Carbon dots derived from citric acid and urea as fluorometric probe for determining melamine contamination in infant formula sample.

Melamine has been intentionally added into food products to increase the protein count at less cost, especially in dairy products for infant resulting in serious adverse effects on health of consumers. Therefore, this study aimed to develop a method to quantify melamine in dairy products based on the change of fluorescent properties of carbon dots (CDs) as sensing probe. CDs with green-fluorescent emission were synthesized from citric acid and urea under microwave irradiation. The synthesized CDs emitted fluorescence at the maximum wavelength of 538 nm with excitation wavelength of 410 nm. Thus, they provided high sensitivity and selectivity on melamine detection by which fluorescent emission of the CDs was increasingly quenched upon increasing melamine concentrations. Optimal conditions for melamine determination using the CDs was under pH 6, volume ratio between CDs and sample of 2:8 and reaction time of 15 min. The developed method provided high precision of melamine determination with less than 5% of %RSD (n = 5), wide detection range from 1.0 to 200.0 ppm, and high sensitivity with limit of detection (LOD) of 0.47 ppm and limit of quantification (LOQ) of 1.56 ppm, which is within the regulated level by the Food and Drug Administration of the United States for melamine in dairy products. Several analytical characterization techniques were conducted to elucidate the reaction mechanism between CDs and melamine, and the hydrogen bonding interaction was proposed.

Synthesis and application of fluorescent N-doped carbon dots/hydrogel composite for Cr(VI) adsorption: Uncovering the ion species transformation and fluorescent quenching mechanism.

A fluorescent composite material fabricated from nitrogen-doped carbon dots with polyvinyl alcohol (PVA)/polyvinylpyrrolidone (PVP)/citric acid (CA) hydrogel was synthesized using a microwave-assisted hydrothermal method. The composite was used as a metal ion sensor and adsorbent to remove chromium (Cr(VI)) from water. The chemical structure and Cr(VI) removal performance of the fluorescent composite films were also characterized. Fluorescent quenching upon Cr(VI) adsorption showed that Cr(VI) binding was attributed to the N-doped carbon dots. The results were confirmed by several analytical techniques, including X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR) and X-ray absorption spectroscopy (XAS). The mechanism of Cr(VI) removal from water by the fluorescent composite film was based on the adsorption and subsequent reduction of N-doped carbon dots within the 3D porous composite film. XPS measurements showed that 53.2% Cr(III) and 46.8% Cr(VI) were present on the composite surface after Cr(VI) adsorption. Moreover, XAS revealed a change in the oxidation state of Cr(VI) to Cr(III) after adsorption and in the Cr-O bond length (1.686 Å to 2.284 Å) after reduction. The Cr(VI) adsorption capacity of the composite film was 4.90 mg g-1 at pH 4 and fit the pseudo-second-order kinetic and Freundlich models. The results of this study could be used as a platform to further apply CDs/HD composites to remove Cr(VI) from water sources.

An application of miniaturized electrochemical sensing for determination of arsenic in herbal medicines.

This study aimed to create a miniaturized electrochemical platform for detecting As(III) contamination in herbal medicines. To reduce the operational steps of modification and determination, only a single drop of mixed standard Au(III) and sample solution is proposed to perform the electrochemical measurements using a screen-printed graphene electrode (SPGE). Square wave anodic stripping voltammetry was employed to integrate the simultaneous modification and determination processes. To perform the measurement, As(III) and Au(III) migrate to the SPGE surface while the reduction potential is held at -0.5 V, forming an Au-As intermetallic alloy. Then, As is stripped off for the electrochemical determination of As(III). The total assay time is less than 3 min. Under suitable conditions, the electrochemical sensing system can detect As(III) at concentrations ranging from 0.1 to 3.0 ppm, with a limit of quantification and limit of detection of 0.1 and 0.03 ppm, respectively. The applicability and accuracy of the proposed sensor were verified by determining As(III) in herbal medicinal samples, and they were found to be in line with the standard method (ICP-OES). The benefits of simple operation, rapid detection, portability, and low cost (<1 USD) make this a more powerful tool for routine monitoring and on-site analysis applications.

Development of a microfluidic membraneless vaporization flow system for trace analysis of arsenic.

