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Min-Zhuo Cheng, Jing Zhang, Dequan Bao, Xiwei Huang
In this communication, we assembled ordered polystyrene (PS) microsphere array as a template with the drop-coating method, and the oxygen plasma was used to etch the template to adjust the spacing between the PS microspheres. Nano-triangular gold array and silver Nano-pyramid array were obtained by ion beam sputtering to deposit precious metal gold and silver. We observed the surface morphology of Au and Au/Ag composite films by scanning electron microscope and characterized the films by X-ray diffraction and Ultraviolet/visible light spectrophotometer...
May 21, 2018: Electrophoresis
Macarena Silva, Carolina Mendiguchía, Carlos Moreno, Pavel Kubáň
Simultaneous electromembrane extraction (EME) of six trace metal cations (Cu2+ , Zn2+ , Co2+ , Ni2+ , Pb2+ , Cd2+ ) from saline samples was investigated. Capillary electrophoresis (CE) with capacitively coupled contactless conductivity detection (C4 D) was used to determine the metals in acceptor solutions due to its excellent compatibility with the minute volumes of acceptor solutions. Bis(2-ethylhexyl)phosphate (DEHPA) was selected as a suitable non-selective modifier for EME transport of target metal cations...
May 21, 2018: Electrophoresis
Rabia Raza, Yu Bai, Huwei Liu
Overexpression of ecto-5'-nucleotidase is found to be linked to cancer progression and other diseases which makes screening of its inhibitors on demand. Unfortunately, there is scarcity of reported ecto-5'-nucleotidase inhibitors because of unavailability of simple, fast and efficient methods to study its inhibition. In this study, we have developed a very fast, reverse polarity capillary electrophoresis based method for screening of ecto-5'-nucleotidase inhibitors. Both EOF and electrophoretic mobility were maintained in same direction by dynamically coating lining of capillary and applying negative voltage, resulting in appearance of both adenosine monophosphate (AMP) and adenosine peaks in less than two minutes...
May 18, 2018: Electrophoresis
Imma Pagano, Anna Lisa Piccinelli, Rita Celano, Luca Campone, Patrizia Gazzerro, Mariateresa Russo, Luca Rastrelli
Artichoke by-products are a suitable source of health-promoting ingredients for the production of dietary supplements and food additives. A pressurized hot water extraction (PHWE) was developed to recover caffeoylquinic acids (CQAs) and flavone glycosides (FLs) from agro-industrial artichoke by-products. The main factors influencing PHWE efficiency and CQA isomerization (temperature, numbers of cycles, modifier and extraction time) were carefully studied and optimized by response surface design. The proposed PHWE procedure provides an exhaustive extraction of CQAs and FLs (recoveries: 93-105 and 90-105%) from artichoke external bracts and leaves of different cultivars (p > 0...
May 18, 2018: Electrophoresis
Mate Szarka, Szabolcs Szilasi, Boglarka Donczo, Daniel Sarkozy, Istvan Rajta, Andras Guttman
On a roundtrip to Mars, astronauts are expectedly exposed to an approximate amount of radiation that exceeds the lifetime limits on Earth. This elevated radiation dose is mainly due to Galactic Cosmic Rays (GCR) and Solar Particle Events (SPE). Specific patterns of the N-glycosylation of human immunoglobulins have already been associated with various ailments such as autoimmune diseases, malignant transformation, chronic inflammation and ageing. The focus of our work was to investigate the effect of low-energy proton irradiation on the IgG N-glycosylation profile with the goal if disease associated changes could be detected during space travel and not altered by space radiation...
May 18, 2018: Electrophoresis
Bolin Zhu, Miaoduo Deng, Yaqi Yao, Jia Yu, Qing Li
Immobilized polysaccharide-based columns showed excellent enantioselectivity in normal phase separation mode. In this work, enantioseparation abilities of four immobilized polysaccharide-derived chiral stationary phases (Chiralpak IA, Chiralpak IB, Chiralpak IC and Chiralpak ID) toward fifteen azole compounds were evaluated. Separation was carried out using n-hexane as mobile phase with ethanol, 1-propanol, 1-butanol and 2-propanol as modifiers. And twelve compounds have achieved baseline separation with the resolutions ranging between 2...
