Search results for: chromatography.

Authors: Anna Vildová, H. Hendrychová, J. Kubeš, L. Tůmová

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The abiotic elicitation is one of the methods for increasing the secondary metabolites production in plant tissue cultures and it seems to be more effective than traditional strategies. This study verified the use of silver nitrate as elicitor to enhance flavonolignans and flavonoid taxifolin production in suspension culture of Sylibum marianum (L.) Gaertn. Silver nitrate in various concentrations (5.887.10-3 mol/L, 5.887.10-4 mol/L, 5.887.10-5 mol/L) was used as elicitor. The content of secondary metabolites in cell suspension cultures was determined by high performance liquid chromatography. The samples were taken after 6, 12, 24, 48, 72 and 168 hours of treatment. The highest content of taxifolin production (2.2 mg.g-1) in cell suspension culture of Silybum marianum (L.) Gaertn. was detected after silver nitrate (5.887.10-4 mol/L) treatment and 72 h application. Flavonolignans such as silybinA, silybin B, silydianin, silychristin, isosilybin A, isosilybin B were not produced by cell suspension culture of S. marianum after elicitor treatment. Our results show that the secondarymetabolites could be released from S. marianum cells into the nutrient medium by changed permeability of cell wall.

Keywords: Silybum marianum (L.) Gaertn., elicitation, silver nitrate, taxifolin.

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Authors: R. Gaceb-Terrak, F. Rahmania

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Takerbucht is the only cultivar of date palm known as being resistant to the bayoud disease, caused by Fusarium oxysporum f. sp. albedinis (F.o.a.). In the aim to understand more about the defense mechanisms implied, we realized phytochemical analyses of this cultivar leaflets and roots and this, for the first time, using gas chromatography-mass spectrometry (GC-MS).The examination of our results shows that fifty-four molecules have been detected, fourteen of which are common to leaflets and roots. This study revealed also the organs' richness in derivatives fatty acids: both saturated and unsaturated are represented mainly by methyl esters of Hexadecanoic and 9,12,15-Octadecatrienoic acids. 1-Dodecanethiol, derivative Dodecanoic acid is only present in roots. It’s of great interest to note that the screening revealed the steroidal saponins abundance, among which Yamogenin acetate and Diosgenin, exclusively detected in Takerbucht. They may play an essential role, in the date palm resistance to the bayoud disease.

Keywords: Analysis by GC-MS, leaflets and roots of resistant date palm to F.o.a., derivatives fatty acids, steroidal saponins.

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Authors: Natarajan. E, Ganapathy Sundaram. E

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Fixed-bed slow pyrolysis experiments of rice husk have been conducted to determine the effect of pyrolysis temperature, heating rate, particle size and reactor length on the pyrolysis product yields. Pyrolysis experiments were performed at pyrolysis temperature between 400 and 600°C with a constant heating rate of 60°C/min and particle sizes of 0.60-1.18 mm. The optimum process conditions for maximum liquid yield from the rice husk pyrolysis in a fixed bed reactor were also identified. The highest liquid yield was obtained at a pyrolysis temperature of 500°C, particle size of 1.18-1.80 mm, with a heating rate of 60°C/min in a 300 mm length reactor. The obtained yield of, liquid, gas and solid were found be in the range of 22.57-31.78 %, 27.75-42.26 % and 34.17-42.52 % (all weight basics) respectively at different pyrolysis conditions. The results indicate that the effects of pyrolysis temperature and particle size on the pyrolysis yield are more significant than that of heating rate and reactor length. The functional groups and chemical compositions present in the liquid obtained at optimum conditions were identified by Fourier Transform-Infrared (FT-IR) spectroscopy and Gas Chromatography/ Mass Spectroscopy (GC/MS) analysis respectively.

Keywords: Slow pyrolysis, Rice husk, Recycling, Biomass.

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Authors: Feng Chia Hsieh, Sheng Kuo Hsieh, Tzyy Rong Jinn

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Characterization and evaluation of the activity of Vespa basalis DPP-IV, which expressed in Spodoptera frugiperda 21 cells. The expression of rDPP-IV was confirmed by SDS–PAGE, Western blot analyses, LC-MS/MS and measurement of its peptidase specificity. One-step purification by Ni-NTA affinity chromatography and the total amount of rDPP-IV recovered was approximately 6.4mg per liter from infected culture medium; an equivalent amount would be produced by 1x109 infected Sf21 insect cells. Through the affinity purification led to highly stable rDPP-IV enzyme was recovered and with significant peptidase activity. The rDPP-IV exhibited classical Michaelis–Menten kinetics, with kcat/Km in the range of 10-500 mM-1×S-1 for the five synthetic substrates and optimum substrate is Ala-Pro-pNA. As expected in inhibition assay, the enzymatic activity of rDPP-IV was significantly reduced by 80 or 60% in the presence of sitagliptin (a DPP-IV inhibitor) or PMSF (a serine protease inhibitor), but was not apparently affected by iodoacetamide (a cysteine protease inhibitor).

