Search results for: gas chromatography

Authors: Ikram Kamal, Mohamed Blaghen

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Biosurfactants are amphiphilic biological compounds consisting of hydrophobic and hydrophilic domains produced extracellularly or as part of the cell membrane by a variety of yeast, bacteria and filamentous fungi. Biosurfactant applications in the environmental industries are promising due to their biodegradability, low toxicity, and effectiveness in enhancing biodegradation and solubilization of low solubility compounds. Currently, the main application is for enhancement of oil recovery and hydrocarbon bioremediation due to their biodegradability and low critical micelle concentration (CMC). The use of biosurfactants has also been proposed for various industrial applications, such as in food additives, cosmetics, detergent formulations and in combinations with enzymes for wastewater treatment. In this study, we have investigated the potential of bacterial strains: Mannheimia haemolytica, Burkholderia cepacia and Serratia ficaria were collected aseptically from the lagoon Marchika (water and soil) in Nador, Morocco; for the production of biosurfactants. This study also aimed to optimize the biosurfactant production process by changing the variables that influence the type and amount of biosurfactant produced by these microorganisms such as: carbon sources and also other physical and chemical parameters such as temperature and pH. Emulsification index, methylene blue test, and thin layer chromatography (TLC) revealed the ability of strains used in this study to produce compounds that could emulsify gasoline. In addition, a GC/MS was used to separate and identify different biosurfactants purified.

Keywords: biosurfactants, Mannheimia haemolytica, biodegradability, Burkholderia cepacia, Serratia ficaria

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Authors: Sumbul Farhat, Pratyay Vaibhav, Sarah Jain, Kapinder Kumar, Archna Kumar

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The Diamondback Moth, Plutella xylostella (Linnaeus), is a major pest of cole crops that causes approximately 50% loss in global production. The utilization of inorganic pesticides is reflected in the development of resistance to this pest. Thus, there is a great need for an eco-friendly, sustainable strategy for the control of this pest. Although this pest, several natural enemies are reported worldwide, none of them can control it efficiently. Therefore, a proposed study is planned to understand the Volatile Organic Compounds (VOCs) mediated signaling interaction mechanism of the plant, pest, and natural enemy. For VOCs collection during different deployment stages of Cabbage POI, Green Ball, Pusa Cabbage, Cabbage Local, Snowball 16, Kanchan Plus, Pusa Meghna, Farm Sona Hybrid F1, and Samridhi F1 Hybrid, the Solid-phase microextraction (SPME) method was employed. Characterization of VOCs was conducted by Gas Chromatography-Mass Spectrometry (GC-MS). The impact of collected VOCs was assessed through Y-Tube Bioassays. The results indicate that the Cabbage variety Green Ball shows maximum repellency for P. xylostella (-100%). The cues present in this variety may be exploited for efficient management of P. xylostella in the cole crop ecosystem.

Keywords: Plutella xylostella, cole crops, volatile organic compounds, GC-MS, Green Ball

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Authors: O. Smara, H. Dendougui, B. Legseir

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Atriplex halimus L. is particularly well adapted to arid and salt-affected areas. In this species, salinity resistance is often attributed to the presence of vesiculated hairs covering leaf surface and containing a large amount of salt. Atriplex halimus L. (Chenopodiaceae) is a perennial shrub native to the Mediterranean basin with excellent tolerance to drought and salinity. The species is present in semiarid to subhumid areas of the north Mediterranean and in arid zones from North Africa and the eastern Mediterranean. The main aim of this study was to identify a medicinal plant used in the Ouargla (Est-southern Algeria) for the treatment of several human pathologies. This plant is an important source for livestock in nitrogenous matter, it is an effective and relatively inexpensive tool in the fight against erosion and desertification and rehabilitation of degraded lands. Phytochemical investigation is applied to the majority of extracts of the powder of the aerial parts of Atriplex halimus L. Different chromatographic methods after liquid-liquid extraction are used; it is the thin layer chromatography (TLC) and paper using multiple systems and chemical revelations. This study followed by an evaluation by the phenol assay the Folin-Ciocalteu method, using gallic acid as a reference for phenols and quercetin for flavonols. Some polar extracts showed an interesting result better than the less polar extracts.

Keywords: Atriples halimus L., chenopodiaceae, flavonoids, phenols

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Authors: Youn Young Shim, Ji Hye Kim, Jae Youl Cho, Martin JT Reaney

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Flaxseed (Linumusitatissimum L.) is gaining popularity in the food industry as a superfood due to its health-promoting properties. Linusorbs (LOs, a.k.a. Cyclolinopeptide) are bioactive compounds present in flaxseed exhibiting potential health effects. The study focused on the effects of processing and storage on the stability of flaxseed-derived LOs added to various bakery products. The flaxseed meal fortified gluten-free (GF) bakery bread was prepared, and the changes of LOs during the bread-making process (meal, fortified flour, dough, and bread) and storage (0, 1, 2, and 4 weeks) at different temperatures (−18 °C, 4 °C, and 22−23 °C) were analyzed by high-performance liquid chromatography-diode array detection. The total oxidative LOs and LO1OB2 were almost kept stable in flaxseed meals at storage temperatures of 22−23 °C, −18 °C, and 4 °C for up to four weeks. Processing steps during GF-bread production resulted in the oxidation of LOs. Interestingly, no LOs were detected in the dough sample; however, LOs appeared when the dough was stored at −18 °C for one week, suggesting that freezing destroyed the sticky structure of the dough and resulted in the release of LOs. The final product, flaxseed meal fortified bread, could be stored for up to four weeks at −18 °C and 4 °C, and for one week at 22−23 °C. All these results suggested that LOs may change during processing and storage and that flaxseed flour-fortified bread should be stored at low temperatures to preserve effective LOs components.

