Exploring the intercalation binding of tiamulin with DNA using multispectroscopy and a molecular modelling approach.

The binding of tiamulin with calf thymus DNA was systematically investigated using multispectroscopy and molecular modelling techniques. For DNA, once tiamulin was added, viscosity (η) and melting temperature (Tm ) both exhibited an uptrend. The fluorescence performance of the tiamulin-DNA complex did not change with the ionic strength changes. The binding constant (Ka ) of tiamulin for single-stranded DNA (ssDNA, 1.48 × 104 M-1 ) was obviously higher than that for double-stranded DNA (dsDNA, 9.51 × 103 M-1 ) at 291 K. The helix structure became looser and the base stack force became stronger for DNA due to the presence of tiamulin as seen from circular dichroic (CD) spectra. The intercalation binding mode of tiamulin with DNA was disclosed. Molecular modelling also revealed tiamulin inserting into the base pairs with the lowest binding free energy of -18.73 kJ mol-1 using van der Waals forces as well as hydrogen bonds.

Assessment on the binding characteristics of residual marbofloxacin in animal-derived food to bovine/human serum albumin by spectroscopy and molecular modelling.

To assess the toxicity of residual marbofloxacin from animal-derived food, the interaction characteristics of marbofloxacin to bovine/human serum albumins (BSA/HSA) were explored using spectroscopic methods combined with molecular modelling. According to fluorescence spectra and time-resolved fluorescence spectra measurements, quenching of BSA/HSA fluorescence induced by marbofloxacin was characterized as static quenching. A 1:1 ground-state complex of marbofloxacin to BSA/HSA was formed with binding constant (Ka ) 1.66 × 104 /9.74 × 103 M-1 at 291 K. The location of marbofloxacin binding at site I within BSA/HSA was clarified by site marker competitive experiments. Molecular modelling demonstrated that the binding region for marbofloxacin to BSA and HSA were at site I with the lowest binding free energies of -22.86 and -21.60 kJ mol-1 , respectively. Hydrogen bonds and van der Waals forces were dominantly involved in the spontaneous binding. Nonradiation energy transferred from BSA and HSA to marbofloxacin, due to the close distance (r0 ) between marbofloxacin and Trp residues of BSA (4.28 nm) and HSA (3.34 nm). As explained by circular dichroism (CD) spectra, an increased BSA/HSA α-helix structure was observed after binding to marbofloxacin. Ultraviolet-visible (UV-vis) and Fourier transform infrared (FT-IR) spectra suggested that conformation of the two proteins was altered by marbofloxacin.

Depicting the binding of furazolidone/furacilin with DNA by multiple spectroscopies, voltammetric as well as molecular docking.

The interaction between DNA and furazolidone/furacillin was investigated using various analytical techniques including spectroscopy and electroanalysis and molecular modelling. With the aid of acridine orange (AO), the fluorescence lifetimes of DNA-AO, DNA-furazolidone/furacillin-AO remained almost the same, which proved that the ground state complex was formed due to furazolidone/furacillin binding with DNA. Circular dichroism spectra and Fourier transform infrared spectroscopy showed that the second structure of DNA changed. Viscosity experiments presented that relative viscosity of DNA was increased with the increasing concentrations of furazolidone and almost unchanged for furacilin. In addition, the results of melting temperature (Tm ), ionic strength, site competition experiments, cyclic voltammetry, and molecular docking all proved the intercalation binding mode for furazolidone and groove binding mode for furacilin. The binding constants (Ka ) obtained from Wolfe-Shimmer equation were calculated as 3.66 × 104 L mol-1 and 3.95 × 104 L mol-1 for furazolidone-DNA and furacilin-DNA, respectively.

A sensitive resonance Rayleigh light scattering method for alpinetin using gold nanorods probes.

