or organically managed cultivars were compared on the basis of their phenolic content, antioxidant capacity, antimicrobial and antimutagenic properties and pesticide loads. Promising results on their biological properties suggest that those extracts would be valuable as food preservatives. The antioxidant capacity of conventional extracts was significantly higher, according to the higher content in catechin, epicatechin and procyanidin B. Pesticide loads did not affect the antimutagenic or antimicrobial properties of the extracts. Both extracts inhibited the growth of Gram-positive foodborne pathogens such as Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium to similar extents. Possibly as a result of higher amounts of quercetin and its derivatives, higher antimicrobial effects against Listeria monocytogenes and Salmonella typhimurium were observed for the organic white grape skin extracts. Conventional or organic extracts did not show remarkable antimutagenic effects when tested against the mutagen IQ by means of the Ames test. Due to the presence of fungicides, the conidial germination of Penicillium expansum, Penicillium chrysogenum and Aspergillus niger, were inhibited by 95% by conventional GSE, while negligible effects were observed with organic grape extracts. The latter, however, showed inhibitory effects against Trichoderma viridie and Aspergillus versicolor.     

Microcalorimetric investigation of the toxic action of ammonium ferric(III) sulfate on the metabolic activity of pure microbes

A series of calorimetric experiments were performed to investigate toxic action of ammonium ferric sulfate (AFS) on Bacillus subtilis, Pseudomonas putida and Candida humicola. The power–time curves of micro-organism metabolism were obtained, and the action of them by addition of AFS was studied. C. humicola, B. subtilis and P. putida were inhibited completely when the concentrations were up to 320.0, 160.0 and 160.0 μg mL⁻¹, respectively. The relationships between growth rate constant (k) and doses of AFS were approximately linear for three microbes, P. putida for 10.0–160.0 μg mL⁻¹ (R = −0.9746), B. subtilis for 0–160.0 μg mL⁻¹ (R = −0.9868) and C. humicola for 10.0–320.0 μg mL⁻¹ (R = −0.9955). The total heat dissipated per milliliter (Q_T) for three microbes remained balance approximately during the lower doses, P. putida and B. subtilis less than the dose of 20.0 μg mL⁻¹, 0.56 ± 0.01 and 0.26 ± 0.01 J mL⁻¹, respectively, C. humicola less than the dose of 40.0 μg mL⁻¹, 0.58 ± 0.03 J mL⁻¹. The biomass and OD₆₀₀ of three micro-organisms growth in the absence of AFS also were obtained. The power–time curve of C. humicola growth coincided with its turbidity curve. It elucidates that microcalorimetric method agreed with the routine microbiology method.     

Comparison of antioxidant activities and total polyphenolic and methylxanthine contents between the unripe fruit and leaves of Ilex paraguariensis A. St. Hil

Ilex paraguariensis is used in Brazil as a stimulating beverage called "mate". Leaves and immature fruit extracts of Ilex paraguariensis were evaluated for their radical scavenging capacity, total methylxanthine and polyphenol contents. Antimicrobial activity of two enriched saponin fractions obtained from the fruits were also evaluated. The radical scavenging activity of the fractioned extracts was determined spectrophotometrically using 1,1-diphenylpicrylhydrazyl free radical (DPPH). The IC50o of L-ascorbic acid, ethyl acetate and n-butanol fractions from the leaves and ethyl acetate fraction from the fruits were 6.48 microg/mL, 13.26 microg/mL, 27.22 microg/mL, and 285.78 microg/mL, respectively. Total methylxanthine content was 1.16 +/- 0.06 mg/g dry weight in the fruits and 8.78 +/- 0.01 mg/g in the leaves. Total polyphenol content varied from 86.82 +/- 3 x 10(-4) to 199.91 +/- 3 x 10(-3) mg/g in leaf fractions and from 54.25 +/- 1 x 10(-3) to 110.36 +/- 4 x 10(-4) mg/g in fruit fractions. Enriched saponin fractions from the fruits showed no antimicrobial activity. To our knowledge, this are the first data available on the antioxidant/antimicrobial activities and polyphenol/methylxanthine contents of Ilex paraguariensis fruits.     

Chemical composition and biological activities of Eruca vesicaria subsp. longirostris essential oils

Context To date, there are no reports to validate the Tunisian traditional and folklore claims of Eruca vesicaria (L) Cav. subsp. longirostris (Brassicaceae) for the treatment of disease.

