Applying resonance Rayleigh scattering and spectrofluorimetric techniques for the selective determination of neomycin sulfate using Congo red as a probe: Applications for wastewater analysis.

Two simple, sensitive, and low-cost fluorescence spectroscopy methods for neomycin (NEO) detection were developed. Both methods were based on the interaction between NEO and Congo red (CR) in acidic buffer medium to form an ion-association complex. The quenching effect of the formed ion-association complex on the fluorescence of CR at 421 nm is a basic principle of fluorescence analysis, whilst the resonance Rayleigh scattering (RRS) method was used to enhance the resonance Rayleigh scattering spectrum at 384 nm by adding NEO. Experimental conditions such as pH, temperature, reaction time, CR concentration, and the ionic strength of the two methods were investigated and optimized. In addition, the effect of common coexisting substances on the method was tested and the results showed good selectivity. The composition of ion-association complexes, the reaction mechanism, and reasons for the enhanced intensity of RRS are discussed. Under optimum conditions, the responses of the fluorescence spectrophotometry and RRS methods showed good linearity with NEO concentrations in the range 0.2-3.0 µg ml-1 and 0.1-3.0 µg ml-1 , respectively. The detection limits of fluorescence spectrophotometry and RRS spectroscopy techniques were 0.02 µg ml-1 and 0.01 µg ml-1 , respectively. Finally, the two methods were applied to the analysis of wastewater and the results were satisfactory.

Colorimetric detection of neomycin sulfate in tilapia based on plasmonic core-shell Au@PVP nanoparticles.

The rapid colorimetric detection of neomycin sulfate has been achieved using polyvinyl pyrrolidone shell coated gold nanoparticle (Au@PVP NPs) sol. We also observed that, the aggregation of Au@PVP NPs, possibly caused by the hydrogen bonds formed between neomycin sulfate and PVP shell, generates a new surface plasmon resonance absorption in the wavelength of 600 ~ 700 nm. The proposed method showed an excellent performance towards the determination of neomycin sulfate in wide linear range from 0.01 ~ 10 µM with a correlation coefficient of 0.99 and low detection limit of 1 nM. After extracted with trichloroacetic acid and treated with hot chloroform, neomycin sulfate in the tilapia fish samples was detected with satisfied recovery. Additionally, the high selectivity of Au@PVP NPs sol towards neomycin sulfate has been achieved even in presence of common interfering agents. This method has the advantages of high sensitivity, rapidity, specificity, low cost and no complicated pretreatment procedure.

Microwave-Assisted Synthesis of Hollow Microspheres with Multicomponent Nanocores for Heavy-Metal Removal and Magnetic Sensing.

The primary advantage of a hollow structure is the likelihood of introducing diverse components in a single particle to achieve multiple missions. Herein, hollow microspheres with multicomponent nanocores (HMMNs) have been prepared based on a template-free strategy via a microwave-assisted hydrothermal treatment of Chlorella. The resulting HMMNs retain the near-spherical hollow morphology and functional groups of the cell wall of Chlorella, obviating the need for templates and chemical modification. The elements (iron, cobalt, calcium, magnesium, chlorine, and phosphorus) naturally present within the Chlorella cells react to form hydroxyapatite/chlorapatite and magnetic nanocores without the need for exogenous chemical reagents. The performances of HMMNs for cadmium ion (Cd2+) removal and antibiotic detection are explored. HMMNs exhibit relatively high adsorbance of Cd2+ (1035.8 mmol/kg) and can be easily recovered by application of an external magnetic field. Ion exchange with Ca2+ and Mg2+ is shown to be the main mechanism of Cd2+ elimination. In addition, HMMNs are a suitable carrier for the construction of a magnetic immunosensor, as demonstrated by the successful development of such an immunosensor with acceptable analytical performance for the detection of neomycin in milk samples. The versatile applications of HMMNs result from their multicomponent nanocores, hollow structure, and the functional groups on their shell. This work not only offers a simple and eco-friendly strategy for the fabrication of novel HMMNs but also provides a valuable advanced material for contaminant detection and heavy-metal removal.

Rapid and sensitive determination of neomycin and kanamycin in measles, mumps, and rubella vaccine via high-performance liquid chromatography-tandem mass spectrometry using modified super-paramagnetic Fe3O4 nanospheres.