A new design of a membraneless vaporization (MBL-VP) unit coupled with a specific flow system is presented for the determination of arsenic at trace levels using a hydride generation process. The MBL-VP unit contains two concentric conical reservoirs, with the outer cone selected as the donor reservoir. The volume of the outer donor reservoir is thereby greater than the acceptor volume, necessary for holding sufficient sample and reagents for the generation of arsine gas by reaction between As(iii) and sodium borohydride under acidic conditions. The arsine gas diffuses into the narrow headspace and is absorbed by an aliquot of 150 µL of mercuric chloride acceptor solution. The resulting reaction produces hydronium ions which is monitored by the absorbance change at 530 nm of the methyl orange indicator added in the acceptor solution. To decrease the detection limit, the aspiration and removal of the donor plug, comprising the sample, borohydride and acid, into and out of the donor cone are repeated several times, while the acceptor solution is kept unchanged. As a result, analysis of arsenic was achieved in the range of 10 to 100 µg L-1 with a detection limit of 8 µg L-1. Application to surface water was investigated. Percent recoveries of spiked surface water samples were in the range of 94-110%. For comparison of total arsenic (As(iii) and As(v)), the results obtained from the developed method are not statistically different from the ICP-OES method.

A Facile Synthesis of Self-Catalytic Hydrogel Films and Their Application as a Wound Dressing Material Coupled with Natural Active Compounds.

A simple and economical method for polyvinyl alcohol/polyvinylpyrrolidone/citric acid (PVA/PVP/CA) hydrogel preparation using microwave-assisted irradiation was presented. The synthesized hydrogels embedded with berberine or chlorogenic acid were investigated as a wound dressing agent. This study showed that the optimum condition for the hydrogel synthesis based on gel fraction and a degree of swelling values was 6:6:3% (w/v) of PVA/PVP/CA under 600 W at 120 °C for 3 min of microwave irradiation. Herbal active compounds, berberine and chlorogenic acid, were loaded onto the hydrogel (4% (w/v)), and both were able to inhibit the growth of Staphylococcus aureus. Additionally, the anti-inflammatory study revealed that 700 µg/mL berberine and 2500 µg/mL chlorogenic acid could inhibit protein degradation equivalent to a 200 µg/mL aspirin solution. The drug release study demonstrated that both compounds showed a more sustained release into PBS than water. The mechanism for the three-dimensional network formation based on esterification and the hydrogen-bonding interaction was also proposed. The ionic liquid-like structure of PVP-CA possibly played an important role in the cross-linking process. In addition, sodium bicarbonate applied to the synthesized hydrogel also had a significant effect in enhancing gel formation. The proposed approach showed a potential of the loaded hydrogels to protect wounds from infection and enhance the healing process.

A simple paper-based approach for arsenic determination in water using hydride generation coupled with mercaptosuccinic-acid capped CdTe quantum dots.

This research aims to develop a simple paper-based device for arsenic detection in water samples where a hydride generation technique coupled with mercaptosuccinic acid-capped CdTe quantum dots (MSA-CdTe QDs) as a detection probe was applied to the detection system. MSA-CdTe QDs were coated on a paper strip, inserted into the cover cap of a reaction bottle, to react with the developed arsine gas. Fluorescent emission of the QDs was quenched upon the presence of arsenic in solutions, whereby only a small amount of the MSA-CdTe QDs was required. The excitation and emission wavelengths for fluorescent detection were 278.5 nm and 548.5 nm, respectively. The proposed system provided a limit of detection of 0.016 mg L-1 and a limit of quantitation of 0.053 mg L-1, and a detection range of 0.05-30.00 mg L-1. In addition, the tolerance level of the detection approach to interference by other vapor-generated species was successfully improved by placing another paper strip coated with a solution of saturated lead acetate in front of the detection paper strip. This developed approach offered a simple and fast, yet accurate and selective detection of arsenic contaminated in water samples. In addition, the mechanism of fluorescent quenching was also proposed.

Removal of Heavy Metal Ions Using Modified Celluloses Prepared from Pineapple Leaf Fiber.