Objective Investigation of the chemical composition antimicrobial and antioxidant activity of essential oils from Eruca longirostris leaves, stems, roots and fruits.

Materials and methods The essential oils of E. longirostris from leaves, stems, roots and fruits were obtained after 4?h of hydrodistillation. Chemical compositions were determined using a combination of GC/FID and GC/MS. The in vitro antimicrobial activity of the volatile constituents of E. longirostris was performed in sterile 96-well microplates against three Gram-positive, four Gram-negative bacteria and one strain as yeast. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration values were reported. Furthermore, the antioxidant activity was evaluated by DPPH and ABTS assays.

Results The main compound for fruits, stems and roots was the erucin (96.6%, 85.3% and 83.7%, respectively), while β-elemene (35.7%), hexahydrofarnesylacetone (23.9%), (E)-β-damascone (15.4%), erucin (10.6%) and α-longipinene (9.6%) constituted the major compounds in the essential oil of the leaves. The experimental results showed that in all tests, essential oil of fruits showed the better antioxidant activity than the others. On the other hand, the oils of stems, fruits and roots showed significant antimicrobial activity with MIC values ranging from 0.125 to 0.31?mg/mL against Candida species, Gram-positive and Gram-negative bacteria, mainly Salmonella enterica.

Conclusions The present results indicate that essential oils of E. longirostris can be used as a source of erucin.     

Protection of extract from leaves of Ardisia compressa against benomyl-induced cytotoxicity and genotoxicity in cultured rat hepatocytes

The potential of the Ardisia compressa extract (EA) was examined regarding its capacity to reduce the cytotoxic effect of benomyl on rat hepatocytes. The protective effect was evaluated by Janus Green dye exclusion method. An approximate 50% cytotoxic effect of benomyl on hepatocytes was observed at 35 μg/ml after 2 hr of incubation. (−)Epigallocatechin 3-gallato (EGCG) and EA decreased the viability of hepatocytes at concentrations above 3 μg/ml and 2.52 μg, equivalent to (+)catechin/ml, respectively. A protective effect against benomyl was observed when hepatocytes were previously exposed to EGCG (3 μg/ml) or EA (2.52 μg, equivalent to (+)catechin/ml) followed by incubation with benomyl (35 μg/ml) alone. When EGCG or EA were in contact with cells, either simultaneously or after pretreatment with benomyl, did not protect hepatocytes. EGCG (1.3×10⁻² μg/ml) or EA (9.8×10⁻² μg, equivalent to (+)catechin/ml) inhibited 57% and 34%, respectively, the unscheduled DNA synthesis (UDS) induced by benomyl at a concentration of 23×10⁻² μ , when both were incubated with hepatocytes prior to benomyl. The simultaneous incubation of benomyl with EGCG or EA did not protect the cell against the genotoxic effect of benomyl. These results indicate that the dried leaves extract of Ardisia compressa protect rat hepatocytes from benomyl-induced cytotoxicity and genotoxicity.     

Investigation of extracts from (Tunisian) Cyperus rotundus as antimutagens and radical scavengers

This study evaluates mutagenic and antimutagenic effects of aqueous, total oligomers flavonoïds (TOF), ethyl acetate and methanol extracts from aerial parts of Cyperus rotundus with the Salmonella typhimurium assay system.

The different extracts showed no mutagenicity when tested withSalmonella typhimurium strains TA98, TA100, TA1535 and TA1538 either with or without the S9 mix. On the other hand, our results showed that all extracts have antimutagenic activity against Aflatoxin B1 (AFB1) in TA100 and TA98 assay system, and against sodium azide in TA100 and TA1535 assay system. TOF, ethyl acetate and methanol extracts exhibited the highest inhibition level of the Ames response induced by the indirect mutagen AFB1. Whereas, ethyl acetate and methanol extracts exhibited the highest level of protection towards the direct mutagen, sodium azide, induced response. In addition to antimutagenic activity, these extracts showed an important free radical scavenging activity towards the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. TOF, ethyl acetate and methanol extracts showed IC₅₀ value of 15, 14 and 20 μg/ml, respectively.

Taken together, our finding showed that C. rotundus exhibits significant antioxidant and antimutagenic activities.     