Keywords: Dipeptidyl-Peptidase IV, Phenylmethylsulfonyl fluoride; Serine protease, Sitagliptin, Vespa basalis

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Authors: Miroslava Šidlovská, Ida Petrovičová, Tomáš Pilka, Branislav Kolena

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Phthalates are ubiquitous environmental pollutants well known because of their endocrine disrupting activity in human organism. The aim of our study was, by biological monitoring, investigate exposure to phthalates of Roma ethnicity group i.e. children and adults from 5 families (n=29, average age 11.8 ± 7.6 years) living in western Slovakia. Additionally, we analysed some associations between anthropometric measures, questionnaire data i.e. socio-economic status, eating and drinking habits, practise of personal care products and household conditions in comparison with concentrations of phthalate metabolites. We used for analysis of urine samples high performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS) to determine concentrations of phthalate metabolites monoethyl phthalate (MEP), mono-n-butyl phthalate (MnBP), mono-iso-butyl phthalate (MiBP), mono(2-ethyl- 5-hydroxyhexyl) phthalate (5OH-MEHP), mono(2-ethyl-5-oxohexyl) phthalate (5oxo-MEHP) and mono(2-etylhexyl) phthalate (MEHP). Our results indicate that ethnicity, lower socioeconomic status and different housing conditions in Roma population can affect urinary concentration of phthalate metabolites.

Keywords: Biomonitoring, ethnicity, human exposure, phthalate metabolites.

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Authors: Ferial M. Abu-Salem, Marwa H. Mahmoud, M. H. El-Kalyoub, A. Y. Gibriel, Azza Abou-Arab

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In order to characterize the soy protein hydrolysate obtained in this study, gel chromatography on Sephadex G-25 was used to perform the separation of the peptide mixture and electrophoresis in SDS-polyacrylamide gel has been employed. Protein hydrolysate gave high antioxidant activities, but didn't give any antimicrobial activities. The antioxidant activities of protein hydrolysate was in the same trend of peptide content which gave high antioxidant activities and high peptide content between fractions 15 to 50. With increasing peptide concentrations, the scavenging effect on DPPH radical increased until about 70%, thereafter reaching a plateau. In compare to different concentrations of BHA, which exhibited higher activity (90%), soybean protein hydrolysate exhibited high antioxidant activities (70%) at a concentration of 1.45 mg/ml at fraction 25. Electrophoresis analysis indicated that, low- MW hydrolysate fractions (F1) appeared, on average, to have higher DPPH scavenging activities than high-MW fractions. These results revealed that soybean peptides probably contain substances that were proton donors and could react with free radicals to convert them to stable diamagnetic molecules. 

Keywords: Antioxidant peptides, hydrolysis, protein hydrolysate, peptide fractions.

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Authors: Said Nurdin, Fatimah A. Misebah, Rosli M. Yunus, Mohd S. Mahmud, Ahmad Z. Sulaiman

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The discarded clam shell waste, fossil and edible oil as biolubricant feedstocks create environmental impacts and food chain dilemma, thus this work aims to circumvent these issues by using activated saltwater clam shell waste (SCSW) as solid catalyst for conversion of Jatropha curcas oil as non-edible sources to ester biolubricant. The characterization of solid catalyst was done by Differential Thermal Analysis-Thermo Gravimetric Analysis (DTATGA), X-Ray Fluorescence (XRF), X-Ray Diffraction (XRD), Brunauer-Emmett-Teller (BET), Field Emission Scanning Electron Microscopy (FESEM) and Fourier Transformed Infrared Spectroscopy (FTIR) analysis. The calcined catalyst was used in the transesterification of Jatropha oil to methyl ester as the first step, and the second stage was involved the reaction of Jatropha methyl ester (JME) with trimethylolpropane (TMP) based on the various process parameters. The formated biolubricant was analyzed using the capillary column (DB-5HT) equipped Gas Chromatography (GC). The conversion results of Jatropha oil to ester biolubricant can be found nearly 96.66%, and the maximum distribution composition mainly contains 72.3% of triester (TE).

Keywords: Conversion, ester biolubricant, Jatropha curcas oil, solid catalyst.

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Authors: N. Zanganeh, M. Zabet

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Lactulose is a synthetic disaccharide which has remarkable applications in food and pharmaceutical fields. Lactulose is not found in nature and it is produced by isomerization reaction of lactose in an alkaline environment. It should be noted that this reaction has a very low yield since significant amount of lactose stays un-reacted in the system. Basically, purification of lactulose is difficult and costly. Previous studies have revealed that solubility of lactose and lactulose are significantly different in ethanol. Considering the fact that solubility is also affected by temperature itself, we investigated the effect of ethanol and temperature on separation process of lactose from the syrup containing lactose and lactulose. For this purpose, a saturated solution containing lactulose and lactose was made at three different temperatures; 25⁰C (room temperature), 31⁰C, and 37⁰C first.  Five samples containing 2g saturated solution was taken and then 2g, 3g, 4g, 5g, and 6g ethanol separately was added to the sampling tubes. Sampling tubes were kept at respective temperatures afterward. The concentration of lactose and lactulose after separation process measured and analyzed by High Performance Liquid Chromatography (HPLC). Results showed that ethanol has such a greater impact than operating temperature on purification process. Also, it was observed that the maximum rate of separation occurred at initial amount of added ethanol.