A sensitive resonance Rayleigh light scattering (RLS) assay for alpinetin was developed based on alpinetin-modified gold nanorods (AuNRs). Alpinetin could interact with AuNRs and formed a new assembly by electrostatic attraction. In pH 7.4 Tris-HCl buffer solution, the assembly of alpinetin-AuNRs showed a sensitive RLS signal. Under optimum conditions, the magnitude of enhanced RLS intensity (ΔIRLS ) was proportional to the concentration of alpinetin over the range 0.027-3.24 µg ml-1 , with a detection limit of 1.79 ng ml-1 (by 3σ). The developed RLS method was successfully applied to the detection of alpinetin in real or synthesized samples. Alpinetin recoveries were 90.4-108.7% with an RSD of 0.82-2.9% (n = 5) for Alpinia katsumadai samples, and 95.1-103.7% with an RSD of 0.28-3.9% (n = 5) for synthesized samples. The results showed that this new approach was convenient, reliable and sensitive.

Investigation of the binding of AuNPs-6-mercaptopurine and the sensitive detection of 6-mercaptopurine using resonance Rayleigh light scattering.

A highly sensitive method for the detection of 6-mercaptopurine (MP) by resonance Rayleigh light scattering (RLS) method was developed. Gold nanoparticles (AuNPs) were synthesized by a modified seed method and characterized using transmission electron microscopy (TEM). AuNPs were bound to MP via covalent bonding to form the MP-AuNPs complex, which increased the RLS intensity of MP at 347 nm (increased by 65.7%). Under optimum conditions, the magnitude of the enhanced RLS intensity for MP-AuNPs was proportional to MP concentration in the range 0.0681-1.702 µg mL-1 . The linear regression equation was represented as follows: ΔIRLS = 9.31 + 82.42c (r = 0.9948). The limit of detection (LOD, 3σ) was 3.32 ng mL-1 . The system was applied successfully to detect MP in pharmaceuticals. MP recoveries were 99.9-101.7% with a relative standard deviation (RSD) (n = 5) of 0.59-0.77% for three synthetic samples, and 97.5-110.0% with an RSD of 0.98-2.10% (n = 5) for tablet samples.

Study on the interactions of mapenterol with serum albumins using multi-spectroscopy and molecular docking.

The interactions of mapenterol with bovine serum albumin (BSA) and human serum albumin (HSA) have been investigated systematically using fluorescence spectroscopy, absorption spectroscopy, circular dichroism (CD) and molecular docking techniques. Mapenterol has a strong ability to quench the intrinsic fluorescence of BSA and HSA through static quenching procedures. At 291 K, the binding constants, Ka, were 1.93 × 10(3) and 2.73 × 10(3) L/mol for mapenterol-BSA and mapenterol-HAS, respectively. Electrostatic forces and hydrophobic interactions played important roles in stabilizing the mapenterol-BSA/has complex. Using site marker competitive studies, mapenterol was found to bind at Sudlow site I on BSA/HSA. There was little effect of K(+), Ca(2+), Cu(2+), Zn(2+) and Fe(3+) on the binding. The conformation of BSA/HSA was changed by mapenterol, as seen from the synchronous fluorescence spectra. The CD spectra showed that the binding of mapenterol to BSA/HSA changed the secondary structure of BSA/HSA. Molecular docking further confirmed that mapenterol could bind to Sudlow site I of BSA/HSA. According to Förster non-radiative energy transfer theory (FRET), the distances r0 between the donor and acceptor were calculated as 3.18 and 2.75 nm for mapenterol-BSA and mapenterol-HAS, respectively.

Characterization of the binding of paylean and DNA by fluorescence, UV spectroscopy and molecular docking techniques.

The interaction of paylean (PL) with calf thymus DNA (ctDNA) was investigated using fluorescence spectroscopy, UV absorption, melting studies, ionic strength, viscosity experiments and molecular docking under simulated physiological conditions. Values for the binding constant Ka between PL and DNA were 5.11 × 10(3) , 2.74 × 10(3) and 1.74 × 10(3) L mol(-1) at 19, 29 and 39°C respectively. DNA quenched the intrinsic fluorescence of PL via a static quenching procedure as shown from Stern-Volmer plots. The relative viscosity and the melting temperature of DNA were basically unchanged in the presence of PL. The fluorescence intensity of PL-DNA decreased with increasing ionic strength. The value of Ka for PL with double-stranded DNA (dsDNA) was larger than that for PL with single-stranded DNA (ssDNA). All the results revealed that the binding mode was groove binding, and molecular docking further indicated that PL was preferentially bonded to A-T-rich regions of DNA. The values for ΔH, ΔS and ΔG suggested that van der Waals forces or hydrogen bonding might be the main acting forces between PL and DNA. The binding distance was determined to be 3.37 nm based on the theory of Förster energy transference, which indicated that a non-radiation energy transfer process occurred. Copyright © 2015 John Wiley & Sons, Ltd.