Since large amounts of pineapple leaves are abandoned after harvest in agricultural areas, the possibility of developing value-added products from them is of interest. In this work, cellulose fiber was extracted from pineapple leaves and modified with ethylenediaminetetraacetic acid (EDTA) and carboxymethyl (CM) groups to produce Cell-EDTA and Cell-CM, respectively, which were then used as heavy metal ion adsorbents. A solution of either lead ion (Pb2+) or cadmium ion (Cd2+) was used as wastewater for the purpose of studying adsorption efficiencies. The adsorption efficiencies of Cell-EDTA and Cell-CM were significantly higher than those of the unmodified cellulose in the pH range 1-7. Maximum adsorptions toward Pb2+ and Cd2+ were, for Cell-EDTA, 41.2 and 33.2 mg g-1, respectively, and, for Cell-CM, 63.4 and 23.0 mg g-1, respectively. The adsorption behaviors of Cell-CM for Pb2+ and Cd2+ fitted well with a pseudo-first-order model, but those of Cell-EDTA for Pb2+ and Cd2+ fitted well with a pseudo-second-order model. All of the adsorption behaviors could be described using the Langmuir adsorption isotherm. Desorption studies of Pb2+ and Cd2+ on both adsorbents using 1 M HCl suggested that regenerability of Cell-EDTA was, for both adsorbates, better than that of Cell-CM. Moreover, adsorption measurements in a mixture of Pb2+ and Cd2+ at various ratios showed that for both adsorbents the adsorption of Pb2+ was higher than that of Cd2+, while the adsorption selectivity for Pb2+ of Cell-CM was greater than that of Cell-EDTA. This study showed that the modified cellulosic adsorbents made from pineapple leaves were able to efficiently adsorb metal ions.

Novel colorimetric assay for paraquat detection on-silica bead using negatively charged silver nanoparticles.

A simple, rapid, sensitive, and economical method based on colorimetry for the determination of paraquat, a widely used herbicide, was developed. Citrate-coated silver nanoparticles (AgNPs) were synthesized as the colorimetric probe. The mechanism of the assay is related to the aggregation of negatively charged AgNPs as induced by positively-charged paraquat resulting from coulombic attraction which causes the color to change from a deep greenish yellow to pale yellow in accordance with the concentrations of paraquat. Silica gel was exploited as the paraquat adsorbent for purification and pre-concentration prior to the direct determination with negatively charged AgNPs without the requirement of the elution step. The validity of the proposed approach was evaluated by spiking standard paraquat in water and plant samples. Recoveries of paraquat in water samples were 93.6% and 95.4% for groundwater and canal water, respectively, while those in plant samples were 89.5% and 86.6% for Chinese cabbage and green apple, respectively,after using the optimized extraction procedure. The absorbance of AgNPs at 400nm was linearly related to the concentration of paraquat over the range of 0.05-50mgL-1, with detection limits of 0.05mgL-1 for water samples, and 0.10mgL-1 for plant samples by naked eye determination.

Simple and fast colorimetric detection of inorganic arsenic selectively adsorbed onto ferrihydrite-coated silica gel using silver nanoplates.

The optical detection for inorganic arsenic (As) semi-quantitative determination is presented by using silver nanoplates (AgNPls). The color of AgNPs is immediately changed in the presence of As(III) and As(V) with the same sensitivity. To improve the selectivity of AgNPls for As detection, ferrihydrite-coated silica gel (SiO2-Fh) was specifically exploited as adsorbent for arsenic prior to As detection by AgNPls. The developed method provides the detection limit of 0.5ppm with the detection range between 0.5ppm and 30.0ppm for As determination observed with naked eye, and allows to determine total inorganic arsenic. This is the first report of As detection approach combining As removal technology together with nanotechnology. This combined technique provides a rapid, sensitive and selective method for monitoring As levels in aqueous samples, and can be employed as a testing field kit to screen arsenic contamination outside of a laboratory.

Structural and mechanistic insight into N-glycan processing by endo-α-mannosidase.

N-linked glycans play key roles in protein folding, stability, and function. Biosynthetic modification of N-linked glycans, within the endoplasmic reticulum, features sequential trimming and readornment steps. One unusual enzyme, endo-α-mannosidase, cleaves mannoside linkages internally within an N-linked glycan chain, short circuiting the classical N-glycan biosynthetic pathway. Here, using two bacterial orthologs, we present the first structural and mechanistic dissection of endo-α-mannosidase. Structures solved at resolutions 1.7-2.1 Å reveal a (ß/α)(8) barrel fold in which the catalytic center is present in a long substrate-binding groove, consistent with cleavage within the N-glycan chain. Enzymatic cleavage of authentic Glc(1/3)Man(9)GlcNAc(2) yields Glc(1/3)-Man. Using the bespoke substrate α-Glc-1,3-α-Man fluoride, the enzyme was shown to act with retention of anomeric configuration. Complexes with the established endo-α-mannosidase inhibitor α-Glc-1,3-deoxymannonojirimycin and a newly developed inhibitor, α-Glc-1,3-isofagomine, and with the reducing-end product α-1,2-mannobiose structurally define the -2 to +2 subsites of the enzyme. These structural and mechanistic data provide a foundation upon which to develop new enzyme inhibitors targeting the hijacking of N-glycan synthesis in viral disease and cancer.

