Search results for: thin-layer chromatography (TLC)

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: Farzaneh Ziaee, Mohammad Ziaee

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N-acetyl-L-cysteine (NAC) is a well-known mucolytic agent, and recently its efficacy has been examined for the prevention and remediation of several diseases such as lung infections caused by Coronavirus. Also, it is administrated as the main antidote in paracetamol overdose and is effective for the treatment of idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD). This medicine is used as an antioxidant to prevent diabetic kidney disease (nephropathy). In this study, a method for the acylation of amino acids is employed to manufacture this drug in a height yield. Regarding this patented path, NAC can be made in a single batch step at ambient pressure and temperature. Moreover, this study offers a technique to make peptide bonds which is of interest for pharmaceutical and medicinal industries. The separation process was undertaken using appropriate solvents to achieve an excellent purification level. The synthesized drug was characterized via proton nuclear magnetic resonance (1H NMR), high-performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FT-IR), elemental analysis, and melting point.

Keywords: N-acetylcysteine, synthesis, mucolytic medication, lung anti-inflammatory, COVID-19, antioxidant, pharmaceutical supplement, characterization

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Authors: Hanane Belayachi, Fadila Nemchi, Amel Belayachi, Sarra Bourahla, Mostefa Belhakem

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In this work, two photocatalytic processes for the degradation of phenol in water are presented. The first one is extensive (EP), which is carried out in a treatment chain of two steps, allowing the adsorption of the pollutant by a naturally activated carbon from the grapes. This operation is followed by a photocatalytic degradation of the residual phenol in the presence of TiO₂. The second process is intensive (IP) and is realized in one step in the presence of a hybrid photocatalytic nanomaterial prepared from naturally activated carbon and TiO₂. The evaluation of the two processes, EP and IP, is based on the analytical monitoring of the initial and final parameters of the water to be treated, i.e., the phenol concentration by liquid phase chromatography (HPLC) and total organic carbon (TOC). For both processes, the sampling was carried out every 10 min for 120 min of treatment time to measure the phenol concentrations. The elimination and degradation rates in the case of the intensive process are better than the extensive process. In both processes, the catechol molecule was detected as an under product of degradation. In the IP case, this intermediate phenol was totally eliminated, and only traces of catechol persisted in the water.

Keywords: photocatalysis, hybrid, activated carbon, phenol

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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 (DTA-TGA), 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, Jatropha curcas oil, ester biolubricant, solid catalyst

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Authors: Mohsina Akhter

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Cancer has become a wide spread disease around the globe and takes many lives every year. Different treatments are being practiced but all have potential side effects with somewhat less specificity towards target sites. Pseudomonas aeruginosa is known to secrete a protein azurin with special anti-cancer function. It has unique cell penetrating peptide comprising of 18 amino acids that have ability to enter cancer cells specifically. Reported function of Azurin is to stabilize p53 inside the tumor cells and induces apoptosis through Bax mediated cytochrome c release from mitochondria. At laboratory scale, we have made recombinant azurin through cloning rpTZ57R/T-azu vector into E.coli strain DH-5α and subcloning rpET28-azu vector into E.coli BL21-CodonPlus (DE3). High expression was ensured with IPTG induction at different concentrations then optimized high expression level at 1mM concentration of IPTG for 5 hours. Purification has been done by using Ni+2 affinity chromatography. We have concluded that azurin can be a remarkable improvement in cancer therapeutics if it produces on a large scale. Azurin does not enter into the normal cells so it will prove a safe and secure treatment for patients and prevent them from hazardous anomalies.

Keywords: azurin, pseudomonas aeruginosa, cancer, therapeutics

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Authors: Maryam Hashemi, Nematollah Razmi, Rasool Madani

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M. paratuberculosis is a slow growing mycobactin dependent mycobacterial species known to be the causative agent of Johne’s disease in all species of domestic ruminants worldwide. JD is characterized by gradual weight loss; decreased milk production. Excretion of the organism may occur for prolonged periods (1 to 2.5 years) before the onset of clinical disease. In recent years, researchers focus on identification a specific antigen of MAP to use in diagnosis test and preparation of effective vaccine. In this paper, for production of polyclonal antibody against proteins of Mycobacterium avium paratuberculosis cell wall a rabbit immunization at a certain time period with antigen. After immunization of the animal, blood samples were collected from the rabbit for producing enriched serum. Antibodies were purified with ion exchange chromatography. For exact measurement of interaction, western blotting test was used and as it is demonstrated in the study, sharp bands appear in nitrocellulose paper and specific bands were 50 and 150 KD molecular weight. These were indicating immunodominant proteins.