Keywords: linum usitatissimum L., flaxseed, linusorb, stability, gluten-free, peptides, cyclolinopeptide

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Authors: N. Laosiripojana, P. Tepamatr

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The influence of bismuth on the water gas shift activities of Pt on ceria was studied. The flow reactor was used to study the activity of the catalysts in temperature range 100-400°C. The feed gas composition contains 5%CO, 10% H₂O and balance N₂. The total flow rate was 100 mL/min. The outlet gas was analyzed by on-line gas chromatography with thermal conductivity detector. The catalytic activities of bimetallic 1%Pt1%Bi/CeO₂ catalyst were greatly enhanced when compared with the activities of monometallic 2%Pt/CeO₂ catalyst. The catalysts were characterized by X-ray diffraction (XRD), Temperature-Programmed Reduction (TPR) and surface area analysis. X-ray diffraction pattern of Pt/CeO₂ and PtBi/CeO₂ indicated slightly shift of diffraction angle when compared with pure ceria. This result was due to strong metal-support interaction between platinum and ceria solid solution, causing conversion of Ce⁴⁺ to larger Ce³⁺. The distortions inside ceria lattice structure generated strain into the oxide lattice and facilitated the formation of oxygen vacancies which help to increase water gas shift performance. The H₂-Temperature Programmed Reduction indicated that the reduction peak of surface oxygen of 1%Pt1%Bi/CeO₂ shifts to lower temperature than that of 2%Pt/CeO₂ causing the enhancement of the water gas shift activity of this catalyst. Pt played an important role in catalyzing the surface reduction of ceria and addition of Bi alter the reduction temperature of surface ceria resulting in the improvement of the water gas shift activity of Pt catalyst.

Keywords: bismuth, platinum, water gas shift, ceria

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Authors: Dalia. G. Aseel

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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: okra leaf curl virus, AV1 gene, sequencing, phylogenetic, cloning, purified protein, genetic diversity and viral proteins

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Authors: Maryam Enteshari, Kooshan Nayebzadeh, Abdorreza Mohammadi

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In this study, the oxidative stability of soybean oil under different storage temperatures (4 and 25˚C) and during 6-month shelf-life was investigated by various analytical methods and headspace-liquid phase microextraction (HS-LPME) coupled to gas chromatography-mass spectrometry (GC-MS). Oxidation changes were monitored by analytical parameters consisted of acid value (AV), peroxide value (PV), p-Anisidine value (p-AV), thiobarbituric acid value (TBA), fatty acids profile, iodine value (IV), and oxidative stability index (OSI). In addition, concentrations of hexanal and heptanal as secondary volatile oxidation compounds were determined by HS-LPME/GC-MS technique. Rate of oxidation in soybean oil which stored at 25˚C was so higher. The AV, p-AV, and TBA were gradually increased during 6 months while the amount of unsaturated fatty acids, IV, and OSI decreased. Other parameters included concentrations of both hexanal and heptanal, and PV exhibited increasing trend during primitive months of storage; then, at the end of third and fourth months a sudden decrement was understood for the concentrations of hexanal and heptanal and the amount of PV, simultaneously. The latter parameters increased again until the end of shelf-time. As a result, the temperature and time were effective factors in oxidative stability of soybean oil. Also intensive correlations were found for soybean oil at 4 ˚C between AV and TBA (r2=0.96), PV and p-AV (r2=0.9), IV and TBA (-r2=0.9), and for soybean oil stored at 4˚C between p-AV and TBA (r2=0.99).

Keywords: headspace-liquid phase microextraction, oxidation, shelf-life, soybean oil

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Authors: Merouani Djilali Redha, F. Abdelmalek, A. A. Addou

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Advanced oxidation processes (AOPs) utilizing Homogenous photocatalysis (Fenton and photo-Fenton reactions), and Heterogeneous photocatalyse (TiO2 and ZnO) were investigated for the degradation of commercial azo dye ‘Orange G’ wastewater. Fenton and photo-Fenton experimental conditions were: Hydrogen peroxide concentration (10-2 M), Ferrous ions concentration (5.10-4 M), pH (2.8 – 3), UV lamp power (6 watt). Adding more ferrous ions enhanced the oxidation rate for the H2O2/Fe2+ and UV/H2O2/Fe2+ processes. The optimum catalyst loading was found 2.0 g.L-1 in our case for both catalysts TiO2 and ZnO. A comparative study of the photocatalytic degradation showed that these two catalysts have a comparable reactivity; it follows a pseudo-first-order kinetics. The degradation trends followed the order: UV365/Fenton > UV365/TiO2 > Solar Fenton > Solar TiO2 > Fenton ~UV365/ZnO. Among AOPs, processes using Fenton type reagent are relatively cheap and easy to operate and maintain. Moreover, UV365/Fenton process has been shown as effective in the treatment of OG dye. Dye was degraded following second-order kinetics. The rate constants was 0,041 .10+6 L.M-1.min-1. The degradation was followed by spectrophotometric method, chemical oxygen demand (COD) measures and high performance liquid chromatography analyses (HPLC). Some aromatic and aliphatic degradation compounds were identified. Degradation of Orange G by UV Fenton mechanism was also proposed.