A simple magnetic dispersive solid-phase extraction (MDSPE) methodology based on mesoporous Fe3O4@ succinic acid nanospheres and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has been developed to determine kanamycin (KNM) and neomycin (NEO) contents in Measles, Mumps, and Rubella (MMR) vaccine products. The monodispersed mesoporous Fe3O4 nanospheres with self-assembled carboxyl terminated shell have been prepared via a simple solvothermal method. These as-synthesized mesoporous Fe3O4 nanospheres showed a high magnetic saturation value (Ms = 46 emu g-1) and large specific surface area (111.12 m2 g-1) which made them potential candidates as sorbents in magnetic solid-phase extraction. The adsorption experimental data fitted well with the Freundlich-Langmuir isotherm and followed a pseudo-second-order kinetic model. Moreover influential parameters on extraction efficiency were investigated and optimized. Under optimal conditions, the limits of detection for KNM and NEO were 1.0 and 0.1 ng mL-1, respectively. Recovery assessments using real samples exhibited recoveries in the range of 96.0 ± 4.3 to 101.5 ± 7.1 %, with relative standard deviations of <10.7% (for intra- day) and <14.6% (for inter- day). The proposed method was successfully applied for different spiked and un-spiked MMR vaccine samples. The presented extraction method provides a fast, selective, robust and practical platform for the detection of KNM and NEO in MMR vaccine samples.

Optical Fiber-Mediated Immunosensor with a Tunable Detection Range for Multiplexed Analysis of Veterinary Drug Residues.

We describe herein a newly developed chemiluminescent optical fiber immunosensor (OFIS) with a tunable detection range for multiplexed analysis of veterinary drug residues with vastly different concentrations in milk samples. The optical fiber probe is used as a carrier of biorecognition element as well as a transducer, enabling a low-cost compact design, which makes this system suitable for cost-effective on-site detection of the target analytes. Importantly, the synergy between modulation of the length of the optical fiber sensing region and the number of fibers allows performing multiplexed immunoassays in an easily controllable manner over a tunable detection range from pg/mL to µg/mL analyte concentrations. By combining the optical fiber sensor with a nanocomplex signal amplification system, a highly sensitive chemiluminescent OFIS system is demonstrated for the multiplexed assaying of veterinary drug residues in milk samples with linear ranges of 10-(2 × 104) pg/mL for chloramphenicol, 0.5-500 ng/mL for sulfadiazine, and 0.1-300 µg/mL for neomycin. This controllable strategy, based on modulation of the fiber probe, provides a versatile platform for multiplexed quantitative detection of both low-abundance and high-abundance targets, which shows great potential for on-site testing in food safety.

Quantification of neomycin in rubella vaccine by off/on metal ion mediated photoluminescence from functionalized graphene quantum dots.

The determination of neomycin sulfate was made using photoluminescent amino-functionalized graphene quantum dots (obtained from hydro-exfoliation of a mixture of citric acid and glutathione). From the several ions tested, Fe3+ was the best mediator to enable an off/on photoluminescence effect used for quantification. The mediation of Fe3+ was found to be crucial as it is responsible for the photoluminescence quenching effect, due to the interaction with quantum dots surface, also having large affinity towards neomycin that removes Fe3+ from the surface of GQDs, consequently, promoting restoration of the original nanomaterial photoluminescence. Such signal restoration was proportional to the neomycin sulfate concentration added. The linearized analytical response covered three orders of magnitude (10-7 to 10-5 mol L-1). The proposed method is an alternative to those requiring labor-intensive procedures for chemical the derivatization of neomycin (due to the lack of chromophore groups in aminoglycosides). The method was successfully tested in the analysis of rubella vaccine containing trace residues of neomycin and in pharmaceutical compositions containing neomycin sulfate after solid phase extraction using an aminoglycoside imprinted polymer to improve selectivity in determinations.

Electrochemiluminecence nanogears aptasensor based on MIL-53(Fe)@CdS for multiplexed detection of kanamycin and neomycin.

A dual gears electrochemiluminecence (ECL) aptasensing strategy for multiple selective determination of kanamycin and neocycin was designed on the basis of the combination of kanamycin and neocycin induced dual gears conversion, the loading platform of metal-organic frameworks (MOFs), surface plasmon resonance (SPR) and ECL resonance energy transfer (ERET) between CdS QDs and AuNPs (or PtNPs). In the absence of target, the dual gears were "off". Then the B1-AuNP (gear B) and aptamer 1-PtNPs acted as signal quenching elements to quench ECL intensity due to ERET process. Upon addition of kanamycin, the aptamer 1-PtNPs were removed from the gear gradually, the ECL was enhanced due to SPR process between AuNPs and CdS QDs. After the incubation of aptamer 2, the dual gears were "off" again and ECL intensity was decreased by ERET process between AuNPs and CdS QDs. In the presence of neomycin, dual gears were "on" again, the ECL signal was enhanced by SPR process between AuNPs and CdS QDs. Under optimal condition, the proposed aptasensor exhibited wide linear ranges of kanamycin (10-10-10-6 M) and neomycin (10-9-10-5 M), and relatively low detection limits to kanamycin (1.7 × 10-11 M) and neomycin (3.5 × 10-10 M). The developed aptasensor realized the multiple ECL detection of kanamycin and neomycin with single luminophore, and was successfully applied to the detection of kanamycin and neomycin in food samples.