Keywords: immunodominant, paratuberculosis, Western blotting, cell wall proteins, protein purification

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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: lactulose, lactose, purification, solubility

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Authors: Devarakonda Ramadevi, Dasari Rambabu, K. Suresh Babu, Battu Ganga Rao, Lakshmi Sirisha Kotikalapudi

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The phytochemical and biological evaluation of the whole plant of Derris scandens is belonging to the family fabaceae. The dried plant of D.scandens was procured from the tirumala. The completely dried powder of the whole plant was taken and ground to a coarse powder which was then subjected to Soxhlet extraction with hexane and chloroform successively for 36 hrs. Chloroform extract was filtered and concentrated by using rotary evaporator an about 100g extract was obtained. The chloroform extract was subjected to column chromatographed over silicagel. From the column chromatography seven compounds were isolated named as osajin, scandinone, scandenone, 4,5,7-tri hydroxy biprenyl isoflavone, derris isoflavone-A, scandenin and isoscandinone. D.scandens resulting in the isolation of seven compounds in the plant was confirmed by spectral data (1H NMR, 13C NMR, ESI-MS and FTIR). The isolated compounds were screened for antioxidant activity, antidiabetic activity, α-glucosidase (inhibitory activity) and anti-bacterial activity. The isolated seven compounds were tested for α-glucosidase inhibitory activity and antioxidant activity. All the seven compounds showed good α-glucosidase inhibitory activity and moderate antioxidant activity.

Keywords: Derris scandens, phytochemical, antioxident, antidiabetic, antibacterial activity

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Authors: Cátia Gonçalves, Teresa Nunes, Inês Pina, Ana Vicente, C. Alves, Felix Charvet, Daniel Neves, A. Matos

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The production of charcoal relies on rudimentary technologies using traditional brick kilns. Charcoal is produced under pyrolysis conditions: breaking down the chemical structure of biomass under high temperature in the absence of air. The amount of the pyrolysis products (charcoal, pyroligneous extract, and flue gas) depends on various parameters, including temperature, time, pressure, kiln design, and wood characteristics like the moisture content. This activity is recognized for its inefficiency and high pollution levels, but it is poorly characterized. This activity is widely distributed and is a vital economic activity in certain regions of Portugal, playing a relevant role in the management of woody residues. The location of the units establishes the biomass used for charcoal production. The Portalegre district, in the Alto Alentejo region (Portugal), is a good example, essentially with rural characteristics, with a predominant farming, agricultural, and forestry profile, and with a significant charcoal production activity. In this district, a recent inventory identifies almost 50 charcoal production units, equivalent to more than 450 kilns, of which 80% appear to be in operation. A field campaign was designed with the objective of determining the composition of the emissions released during a charcoal production cycle. A total of 30 samples of particulate matter and 20 gas samples in Tedlar bags were collected. Particulate and gas samplings were performed in parallel, 2 in the morning and 2 in the afternoon, alternating the inlet heads (PM₁₀ and PM₂.₅), in the particulate sampler. The gas and particulate samples were collected in the plume as close as the emission chimney point. The biomass (dry basis) used in the carbonization process was a mixture of cork oak (77 wt.%), holm oak (7 wt.%), stumps (11 wt.%), and charred wood (5 wt.%) from previous carbonization processes. A cylindrical batch kiln (80 m³) with 4.5 m diameter and 5 m of height was used in this study. The composition of the gases was determined by gas chromatography, while the particulate samples (PM₁₀, PM₂.₅) were subjected to different analytical techniques (thermo-optical transmission technique, ion chromatography, HPAE-PAD, and GC-MS after solvent extraction) after prior gravimetric determination, to study their organic and inorganic constituents. The charcoal production cycle presents widely varying operating conditions, which will be reflected in the composition of gases and particles produced and emitted throughout the process. The concentration of PM₁₀ and PM₂.₅ in the plume was calculated, ranging between 0.003 and 0.293 g m⁻³, and 0.004 and 0.292 g m⁻³, respectively. Total carbon, inorganic ions, and sugars account, in average, for PM10 and PM₂.₅, 65 % and 56 %, 2.8 % and 2.3 %, 1.27 %, and 1.21 %, respectively. The organic fraction studied until now includes more than 30 aliphatic compounds and 20 PAHs. The emission factors of particulate matter to produce charcoal in the traditional kiln were 33 g/kg (wooddb) and 27 g/kg (wooddb) for PM₁₀ and PM₂.₅, respectively. With the data obtained in this study, it is possible to fill the lack of information about the environmental impact of the traditional charcoal production in Portugal. Acknowledgment: Authors thanks to FCT – Portuguese Science Foundation, I.P. and to Ministry of Science, Technology and Higher Education of Portugal for financial support within the scope of the project CHARCLEAN (PCIF/GVB/0179/2017) and CESAM (UIDP/50017/2020 + UIDB/50017/2020).