Keywords: AOPs, homogeneous catalysis, heterogeneous catalysis, acid orange 10, hydroxyl radical

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Authors: N. Lovanh, J. Loughrin, G. Ruiz-Aguilar

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Animal wastes can serve as the feedstock for biogas production (mainly methane) that could be used as alternative energy source. The green energy derived from animal wastes is considered to be carbon neutral and offsetting those generated from fossil fuels. In this study, an evaluation of system parameters on methane production from anaerobic digesters utilizing poultry rendering plant wastewater was carried out. Anaerobic batch reactors and continuous flow system subjected to different operation conditions (i.e., flow rate, temperature, and etc.) containing poultry rendering wastewater were set up to evaluate methane potential from each scenario. Biogas productions were sampled and monitored by gas chromatography and photoacoustic gas analyzer over six months of operation. The results showed that methane productions increased as the temperature increased. However, there is an upper limit to the increase in the temperature on the methane production. Flow rates and type of systems (batch vs. plug-flow regime) also had a major effect on methane production. Constant biogas production was observed in plug-flow system whereas batch system produced biogas quicker and tapering off toward the end of the six-month study. Based on these results, it is paramount to consider operating conditions and system setup in optimizing biogas production from agricultural wastewater.

Keywords: anaerobic digestion, methane, poultry rendering wastewater, biotechnology

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Authors: Saidu Garba Okereafor Stella

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The presence of 15 organochlorine pesticides residue was assessed from 29 different fruits and fruit juice samples from selected farms in Kaduna and Niger States using the quick easy cheap effective rugged and safe (QuEChERS), followed by gas chromatography-tandem mass spectrometry (GC-MS/MS). The results showed the presence of varying concentrations of ten (10) organochlorine pesticide residues in all the samples with Endrin ketone showing the highest concentration in 3 samples from Kaduna (guava juice 1 and 2 0.099 to 0.145 mg/kg) and Niger States (orange juice J19 0.102 mg/kg). The heptachlor was detected at high concentration in 11 samples, 7 samples from Kaduna State (mango juice 0.011 mg/kg, Washington orange 0.014 mg/kg, Valencia orange fruit 0.020 mg/kg, orange juice 0.011, white guava fruit 0.024 mg/kg, guava juice 0.023 mg/kg, guava juice 2 0.024 mg/kg) and 4 samples from (mango juice 1 0.015 mg/kg, pineapple juice 1 0.0120 mg/kg pineapple juice 2 011 mg/kg and mix juice 2 0.012 mg/kg) from Niger State. Dieldrine and endosulfansulfate were detected at high levels in one sample each from Niger (guava fruit 0.019 mg/kg and mixed juice1 0.011mg/kg), respectively. However, all were above the maximum residue limits (MRLs) set by WHO/FAO which suggest that people consuming these type of contaminated fruits and fruits juices may contact diseases associated with those organochlorine pesticides residue. Minute concentrations of other organochlorines (α- BHC, δ- BHC, β- BHC, Lindane, and p’p DDT) ranged from 0.003 to 0.015 were recorded below the MRLs.

Keywords: fruits and fruits juices, organochlorine pesticide residue, comparative studies, gc-ms spectrophometer

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Authors: Bilal Murtaza, Xiaoyu Li, Liming Dong, Muhammad Kashif Saleemi, Gen Li, Bowen Jin, Lili Wang, Yongping Xu

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Fusarium spp. produce numerous mycotoxins, such as zearalenone (ZEN), deoxynivalenol (DON), and its acetylated compounds, 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON) (15-ADON). In a co-culture system, the soil-derived Escherichia marmotae strain degrades ZEN and DON into 3-keto-DON and DOM-1 via enzymatic deep-oxidation. When pure mycotoxins were subjected to Escherichia marmotae in culture flasks, degradation, and detoxification were also attained. DON and ZEN concentrations, ambient pH, incubation temperatures, bacterium concentrations, and the impact of acid treatment on degradation were all evaluated. The results of the ELISA and high-performance liquid chromatography-electrospray ionization-high resolution mass spectrometry (HPLC-ESI-HRMS) tests demonstrated that the concentration of mycotoxins exposed to Escherichia marmotae was significantly lower than the control. ZEN levels were reduced by 43.9%, while zearalenone sulfate ([M/z 397.1052 C18H21O8S1) was discovered as a derivative of ZEN converted by microbes to a less toxic molecule. Furthermore, Escherichia marmotae appeared to metabolize DON 35.10% into less toxic derivatives (DOM-1 at m/z 281 of [DON - O]+ and 3-keto-DON at m/z 295 of [DON - 2H]+). These results show that Escherichia marmotae can reduce Fusarium mycotoxins production, degrade pure mycotoxins, and convert them to less harmful compounds, opening up new possibilities for study and innovation in mycotoxin detoxification.