Design of a simple paper-based colorimetric biosensor using polydiacetylene liposomes for neomycin detection.

We developed a paper-based analytical device (µPAD) combined with self-signaling polydiacetylene (PDA) liposomes for convenient visual neomycin detection. The simple dot array type of µPAD was fabricated by the wax printing technique, and the PDA liposomes in the aqueous solution were facilely immobilized onto the hydrophilic dot region of the paper substrate. We found that, when the PDA liposomes were inserted to the paper matrix, the stability of the PDA liposomes can be significantly enhanced by adding a hydrophilic reagent such as polyvinyl alcohol and glycerol to the liposome solution. In particular, polyvinyl alcohol (PVA) provides the best stabilization among the various hydrophilic reagents tested in this contribution, and the enhanced stability sharply increased the sensitivity of the PDA liposomes in the paper matrix. Based on the above results, we successfully detected neomycin through both naked-eye observation and fluorescence measurement of PDA signals. The detection limit was 1 ppm and was selective to non-aminoglycoside antibiotics.

Monitoring of neomycin sulfate antibiotic in microbial fuel cells.

Indirect detection and quantification of the neomycin sulfate antibiotic was accomplished in microbial fuel cells. Performance of the microbial fuel cells was examined on the basis of the following parameters; voltage generation, power density, current density and coulombic efficiencies. Removal of neomycin sulfate was monitored using LC-MS/MS in parallel with chemical oxygen demand and total carbohydrate removal. While neomycin sulfate was partially degraded, microbial fuel cell performance appeared to be affected and eventually inhibited by neomycin sulfate on a concentration-based fashion. In order to further examine the neomycin sulfate bio-sensing activity of the microbial fuel cell, a computational chemistry approach was used to obtain the information about the highest occupied molecular orbital-lowest unoccupied molecular orbital energy values of outer electron orbitals, their distribution, and ionization potentials (IPs). The results showed that electroactive bio-film-based MFCs can be used for sensitive detection of neomycin sulfate found in wastewaters.

Rapid analysis of neomycin in cochlear perilymph of guinea pigs using disposable SPE cartridges and high performance liquid chromatography-tandem mass spectrometry.

Irreversible hearing loss induced by aminoglycoside in human through local or systemic administration route negatively impacts quality of life. The aim of this work was to develop and validate an analytical method suitable for the detection and quantification of neomycin in cochlear perilymph of guinea pig after local application. The SupelMIP SPE column was used for the pre-treatment of matrix. Chromatographic separation was conducted by a reversed phase ODS column (100 × 2.1 mm, 3 µm) at 40 °C in gradient mode with 0.2‰ (v/v) HFBA in water and 0.2‰ (v/v) HFBA in acetonitrile as mobile phase, at a flow rate of 0.30 mL/min, with retention time of 3.50 and 3.62 min for internal standard tobramycin and analyte neomycin, respectively. The MS was performed with positive ionization mode, with data acquisition in Multi Reaction Monitor (MRM) mode. This method was proved to be specific, accurate (97.1-115% of nominal values) and precise (CV% < 15%). Calibration curves for matrix matched standard of neomycin ranged from 1.25 to 200 µg/mL, with LOD and LLOQ of 0.625 and 1.25 µg/mL in blank matrix. The matrix effect was corrected to (-0.1) - 1.33 by adding internal standard. The relative SPE recovery values were ≥98.9% in low, medium and high QC samples. Neomycin in matrix proved to be stable under room temperature - and -20 °C, or under three freeze-thawing cycles, or under processing as well. Finally, the proposed method was successfully applied to a toxicokinetics study of neomycin in perilymph after round window membrane (RWM) administration, which was in accordance with threshold shift of auditory brainstem response (ABR) test related to hearing loss.

A conjugated microporous polymer based visual sensing platform for aminoglycoside antibiotics in water.