Keywords: brick kilns, charcoal, emission factors, PAHs, total carbon

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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: He Yuhai, Ahmad Ziad Bin Sulaiman

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Tongkat Ali, or Eurycoma longifolia, is a traditional Malay and Orang Asli herb used as aphrodisiac, general tonic, anti-Malaria, and anti-Pyretic. It has been recognized as a cashcrop by Malaysia due to its high value for the pharmaceutical use. In Tongkat Ali, eurycomanone, a quassinoid is usually chosen as a marker phytochemical as it is the most abundant phytochemical. In this research, ultrasound and enzyme were used to enhance the extraction of Eurycomanone from Tongkat Ali. Ultrasonic assisted extraction (USE) enhances extraction by facilitating the swelling and hydration of the plant material, enlarging the plant pores, breaking the plant cell, reducing the plant particle size and creating cavitation bubbles that enhance mass transfer in both the washing and diffusion phase of extraction. Enzyme hydrolyses the cell wall of the plant, loosening the structure of the cell wall, releasing more phytochemicals from the plant cell, enhancing the productivity of the extraction. Possible effects of ultrasound on the activity of the enzyme during the hydrolysis of the cell wall is under the investigation by this research. The extracts was analysed by high performance liquid chromatography for the yields of Eurycomanone. In this whole process, the conventional water extraction was used as a control of comparing the performance of the ultrasound and enzyme assisted extraction.

Keywords: ultrasound, enzymatic, extraction, Eurycoma longifolia

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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: Jelena Filipovic, Milenko Kosutic

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The dynamic way of life has the tendency to simplify and decrease preparing healthy, quick, cheap and safe meals. Spelt pasta is meeting most of these goals. Contrary to bread, pasta can be stored a long time without deterioration in flavour, odour and usability without losing quality. This paper deals with the chemical composition and content of fatty acids in flaxseed and spelt flour. Ratio of essential fatty acids ω-6/ω-3 is also analysed in spelt pasta and pasta with 0%, 10% and 20% flaxseed flour. Gas chromatography with mass spectrometry is used for carrying out a quantitative analysis of flaxseed flour, spelt flour and pasta liposoluble extracts. Flaxseed flour has a better fatty acid profile than spelt flour, with low levels of saturated fat (approximately 9g/100g), high concentration of linolenic acid (57g/100g) and lower content of linoleic acid (16g/100g), as well as superior ω-6/ω-3 ratio that is 1:4. Flaxseed flour in the share of 10% and 20% in spelt pasta positively contributes to the essential fatty acids daily intake recommended by nutritionists and the improvement of ω-6/ω-3 ratio (6,7:1 and 1:1.2). This paper points out that investigated pasta with flaxseed is a new product with improved functional properties due to high level of ω-3 fatty acids and it is acceptable for consumers in regard to sensory properties.

Keywords: flaxseed, spelt, fatty acids, ω-3/ω-6 ratio, pasta

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Authors: Reda Dzingelevičienė, Vytautas Abromaitis, Nerijus Dzingelevičius, Kęstutis Baranauskis, Saulius Raugelė, Malgorzata Mlynska-Szultka, Sergej Suzdalev, Reza Pashaei, Sajjad Abbasi, Boguslaw Buszewski

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A unique hybrid technology was developed for the removal of COVID-19 specific contaminants from wastewater. Reactor testing was performed using model water samples contaminated with COVID-19 pharmaceuticals and microplastics. Different hydraulic retention times, concentrations of pollutants and dissolved ozone were tested. Liquid Chromatography-Mass Spectrometry, solid phase extraction, surface area and porosity, analytical tools were used to monitor the treatment efficiency and remaining sorption capacity of the spent adsorbent. The combination of advanced oxidation and adsorption processes was found to be the most effective, with the highest 90-99% and 89-95% molnupiravir and microplastics contaminants removal efficiency from the model wastewater. The research has received funding from the European Regional Development Fund (project No 13.1.1-LMT-K-718-05-0014) under a grant agreement with the Research Council of Lithuania (LMTLT), and it was funded as part of the European Union’s measure in response to the COVID-19 pandemic.