Keywords: mycotoxins, zearalenone, deoxynivalenol, bacterial degradation

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Authors: Renata Adriana Labanca, Gabriela Rezende Costa, Nilton de Oliveira Couto e Silva, José Marcos Gontijo Mandarino, Rodrigo Santos Leite, Nilson César Castanheira Guimarães, Roberto Gonçalves Junqueira

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The objective of this study was to evaluate the differences in composition between six brands of conventional soybean and six genetically modified cultivars (GM), all of them from Minas Gerais State, Brazil. We focused on the isoflavones profile and mineral content questioning the substantial equivalence between conventional and GM organisms. The statement of compliance label for conventional grains was verified for the presence of genetic modified genes by real time polymerase chain reaction (PCR). We did not detect the presence of the 35S promoter in commercial samples, indicating the absence of transgene insertion. For mineral analysis, we used the method of inductively coupled plasma-optical emission spectrometry (ICP-OES). Isoflavones quantification was performed by high performance liquid chromatography (HPLC). The results showed no statistical difference between the conventional and transgenic soybean groups concerning isoflavone content and mineral composition. The concentration of potassium, the main mineral component of soy, was the highest in conventional soybeans compared to that in GM soy, while GM samples presented the highest concentrations of iron.

Keywords: glycine max, genetically modified organism, bioactive compounds, ICP-OES, HPLC

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Authors: Atifa Mushtaq, Tanweer Khaliq, Hafiz Alam Sher, Asia Farid, Anila Kanwal, Maliha Sarfraz

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The study was designed to assess the various pharmacokinetic parameters of a commercially available clarithromycin Tablet (Klaricid® 250 mg Abbot, Pakistan) in plasma sample of healthy adult female volunteers by applying a rapid, sensitive and accurate HPLC-UV analytical method. The human plasma samples were evaluated by using an isocratic High Performance Liquid Chromatography (HPLC) system of Sykam consisted of a pump with a column C18 column (250×4.6mn, 5µm) UV-detector. The mobile phase comprises of potassium dihydrogen phosphate (50 mM, pH 6.8, contained 0.7% triethylamine), methanol and acetonitrile (30:25:45, v/v/v) was delivered with injection volume of 20µL at flow rate of 1 mL/min. The detection was performed at λmax 275 nm. By applying this method, important pharmacokinetic parameters Cmax, Tmax, Area under curve (AUC), half-life (t1/2), , Volume of distribution (Vd) and Clearance (Cl) were measured. The parameters of pharmacokinetics of clarithromycin were calculated by software (APO) pharmacological analysis. Maximum plasma concentrations Cmax 2.78 ±0.33 µg/ml, time to reach maximum concentration tmax 2.82 ± 0.11 h and Area under curve AUC was 20.14 h.µg/ml. The mean ± SD values obtained for the pharmacokinetic parameters showed a significant difference in pharmacokinetic parameters observed in previous literature which emphasizes the need for dose adjustment of clarithromycin in Pakistani population.

Keywords: Pharmacokinetc, Clarothromycin, HPLC, Pakistan

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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: Mariane G. Tadros, Shahira M. Ezzat, Maha M. Salama, Mohamed A. Farag

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In this study, the in vitro anticholinesterase activity of the volatile oils of both O. basilicum and O. africanum was investigated and both samples showed significant activity. As a result, the major constituents of the two oils were isolated using several column chromatography. Linalool, 1,8-cineol and eugenol were isolated from the volatile oil of O. basilicum and camphor was isolated from the volatile oil of O. africanum. The anticholinesterase activity of the isolated compounds were also evaluated where 1,8-cineol showed the highest inhibitory activity followed by camphor. To confirm these activities, learning and memory enhancing effects were tested in mice. Memory impairment was induced by scopolamine, a cholinergic muscarinic receptor antagonist. Anti-amnesic effects of both volatile oils and their terpenoids were investigated by the passive avoidance task in mice. We also examined their effects on brain acetylcholinesterase activity. Results showed that scopolamine-induced cognitive dysfunction was significantly attenuated by administration of the volatile oils and their terpenoids, eugenol and camphor, in the passive avoidance task and inhibited brain acetylcholinesterase activity. These results suggest that O. basilicum and O. africanum volatile oils can be good candidates for further studies on Alzheimer’s disease via their acetylcholinesterase inhibitory actions.

Keywords: Ocimum baselicum, Ocimum africanum, GC/MS analysis, anticholinesterase

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Authors: Taruna Katyal Arora, Geeta Kumari, Hari Shankar, Neelima Mishra

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Substandard and counterfeit antimalarials is a major problem in malaria endemic areas. The availability of counterfeit/ substandard medicines is not only decreasing the efficacy in patients, but it is also one of the contributing factors for developing antimalarial drug resistance. Owing to this, a pilot study was conducted to survey quality of drugs collected from different malaria endemic areas of India. Artesunate+Sulphadoxine-Pyrimethamine (AS+SP), Artemether-Lumefantrine (AL), Chloroquine (CQ) tablets were randomly picked from public health facilities in selected states of India. The quality of antimalarial drugs from these areas was assessed by using Global Pharma Health Fund Minilab test kit. This includes physical/visual inspection and disintegration test. Thin-layer chromatography (TLC) was carried out for semi-quantitative assessment of active pharmaceutical ingredients. A total of 45 brands, out of which 21 were for CQ, 14 for AL and 10 for AS+SP were tested from Uttar Pradesh (U.P.), Mizoram, Meghalaya and Gujrat states. One out of 45 samples showed variable disintegration and retension factor. The variable disintegration and retention factor which would have been due to substandard quality or other factors including storage. However, HPLC analysis confirms standard active pharmaceutical ingredient, but may be due to humid temperature and moisture in storage may account for the observed result.