Keywords: Ethanol, lactose, lactulose syrup, purification.

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Authors: S. Zolghadri, H. Yousefnia, A. R. Jalilian

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Bone metastases are observed in a wide range of cancers leading to intolerable pain. While early detection can help the physicians in the decision of the type of treatment, various radiopharmaceuticals using phosphonates like ⁶⁸Ga-EDTMP have been developed. In this work, due to the importance of absorbed dose, human absorbed dose of this new agent was calculated for the first time based on biodistribution data in Wild-type rats. ⁶⁸Ga was obtained from ⁶⁸Ge/⁶⁸Ga generator with radionuclidic purity and radiochemical purity of higher than 99%. The radiolabeled complex was prepared in the optimized conditions. Radiochemical purity of the radiolabeled complex was checked by instant thin layer chromatography (ITLC) method using Whatman No. 2 paper and saline. The results indicated the radiochemical purity of higher than 99%. The radiolabelled complex was injected into the Wild-type rats and its biodistribution was studied up to 120 min. As expected, major accumulation was observed in the bone. Absorbed dose of each human organ was calculated based on biodistribution in the rats using RADAR method. Bone surface and bone marrow with 0.112 and 0.053 mSv/MBq, respectively, received the highest absorbed dose. According to these results, the radiolabeled complex is a suitable and safe option for PET bone imaging.

Keywords: Absorbed dose, EDTMP, 68Ga, rats.

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Authors: Esther T. Akinlabi, Milan Isvarial, Stephen A. Akinlabi

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Today, canines are still used effectively in acceleration detection situation. However, this method is becoming impractical in modern age and a new automated replacement to the canine is required. This paper reports the design of an innovative accelerant detector. Designing an accelerant detector is a long process as is any design process; therefore, a solution to the need for a mobile, effective accelerant detector is hereby presented. The device is simple and efficient to ensure that any accelerant detection can be conducted quickly and easily. The design utilizes Ultra Violet (UV) light to detect the accelerant. When the UV light shines on an accelerant, the hydrocarbons in the accelerant emit florescence. The advantages of using the UV light to detect accelerant are also outlined in this paper. The mobility of the device is achieved by using a Direct Current (DC) motor to run tank tracks. Tank tracks were chosen as to ensure that the device will be mobile in the rough terrain of a fire site. The materials selected for the various parts are also presented. A Solid Works Simulation was also conducted on the stresses in the shafts and the results are presented. This design is an innovative solution which offers a user friendly interface. The design is also environmentally friendly, ecologically sound and safe to use.

Keywords: Accelerant detector, Canines, Gas Chromatography- Mass Spectrometry (GC-MS), Ultra Violet light.

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Authors: B. Kabane, G. G. Redhi

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An ammonium based ionic liquid (methyltrioctylammonium chloride) [N_{8 8 8 1}] [Cl] was investigated as an extraction potential solvent for volatile organic solvents (in this regard, solutes), which includes alkenes, alkanes, ketones, alkynes, aromatic hydrocarbons, tetrahydrofuran (THF), alcohols, thiophene, water and acetonitrile based on the experimental activity coefficients at infinite THF measurements were conducted by the use of gas-liquid chromatography at four different temperatures (313.15 to 343.15) K. Experimental data of activity coefficients obtained across the examined temperatures were used in order to calculate the physicochemical properties at infinite dilution such as partial molar excess enthalpy, Gibbs free energy and entropy term. Capacity and selectivity data for selected petrochemical extraction problems (heptane/thiophene, heptane/benzene, cyclohaxane/cyclohexene, hexane/toluene, hexane/hexene) were computed from activity coefficients data and compared to the literature values with other ionic liquids. Evaluation of activity coefficients at infinite dilution expands the knowledge and provides a good understanding related to the interactions between the ionic liquid and the investigated compounds.

Keywords: Separation, activity coefficients, ionic liquid, methyltrioctylammonium chloride, capacity.

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Authors: P. Martínez-Nieto, M. Vanegas-Hoyos, M. Zapata-Pineda, J. Robles-Camargo

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The aquatic plants are a promising renewable energy resource. Lake Fúquene polluting macrophytes, water hyacinth (Eichhornia crassipes C. Mart.) and Brazilian elodea (Egeria densa Planch.), were saccharifiedby different treatments and fermented to ethanol by native yeasts. Among the tested chemical and biological methods for the saccharification, Pleurotus ostreatus at 10% (m/v) was chosen as the best pre-treatment in both macrophytes (P<0.01). Subsequently 49 yeasts were isolated from Lake Fúquene and nine strains were selected, which presented the highest precipitates characteristic of ethanol in the iodoform test. The fermentations from water hyacinth and Brazilian elodea hydrolysates using these yeasts produced ethanol at a rate between 0.38 to 0.80gL-1h-1 and 0.15 to 0.27gL-1h-1 respectively. The ethanol presence was confirmed by gas chromatography–mass spectrometry. The nine yeasts chosen were preliminarily identified as belonging to the genera Candida spp., Brettanomyces sp. and Hansenula spp.