Development and application of mass spectrometric methods for the analysis of progranulin N-glycosylation.

PGRN is a modular protein with 7 1/2 repeats of the granulin domain separated by short spacer sequences. Elevated expression of PGRN is associated with cancer growth, while mutations of PGRN cause frontotemporal lobar degeneration (FTLD), an early onset form of dementia. PGRN is a glycoprotein, containing five N-glycosylation consensus sequons, three of which fall within granulin domains. A method tailored to enable detailed analysis of the PGRN oligosaccharides and glycopeptides has been developed. The approach involves in-gel deglycosylation using peptide-N-glycosidase F (PNGase F) followed by permethylation of the released oligosaccharides. Permethylation was applied for rapid sample clean-up and to improve sensitivity of MS detection and mass spectrometric fragmentation. Reversed-phase monolithic LC-ESI-MS/MS was used for analysis of permethylated oligosaccharides, enabling structural characterization of released N-linked glycans in one chromatographic run. In-gel tryptic digestion was further applied to the gel pieces containing deglycosylated protein, for N-glycosylation site determination. In addition, glycopeptides were produced using in-solution pronase digestion to identify species of N-glycan attached at particular sites. The method developed was applied to progranulin (PGRN) to characterize the structures of the released glycans and to identify the sites of glycosylation. Glycosylation of four out of five potential PGRN N-glycosylation consensus sites was demonstrated (the final one remains undetermined), with one of the four observed to be partially occupied. Two of the observed glycosylation sites occur within granulin domains, which may have important implications for understanding the structural basis of PGRN action.

Nodulation gene mutants of Mesorhizobium loti R7A-nodZ and nolL mutants have host-specific phenotypes on Lotus spp.

Rhizobial Nod factors induce plant responses and facilitate bacterial infection, leading to the development of nitrogen-fixing root nodules on host legumes. Nodule initiation is highly dependent on Nod-factor structure and, hence, on at least some of the nodulation genes that encode Nod-factor production. Here, we report the effects of mutations in Mesorhizobium loti R7A nodulation genes on nodulation of four Lotus spp. and on Nod-factor structure. Most mutants, including a DeltanodSDeltanolO double mutant that produced Nod factors lacking the carbamoyl and possibly N-methyl groups on the nonreducing terminal residue, were unaffected for nodulation. R7ADeltanodZ and R7ADeltanolL mutants that produced Nod factors without the (acetyl)fucose on the reducing terminal residue had a host-specific phenotype, forming mainly uninfected nodule primordia on Lotus filicaulis and L. corniculatus and effective nodules with a delay on L. japonicus. The mutants also showed significantly reduced infection thread formation and Nin gene induction. In planta complementation experiments further suggested that the acetylfucose was important for balanced signaling in response to Nod factor by the L. japonicus NFR1/NFR5 receptors. Overall the results reveal differences in the sensitivity of plant perception with respect to signaling leading to root hair deformation and nodule primordium development versus infection thread formation and rhizobial entry.

Denaturing and non-denaturing microsolution isoelectric focussing to mine the metalloproteome.

Metals bound to proteins play essential roles in living systems. Elements such as phosphorus, selenium and iodine are commonly covalently linked to proteins while others are non-covalently complexed. Thus, the identification and characterization of the metal-protein complexes require a careful hyphenation of techniques able to separate and detect the intact binding complexes with both high resolution and high sensitivity. This study has investigated for the first time the potential of microsolution isoelectric focussing to separate a mixture of metal-binding protein standards under well-established denaturing conditions and a novel non-denaturing separation protocol has also been developed. SEC-ICP-MS analysis was used to evaluate the ability of the two separation procedures to separate and maintain the integrity of standard metal-protein complexes. Microsolution isoelectric focussing under denaturing conditions separates the metalloprotein mixtures with high resolution, although the stability of the complexes is affected. Microsolution isoelectric focussing under our newly developed non-denaturing conditions shows a lower degree of resolution, although the stability of the metal-protein complexes is preserved. The applicability of the two procedures to a biological metalloproteome has also been evaluated.