ZIF-8-wrapped AgNPs modified with ß-cyclodextrin for sensitive detection of thiacloprid and imidacloprid by SERS technology.

The high efficient surface-enhanced Raman scatterring (SERS) methods to detect thiacloprid and imidacloprid were established using ZIF-8-wrapped Ag nanoparticles (AgNPs) modified with ß-cyclodextrin (ß-CD). The substrate of ZIF-8/ß-CD@AgNPs was characterized by ultraviolet visible spectra (UV-vis), thermogravimetric analysis (TGA), X-ray diffraction (XRD), transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM). The interaction between the substrate and thiacloprid/imidacloprid was also explored. The optimum measurement conditions were obtained by response surface model based on single-factor experiments. Enhancement factors (EFs) of thiacloprid and imidacloprid were respectively 2.29 × 106 and 2.60 × 106. A good linearity between the scattering intensity and the concentration of thiacloprid/imidacloprid within 3-1000 nmol L-1/6-400 nmol L-1 was established. The interference experiments indicated that the methods had good selectivity. The SERS methods were successfully applied to detect thiacloprid and imidacloprid in several vegetables samples. The recoveries ranged from 95.5 % to 105 % (n = 5). The detection limits (LODs) (S/N = 3) for thiacloprid and imidacloprid were 1.50 and 0.83 nmol L-1, respectively.

Sensitive detection of dextromethorphan hydrobromide based on portable Raman spectrometer and CuO@AgNPs nano composite SERS substrate.

A highly sensitive surface-enhanced Raman spectroscopy approach was established for the detection of dextromethorphan hydrobromide (DXM) utilizing nano CuO modified silver nanoparticles (CuO@AgNPs) as substrate. Ultraviolet visible spectra (UV-vis), X-ray diffraction (XRD) and transmission electron microscopy (TEM) characterized the synthesized CuO@AgNPs. UV-vis and fourier transform infrared (FT-IR) were adopted to investigate the interaction between DXM and CuO@AgNPs. The optimal experimental conditions (the dosages of CuO@AgNPs and NaCl as well as mixing time) were explored. The enhancement factor (EF) was 1.71 × 106. The linear relationship between SERS intensity and the concentration of DXM in the range of 67 - 1000 nmol L-1 was obtained as ISERS = 25.81 c + 40398.77, and the limit of detection (LOD) was 2.12 nmol L-1 (S/N = 3). The interference of K+, Mg2+, Zn2+, Ca2+, Cu2+, Fe3+, glucose, HSA, L-tryptophan, soluble starch and ibuprofen were investigated. The method was successfully applied to test DXM in serum samples. The recovery was 99.06% - 101.51% with the relative standard deviation (RSD) of 0.74% - 3.87%.

Silver nanoparticles modified with MoS2 and ß-cyclodextrin as SERS substrate for rapid determination of cysteamine hydrochloride in meat products.

An efficient Surface-enhanced Raman scattering (SERS) method for the detection of cysteamine hydrochloride (CSH) was developed by synthesizing a composite substrate comprising silver nanoparticles (AgNPs) functionalized with MoS2 and ß-cyclodextrin (ß-CD). The enhanced Raman signals of CSH by ß-CD/MoS2/AgNPs substrate were the contribution of electromagnetic enhancement (EM) as well as chemical enhancement (CM), and the enhancement factor (EF) can reach up to 3.11 × 106 (peak at 633 cm-1). Various instrumental techniques were used to characterize the substrate, such as X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM), high-angle annular dark field scanning transmission electron microscopy (HAADF-STEM) and ultraviolet visible (UV-vis). The binding of ß-CD/MoS2/AgNPs and CSH was confirmed by UV-vis and Fourier transform infrared (FT-IR). The optimal experimental conditions were determined by single factor experiments as well as response surface model. The influences of different metal ions and analogous drugs on the detection of CSH were investigated. Under optimum conditions, a good linear correlation (R = 0.9997) was established for CSH in the range of 10.00-1000.00 nmol/L, and the limit of detection (LOD) was as low as 0.78 nmol/L (S/N = 3). The contents of CSH in meat samples were detected. The recovery was 96.6-103.1 %, and the relative standard deviation (RSD) of the measurement was 0.7-3.9 % (n = 7).