Keywords: Bio-ethanol, Chemical hydrolysis, Invasive aquatic macrophytes, Native yeasts fermenting, P. ostreatus

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Authors: S.S. Abdullah, S. Yusup, M.M. Ahmad, A. Ramli, L. Ismail

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This paper aims to study decomposition behavior in pyrolytic environment of four lignocellulosic biomass (oil palm shell, oil palm frond, rice husk and paddy straw), and two commercial components of biomass (pure cellulose and lignin), performed in a thermogravimetry analyzer (TGA). The unit which consists of a microbalance and a furnace flowed with 100 cc (STP) min-1 Nitrogen, N2 as inert. Heating rate was set at 20⁰C min-1 and temperature started from 50 to 900⁰C. Hydrogen gas production during the pyrolysis was observed using Agilent Gas Chromatography Analyzer 7890A. Oil palm shell, oil palm frond, paddy straw and rice husk were found to be reactive enough in a pyrolytic environment of up to 900°C since pyrolysis of these biomass starts at temperature as low as 200°C and maximum value of weight loss is achieved at about 500°C. Since there was not much different in the cellulose, hemicelluloses and lignin fractions between oil palm shell, oil palm frond, paddy straw and rice husk, the T-50 and R-50 values obtained are almost similar. H2 productions started rapidly at this temperature as well due to the decompositions of biomass inside the TGA. Biomass with more lignin content such as oil palm shell was found to have longer duration of H2 production compared to materials of high cellulose and hemicelluloses contents.

Keywords: biomass, decomposition, hydrogen, lignocellulosic, thermogravimetry

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Authors: Mahmood Ahmad, Ghulam Murtaza, Sonia Khiljee, Muhammad Asadullah Madni

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A new reverse phase-high performance liquid chromatography (RP-HPLC) method with fluorescent detector (FLD) was developed and optimized for Norfloxacin determination in human plasma. Mobile phase specifications, extraction method and excitation and emission wavelengths were varied for optimization. HPLC system contained a reverse phase C18 (5 μm, 4.6 mm×150 mm) column with FLD operated at excitation 330 nm and emission 440 nm. The optimized mobile phase consisted of 14% acetonitrile in buffer solution. The aqueous phase was prepared by mixing 2g of citric acid, 2g sodium acetate and 1 ml of triethylamine in 1 L of Milli-Q water was run at a flow rate of 1.2 mL/min. The standard curve was linear for the range tested (0.156–20 μg/mL) and the coefficient of determination was 0.9978. Aceclofenac sodium was used as internal standard. A detection limit of 0.078 μg/mL was achieved. Run time was set at 10 minutes because retention time of norfloxacin was 0.99 min. which shows the rapidness of this method of analysis. The present assay showed good accuracy, precision and sensitivity for Norfloxacin determination in human plasma with a new internal standard and can be applied pharmacokinetic evaluation of Norfloxacin tablets after oral administration in human.

Keywords: Norfloxacin, Aceclofenac sodium, Methodoptimization, RP-HPLC method, Fluorescent detection, Calibrationcurve.

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Authors: Myagmarsuren Soronzonjav, N. Togtokhbayar, L. Davaahuu, B. Minjigdorj, Seong Gu Hwang

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The number of natural or organic product consumers is increased in recent years and this healthy demand pushes to increase usage of healthy meat. At the same time, consumers pay more attention on the healthy fat, especially on unsaturated fatty acids. These long chain carbohydrates reduce heart diseases, improve memory and eye sight and activate the immune system. One of the important issues to be solved for our Mongolia’s food security is to provide healthy, fresh, widely available and cheap meat for the population. Thus, an importance of the Selenge breed meat production is increasing in order to supply the quality meat food security since the Selenge breed cattle are rapidly multiplied, beneficial in term of income, the same quality as Mongolian breed, and well digested for human body. We researched the lipid, unsaturated and saturated fatty acid contents of meat of Selenge breed younger cattle by their muscle types. Result of our research reveals that 11 saturated fatty acids are detected. For the content of palmitic acid among saturated fatty acids, 23.61% was in the sirloin meat, 24.01% was in the round and chuck meat, and 24.83% was in the short loin meat.

Keywords: Chromatogram, gas chromatography, organic resolving, saturated and unsaturated fatty acids.

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Authors: S. Techaoei, K. Jarmkom, P. Eakwaropas, W. Khobjai

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The objective of this research was focused on investigating in vitro antimicrobial activity of Phellinus linteus fruiting body extracts on Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Phellinus linteus fruiting body was extracted with ethanol and ethyl acetate and was vaporized. The disc diffusion assay was used to assess antimicrobial activity against tested bacterial strains. Primary screening of chemical profile of crude extract was determined by using thin layer chromatography. The positive control and the negative control were used as erythromycin and dimethyl sulfoxide, respectively. Initial screening of Phellinus linteus crude extract with the disc diffusion assay demonstrated that only ethanol had greater antimicrobial activity against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. The MIC assay showed that the lower MIC was observed with 0.5 mg/ml of Pseudomonas aeruginosa and Methicillin-resistant Staphylococcus aureus and 0.25 mg/ml. of Escherichia coli and Staphylococcus aureus, respectively. TLC chemical profile of extract was represented at R_f ≈ 0.71-0.76.