Keywords: antimalarial medicines, counterfeit, substandard, TLC

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Authors: Yen-An Tsai, Ching-Ling Lin, Jia-Woei Hou, Mei-Lien Chen

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Endocrine-disrupting chemicals (EDCs) adversely affect the endocrine system. Phthalates, also called phthalic acid esters (PAEs), are manmade chemicals that are used as stabilizing agents in personal care products such as perfumes, lotions, and cosmetics. The aim was to explore whether PAEs exposure was associated with central precocious puberty (CPP) or constitutional delays in growth (CDGP). This case-control study included 48 female with CPP, 37 male with constitutional delays in growth, and 127 normal children and was conducted from December 2011 to August 2014. All participants completed a structured questionnaire regarding socio-demographic characteristics, lifestyle, and secondary sexual characteristics. The analytical method was based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) with isotope dilution for the quantitative detection of several phthalate metabolites in human urine. The risk of CPP with mep, mnbp, LMW >50th percentile were higher than those with 50th percentile were higher than those with <50 percentile in model 2. In model 1, we only found higher CDGP risk in mep, mnbp, and ΣPAEs. It shows that high phthalate exposure may associate with CDGP. In this case-control study, we found PAEs exposure was associated with central precocious puberty (CPP) or constitutional delays in growth.

Keywords: phthalates, puberty, delays, growth

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Authors: Jeong Woo Kang, MiYoung Baek, Ki-Suk Kim, Kwang-Jick Lee, ByungJae So

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Introduction: A fast, easy and sensitive detection method was developed and validated by liquid chromatography tandem mass spectrometry for the determination of marbofloxacin in pig plasma which was further applied to study the pharmacokinetics of marbofloxacin. Materials and Methods: The plasma sample (500 μL) was mixed with 1.5 ml of 0.1% formic acid in MeCN to precipitate plasma proteins. After shaking for 20 min, The mixture was centrifuged at 5,000 × g for 30 min. It was dried under a nitrogen flow at 50℃. 500 μL aliquot of the sample was injected into the LC-MS/MS system. Chromatographic analysis was carried out mobile phase gradient consisting 0.1% formic acid in D.W. (A) and 0.1% formic acid in MeCN (B) with C18 reverse phase column. Mass spectrometry was performed using the positive ion mode and the selected ion monitoring (MRM). Results and Conclusions: The method validation was performed in the sample matrix. Good linearities (R2>0.999) were observed and the quantified average recoveries of marbofloxacin were 87 - 92% at level of 10 ng g-1 -100 ng g-1. The percent of coefficient of variation (CV) for the described method was less than 10 % over the range of concentrations studied. The limits of detection (LOD) and quantification (LOQ) were 2 and 5 ng g-1, respectively. This method has also been applied successfully to pharmacokinetic analysis of marbofloxacin after intravenous (IV), intramuscular (IM) and oral administration (PO). The mean peak plasma concentration (Cmax) was 2,597 ng g-1at 0.25 h, 2,587 ng g-1at 0.44 h and 2,355 ng g-1at 1.58 h for IV, IM and PO, respectively. The area under the plasma concentration-time curve (AUC0–t) was 24.8, 29.0 and 25.2 h μg/mL for IV, IM and PO, respectively. The elimination half-life (T1/2) was 8.6, 13.1 and 9.5 for IV, IM and PO, respectively. Bioavailability (F) of the marbofloxacin in pig was 117 and 101 % for IM and PO, respectively. Based on these result, marbofloxacin does not have any obstacles as therapeutics to develop the oral formulations such as tablets and capsules.

Keywords: marbofloxacin, LC-MS/MS, pharmacokinetics, chromatographic

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Authors: Waranya Imprasittichai, Patsarawadee Paojinda, 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. In this study, we constructed gene fusions of Plasmodium falciparum OMPDC-OPRT (1,836 bp) in pTrcHisA vector and expressed as an 6xHis-tag bifunctional protein in three Escherichia coli strains (BL21, Rosetta, TOP10) at 18 °C, 25 °C and 37 °C. The recombinant bifunctional protein was partially purified by Ni-Nitrilotriacetic acid-affinity chromatography. Specific activities of OPRT and OMPDC domains in the bifunctional enzyme expressed in E. coli TOP10 cells were approximately 3-4-fold higher than those in BL21 cells. There were no enzymatic activities when the construct vector expressed in Rosetta cells. Maximal expression of the fused gene was observed at 18 °C and the bifunctional enzyme had specific activities of OPRT and OMPDC domains in a ratio of 1:2. These results provide greater yields and better catalytic activities of the bifunctional OMPDC-OPRT enzyme for further purification and kinetic study.