Efficient detection of aklomide/nitromide based on a sensor of MPA capped CdTe QDs using fluorescence quenching method.

Based on the fluorescence sensor of 3-Mercaptopropionic acid (MPA) capped CdTe quantum dots (QDs), two novel detection methods for aklomide and nitromide were developed. The MPA-CdTe QDs were synthesized and characterized by transmission electron microscopy (TEM), thermogravimetric analysis (TGA), X-ray diffraction (XRD), ultraviolet visible (UV-vis) and fluorescence (FL). The quenchings were all static. The binding constants (Ka) at different temperatures were obtained. Electrostatic forces were the main forces for the two bindings. For the detection of aklomide and nitromide, under the optimal conditions, the effects of some metal ions, glucose, bovine serum albumin (BSA) and congeneric drug on the determination were explored. The standard equations were established and the limits of detection (LOD) were 0.0215 and 0.0388 µg mL-1 (3S0/S), repectively. The methods were applied to analyse the samples of chicken and duck, the recoveries were 99.41 % - 101.24 % with RSDs of 0.29 % - 1.19 % (n = 5).

Rapid determination of isepamicin by using SERS based on BSA-protected AgNPs modified by α-Fe2O3.

A surface-enhanced Raman spectroscopy (SERS) method for the determination of isepamicin (ISE) using silver nanoparticles (AgNPs) protected by bovine serum albumin (BSA) and modified by α-Fe2O3 as an efficient substrate was established. The synthesized substrate was characterized and verified by transmission electron microscopy (TEM), ultraviolet visible spectroscopy (UV-vis), and fourier transform infrared spectroscopy (FT-IR). The conditions affecting the Raman signal of ISE were optimized by single factor and response surface experiments. Under optimized conditions, a standard curve ISERS = 43.08c + 63598.69 (c: nmol/L) with a linear relationship (r = 0.9976) was established between the SERS intensity and ISE concentration in the range of 20.00 - 2000.00 nmol/L. The limit of detection (LOD) for ISE was 16.58 nmol/L (S/N = 3). The recovery of ISE in the samples was 96.29 % - 104.12 %, with relative standard deviation (RSD) was 1.53 % - 3.43 % (n = 5). The SERS method was reliable and satisfactory for the quantitative analysis of ISE at low concentration.

A sensitive surface-enhanced Raman spectroscopy detection for gentamicin and tobramycin using γ-Al2O3-modified silver nanoparticles coated with bovine serum albumin as substrate.

Two aminoglycoside antibiotics (AGs), gentamicin (GEN) and tobramycin (TOB), have good antibacterial activity against most pseudomonas aeruginosa and staphylococcus. The molecular structure of these drugs lack chromogenic groups, which brings challenges to their detection. In this project, the detecting methods for GEN and TOB utilizing surface-enhanced Raman spectroscopy (SERS) based on γ-Al2O3-modified silver nanoparticles (AgNPs) coated with bovine serum albumin (BSA) were established. The enhancement factors (EFs) of GEN and TOB were 2.44 × 105 and 2.67 × 106, respectively. The transmission electron microscopy (TEM), X-ray diffraction (XRD), thermogravimetric analysis (TGA) and spectrophotometric techniques were used to characterize the substrate and the combination of the substance with drugs. The pH, the addition amounts for the substrate and coagulant, as well as the mixing time were optimized. On the basis of single factor experiments, a more scientific response surface model was established. The concentrations of GEN and TOB showed good linear relationships with their Raman signals in the ranges of 6.67 × 10-8 - 2.00 × 10-6 and 6.67 × 10-9 - 3.00 × 10-7 mol L-1 respectively. The limits of detection (LODs) were 11.88 and 1.26 nmol L-1 for GEN and TOB, respectively. The methods were used successfully for the samples determination of the two AGs in commercial drugs and meat products.