May 18, 2018: Electrophoresis
Nerea Muñoz-Almagro, Fabián Rico-Rodriguez, Peter J Wilde, Antonia Montilla, Mar Villamiel
An optimisation of temperature, time and extracting agent concentration of pectin extraction from sunflower heads using sodium citrate and nitric acid (SP-SC and SP-NA) was carried out. At optimal conditions, the yield of extraction with nitric acid (SPO-NA) was 2-fold greater than the corresponding with sodium citrate (SPO-SC) (14.3 vs 7.7%, respectively). Regarding pectin structure, the galacturonic acid (GalA) content in both, SPO-SC and SPO-NA, was similar (∼85%). However, SPO-NA showed lower molecular weight (Mw) (88...
May 18, 2018: Electrophoresis
Alexandre Goyon, Yannis Nicolas Francois, Olivier Colas, Alain Beck, Jean Luc Veuthey, Davy Guillarme
The determination of mAb critical quality attributes (CQA) is crucial for their successful application in health diseases. A generic CZE method was developed for the high-resolution separation of various mAb charge variants, which are often recognized as important CQA. A dynamic coating of the capillary was obtained with polyethylene oxide (PEO), whereas Bis-Tris allowed the analysis of mAbs under native conditions at pH 7.0. The effect of PEO and Bis-Tris concentrations, as well as the nature of the acidic counter ion on the method performance were systematically studied...
May 17, 2018: Electrophoresis
Nathália Soares Simões, Hanna Leijoto de Oliveira, Ricky Cássio Santos da Silva, Leila Suleimara Teixeira, Thaís Lorenna Souza Sales, Whocely Victor de Castro, Maria José Nunes de Paiva, Cristina Sanches, Keyller Bastos Borges
In this work a hollow mesoporous structured molecularly imprinted polymer was synthetized and used as adsorbent in pipette-tip solid-phase extraction for the determination of lamivudine (3TC), zidovudine (AZT) and efavirenz (EFZ) from plasma of human immunodeficiency virus (HIV) infected patients by high-performance liquid chromatography (HPLC). All parameters that influence the recovery of the pipette tip based on hollow mesoporous molecularly imprinted polymer solid-phase extraction (PT-HM-MIP-SPE) method were systematically studied and discussed in detail...
May 17, 2018: Electrophoresis
Sarah Seashols-Williams, Raquel Green, Denise Wohlfahrt, Angela Brand, Antonio Limjuco Tan-Torres, Francy Nogales, J Paul Brooks, Baneshwar Singh
Sequencing and classification of microbial taxa within forensically relevant biological fluids has the potential for applications in the forensic science and biomedical fields. The quantity of bacterial DNA from human samples is currently estimated based on quantity of total DNA isolated. This method can miscalculate bacterial DNA quantity due to the mixed nature of the sample, and consequently library preparation is often unreliable. We developed an assay that can accurately and specifically quantify bacterial DNA within a mixed sample for reliable 16S ribosomal DNA (16S rDNA) library preparation and high throughput sequencing (HTS)...
May 17, 2018: Electrophoresis
Qing-Xia Zhang, Meng Yang, Ya-Jiao Pan, Jing Zhao, Bao-Wang Qu, Feng Cheng, Ya-Ran Yang, Zhang-Ping Jiao, Li Liu, Jiang-Wei Yan
Massively parallel sequencing (MPS) has been used in forensic genetics in recent years owing to several advantages, such as MPS can provide precise descriptions of the repeat allele structure and variation in the repeat-flanking regions, increasing the discriminating power among loci and individuals. However, it cannot be fully utilized unless sufficient population data is available for all loci. Thus, there is a pressing need to perform population studies providing a basis for the introduction of MPS into forensic practice...