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

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Authors: Lila Boulekbache-Makhlouf, Kahina Djaoud, Myriam Tazarourte, Samir Hadjal, Khodir Madani

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In Algeria, important quantities of secondary date variety (Phoenix dactylifera L.) are generated in each campaign; their chemical composition is similar to that of commercial dates. The present work aims to valorize this common date variety (Degla-Beida) which is often poorly exploited. In this context, we tried to prepare syrup from the secondary date variety and to evaluate the effect of conventional extraction (CE) or water bath extraction (WBE) and alternative extraction (microwaves assisted extraction (MAE), and ultrasounds assisted extraction (UAE)) on its total sugar content (TSC), using response surface methodology (RSM). Then, the analysis of individual sugars was performed by high-performance liquid chromatography (HPLC). Maximum predicted TSC recoveries under the optimized conditions for MAE, UAE and CE were 233.248 ± 3.594 g/l, 202.889 ± 5.797 g/l, and 233.535 ± 5.412 g/l, respectively, which were close to the experimental values: 233.796 ± 1.898 g/l; 202.037 ± 3.401 g/l and 234.380 ± 2.425 g/l. HPLC analysis revealed high similarity in the sugar composition of date juices obtained by MAE (60.11% sucrose, 16.64% glucose and 23.25% fructose) and CE (50.78% sucrose, 20.67% glucose and 28.55% fructose), although a large difference was detected for that obtained by UAE (0.00% sucrose, 46.94% glucose and 53.06% fructose). Microwave-assisted extraction was the best method for the preparation of date syrup with an optimal recovery of total sugar content. However, ultrasound-assisted extraction was the best one for the preparation of date syrup with high content of reducing sugars.

Keywords: dates, extraction, RSM, sugars, syrup

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Authors: Sidali Khedidji, Noureddine Yassaa, Riad Ladji

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In this study, n-alkanes which are hazardous for the environment and human health were investigated in Sour El Ghozlane suburban atmosphere at a sampling point from April 2013 to Mai 2013. Ambient concentration measurements of n-Alkanes were carried out at a regional study of the cement industry in Sour El Ghozlane. During sampling, the airborne particulate matter was enriched onto PTFE filters by using a two medium volume samplers with or without a size-selective inlet for PM10 and TSP were used and each sampling period lasted approximately 24 h. The organic compounds were characterized using gas chromatography coupled with mass spectrometric detection (GC-MS). Total concentrations for n-Alkanes recorded in Sour El Ghozlane suburban ranged from 42 to 69 ng m-3. Gravimeter method was applied to the black smoke concentration data for Springer seasons. The 24 h average concentrations of n-alkanes contain the PM10 and TSP of Sour El Ghozlane suburban atmosphere were found in the range 0.50–7.06 ng/m3 and 0.29–6.97 ng/m3, respectively, in the sampling period. Meteorological factors, such as (relative humidity and temperature) were typically found to be affecting PMs, especially PM10. Air temperature did not seem to be significantly affecting TSP and PM10 mass concentrations. The guide value fixed by the European Community, 40 μg/m3 was not to exceed 35 days, was exceeded in some samples. However, it should be noted that the value limit fixed by the Algerian regulations 80 μg/m3 has been exceeded in 1 sampler during the period study.

Keywords: n-alkanes, PM10, TSP, particulate matter, cement industry

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Authors: Ishmael Ntlhamu

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In Limpopo Province, the use of herbal concoctions is becoming very popular. These concoctions are claimed to be capable of treating ulcers, diabetes, certain STDs, blood cleansing, and many more types of diseases. The aim of this study was to evaluate the phytochemical composition, evaluate the pharmacological effects and consumption safety in herbal concoctions to treat various kinds of ailments in Limpopo. The concoctions were extracted with 80% acetone. Microorganisms in the concoctions were identified using the Vitek 2 compact system. Qualitative phytochemical analysis was determined using standard chemical tests and thin layer chromatography (TLC). Total polyphenol content was quantified. Antioxidant activity was quantified using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay and ferric reducing power. Antimicrobial activities were determined using a broth micro-dilution assay and bioautography. Cell viability assay was used to determine the cytotoxicity. Results showed that concoctions had antioxidant activity. Presence of different phytoconstituents was observed. Isolated microorganisms were identified as Burkholderia pseudomallei, Staphylococcus vitulimus, Enterococcus columbae, Kocuria kristanae, Staphylococcus intermedius, Cryptococcus laurenti. and Burkholderia pseudomallei (highly pathogenic). Therefore, phytochemicals prove that the concoctions can heal as the antimicrobial tests also displayed activity. Moreover, the concoctions did not exhibit cytotoxic effects. However, contaminants raise concerns, not only for consumer safety but also the quality of herbal concoctions available as part of the traditional medicinal practice in Limpopo.