Keywords: Staphylococcus aureus, Phellinus linteus, methicillin-resistant Staphylococcus aureus, antimicrobial activity, Escherichia coli.

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Authors: A. Mandić, M. Sakač, A. Mišan, B. Šojić, L. Petrović, I. Lončarević, B. Pajin, I. Sedej

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Aldehydes as secondary lipid oxidation products are highly specific to the oxidative degradation of particular polyunsaturated fatty acids present in foods. Gas chromatographic analysis of those volatile compounds has been widely used for monitoring of the deterioration of food products. Developed static headspace gas chromatography method using flame ionization detector (SHS GC FID) was applied to monitor the aldehydes present in processed foods such as bakery, meat and confectionary products.

Five selected aldehydes were determined in samples without any sample preparation, except grinding for bakery and meat products. SHS–GC analysis allows the separation of propanal, pentanal, hexanal, heptanal and octanal, within 15min. Aldehydes were quantified in fresh and stored samples, and the obtained range of aldehydes in crackers was 1.62±0.05 – 9.95±0.05mg/kg, in sausages 6.62±0.46 – 39.16±0.39mg/kg; and in cocoa spread cream 0.48±0.01 – 1.13±0.02mg/kg. Referring to the obtained results, the following can be concluded, proposed method is suitable for different types of samples, content of aldehydes varies depending on the type of a sample, and differs in fresh and stored samples of the same type.

Keywords: Lipid oxidation, aldehydes, crackers, sausage, cocoa cream spread.

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Authors: Tanwarat Kajsongkram, Saowalux Rotamporn, Sirinat Limbunruang, Sirinan Thubthimthed

Abstract:

High Performance Liquid Chromatography (HPLC) method was developed and validated for simultaneous estimation of 6-Gingerol(6G) and 6-Shogaol(6S) in joint pain relief gel containing ginger extract. The chromatographic separation was achieved by using C18 column, 150 x 4.6mm i.d., 5μ Luna, mobile phase containing acetonitrile and water (gradient elution). The flow rate was 1.0 ml/min and the absorbance was monitored at 282 nm. The proposed method was validated in terms of the analytical parameters such as specificity, accuracy, precision, linearity, range, limit of detection (LOD), limit of quantification (LOQ), and determined based on the International Conference on Harmonization (ICH) guidelines. The linearity ranges of 6G and 6S were obtained over 20- 60 and 6-18 μg/ml respectively. Good linearity was observed over the above-mentioned range with linear regression equation Y= 11016x- 23778 for 6G and Y = 19276x-19604 for 6S (x is concentration of analytes in μg/ml and Y is peak area). The value of correlation coefficient was found to be 0.9994 for both markers. The limit of detection (LOD) and limit of quantification (LOQ) for 6G were 0.8567 and 2.8555 μg/ml and for 6S were 0.3672 and 1.2238 μg/ml respectively. The recovery range for 6G and 6S were found to be 91.57 to 102.36 % and 84.73 to 92.85 % for all three spiked levels. The RSD values from repeated extractions for 6G and 6S were 3.43 and 3.09% respectively. The validation of developed method on precision, accuracy, specificity, linearity, and range were also performed with well-accepted results.

Keywords: Ginger, 6-gingerol, HPLC, 6-shogaol.

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Authors: Bahdja Guerfi, N. Hadhoum, I. Azouz, M. Bendoumia, S. Bouafia, F. Z. Hadjadj Aoul

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Background: 'Penicillin V' is a narrow, bactericidal antibiotic of the beta-lactam family of the naturally occurring penicillin group. It is limited to infections due to the germs defined as sensitive. The objective of this work was to identify and to characterize Penicillin V acid and its related compounds by High-performance liquid chromatography (HPLC). Methods: Firstly phenoxymethylpenicillin was identified by an infrared absorption. The organoleptic characteristics, pH, and determination of water content were also studied. The dosage of Penicillin V acid active substance and the determination of its related compounds were carried on waters HPLC, equipped with a UV detector at 254 nm and Discovery HS C18 column (250 mm X 4.6 mm X 5 µm) which is maintained at room temperature. The flow rate was about 1 ml per min. A mixture of water, acetonitrile and acetic acid (65:35:01) was used as mobile phase for phenoxyacetic acid ‘impurity B' and a mixture of water, acetonitrile and acetic acid (650:150:5.75) for the assay and 4-hydroxypenicillin V 'impurity D'. Results: The identification of Penicillin V acid active substance and the evaluation of its chemical quality showed conformity with USP 35th edition. The Penicillin V acid content in the raw material is equal to 1692.22 UI/mg. The percentage content of phenoxyacetic acid and 4-hydroxypenicillin V was respectively: 0.035% and 0.323%. Conclusion: Through these results, we can conclude that the Penicillin V acid active substance tested is of good physicochemical quality.

Keywords: Penicillin V acid, characterization, related substances, HPLC.