Keywords: adsorption, hybrid reactor system, pharmaceuticals-microplastics, wastewater

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Authors: Abdelkader Basli, Jean-Claude Delaunay, Eric Pedrot, Jean-Michel Mérillon, Jean-Pierre Monti, Khodir Madani, Mohamed Chibane, Tristan Richard

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The antioxidant and antimicrobial activities of Origanum glandulosum collected in Algeria have been studied. Extract was prepared from aerial part of endemic Algerian oregano. The produced extract has been characterized in terms of total phenols (using Folin method), total flavonoid, antioxidant activities (using the DPPH radical scavenging method and ORAC assay) and microbial activity against four bacteria: Streptococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae one yeast: Candida albicans and one fungi: Aspergillus niger. The results pointed the antioxidant activities of the extract of O. glandulosum and antimicrobial activities against all bacteria and C. Candida, but no effect on A. niger. High performance liquid chromatography combined with mass spectrometry (LC-MS) and nuclear magnetic resonance (LC-NMR) were used to separate and identify the major compounds present in the oregano extract. Rosmarinic acid, globoidnan A and B, lithospermic acid B and three flavonoids were identified.

Keywords: origanum glandulosum, antioxidant, microbial activity, polyphenol, LC-MS, LC-NMR

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Authors: Saleh H. Salmen, Sulaiman Ali Alharbi, Hany M. Yehia, Mohammad A. Khiyami, Milton Wainwright, Naiyf S. Alharbi, Arunachalam Chinnathambi

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An antifungal protein synthesized by Bacillus cereus has been partially purified by the use of ammonium sulfate precipitation and Sephadex-G-200 column chromatography. The protein was produced from Bacillus cereus grown in potato Dextrose Broth Medium (PDB) at 30 ºC for 3 days at 100 rpm. The protein showed antagonistic effect against some fungi and yeasts. Crude extract from medium and semi-purified protein were tested in vitro against both fungi and yeasts using the disc diffusion method in order to detect the inhibitory effect of the protein. Zones of inhibition of the following diameter were found (mm) were Alternaria alternate (28), Rhodotorula glutinis (20), Fusarium sp. (16), Rhizopus sp. (15), Penicillium digitatum (13), Mucor sp. (13) and Aspergillus niger (10). The isolated protein was found to have a molecular weight of ~35kDa by sodium deodecyl sulfate-poly acrylamide gel electrophoresis. The data showed that the protein of Bacillus cereus has antifungal activity, a fact which points to the possibility of using it as a bio-control agent against some fungi, findings which emphasize the potential role of B. cereus as an important bio-control agent.

Keywords: bacillus cereus, ~35kDa protein, molds, yeasts

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Authors: Vikas Sharma, V. K. Dixit

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The roots of Anacyclus pyrethrum DC (AP) (Asteraceae) are frequently used in traditional medicine as Vajikarana Rasayana. An ethanolic extract of root of Anacyclus pyrethrum demonstrated its potential to enhance the sexual behaviour of male rats, with a dose dependent effect on sperm count and androgens concentration. Phytochemical analysis of ethanolic extract of Anacyclus pyrethrum revealed that it is rich in N-alkylamide. This study therefore sought to assess characterization of ethanolic extract of Anacyclus pyrethrum root. Root extract was performed using a gradient reversed phase high performance liquid chromatography/UV/electrospray ionization ion trap mass spectrometry (HPLC/ESI-MS) method on an embedded polar column. MS1 and MS2 fragmentation data were used for identification purposes, while UV was used for quantification. Thirteen N-alkylamides (five N-isobutylamides, three N-methyl isobutylamides, four tyramides, and one 2-phenylethylamide) were detected. Five of them identified as undeca-2E,4E-diene-8,10-diynoic acid N-methyl isobutylamide, tetradeca-2E,4E-diene-8,10-diynoic acid tyramide, deca-2E,4E-dienoic acid N-methyl isobutylamide, tetradeca-2E,4E,XE/Z-trienoic acid tyramide and tetradeca-2E,4E,8Z,10Z-tetraenoic isobutylamide are novel compounds, which have never been identified in Anacyclus pyrethrum.

Keywords: Anacyclus pyrethrum (Asteraceae), LC-MS plant profiling, N-alkylamides, pellitorine, anacycline

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Authors: Ozcan Baris Citil

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The aim of the present study is the determination of the effects of variations in the proximate analysis, cholesterol content and fatty acid compositions of Helix aspersa. Garden snails (Helix aspersa) were picked up by hand from the Central Anatolia Region of Turkey, in autumn (November) in 2015. Fatty acid methyl esters (FAMEs) and cholesterol analysis were analyzed by gas chromatography (GC). The protein contents of snail muscle were determined with Kjeldahl distillation units. Statistical comparisons were made by using SPSS Software (version 16.0). Thirty different fatty acids of different saturation levels were detected. As the predominant fatty acids, stearic acid (C18:0), oleic acid (C18:1ω9), linoleic acid (C18:2ω6), palmitic acid (C16:0), arachidonic acid (C20:4ω6), eicosadienoic acid (C20:2) and linolenic acid (C18:3ω3) were found in Helix aspersa. Palmitic acid (C16:0) was identified as the major SFA in autumn. Linoleic acid (C18:2ω6), eicosadienoic acid (C20:2) and arachidonic acid (C20:4ω6) have the highest levels among the PUFAs. In the present study, ω3 were found 5.48% in autumn. Linolenic acid and omega-3 fatty acid amounts in the autumn decreased significantly but cholesterol content was not affected in Helix aspersa in autumn (November) in 2015.