A fluorescence nanoprobe of N-Acetyl-L-Cysteine capped CdTe QDs for sensitive detection of nitrofurazone.

A method was established for detecting the content of nitrofurazone (NFZ) by fluorescence quenching of N-Acetyl-L-Cysteine (NAC) coated cadmium telluride quantum dots (CdTe QDs). By means of transmission electron microscopy (TEM) and multispectral methods such as fluorescence and ultraviolet visible spectra (UV-vis), the synthesized CdTe QDs were characterized. The quantum yield (φ) of CdTe QDs was measured as 0.33 by reference method. The CdTe QDs had a better stability, the RSD of fluorescence intensity was 1.51% in three months. NFZ quenching the emission light of CdTe QDs was observed. The analyses of Stern-Volmer and time-resolved fluorescence suggested the quenching was static. The binding constants (Ka) between NFZ and CdTe QDs were 1.14 × 104 (293 K), 0.74 × 104 (303 K) and 0.51 × 104 (313 K) L mol-1. The hydrogen bond or van der Waals force was the dominated binding force between NFZ and CdTe QDs. The interaction was further characterized by UV-vis absorption as well as Fourier transform infrared spectra (FT-IR). Using fluorescence quenching effect, a quantitative determination of NFZ was carried out. The optimal experimental conditions were studied and determined as following: pH was 7 and contact time was 10 min. The effects of reagent addition sequence, temperature and the foreign substances including some metals (Mg2+; Zn2+; Ca2+; K+; Cu2+), glucose, bovine serum albumin (BSA) and furazolidone on the determination were studied. There was a high correlation between the concentration of NFZ (0.40 - 39.63 µg mL-1) and F0/F with the standard curve F0/F = 0.0262c + 0.9910 (r = 0.9994). The detection limit (LOD) reached 0.04 µg mL-1 (3S0/S). The contents of NFZ in beef and bacteriostatic liquid were detected. The recovery of NFZ was 95.13% - 103.03% and RSD was 0.66% - 1.37% (n = 5).

Sensitive surface-enhanced Raman spectroscopy (SERS) determination of nitrofurazone by ß-cyclodextrin-protected AuNPs/γ-Al2O3 nanoparticles.

A novel surface-enhanced Raman spectroscopy (SERS) method for the determination of nitrofurazone was developed using AuNPs/γ-Al2O3 nanoparticles protected by ß-cyclodextrin (ß-CD) as substrate prepared in our lab. The optimum experimental conditions were obtained from single factor procedure and response surface modeling. A linear relationship (ISERS = 508.96c + 31987.87, c: nmol L-1, R2 = 0.996) between SERS intensity and the concentration of nitrofurazone in the range of 3.3 - 667.0 nmol L-1 was established, the limit of detection (LOD) was found at nmol L-1 level (0.37 nmol L-1 by 3S0/S). The selectivity for the method was studied by the influences of foreign substances on the determination. The recoveries and RSD (n = 5) for the six meat samples were 95.1 % - 104.5% and 2.4 % - 4.8% respectively, which suggesting that the new SERS method was successfully to detecting nitrofurazone.

Multispectral and molecular modeling investigations on the binding behaviors of two anticoccidials with serum albumins.

The interaction properties of monensin/clopidol with bovine/human serum albumin (BSA/HSA) were determined via multispectral together with molecular modeling techniques in the report. Fluorescence quenching spectra at different temperatures and fluorescence lifetime determination demonstrated that the fluorescence quenching belonged to a static quenching type. In the case of monensin-BSA, clopidol-BSA, monensin-HSA and clopidol-HSA, the binding constants Ka (291 K) were 5.42 × 104, 4.96 × 104, 3.22 × 104 and 2.99 × 104 M-1, respectively; the binding distances r0 were 1.88, 2.53, 2.19 and 2.02 nm, respectively. Monensin and clopidol bound strongly with BSA/HSA with binding free energies equal to -26.37/-25.11 and -26.11/-24.93 kJ mol-1, respectively. The spontaneous binding process was dominated by hydrogen bonds and van der Waals forces as reflected in thermodynamic parameters analyses. Synchronous, CD, FTIR and UV-vis spectra assays confirmed that serum albumins conformations were altered. Using competitive experiment, monensin/clopidol was observed to bind at site I of serum albumins, which were reconfirmed by the results of molecular modeling.Communicated by Ramaswamy H. Sarma.