A donor-acceptor based conjugated microporous polymer, PER@NiP-CMOP-1, has been synthesized which can achieve highly sensitive stereo-specific "Turn ON" biosensing of an aminoglycoside up to the ppb level. The coordination-driven inhibition of photo-induced electron transfer (d-PET) for d-electrons and the rotational freezing are the key factors for the recovery of the emission.

Rapid determination of neomycin in biological samples using fluorescent sensor based on quantum dots with doubly selective binding sites.

A new analytical method was developed to detect neomycin in complex biological samples using molecularly imprinted polymer to construct an optical sensor. Fluorescent neomycin-imprinted polymers (fMIPs) containing both imprinted cavity and boronate affinity site were synthesized on the surface of silica-modified quantum dots. The fMIPs exhibited high selectivity to neomycin by having two binding sites for the target analyte. Neomycin analogues (competing for imprinted cavity) and D-glucose (competing for the boronate affinity site) did not affect the selectivity of the fMIPs. When combined with a fluorescent microplate reader, the obtained fMIP sensor displayed a linear concentration-dependent fluorescence quenching in response to neomycin in the range of 2-1000 µg/L, with a limit of detection as 0.16 µg/L. The fMIP sensor was able to detect trace neomycin in biological samples accurately after simple sample pre-treatment. The sensitivity of the fMIP sensor was higher than HPLC equipped with a fluorescence detector. The fMIP sensor containing the doubly selective binding sites provides a selective, sensitive, accurate, and high through-put approach for neomycin monitoring.

A SERS-based multiple immuno-nanoprobe for ultrasensitive detection of neomycin and quinolone antibiotics via a lateral flow assay.

The authors describe an ultrasensitive method for simultaneous detection of neomycin (NEO) and quinolones antibiotics (QNS). It is based on the use of (a) two immuno-nanoprobes (a probe for NEO and a probe for QNS), (b) surface-enhanced Raman scattering (SERS) detection, and (c), a portable lateral flow assay (LFA). The two probes consist of gold nanoparticles (AuNPs) conjugated to the Raman active molecule 4-aminothiophenol (PATP), and to monoclonal antibody against NEO (NEO mAb) or against NOR (NOR mAb). Quantitative detection of NEO and QNS was realized via SERS of the PATP-coated AuNPs captured in the test line of a LFA. Under optimized condition, the visual limits of LFA are 10 ng·mL-1 for NEO and 200 ng·mL-1 for NOR, and with LODs down to 0.37 pg·mL-1 and 0.55 pg·mL-1 by using SERS. The NEO test line is not interfered by the NEO analogues gentamycin, streptomycin and tobramycin, but the NOR test line suffers from different degrees of cross-reactivity (CR) to 12 common other QNS, the CRs ranging from 1.5% to 136%. The recoveries of NEO and NOR from spiked milk samples ranged between 86% and 121%, with relative standard deviations (RSD) from 3% to 6%. The method is highly sensitive, accurate and effective. It may be applied to simultaneous detection of NEO and 8 QNS, including NOR, enoxacin, ciprofloxacin, ofloxacin, fleroxacin, marbofloxacin, enrofloxacin, and pefloxacin. Graphical abstract Schematic of a lateral flow assay (LFA) based on an indirect competitive model. By using two test lines, the LFA can detect the neomycin and quinolones antibiotics simultaneously. Based on the surface-enhanced Raman scattering (SERS), the LFA shows high sensitivity to antibiotics with low limit of detection.

Genetic manipulation of Fonsecaea pedrosoi using particles bombardment and Agrobacterium mediated transformation.

Fonsecaea pedrosoi, a melanized fungal pathogen that causes Chromoblastomycosis, a human disease with a worldwide distribution. Biolistic is a widely used technique for direct delivery of genetic material into intact cells by particles bombardment. Another well-established transformation method is Agrobacterium-mediated transformation (ATMT), which involves the transfer of a T-DNA from the bacterium to the target cells. In F. pedrosoi there are no reports of established protocols for genetic transformation, which require optimization of physical and biological parameters. In this work, intact conidia of F. pedrosoi were particle bombarded and subjected to ATMT. In addition, we proposed hygromycin B, nourseothricin and neomycin as dominant selective markers for F. pedrosoi and vectors were constructed. We tested two parameters for biolistic: the distance of the particles to the target cells and time of cells recovery in nonselective medium. The biolistic efficiency was 37 transformants/µg of pFpHYG, and 45 transformants/µg of pAN7.1. Transformants expressing GFP were successfully obtained by biolistic. A co-culture ratio of 10: 1 (bacterium: conidia) and co-incubation time of 72 h yielded the largest number of transformants after ATMT. Southern blot analysis showed the number of foreign DNA insertion into the genome is dependent upon the plasmid used to generate the mutants. This work describes for the first time two efficient methods for genetic modification of Fonsecaea and these results open new avenues to better understand the biology and pathogenicity of the main causal agent of this neglected disease.