Keywords: Helix aspersa, fatty acid, SFA, PUFA, cholesterol

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Authors: Pannaga Pavan Jutur

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Parachlorella kessleri, a marine unicellular green alga belonging to class Trebouxiophyceae, accumulates large amounts of oil, i.e., lipids under nutrient-deprived (-N, -P, and -S) conditions. Understanding their metabolic imprints is important for elucidating the physiological mechanisms of lipid accumulations in this microalga subjected to nutrient deprivation. Metabolic and lipidomic profiles were obtained respectively using gas chromatography-mass spectrometry (GC-MS) of P. kessleri under nutrient starvation (-N, -P and -S) conditions. Relative quantities of more than 100 metabolites were systematically compared in all these three starvation conditions. Our results demonstrate that in lipid metabolism, the quantities of neutral lipids increased significantly followed by the decrease in other metabolites involved in photosynthesis, nitrogen assimilation, etc. In conclusion, the metabolomics and lipidomic profiles have identified a few common metabolites such as citric acid, valine, and trehalose to play a significant role in the overproduction of oil by this microalga subjected to nutrient deprivation. Understanding the entire system through untargeted metabolome profiling will lead to identifying relevant metabolites involved in the biosynthesis and degradation of precursor molecules that may have the potential for biofuel production, aiming towards the vision of tomorrow’s bioenergy needs.

Keywords: algae, biofuels, nutrient stress, omics

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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, methyltrioctylammonium chloride, ionic liquid, capacity

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Authors: Abdulmannan Fadel

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Arabinoxylan is a non-starch polysaccharide (NSP), which is one of the most important polysaccharides contained within cereal grains. Wheat endosperm pentosan and rice bran contain a significant amount of arabinoxylan (7% in rice bran and 10-12% in wheat endosperm pentosan). Several methods have been used for arabinoxylan extraction with varying degrees of success e.g. enzymatic and alkaline treatment. Yet, the use of extrusion alone as a pre-treatment to increase the yield and reduce the molecular weight in wheat endosperm pentosan and rice bran has not been investigated. The samples (wheat pentosan and rice bran) were extruded using a Twin-screw extruder at a range of screw speeds (80 and 160 rpm) and barrel temperatures range (80 to 140°C) with a throughput of 30 Kg hr-1 and moisture content of 25%. Arabinoxylans were extracted with water and the extraction yield and molecular weight was determined using size exclusion high-pressure liquid chromatography system. It was found that increasing screw speed from 80 rpm to 160 rpm, did not effect the extraction yield (p < 0.05) of arabinoxylan from either the wheat endosperm pentosan or the rice bran. However, the molecular weight of the extracted arabinoxylans from pentosan was found to decrease with increasing screw speed in wheat endosperm pentosan. These low molecular weight arabinoxylans have been suggested as immunomodulators.

Keywords: arabinoxylans, extrusion, wheat endosperm pentosan, rice bran

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Authors: Nirmala Deenadayalu, Kwanele B. Mazibuko, Lethiwe D. Mthembu

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Sugarcane bagasse is the residue obtained after the extraction of sugar from the sugarcane. The main aim of this work was to produce catechol from sugarcane bagasse. The optimization of catechol production was investigated using a Box-Behnken design of experiments. The sugarcane bagasse was heated in a Parr reactor at a set temperature. The reactions were carried out at different temperatures (100-250) °C, catalyst loading (1% -10% KOH (m/v)) and reaction times (60 – 240 min) at 17 bar pressure. The solid and liquid fractions were then separated by vacuum filtration. The liquid fraction was analyzed for catechol using high-pressure liquid chromatography (HPLC) and characterized for the functional groups using Fourier transform infrared spectroscopy (FTIR). The optimized condition for catechol production was 175 oC, 240 min, and 10 % KOH with a catechol yield of 79.11 ppm. Since the maximum time was 240 min and 10 % KOH, a further series of experiments were conducted at 175 oC, 260 min, and 20 % KOH and yielded 2.46 ppm catechol, which was a large reduction in catechol produced. The HPLC peak for catechol was obtained at 2.5 min for the standards and the samples. The FTIR peak at 1750 cm⁻¹ was due to the C=C vibration band of the aromatic ring in the catechol present for both the standard and the samples. The peak at 3325 cm⁻¹ was due to the hydrogen-bonded phenolic OH vibration bands for the catechol. The ANOVA analysis was also performed on the set of experimental data to obtain the factors that most affected the amount of catechol produced.