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Authors: Isam A. Mohamed Ahmed, Jiro Arima, Tsuyoshi Ichiyanagi, Emi Sakuno, Nobuhiro Mori

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The aim of this study was to screen for microorganism that able to utilize 3-N-trimethylamino-1-propanol (homocholine) as a sole source of carbon and nitrogen. The aerobic degradation of homocholine has been found by a gram-positive Rhodococcus sp. bacterium isolated from soil. The isolate was identified as Rhodococcus sp. strain A4 based on the phenotypic features, physiologic and biochemical characteristics, and phylogenetic analysis. The cells of the isolated strain grown on both basal-TMAP and nutrient agar medium displayed elementary branching mycelia fragmented into irregular rod and coccoid elements. Comparative 16S rDNA sequencing studies indicated that the strain A4 falls into the Rhodococcus erythropolis subclade and forms a monophyletic group with the type-strains of R. opacus, and R. wratislaviensis. Metabolites analysis by capillary electrophoresis, fast atom bombardment-mass spectrometry, and gas chromatography- mass spectrometry, showed trimethylamine (TMA) as the major metabolite beside β-alanine betaine and trimethylaminopropionaldehyde. Therefore, the possible degradation pathway of trimethylamino propanol in the isolated strain is through consequence oxidation of alcohol group (-OH) to aldehyde (-CHO) and acid (-COOH), and thereafter the cleavage of β-alanine betaine C-N bonds yielded trimethylamine and alkyl chain.

Keywords: Homocholine, 3-N-trimethylamino-1-propanol, Quaternary ammonium compounds, 16S rDNA gene sequence.

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Authors: M. Ramezani, S. Ghaderifard, HR. Monsef-Esfahani, S. Nasri

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The aim of this study is evaluating the antinociceptive and anti-inflamatory activity of Geum kokanicum. After determination total extract LD50, different doses of extract were chosen for intrapritoneal injections. In inflammation test, male NMRI mice were divided into 6 groups: control (normal saline), positive control (Dexamethasone 15mg/kg), and total extract (0.025, 0.05, 0.1, and 0.2 gr/kg). The inflammation was produced by xyleneinduced edema. In order to evaluate the antinociceptive effect of total extract, formalin test was used. Mice were divided into 6 groups: control, positive control (morphine 10mg/kg), and 4 groups which received total extract. Then they received Formalin. The animals were observed for the reaction to pain. Data were analyzed using One-way ANOVA followed by Tukey-Kramer multiple comparison test. LD50 was 1 gr/kg. Data indicated that 0.5,0.1 and 0.2 gr/kg doses of total extract have particular antinociceptive and antiinflammatory effects in a comparison with control (P<0.001). The most effective dose was 0.2 gr/kg which did not show any significant difference in a comparison with positive control. Results indicated that total extract can inhibit nociception in the first and second phase. The antinociceptive effects in high doses are the same as morphine as a strong analgesic substance. TLC chromatography indicated presence of steroids and triterpenoids in this plant. The effects of extract may be related to presence of these compounds.

Keywords: Anti-inflammatory, Antinociceptive, Geum kokanicum, Mice.

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Authors: Dalia. G. Aseel

Abstract:

Begomoviruses are economically important plant viruses that infect dicotyledonous plants and exclusively transmitted by the whitefly Bemisia tabaci. Here, replicative form was isolated from Okra, Cotton, Tomato plants and whitefly infected with Begomoviruses. Using coat protein specific primers (AV1), the viral infection was verified with amplicon at 450 bp. The sequence of OLCuV-AV1 gene was recorded and received an accession number (FJ441605) from Genebank. The phylogenetic tree of OLCuV was closely related to Okra leaf curl virus previously isolated from Cameroon and USA with nucleotide sequence identity of 92%. The protein purification was carried out using His-Tag methodology by using Affinity Chromatography. The purified protein was separated on SDS-PAGE analysis and an enriched expected size of band at 30 kDa was observed. Furthermore, RAPD and SDS-PAGE were used to detect genetic variability between different hosts of okra leaf curl virus (OLCuV), cotton leaf curl virus (CLCuV), tomato yellow leaf curl virus (TYLCuV) and the whitefly vector. Finally, the present study would help to understand the relationship between the whitefly and different economical crops in Egypt.

Keywords: Begomovirus, AV1 gene, sequence, cloning, whitefly, okra, cotton, tomato, RAPD, phylogenetic tree and SDS-PAGE.

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Authors: D. Pujimulyani, S. Raharjo, Y. Marsono, U. Santoso

Abstract:

Background and objectives: Most of the agricultural products are processed by blanching. Blanching can increase antioxidant activity in white saffron products. The objective of this research were to determine antioxidant activity, to identify, and to measure changes in phenolic substances of fresh and blanched white saffron rhizomes (Curcuma mangga Val.). Methods: White saffron rhizomes were peeled, washed and blanched in boiling water containing 0% or 0.05% citric acid solution for 5 and 10 minutes. Samples were extracted using methanol, rotaevaporated, and freezedried. Dried extract was determined antioxidant activity by DPPH method, identified and quantified for the phenolic substances by High Performance Liquid Chromatography (HPLC) equipped with coloumn C₁₈ and Photodiode-array detector (PAD). Result: This research showed that the quantity of the 6 phenolic substances identified in blanched white saffron in citric acid solution increased significantly compared to that of the non-blanched. Blanching white saffron in 0.05% citric acid media for 5 minutes increased its antioxidant activity, and total phenolic content. Conclusions: The identified phenolic substances of white saffron were Gallic Acid (GA), Catechin (C), Epicatechin (EC), Epigallocatechin (EGC), Epigallocatechingallat (EGCG) and Gallocatechingallat (GCG). The blanched white saffron contained C and EGCG significantly higher than that of fresh rhizomes.