Keywords: antimicrobials, concoctions, cytotoxicity, phytochemicals

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Authors: Anjana Sagar

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Arsenic poisoned ground water is a major concern in South Asia. The arsenic enters the food chain not only through drinking but also by using arsenic polluted water for irrigation. Arsenic is highly toxic in its inorganic forms; however, organic forms of arsenic are comparatively less toxic. In terrestrial plants, inorganic form of arsenic is predominantly found; however, we found that significant proportion of organic arsenic was present in root and shoot of a staple legume, chickpea (Cicer arientinum L) plants. Chickpea plants were raised in pot culture on soils spiked with arsenic ranging from 0-70 mg arsenate per Kg soil. Total arsenic concentrations of chickpea shoots and roots were determined by inductively coupled plasma-mass-spectrometry (ICP-MS) ranging from 0.76 to 20.26, and 2.09 to 16.43 µg g⁻¹ dry weight, respectively. Information on arsenic species was acquired by methanol/water extraction method, with arsenic species being analyzed by high-performance liquid chromatography (HPLC) coupled with ICP-MS. Dimethylarsinic acid (DMA) was the only organic arsenic species found in amount from 0.02 to 3.16 % of total arsenic shoot concentration and 0 to 6.93 % of total arsenic root concentration, respectively. To investigate the source of the organic arsenic in chickpea plants, arsenic species in the rhizosphere of soils of plants were also examined. The absence of organic arsenic in soils would suggest the possibility of formation of DMA in plants. The present investigation provides useful information for better understanding of distribution of arsenic species in terrestrial legume plants.

Keywords: arsenic, arsenic speciation, dimethylarsinic acid, organoarsenic

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Authors: Sandra Tapin-Lingua, Katia Ruel, Jean-Paul Joseleau, Daouia Messaoudi, Olivier Fahy, Michel Petit-Conil

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The preservative efficacy of organic biocides is strongly related to their capacity of penetration and retention within wood tissues. The specific detection of the pyrethroid insecticide is currently obtained after extraction followed by chemical analysis by chromatography techniques. However visualizing the insecticide molecule within the wood structure requires specific probes together with microscopy techniques. Therefore, the aim of the present work was to apply a new methodology based on antibody-antigen recognition and electronic microscopy to visualize directly pyrethroids in the wood material. A polyclonal antibody directed against cypermethrin was developed and implement it on Pinus sylvestris wood samples coated with technical cypermethrin. The antibody was tested on impregnated wood and the specific recognition of the insecticide was visualized in transmission electron microscopy (TEM). The immunogold-TEM assay evidenced the capacity of the synthetic biocide to penetrate in the wood. The depth of penetration was measured on sections taken at increasing distances from the coated surface of the wood. Such results correlated with chemical analyzes carried out by GC-ECD after extraction. In addition, the immuno-TEM investigation allowed visualizing, for the first time at the ultrastructure scale of resolution, that cypermethrin was able to diffuse within the secondary wood cell walls.

Keywords: cypermethrin, insecticide, wood penetration, wood retention, immuno-transmission electron microscopy, polyclonal antibody

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Authors: Samia Khadhar, Anis Chekirbene, Nouha Khiari, Amira Mabrouki

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The persistent organic pollutants (POPs) in aquatic environments are one of the most worrying problems for environmental sustainability and human health because of their carcinogenic and toxic characteristics. Human anthropogenic actions (wastewater discharges, agricultural and industrial activities) without prior treatment are the main cause of this organic pollution. Oued Madjerda is considered the most important river in Tunisia, hence the importance of assessing the level of organic pollution of water and sediments, taking into account the anthropogenic stress exerted on this river. Water and sediment samples were taken from the middle and lower valley of the Medjerda to determine the state of contamination by 7PCBs, priority 15PAHs, and pesticides. The analysis was performed by gas chromatography (GC) and liquid phase coupled to HPLC MS-MS mass spectroscopy. The results showed that for the waters, the total PAH and PCB contents vary respectively from 0.0023 to 7.72 mg/l and from 0.0001 to 0.179 mg/l. In surface sediments 0 to 15 cm, 7PCB levels vary from 1,112 to 110,062 µg/kg-1. In this study, we tried to determine the concentration of 96 pesticides in surface sediments; analyzes showed the presence of Buprofezin, propamocarb-HCl, hexaconazole, flutriafol, quinalphos, and tebufenpyrad with concentrations varying from 1.06 to 2.388 µg/kg-1. The pace of the spatial variation confirms the impact of wastewater discharged on the quality of water and sediments despite the perennial aspect of the river.

Keywords: Wadi Madjerda, organic pollution, water and sediments surface, anthropics stress

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Authors: Mateen Abbas, Abdul Muqeet Khan, Sadia Bashir, Muhammad Awais Khalid, Aamir Ghafoor, Zara Hussain, Mashal Shahid

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The irrational use of insecticides in everyday life has drawn attention worldwide towards its harmful effects. To mitigate the toxic effects of insecticides to humans, present study was planned on the degradation/detoxification of the neonicotinoid insecticides including imidacloprid and acetamiprid. Biocarbon of fruit peels (Banana & Watermelon) and biochar (activated or non-activated) of rice husk was utilized as adsorbents for degradation of selected pesticides. Both activated and non-activated biochar were prepared for treatment and then applied in different concentrations (0.5 to 2.0 ppm) and dosage (1.0 to 2.5g) to insecticides (Acetamiprid & Imidacloprid) as well as studied at different times (30-120 minutes). Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) coupled with Photodiode array detector was used to quantify the insecticides. Results depicted that activated biochar of rice husk minimized the 73% concentrations of both insecticides however, watermelon activated biocarbon degraded 72% of imidacloprid and 56% of acetamiprid. Results proved the efficiency of the method employed and it was also inferred that high concentration of biocarbon resulted in larger percentage of degradation. The applied method is cheaper, easy and accessible that can be used to minimize the pesticide residues in animal feed. Degradation using biochar proved significant degradation, eco-friendly and economic method to reduce toxicity of insecticides.