May 17, 2018: Electrophoresis
Julia Kahle, Hermann Wätzig
Biopharmaceuticals are increasingly present in pharmaceutical therapy. These highly complex molecules are still challenging for development and quality control. Charge heterogeneity is next to size heterogeneity, glycan analysis or peptide mapping one main aspect of their thorough characterization. This review focuses on the charge-based analysis of biopharmaceuticals through capillary electrophoresis. An overview is given about the three separation techniques cIEF, icIEF and CZE for the analyses of charge variants, emphasizing assays and their precision...
May 17, 2018: Electrophoresis
Proteinase pattern of honeybee prepupae from healthy and American Foulbrood infected bees investigated by zymography: The numbered spots (figure 4) were excised from the gels and they were individually transferred to a 1.5 mL microcentrifuge tube. Spots were destained and proteins digested as reported by Dani et al. 40. Each peptide mixture was submitted to capillary-LC-μESI-MS/MS analysis on an Ultimate 3000 HPLC (Dionex, San Donato Milanese, Milano, Italy) coupled to a LTQ Orbitrap mass spectrometer (Thermo Fisher, Bremen, Germany). Peptides were concentrated on a precolumn cartridge PepMap100 C18 (300 μm i.d. × 5 mm, 5 μm, 100 Å, LC Packings Dionex) and then eluted on a homemade capillary column packed with Aeris Peptide XB-C18 phase (180 μm i.d. × 15 cm, 3.6 μm, 100 Å, Phenomenex, Torrance, CA, USA) at 1 μl/min. The loading mobile phases were: 0.1% TFA in H 2 O (phase A) and 0.1% TFA in CH 3 CN (phase B). The elution mobile phases composition was: H 2 O 0.1% formic acid/CH 3 CN 97/3 (phase A) and CH 3 CN 0.1% formic acid/ H 2 O 97/3 (phase B). The elution program was: 0 min, 4% B; 10 min, 40% B; 30 min, 65% B; 35 min, 65% B; 36 min, 90% B; 40 min, 90% B; 41 min, 4%B; 60 min, 4% B. Mass spectra were acquired in positive ion mode, setting the spray voltage at 1.8 kV, the capillary voltage and temperature respectively at 45 V and 200°C, and the tube lens at 130 V. Data were acquired in data-dependent mode with dynamic exclusion enabled (repeat count 2, repeat duration 15 s, exclusion duration 30 s); survey MS scans were recorded in the Orbitrap analyzer in the mass range 300-2000 m/z at a 15,000 nominal resolution at m/z = 400; then up to three most intense ions in each full MS scan were fragmented (isolation width 3 m/z, normalized collision energy 30) and analyzed in the IT analyzer. Monocharged ions did not trigger MS/MS experiments.
Antonio Felicioli, Barbara Turchi, Filippo Fratini, Matteo Giusti, Roberta Nuvoloni, Francesca Romana Dani, Simona Sagona
American foulbrood disease (AFB) is the main devastating disease that affects honeybees' brood, caused by Paenibacillus larvae. The trend of the research on AFB has addressed the mechanisms by which P. larvae bacteria kill honeybee larvae. Since prepupae could react to the infection of AFB by increasing protease synthesis, the aim of this work was to compare protease activity in worker prepupae belonging to healthy colonies and to colonies affected by AFB. This investigation was performed by zymography. In gel, proteolytic activity was observed in prepupae extracts belonging to the healthy colonies...
May 15, 2018: Electrophoresis
Markus Islinger, Robert Wildgruber, Alfred Völkl
Free-flow electrophoresis (FFE) exploits differences in the overall charge of bio-particles separating cells, organelles, macromolecules, ions etc. according to their distinct electrophoretic mobility and isoelectric point (pI) values. Indeed, around a neutral pH organelles usually exhibit a negative surface charge, migrating in an electric field from the cathode towards the anode. Since its introduction more than five decades ago by Barrollier et al. (1958) and Hannig (1961), FFE has become an established analytical and preparative separation method for the isolation of a variety of organelles...