Knowledge mapping of Opuntia Milpa Alta since 1998: A scientometric analysis

建立超高效液相色谱串联四极杆静电场轨道阱质谱方法(UPLC-QE-Orbitrap-MS)对米邦塔仙人掌的叶状茎及果实化学成分进行表征,并对其抗氧化活性进行考察,初步探讨成分与活性的关系。

选择UPLC HSS T3 C₁₈ (2.1 mm×10 mm,1.8 μm)色谱柱,以乙腈-0.1%甲酸水为流动相,梯度洗脱,流速为0.2 mL·min^–1;采用加热电喷雾离子源,正、负离子模式分别进行一级、二级质谱数据采集;采用数据库和文献报道对米邦塔仙人掌叶状茎及果实的化学成分进行表征。采用DPPH、羟自由基(·OH)和超氧阴离子自由基(·${\\mathrm{O}}^{-}_{{2}} $)清除能力测试方法,对米邦塔仙人掌叶状茎及果实的抗氧化活性进行考察。

从米邦塔仙人掌的叶状茎及果实中共表征出39种化合物,其中酚类22个,黄酮类13个,其他成分4个。米邦塔仙人掌叶状茎及果实均具有一定的抗氧化能力,米邦塔仙人掌果实对DPPH的清除能力强于叶状茎,而叶状茎对·OH清除能力稍强于果实,叶状茎皮和果皮清除能力相当,果汁对·${\\mathrm{O}}^{-}_{{2}} $清除能力最强,米邦塔仙人掌的抗氧化活性与其富含的酚酸类成分密切相关。

本研究建立了一种快速、准确、可靠的米邦塔仙人掌茎果化学成分的鉴别方法,为米邦塔仙人掌资源的综合开发利用提供了科学依据。

     

Enantioselective LC analysis of synephrine in natural products on a protein-based chiral stationary phase

An enantioselective LC method with photodiode array detection (PAD) was developed for the enantioseparation of (±)-synephrine from C. aurantium L. var. amara fruits and phytotherapic derivatives by using a protein-based chiral stationary phase with cellobiohydrolase as the chiral selector (Chiral-CBH). Analyses were carried out on a Chiral-CBH column (100 × 4.0 mm i.d., 5 μm), with a mobile phase consisting of 2-propanol (5%, w/w) in sodium phosphate buffer (pH 6.0; 10 mM) and disodium EDTA (50 μM). The flow rate was 0.8 mL/min. Detection was set at 225 nm. To identify the order of elution, the racemate was resolved by the preparation of suitable diastereoisomeric salts with antipodes of appropriate organic acids.

Isolation of synephrine from C. aurantium fruits and phytoproducts was performed by solid-phase extraction (SPE) with a strong cation-exchange phase.

The method developed was validated and was found to be linear in the 0.40–40.14 μg/mL range (r² = 1.000, P < 0.0001) for both synephrine enantiomers. The limit of detection (LOD) for each enantiomer was 0.04 μg/mL. The limit of quantification (LOQ) for each enantiomer was 0.13 μg/mL. Intra-day precision (calculated as %R.S.D.) ranged from 0.03 to 0.24% for (−)-synephrine and from 0.03 to 0.35% for (+)-synephrine. Inter-day precision (calculated as %R.S.D.) ranged from 0.07 to 1.45% for (−)-synephrine and from 0.06 to 1.26% for (+)-synephrine. Intra- and inter-day accuracies (calculated as %recovery) were in the ranges of 97.4–100.6 and 98.0–101.6% for (−)-synephrine, and in the ranges 97.0–101.5 and 98.1–102.8% for (+)-synephrine.

The results of the application of the method to the analysis of C. aurantium samples showed that (−)-synephrine was the main component. (+)-Synephrine was not detected in C. aurantium fruits and was present in low concentration in the phytoproducts.     

Protective effects of dried flower extracts of Hibiscus sabdariffa L. against oxidative stress in rat primary hepatocytes