Keywords: White saffron, antioxidant activity, blanching, phenolic.

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Authors: Sarkar M. A. Kawsar, Sarkar M. A. Mamun, Md S. Rahman, Hidetaro Yasumitsu, Yasuhiro Ozeki

Abstract:

Lectins have a good scope in current clinical microbiology research. In the present study evaluated the antimicrobial activities of a D-galactose binding lectin (PnL) was purified from the annelid, Perinereis nuntia (polychaeta) by affinity chromatography. The molecular mass of the lectin was determined to be 32 kDa as a single polypeptide by SDS-PAGE under both reducing and non-reducing conditions. The hemagglutinating activity of the PnL showed against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-Gal, GalNAc, Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro antibacterial screening studies against 11 gram-positive and gram-negative microorganisms. From the screening results, it was revealed that PnL exhibited significant antibacterial activity against gram-positive bacteria. Bacillus megaterium showed the highest growth inhibition by the lectin (250 μg/disc). However, PnL did not inhibit the growth of gram-negative bacteria such as Vibrio cholerae and Pseudomonas sp. PnL was also examined for in vitro antifungal activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited the mycelial growth of Alternaria alternata (24.4%). These results indicate that future findings of lectin applications obtained from annelids may be of importance to life sciences.

Keywords: Perinereis nuntia, Lectin, Inhibition zone, Mycelial growth.

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Authors: Z. Honarvar, M. Farhoodi, M. R. Khani, S. Shojaee-Aliabadi

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The purpose of the present study was to evaluate carboxymethyl cellulose (CMC) coated polypropylene (PP) films containing Zataria multiflora (ZEO) essential oils (4%) as an antimicrobial packaging for chicken breast stored at 4 °C. To increase PP film hydrophilicity, it was treated by atmospheric cold plasma prior to coating by CMC. Then, different films including PP, PP/CMC, PP/CMC containing 4% of ZEO were used for the chicken meat packaging in vapor phase. Total viable count and pseudomonads population and oxidative (TBA) changes of the chicken breast were analyzed during shelf life. Results showed that the shelf life of chicken meat kept in films containing ZEO improved from three to nine days compared to the control sample without any direct contact with the film. Study of oxygen barrier properties of bilayer film without essential oils (0.096 cm³ μm/m² d kPa) in comparison with PP film (416 cm³ μm/m² d kPa) shows that coating of PP with CMC significantly reduces oxygen permeation of the obtained packaging (P<0.05), which reduced aerobic bacteria growth. Chemical composition of ZEO was also evaluated by gas chromatography–mass spectrometry (GC–MS), and this shows that thymol was the main antimicrobial and antioxidant component of the essential oil. The results revealed that PP/CMC containing ZEO has good potential for application as active food packaging in indirect contact which would also improve sensory properties of product.

Keywords: Shelf life, chicken breast, polypropylene, carboxymethyl cellulose, essential oil.

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Authors: Sarkar M. A. Kawsar, Sarkar M. A. Mamun, Md S. Rahman, Hidetaro Yasumitsu, Yasuhiro Ozeki

Abstract:

The present study has been taken to explore the screening of in vitro antimicrobial activities of D-galactose-binding sponge lectin (HOL-30). HOL-30 was purified from the marine demosponge Halichondria okadai by affinity chromatography. The molecular mass of the lectin was determined to be 30 kDa with a single polypeptide by SDS-PAGE under non-reducing and reducing conditions. HOL-30 agglutinated trypsinized and glutaraldehydefixed rabbit and human erythrocytes with preference for type O erythrocytes. The lectin was subjected to evaluation for inhibition of microbial growth by the disc diffusion method against eleven human pathogenic gram-positive and gram-negative bacteria. The lectin exhibited strong antibacterial activity against gram-positive bacteria, such as Bacillus megaterium and Bacillus subtilis. However, it did not affect against gram-negative bacteria such as Salmonella typhi and Escherichia coli. The largest zone of inhibition was recorded of Bacillus megaterium (12 in diameter) and Bacillus subtilis (10 mm in diameter) at a concentration of the lectin (250 μg/disc). On the other hand, the antifungal activity of the lectin was investigated against six phytopathogenic fungi based on food poisoning technique. The lectin has shown maximum inhibition (22.83%) of mycelial growth of Botrydiplodia theobromae at a concentration of 100 μg/mL media. These findings indicate that the lectin may be of importance to clinical microbiology and have therapeutic applications.

Keywords: Antibacterial, Halichondria okadai, Inhibition zone, Lectin.