Keywords: insecticides, acetamiprid, imidacloprid, biochar, HPLC

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Authors: Julius Ilawe Osayi, Peter Osifo

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Solid waste disposal, such as used tires is a global challenge as well as energy crisis due to rising energy demand amidst price uncertainty and depleting fossil fuel reserves. Therefore, the effectiveness of pyrolysis as a disposal method that can transform used tires into liquid fuel and other end-products has made the process attractive to researchers. Although used tires have been converted to liquid fuel using pyrolysis, there is the need to improve on the liquid fuel properties. Hence, this paper reports the investigation of zeolite NaY synthesized from kaolin, a locally abundant soil material in the Benin metropolis as a suitable catalyst and its effect on the properties of pyrolytic oil produced from used tires. The pyrolysis process was conducted for a range of 1 to 10 wt.% of catalyst concentration to used tire at a temperature of 600 oC, a heating rate of 15oC/min and particle size of 6mm. Although no significant increase in pyrolytic oil yield was observed compared to the previously investigated non-catalytic pyrolysis of a used tire. However, the Fourier transform infrared (FTIR), Nuclear Magnetic Resonance (NMR); and Gas chromatography-mass spectrometry (GC-MS) characterization results revealed the pyrolytic oil to possess an improved physicochemical and fuel properties alongside valuable industrial chemical species. This confirms the possibility of transforming kaolin into a catalyst suitable for improved fuel properties of the liquid fraction obtainable from thermal cracking of hydrocarbon materials.

Keywords: catalytic pyrolysis, fossil fuel, kaolin, pyrolytic oil, used tyres, Zeolite NaY

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Authors: Candra Irawan, David Yudianto, Ahsanu Nadiyya, Dewi Anna Br Sitepu, Hanafi, Erna Styani

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Jelly drink was produced from a combination of both natural and synthetic materials, such as acesulfame potassium (acesulfame-K) as synthetic sweetener material. Acesulfame-K content in jelly drink could be determined by High-Performance Liquid Chromatography (HPLC), but this method needed validation due to having a change on the reagent addition step which skips the carrez addition and comparison of mix mobile phase (potassium dihydrogen phosphate and acetonitrile) with ratio from 75:25 to 90:10 to be more efficient and cheap. This study was conducted to evaluate the performance of determination method for acesulfame-K content in the jelly drink by HPLC. The method referred to Deutsches Institut fur Normung European Standard International Organization for Standardization (DIN EN ISO):12856 (1999) about Foodstuffs, Determination of acesulfame-K, aspartame and saccharin. The result of the correlation coefficient value (r) on the linearity test was 0.9987 at concentration range 5-100 mg/L. Detection limit value was 0.9153 ppm, while the quantitation limit value was 1.1932 ppm. The recovery (%) value on accuracy test for sample concentration by spiking 100 mg/L was 102-105%. Relative Standard Deviation (RSD) value for precision and homogenization tests were 2.815% and 4.978%, respectively. Meanwhile, the comparative and stability tests were tstat (0.136) < ttable (2.101) and |µ1-µ2| (1.502) ≤ 0.3×CV Horwitz. Obstinacy test value was tstat < ttable. It can be concluded that the HPLC  method for the determination of acesulfame-K in jelly drink product by HPLC has been valid and can be used for analysis with good performance.

Keywords: acesulfame-K, jelly drink, HPLC, validation

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Authors: Juan Jose Filgueira, Daniela Londono-Serna, Liliana Maria Hoyos

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Carnation (Dianthus caryophyllus) is one of the most important products of exportation in the floriculture industry worldwide. Fusariosis is the disease that causes the highest losses on farms, in particular the one produced by Fusarium oxysporum f.sp. dianthi, called vascular wilt. Gene identification and metabolic routes of the genes that participate in the building of the plant response to Fusarium are some of the current targets in the carnation breeding industry. The techniques for the identifying of resistant genes in the plants, is the analysis of the transcriptome obtained during the host-pathogen interaction. In this work, we report the cell transcriptome of different varieties of carnation that present differential response from Fusarium oxysporum f.sp. dianthi attack. The cells of the different hybrids produced in the outbreeding program were cultured in vitro and elicited with the parasite in a dual culture. The isolation and purification of mRNA was achieved by using affinity chromatography Oligo dT columns and the transcriptomes were obtained by using Illumina NGS techniques. A total of 85,669 unigenes were detected in all the transcriptomes analyzed and 31,000 annotations were found in databases, which correspond to 36.2%. The library construction of genic expression techniques used, allowed to recognize the variation in the expression of genes such as Germin-like protein, Glycosyl hydrolase family and Cinnamate 4-hydroxylase. These have been reported in this study for the first time as part of the response mechanism to the presence of Fusarium oxysporum.

Keywords: Carnation, Fusarium, vascular wilt, transcriptome

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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

Procedia PDF Downloads 278