Dried flower extracts of Hibiscus sabdarrifa L., a local soft drink material and medical herb, was found to possess antioxidant activity in the present study. In the preliminary studies, antioxidant potential of three fractions of the ethanol crude extract (HS-C: chloroform-soluble fraction; HS-E: ethyl acetate soluble fraction; HS-R: residual fraction) obtained from the dried flowers of Hibiscus sabdarrifa L. were evaluated by their capacity of quenching 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and inhibiting xanthine oxidase (XO) activity. HS-E showed the greatest capacity of scavenging free radical (EC₅₀ = 0.017 mg/ml), and HS-C showed the strongest inhibitory effect on XO activity (EC₅₀ = 0.742 mg/ml). Furthermore, antioxidant bioactivities of these crude extracts were investigated using a model of tert-butyl hydroperoxide (t-BHP)-induced oxidative damage in rat primary hepatocytes. All fractions were found to inhibit significantly the unscheduled DNA synthesis (UDS) induced by t-BHP at a concentration of 0.20 mg/ml. HS-C and HS-E also decreased the leakage of lactate dehydrogenase (LDH) and the formation of malondialdehyde (MDA) induced by t-BHP (1.5 m ) considerably at a concentration of 0.10 and 0.20 mg/ml in the rat primary hepatocyte cultures. These results indicated that the dried flower extracts (HS-C and HS-E) of H. sabdarrifa L. protect rat hepatocytes from t-BHP-induced cytotoxicity and genotoxicity by different mechanisms.     

HPLC separation of naringin, neohesperidin and their C-2 epimers in commercial samples and herbal medicines

Flavanone glycosides, such as naringin and neohesperidin, are distributed in some Citrus species and have a chiral center in the C-2 position of the flavanone moiety. Naringin and neohesperidin (2S-form) were separated from the corresponding C-2 epimers (2R-epi-form) by normal-phase HPLC using a polysaccaride-derived chiral stationary phases (CSPs), CHIRALPAK^® IB. The analyses of commercial samples of naringin revealed that the relative ratios of naringin to the C-2 epimer were 29–89%. In the case of a commercial sample of neohesperidin, the relative ratio of the neohesperidin (2S-form) is 84%.

The HPLC application to Citrus species used as crude drugs in Japan (Kijitsu, Kikoku and Tohi) showed that the relative ratios of naringin to the C-2 epimer were 75–93% in Kijitsu, 74–79% in Kikoku and 54–64% in Tohi. However, there is a quite small ratio of the (2R)-epi-neohesperidin in Citrus. This result suggested that the averages of relative ratio of (2S)-naringin in Citrus species reduced according to the maturity of fruits (Kijitsu < Kikoku < Tohi). Since the relative ratios of (2S)-naringin of dry extracts of 5 Kampo formulations (including Kijitsu or Kikoku) decreased to 42–54%, the conversion from naringin to the (2R)-epimer might be enhanced during the decoction process of the formulations.     

Phytochemical contents and biological evaluation of Ruta chalepennsis L. growing in Saudi Arabia

Phytochemical screening of Ruta chalepensis L. exhibited the presence of different chemical groups. The dried aerial parts of the plant was total extracted by ethanol and successively using chloroform, ethyl acetate and Butanol, out of the successive extracts four compounds namely, scopletin, kaempferol, quercetin, quercetin 3-O-α-L-rhamno glucopyranosyl (Rutin) were isolated and biological evaluations. Total ethanol and successive extracts; chloroform, ethyl acetate and Butanol were produced excellent antimicrobial activities against gram negative bacteria, gram positive bacteria and fungi. Ethyl acetate extract was the best for inhibition of the microorganism’s growth. All extracts (total ethanol, and successive extracts) showed DPPH radical scavenging activity in a concentration–dependent manner. The best antioxidant activity was obtained by ethyl acetate & n-butanol extract (94.28%, IC₅₀?=?56.6?µg/ml). Also All extracts (total ethanol, and successive extracts) showed anticoagulant activity at higher concentration with prolonged clotting time 6:30 and 4:30?s at 10?mg/ml concentrations, respectively.     

Evernia prunastri and Pseudoevernia furfuraceae lichens and their major metabolites as antioxidant,antimicrobial and anticancer agents

The aim of this study is to investigate chemical composition of acetone extracts of the lichens Evernia prunastri and Pseudoevernia furfuraceae and in vitro antioxidant, antimicrobial, and anticancer activities of these extracts and some their major metabolites. HPLC–UV method was used for identification of secondary metabolites. Antioxidant activity was evaluated by free radical scavenging, superoxide anion radical scavenging, reducing power and determination of total phenolic compounds. As a result of the study physodic acid had largest antioxidant activities. Total content of phenol in extracts was determined as pyrocatechol equivalent. The antimicrobial activity was estimated by determination of the minimal inhibitory concentration by the broth microdilution method. The most active was also physodic acid. Anticancer activity was tested against FemX (human melanoma) and LS174 (human colon carcinoma) cell lines using MTT method.