A simple and selective resonance Rayleigh scattering-energy transfer spectral method for determination of trace neomycin sulfate using Cu2O particle as probe.

The stable Cu2O nanocubic (Cu2ONC) sol was prepared, based on graphene oxide (GO) catalysis of glucose-Fehling's reagent reaction, and its absorption and resonance Rayleigh scattering (RRS) spectra, transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS) were examined. Using the as-prepared Cu2ONC as RRS probe, and coupling with the neomycin sulfate (NEO) complex reaction, a new, simple, sensitive and selective RRS-energy transfer (RRS-ET) method was established for detection of neomycin sulfate, with a linear range of 1.4-112µM and a detection limit of 0.4µM. The method has been applied to the detection of neomycin sulfate in samples with satisfactory results.

Colorimetric Microtiter Plate Assay of Polycationic Aminoglycoside Antibiotics in Culture Broth Using Amaranth.

We present a colorimetric method for the detection of aminoglycoside antibiotics such as neomycin (NEO) using a reddish anionic dye, amaranth (AR3-). Under acidic conditions, at which NEO exists in fully protonated form (NEOH6+), the AR3- anion associates with the NEOH6+ cation to form a precipitate, NEOH(AR)2. The precipitate was soluble in a buffer solution of pH 8.5, yielding a reddish solution with an absorption maximum at around 520 nm. Tobramycin and gentamycin, which exist as pentavalent cations under acidic conditions, gave almost the same results. On the other hand, kanamycin, amikacin and streptomycin, which would exist as tri- and tetravalent cations, were not precipitated. Thus, the AR3- anion could be considered to be an analytical reagent for specific aminoglycosides with polycationic functionality. However, since the precipitation reaction was considerably affected by other anions, a separation method using the tetraphenylborate anion was employed as a pretreatment. The separation method involves precipitating the polycationic aminoglycosides with the tetraphenylborate anion, washing the precipitate with acetonitrile, and re-precipitating the aminoglycosides as hydrochloride salts. Thus, the present method was applied to a microtiter plate assay of the products in an NEO-producing culture broth.

Recent progress in stimuli-induced polydiacetylenes for sensing temperature, chemical and biological targets.

Polydiacetylenes (PDAs) have received increasing attention as smart materials owing to their unique properties. Upon addition of various stimuli, blue PDAs can undergo a colorimetric transition from blue to red along with a change from non-fluorescent to fluorescent. The optical changes can be readily detected by the naked eye and by using absorption and fluorescence spectrometers. These properties make PDAs excellent materials for use in platforms for sensing chemical or biological targets. In recent years, a number of biosensors and chemosensors based on the optical responses of polydiacetylenes have been reported. In this review, recent advances made in this area were discussed following a format based on different cognizing targets, including temperature, metal ions, anions, surfactants, amines, water, gas, sugars, hydrocarbons, neomycin, heparin, virus, enzymes, bacteria, and cancers. Emphasis is given to the methods used to prepare PDA sensing systems as well as their sensing performance.

What a Difference an OH Makes: Conformational Dynamics as the Basis for the Ligand Specificity of the Neomycin-Sensing Riboswitch.

To ensure appropriate metabolic regulation, riboswitches must discriminate efficiently between their target ligands and chemically similar molecules that are also present in the cell. A remarkable example of efficient ligand discrimination is a synthetic neomycin-sensing riboswitch. Paromomycin, which differs from neomycin only by the substitution of a single amino group with a hydroxy group, also binds but does not flip the riboswitch. Interestingly, the solution structures of the two riboswitch-ligand complexes are virtually identical. In this work, we demonstrate that the local loss of key intermolecular interactions at the substitution site is translated through a defined network of intramolecular interactions into global changes in RNA conformational dynamics. The remarkable specificity of this riboswitch is thus based on structural dynamics rather than static structural differences. In this respect, the neomycin riboswitch is a model for many of its natural counterparts.

Highly sensitive turn-on biosensors by regulating fluorescent dye assembly on liposome surfaces.

We developed a new self-signaling sensory system built on phospholipid liposomes having H-aggregated R6G dyes on their surface. Selective molecular recognition of a target by the phospholipid displaces R6G from the liposome surface to turn on fluorescence signal. Selective and sensitive detection of neomycin down to 2.3 nM is demonstrated.