Keywords: catechol, sugarcane bagasse, lignin, hydrothermal liquefaction

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Authors: L. Bouzina, A. Bensafi, M. Duval, C. Mathis, M. Rawiso

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We study the stability and structural properties of mixtures of model nanoparticles and non-adsorbing polymers in the 'protein limit', where the size of polymers exceeds the particle size substantially. We have synthesized in institute (Charles Sadron Strasbourg) model nanoparticles by coating fullerene C60 molecules with low molecular weight polystyrene (PS) chains (6 PS chains with a degree of polymerization close to 25 and 50 are grafted on each fullerene C60 molecule. We will present a Small Angle Neutron scattering (SANS) study of Tetrahydrofuran (THF) solutions involving long polystyrene (PS) chains and fullerene (C60) nanoparticles. Long PS chains and C60 nanoparticles with different arm lengths were synthesized either hydrogenated or deuteriated. They were characterized through Size Exclusion Chromatography (SEC) and Quasielastic Light Scattering (QLS). In this way, the solubility of the C60 nanoparticles in the usual good solvents of PS was controlled. SANS experiments were performed by use of the contrast variation method in order to measure the partial scattering functions related to both components. They allow us to obtain information about the dispersion state of the C60 nanoparticles as well as the average conformation of the long PS chains. Specifically, they show that the addition of long polymer chains leads to the existence of an additional attractive interaction in between soft nanoparticles.

Keywords: fulleren nanoparticles, polymer, small angle neutron scattering, solubility

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Authors: Mohsen Danesh Mesgaran, Ali Hodjatpanah Montazeri, Alireza Vakili, Mansoor Tahmasbei

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To compare peppermint essential oil versus a mixture of formic and propionic acids a study was conducted to their effects on volatile fatty acid proportion and VFA score of corn silage. Chopped whole crop corn (control) was treated with peppermint essential oil (240 mg kg-1 DM) or a mixture of formic and propionic acids (2:1) at 0.4% of fresh forage weight, and ensiled for 30 days. Then, silage extract was provided and the concentration of each VFA was determined using gas chromatography. The VFA score was calculated according to the patented formula proposed by Dairy One Scientific Committee. The score is calculated based on the positive impact of lactic and acetic acids versus the negative effect of butyric acid to achieve a single value for evaluating silage quality. The essential oil declined pH and increased the concentration of lactic and acetic acids in the silage extract. All corn silages evaluated in this study had a VFA score between 6 through 8. However, silage with peppermint essential oils had lower volatile fatty acids score than those of the other treatments. Both of applied additives caused a significant improvement in silage aerobic stability.

Keywords: peppermint, essential oil, corn silage, VFA (volatile fatty acids)

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Authors: Rasha Saad, Jean Frederic Weber, Fatimah Bebe, Sadia Sultan

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The goal of study was to screen the effects of candesartan and four endophytic fungi for their potential in microbial biotransformation. In this experiment, four types of unidentified fungi with the codes of TH2L1, TH2R10, TH1P35 and TH1S46 were used in screening process by MECFUS (Microtiter plate, Elicitors, Combination, Freeze-drying, UHPLC, Statistical analysis) protocol. The experiment was carried out by using 96-well microtiter plate (MTP) with different media and elicitors. Various media with two concentrations of Potato Dextrose Broth (PDB) and elicitors used were to induce the production of secondary metabolites from the fungi as well as the biotransformation of the drug compound. After incubation, cultures were extracted by freeze drying method and finally analyzed by ultra-High performance Liquid Chromatography (uHPLC). The extracts analyzed by uHPLC followed by LC/Ms, demonstrated the presence of biotransformation products from the drug compound and elicitation of the secondary metabolism from the fungi by the occurrence of the additional peaks. From the four fungi, TH1S46 showed highly potential produced secondary metabolites as well as the biotransformation of candesartan. For other fungi, they responded when candesartan was introduced. Moreover, the additional peaks produced in uHPLC need to be further investigation by using LC-MS or NMR.