Study on the binding of sulfaclozine sodium monohydrate with bovine and human serum albumins using multi-spectroscopy and molecular docking.

The interactions of sulfaclozine sodium monohydrate (SSM) with bovine and human serum albumins (BSA and HSA) were studied by multi-spectroscopy and molecular docking technique. Stern-Volmer analysis and fluorescence lifetime measurements suggested the quenching processes were static. According to the Fluorescence resonance energy transfer (FRET) theory, the binding distances were obtained indicating SSM interacted with BSA/HSA along with non-radiation energy conversion. Electrostatic attraction was the main force in keeping the stability of the compound based on thermodynamic parameters. Circular dichroism (CD), synchronous fluorescence and Fourier Transform infrared (FT-IR) spectra embodied the secondary structures of serum albumins were transformed by SSM. The site marker competitive and molecular docking measurements testified SSM bound to BSA/HSA at site I. In conclusion, the secondary structures of BSA/HSA were changed by SSM in the static fluorescence quenching processes with the non-radiation energy conversion. The binding sites were all located at site I and electrostatic attraction was the main force for the new compound. Communicated by Ramaswamy H. Sarma.

Rapid determination of marbofloxacin by surface-enhanced Raman spectroscopy of silver nanoparticles modified by ß-cyclodextrin.

A novel surface enhanced-Raman spectroscopy (SERS) assay for marbofloxacin was developed based on ß-cyclodextrin-modified silver nanoparticles (ß-CD-AgNPs). The marbofloxacin could interact with ß-CD-AgNPs and a new assembly was formed by AgN covalent bond. This assembly was characterized by the spectra of FT-IR and UV-vis. The optimal measurement conditions were studied in detail. In 0.033 mol L-1 HCl solution, marbofloxacin had a sensitive SERS signal at 806 cm-1. The enhancement factor (EF) was 2.11 × 107. There was a good linear correlation between the concentration of marbofloxacin and SERS intensity: the linear range was 0.003-0.03 µmol L-1 (r2 = 0.996). The limit of detection (LOD) (S/N = 3) was 1.7 nmol L-1 (S/N = 3). Moreover, the influence of some interferences including Cu2+, K+, Zn2+, Ca2+, Na+, Mg2+, glucose and tiamulin on the determination were studied. The developed SERS method was used to detect the content of marbofloxacin in chicken and duck, the recovery was 101.3%-103.1% with RSD 4.07%-6.83%.

Spectral and molecular docking studies on the interaction of three flavonoids with bovine serum albumin.

Communicated by Ramaswamy H. Sarma.

Multispectral and molecular docking investigations on the interaction of primethamine/trimethoprim with BSA/HSA.

Primethamine (PMA) and trimethoprim (TMP) were investigated as traditional coccidiostats on the binding of bovine serum albumin (BSA) and human serum albumin (HSA) by multispectral and molecular docking techniques. The Stern-Volmer plots and time-resolved fluorescence measurement declared that PMA/TMP quenching the intrinsic fluorescence of BSA/HSA was static quenching process. The binding constants (Ka) and binding sites (n) were calculated at different temperatures. Meanwhile, thermodynamic parameters showed electrostatic forces played a leading role in the interaction of PMA/TMP with BSA/HSA. Some metal ions such as K+, Mg2+, Cu2+, Ca2+, Zn2+ and Fe3+ had no effects on the binding system. The UV-vis absorption spectra confirmed that the interaction between PMA/TMP and BSA/HSA did happen. The analyses of synchronous fluorescence, FT-IR and circular dichroism spectra illustrated that PMA/TMP changed the secondary structures of BSA/HSA. According to Förster non-radiative energy transfer theory, the binding distance between PMA/TMP and BSA/HSA was calculated. The binding location of PMA/TMP to BSA/HSA was identified as sub-domain IIA, which was further confirmed by molecular docking.Communicated by Ramaswamy H. Sarma.