May 15, 2018: Electrophoresis
Yaqi Yao, Yu Zhao, Xue Zhang, Jia Yu, Xingjie Guo
A method consisting of cation-selective exhaustive injection and sweeping (CSEI-sweeping) as online preconcentration followed by a cyclodextrin modified electrokinetic chromatography (CDEKC) enantioseparation has been developed for the simultaneous determination of two brompheniramine enantiomers in rat plasma. In this method, analytes were electrokinetically injected at a voltage of 8 kV for 80 s in a fused-silica capillary. Prior to the injection, the capillary was rinsed with 50 mM phosphate buffer pH 3...
May 13, 2018: Electrophoresis
Xuan Liu, Xu Yan, Jinfeng Bi, Jianing Liu, Mo Zhou, Xinye Wu, Qinqin Chen
White-flesh guava is widely planted in tropical or subtropical areas of Southeast Asia. Despite of folk statements on specific function, few researches are focused on the description of its plant secondary metabolites. In the present work, contents of total phenolics and flavonoids as well as antioxidant activity from different parts (peel, flesh and seed) were determined. The constituents of ethanol extracts were characterized by HPLC-QTOF-MS. A total of 69 phenolic compounds as well as 9 polar compounds were detected, with flavonoids, hydrolyzable tannins, phenolic acid derivatives and benzophenones of the four predominant phenolic compounds...
May 13, 2018: Electrophoresis
Roshanak Aslebagh, Devika Channaveerappa, Kathleen F Arcaro, Costel C Darie
Breast Cancer (BC) remains a major cause of mortality, and early detection is considered important for reducing BC-associated deaths. Early detection of BC is challenging in young women, due to the limitations of mammography on the dense breast tissue of young women. We recently reported results of a pilot proteomics study, using one-dimensional polyacrylamide gel electrophoresis (1D-PAGE) and mass spectrometry (MS) to investigate differences in milk proteins from women with and without BC. Here, we applied two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MS to compare the protein pattern in milk from the breasts of a single woman who was diagnosed with BC in one breast 24 months after donating her milk...
May 13, 2018: Electrophoresis
Ozan Nazim Ciftci, Jessica Cahyadi, Selma E Guigard, Marleny D A Saldaña
Malaria is a high priority life-threatening public health concern in developing countries, and therefore there is a growing interest to obtain artemisinin for the production of artemisinin-based combination therapy products. In this study, artemisinin was extracted from the Artemisia annua L. plant using supercritical carbon dioxide (SC-CO2 ) modified with ethanol. Response surface methodology (RSM) based on central composite rotatable design (CCRD) was employed to investigate and optimize the extraction conditions of pressure (9...
May 13, 2018: Electrophoresis
Rebecca Richards, Jayshree Patel, Kate Stevenson, SallyAnn Harbison
Epigenetics is an emerging area of interest in forensic science. DNA methylation, a type of epigenetic modification, can be applied to chronological age estimation, identical twin differentiation and body fluid identification. However, there is not yet an agreed, established methodology for targeted detection and analysis of DNA methylation markers in forensic research. Recently a massively parallel sequencing-based approach has been suggested. The use of massively parallel sequencing is well established in clinical epigenetics and is emerging as a new technology in the forensic field...
May 11, 2018: Electrophoresis
Antonio Alonso, Pedro A Barrio, Petra Müller, Steffi Köcher, Burkhard Berger, Pablo Martin, Martin Bodner, Sascha Willuweit, Walther Parson, Lutz Roewer, Bruce Budowle
The current state of validation and implementation strategies of MPS technology for the analysis of STR markers for forensic genetics use is described, covering the topics of the current catalogue of commercial MPS-STR panels, leading MPS-platforms, and MPS-STR data analysis tools. In addition, the developmental and internal validation studies carried out to date to evaluate reliability, sensitivity, mixture analysis, concordance, and the ability to analyze challenged samples are summarized. The results of various MPS-STR population studies that showed a large number of new STR sequence variants that increase the power of discrimination in several forensically-relevant loci are also presented...
May 11, 2018: Electrophoresis
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