Keywords: biotransformation, candesartan, endophytes, secondary metabolites

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Authors: Patsarawadee Paojinda, Waranya Imprasittichai, Sudaratana R. Krungkrai, Nirianne Marie Q. Palacpac, Toshihiro Horii, Jerapan Krungkrai

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Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. Here, we produced gene fusions of Plasmodium falciparum OMPDC-OPRT and expressed the bifunctional protein in Escherichia coli. The enzyme was purified to homogeneity using affinity and anion-exchange chromatography, exhibited enzymatic activities and functioned as a dimer. The activities, although unstable, can be stabilized by its substrate and product during purification and long-term storage. Furthermore, the enzyme expressed a perfect catalytic efficiency (kcat/Km). The kcat was selectively enhanced up to 3 orders of magnitude, while the Km was not much affected and remained at low µM levels when compared to the monofunctional enzymes. The fusion of the two enzymes, creating a “super-enzyme” with perfect catalytic power and more flexibility, reflects cryptic relationship of enzymatic reactivaties and metabolic functions on molecular evolution.

Keywords: bifunctional enzyme, orotate phosphoribosyltransferase, orotidine 5'-monophosphate decarboxylase, plasmodium falciparum

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Authors: Malith Premarathne, Shehan Bandara, Kaushalya G. Batawala, Thilini U. Ariyadasa

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The rapid depletion of fossil fuel supplies and the emission of carbon dioxide by their continued combustion have paved the way for increased production of carbon-neutral biodiesel from naturally occurring oil sources. The high biomass growth rate and lipid production of microalgae make it a viable source for biodiesel production compared to conventional feedstock. In Sri Lanka, the production of biodiesel by employing indigenous microalgae species is at its emerging stage. This work was an attempt to compare the various pre-treatment methods before extracting lipids such as autoclaving, microwaving and sonication. A mixed culture of microalgae predominantly consisting of Chlorella sp. was obtained from Beire Lake which is an algae rich, organically polluted water body located in Colombo, Sri Lanka. After each pre-treatment method, a standard solvent extraction using Bligh and Dyer’s method was used to compare the total lipid content in percentage dry weight (% dwt). The fatty acid profiles of the oils extracted with each pretreatment method were analyzed using gas chromatography-mass spectrometry (GC-MS). The properties of the biodiesels were predicted by Biodiesel Analyzer© Version 1.1, in order to compare with ASTM 6751-08 biodiesel standard.

Keywords: biodiesel, lipid extraction, microalgae, pre-treatment

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Authors: D. S. Fardhyanti, M. Megawati, H. Prasetiawan, U. Mediaty

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Currently, fossil fuel is supplying most of world’s energy resources. However, fossil fuel resources are depleted rapidly and require an alternative energy to overcome the increasing of energy demands. Bio-oil is one of a promising alternative renewable energy resources which is converted from biomass through pyrolysis or fast pyrolysis process. Bio-oil is a dark liquid fuel, has a smelling smoke and usually obtained from sugar cane, wood, coconut shell and any other biomass. Sugar cane content analysis showed that the content of oligosaccharide, hemicellulose, cellulose and lignin was 16.69%, 25.66%, 51.27% and 6.38% respectively. Sugar cane is a potential sources for bio-oil production shown by its high content of cellulose. In this study, production of bio-oil from sugar cane bagasse was investigated via fast pyrolysis reactor. Fast pyrolysis was carried out at 500 °C with a heating rate of 10 °C and 1 hour holding time at pyrolysis temperature. Physical properties and chemical composition of bio-oil were analyzed. The viscosity, density, calorific value and molecular weight of produced bio-oil was 3.12 cp, 2.78 g/cm3, 11,048.44 cals/g, and 222.67 respectively. The Bio-oil chemical composition was investigated using GC-MS. Percentage value of furfural, phenol, 3-methyl 1,2-cyclopentanedione, 5-methyl-3-methylene 5-hexen-2-one, 4-methyl phenol, 4-ethyl phenol, 1,2-benzenediol, and 2,6-dimethoxy phenol was 20.76%, 16.42%, 10.86%, 7.54%, 7.05%, 7.72%, 5.27% and 6.79% respectively.