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Authors: Zs. Csanádi, A. Szabó Nagy, J. Szabó, J. Erdős

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Atmospheric aerosols have several important environmental impacts and health effects in point of air quality. Monitoring the PM10-bound polycyclic aromatic hydrocarbons (PAHs) could have important environmental significance and health protection aspects. Benzo(a)pyrene (BaP) is the most relevant indicator of these PAH compounds. In Hungary, the Hungarian Air Quality Network provides air quality monitoring data for several air pollutants including BaP, but these data show only the annual mean concentrations and maximum values. Seasonal variation of BaP concentrations comparing the heating and non-heating periods could have important role and difference as well. For this reason, the main objective of this study was to assess the annual concentration and seasonal variation of BaP associated with PM10 in the ambient air of Northwestern Hungary seven different sampling sites (six urban and one rural) in the sampling period of 2008–2013. A total of 1475 PM10 aerosol samples were collected in the different sampling sites and analyzed for BaP by gas chromatography method. The BaP concentrations ranged from undetected to 8 ng/m3 with the mean value range of 0.50-0.96 ng/m3 referring to all sampling sites. Relatively higher concentrations of BaP were detected in samples collected in each sampling site in the heating seasons compared with non-heating periods. The annual mean BaP concentrations were comparable with the published data of the other Hungarian sites.

Keywords: Air quality, benzo(a)pyrene, PAHs, polycyclic aromatic hydrocarbons.

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Authors: Eva Tvrdá, Barbora Babečková, Michal Ďuračka, Róbert Kirchner, Július Árvay

Abstract:

This study was designed to assess the in vitro effects of Origanum vulgare L. (oregano) extract on the testicular steroidogenesis. We focused on identifying major biomolecules present in the oregano extract, as well as to investigate its in vitro impact on the secretion of cholesterol, testosterone, dehydroepiandrosterone and androstenedione by murine testicular fragments. The extract was subjected to high performance liquid chromatography (HPLC) which identified cyranosid, daidzein, thymol, rosmarinic and trans-caffeic acid among the predominant biochemical components of oregano. For the in vitro experiments, testicular fragments from 20 sexually mature Institute of Cancer Research (ICR) mice were incubated in the absence (control group) or presence of the oregano extract at selected concentrations (10, 100 and 1000 μg/mL) for 24 h. Cholesterol levels were quantified using photometry and the hormones were assessed by ELISA (Enzyme-Linked Immunosorbent Assay). Our data revealed that the release of cholesterol and androstenedione (but not dehydroepiandrosterone and testosterone) by the testicular fragments was significantly impacted by the oregano extract in a dose-dependent fashion. Supplementation of the extract resulted in a significant decline of cholesterol (P < 0.05 in case of 100 μg/mL; P < 0.01 with respect 100 μg/mL extract), as well as androstenedione (P < 0.01 with respect to 100 and 1000 μg/mL extract). Our results suggest that the biomolecules present in Origanum vulgare L. could exhibit a dose-dependent impact on the secretion of male steroids, playing a role in the regulation of testicular steroidogenesis.

Keywords: Mice, Origanum vulgare L., steroidogenesis, testes.

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Authors: O. B. Citil, V. Sariyel, M. Akoz

Abstract:

In this study, total fatty acid composition of muscle lipids of Cyprinus carpio L. living in Suğla Dam Lake, Altinapa Dam Lake, Eğirdir Lake and Burdur Lake were determined using GC. During this study, for the summer season of July was taken from each region of the land and they were stored in deep-freeze set to -20 degrees until the analysis date. At the end of the analyses, 30 different fatty acids were found in the composition of Cyprinus carpio L. which lives in different lakes. Cyprinus carpio Suğla Dam Lake of polyunsaturated fatty acids (PUFAs), were higher than other lakes. Cyprinus carpio L. was the highest in the major SFA palmitic acid. Polyunsaturated fatty acids (PUFA) of carp, the most abundant fish species in all lakes, were found to be higher than those of saturated fatty acids (SFA) in all lakes. Palmitic acid was the major SFA in all lakes. Oleic acid was identified as the major MUFA. Docosahexaenoic acid (DHA) was the most abundant in all lakes. ω3 fatty acid composition was higher than the percentage of the percentage ω6 fatty acids in all lake. ω3/ω6 rates of Cyprinus carpio L. Suğla Dam Lake, Altinapa Dam Lake, Eğirdir Lake and Burdur Lake, 2.12, 1.19, 2.15, 2.87, and 2.82, respectively. Docosahexaenoic acid (DHA) was the major PUFA in Eğirdir and Burdur lakes, whereas linoleic acid (LA) was the major PUFA in Altinapa and Suğla Dam Lakes. It was shown that the fatty acid composition in the muscle of carp was significantly influenced by different lakes.

Keywords: Chromatography, Cyprinus carpio L., fatty acid composition.

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Authors: Zakaria Boual, Abdellah Kemassi, Toufik Chouana, Philippe Michaud, Mohammed Didi Ould El Hadj

Abstract:

In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% is neutral sugar and the rest 16.25±1.62% is uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructooligosaccharide (RP95). The oligosaccharides concentration was 1g/L of Man, Rogosa, Sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

Keywords: Zizyphus lotus, polysaccharides, characterization, prebiotic effects.

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