Keywords: bio-oil, pyrolysis, bagasse, sugar cane, gas chromatography-mass spectroscopy

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Authors: Patrizia Firmani, Sara Perucchini, Irene Rapone, Raffella Borrelli, Stefano Chiaberge, Manuela Grande, Rosamaria Marrazzo, Alberto Savoini, Andrea Siviero, Silvia Spera, Fabio Vago, Davide Deriu, Sergio Fanutti, Alessandro Oldani

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According to the Renewable Energy Directive II, the use of palm oil in diesel will be gradually reduced from 2023 and should reach zero in 2030 due to the deforestation caused by its production. Eni aims at finding alternative feedstocks for its biorefineries to eliminate the use of palm oil by 2023. Therefore, the ideal vegetable oils to be used in bio-refineries are those obtainable from plants that grow in marginal lands and with low impact on food-and-feed chain; hence, Eni research is studying the possibility of using oleaginous seeds, such as castor, croton, and cotton, to extract the oils to be exploited as feedstock in bio-refineries. To verify their suitability for the upgrading processes, an analytical protocol for their characterization has been drawn up and applied. The analytical characterizations include a step of water and ashes content determination, elemental analysis (CHNS analysis, X-Ray Fluorescence, Inductively Coupled Plasma - Optical Emission Spectroscopy, ICP– Mass Spectrometry), and total acid number determination. Gas chromatography coupled to flame ionization detector (GC-FID) is used to quantify the lipid content in terms of free fatty acids, mono-, di- and triacylglycerols, and fatty acids composition. Eventually, Nuclear Magnetic Resonance and Fourier Transform-Infrared spectroscopies are exploited with GC-MS and Fourier Transform-Ion Cyclotron Resonance to study the composition of the oils. This work focuses on the GC-FID analysis of the lipid fraction of these oils, as the main constituent and of greatest interest for bio-refinery processes. Specifically, the lipid component of the extracted oil was quantified after sample silanization and transmethylation: silanization allows the elution of high-boiling compounds and is useful for determining the quantity of free acids and glycerides in oils, while transmethylation leads to a mixture of fatty acid esters and glycerol, thus allowing to evaluate the composition of glycerides in terms of Fatty Acids Methyl Esters (FAME). Cotton oil was extracted from cotton oilcake, croton oil was obtained by seeds pressing and seeds and oilcake ASE extraction, while castor oil comes from seed pressing (not performed in Eni laboratories). GC-FID analyses reported that the cotton oil is 90% constituted of triglycerides and about 6% diglycerides, while free fatty acids are about 2%. In terms of FAME, C18 acids make up 70% of the total and linoleic acid is the major constituent. Palmitic acid is present at 17.5%, while the other acids are in low concentration (<1%). Both analyzes show the presence of non-gas chromatographable compounds. Croton oils from seed pressing and extraction mainly contain triglycerides (98%). Concerning FAME, the main component is linoleic acid (approx. 80%). Oilcake croton oil shows higher abundance of diglycerides (6% vs ca 2%) and a lower content of triglycerides (38% vs 98%) compared to the previous oils. Eventually, castor oil is mostly constituted of triacylglycerols (about 69%), followed by diglycerides (about 10%). About 85.2% of total FAME is ricinoleic acid, as a constituent of triricinolein, the most abundant triglyceride of castor oil. Based on the analytical results, these oils represent feedstocks of interest for possible exploitation as advanced biofuels.

Keywords: analytical protocol, biofuels, biorefinery, gas chromatography, vegetable oil

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Authors: Hee-Jae Suh, Kyung-Su Kim, Min-Ji Kim, Yeon-Seong Jeong, Ok-Hwan Lee, Jae-Wook Shin, Hyang-Sook Chun, Chan Lee

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Canthaxanthin is keto-carotenoid produced from beta-carotene and it has been approved to be used in many countries as a food coloring agent. Canthaxanthin has been analyzed using High Performance Liquid Chromatography (HPLC) system with various ways of pretreatment methods. Four official methods for verification of canthaxanthin at FSA (UK), AOAC (US), EFSA (EU) and MHLW (Japan) were compared to improve its analytical and the pretreatment method. The Linearity, the limit of detection (LOD), the limit of quantification (LOQ), the accuracy, the precision and the recovery ratio were determined from each method with modification in pretreatment method. All HPLC methods exhibited correlation coefficients of calibration curves for canthaxanthin as 0.9999. The analysis methods from FSA, AOAC, and MLHW showed the LOD of 0.395 ppm, 0.105 ppm, and 0.084 ppm, and the LOQ of 1.196 ppm, 0.318 ppm, 0.254 ppm, respectively. Among tested methods, HPLC method of MHLW with modification in pretreatments was finally selected for the analysis of canthaxanthin in lab, because it exhibited the resolution factor of 4.0 and the selectivity of 1.30. This analysis method showed a correlation coefficients value of 0.9999 and the lowest LOD and LOQ. Furthermore, the precision ratio was lower than 1 and the accuracy was almost 100%. The method presented the recovery ratio of 90-110% with modification in pretreatment method. The cross-validation of coefficient variation was 5 or less among tested three institutions in Korea.

Keywords: analytic method, canthaxanthin, food colorants, pretreatment method

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