Tianjiao Wang

Investigation on the interactions of clenbuterol to bovine serum albumin and lysozyme by molecular fluorescence technique.

Clenbuterol interacting with bovine serum albumin (BSA) or lysozyme (LYS) in physiological buffer (pH 7.4) was investigated by the fluorescence spectroscopy and UV-vis absorption spectroscopy. The results indicated that clenbuterol quenched the intrinsic fluorescence of BSA and LYS via a static quenching procedure. The binding constants of clenbuterol with BSA and LYS were 1.16×10(3) and 1.49×10(3) L mol(-1) at 291 K. The values of ΔH and ΔS implied that hydrophobic and electrostatic interaction played a major role in stabilizing the complex (clenbuterol-BSA or clenbuterol-LYS). In the presence of Fe2+, Fe3+, Cu2+, Mg2+, Ca2+, or Zn2+, the binding constants of clenbuterol to BSA or LYS had no significant differences. The distances between the donor (BSA or LYS) and acceptor (clenbuterol) were 2.61 and 2.19 nm for clenbuterol-BSA and clenbuterol-LYS respectively. Furthermore, synchronous fluorescence spectrometry was used to analyze the conformational changes of BSA and LYS.

An investigation on the interaction of DNA with hesperetin/apigenin in the presence of CTAB by resonance Rayleigh light scattering technique and its analytical application

Two new systems for measuring DNA at nanogram levels by a resonance Rayleigh light scattering (RLS) technique with a common spectrofluorometer were proposed. In the presence of cetyltrimethylammonium bromide (CTAB), the interaction of DNA with hesperetin and apigenin (two effective components of Chinese herbal medicine) could enhance RLS signals with the maximum peak at 363 and 433 nm respectively. The enhanced intensity of RLS was directly proportional to the concentration of DNA in the range of 0.022-4.4 μg mL(-1) for DNA-CTAB-hesperetin system and 0.013-4.4 μg mL(-1) for DNA-CTAB-apigenin system. The detection limit was 2.34 ng mL(-1) and 2.97 ng mL(-1) respectively. Synthetic samples were measured satisfactorily. The recovery of DNA-CTAB-hesperetin system was 97.3-101.9% and that of DNA-CTAB-apigenin system was 101.2-109.5%.

In vitro studies on the behavior of salmeterol xinafoate and its interaction with calf thymus DNA by multi-spectroscopic techniques

The salmeterol xinafoate (SX) binding to calf thymus DNA in vitro was explored by fluorescence, resonance light scattering (RLS), UV-vis absorption, as well as viscometry, ionic strength effect and DNA melting techniques. It was found that SX could bind to DNA weakly, and the binding constants (Ka) were determined as 8.52×10(3), 8.31×10(3) and 6.14×10(3) L mol(-1) at 18, 28 and 38°C respectively. When bound to DNA, SX showed fluorescence quenching in the fluorescence spectra and hyperchromic effect in the absorption spectra. Stern-Volmer plots revealed that the quenching of fluorescence of SX by DNA was a static quenching. Furthermore, the relative viscosity and melting temperature of DNA solution were hardly influenced by SX, while the fluorescence intensity of SX-DNA was observed to decrease with the increasing ionic strength of system. Also, the binding constant between SX and double stranded DNA (dsDNA) was much weaker than that between SX and single stranded DNA (ssDNA). All these results suggested that the binding mode of SX to DNA should be groove binding. The obtained thermodynamic parameters indicated that electrostatic force might play a predominant role in SX binding to DNA. The quantum yield (φ) of SX was measured as 0.13 using comparative method. Based on the Förster resonance energy transfer theory (FRET), the binding distance (r0) between the acceptor and donor was calculated as 4.10 nm.

Using gold nanoparticles as probe for detection of salmeterol xinafoate by resonance Rayleigh light scattering

The paper explores the method of determination of salmeterol xinafoate at nanogram level with gold nanoparticles (AuNPs) probe, to measure the intensity of resonance Rayleigh light scattering (RLS) by a common spectrofluorometer. The RLS intensity of salmeterol xinafoate was greatly enhanced by AuNPs, with the maximum scattering peak at 357 nm. The salmeterol xinafoate was determined basing on the binding of salmeterol xinafoate to AuNPs by electrostatic adsorption. Under the optimum conditions, the enhanced RLS intensity was directly proportional to the concentration of salmeterol xinafoate in the range of 0.054-6.038 μg mL(-1) with a good linear relationship (r=0.9928). The limit of detection (LOD) was 9.48 ng mL(-1). The interference tests were performed carefully. With the proposed method, the synthetic samples were analyzed satisfactorily, the recovery and RSD were 102.5-103.0% and 0.67-1.0% respectively.

Binding characteristics of salbutamol with DNA by spectral methods

Salbutamol interacting with deoxyribonucleic acid (DNA) was examined by fluorescence, UV absorption, viscosity measurements, and DNA melting techniques. The binding constants and binding sites were obtained at different temperatures by fluorescence quenching. The Stern-Volmer plots showed that the quenching of fluorescence of salbutamol by DNA was a static quenching. To probe the binding mode, various analytical methods were performed and the results were as follows: hyperchromic effect was shown in the absorption spectra of salbutamol upon addition of DNA; there was no appreciable increase in melting temperature of DNA when salbutamol was presented in DNA solution; the fluorescence intensity of salbutamol-DNA decrease with the increasing ionic strength; the relative viscosity of DNA did not change in the presence of salbutamol; the binding constant of salbutamol with double strand DNA (dsDNA) was much higher than that of it with single strand DNA (ssDNA). All these results indicated that the binding mode of salbutamol to DNA should be groove binding. The thermodynamic parameters suggested that hydrogen bond or van der Waals force might play an important role in salbutamol binding to DNA. According to the Förster energy transference theory, the binding distance between the acceptor and donor was 3.70 nm.

The analytical application and spectral investigation of DNA-CPB-emodin and sensitive determination of DNA by resonance Rayleigh light scattering technique.

A new sensitive DNA probe containing cetylpyridinium bromide (CPB) and emodin (an effective component of Chinese herbal medicine) was developed using the resonance Rayleigh light scattering (RLS) technique. A novel assay was first developed to detect DNA at nanogram level based on the ternary system of DNA-CPB-emodin. The RLS signal of DNA was enhanced remarkably in the presence of emodin-CPB, and the enhanced RLS intensity at 340.0 nm was in direct proportion to DNA concentration in the range of 0.01-2.72 μg mL(-1) with a good linear relationship. The detection limit was 1.5 ng mL(-1). Three synthetic DNA samples were measured obtaining satisfactory results, the recovery was 97.6-107.3%.

The resonance Rayleigh light scattering spectral investigation on the interaction of DNA with camellia sinensis in the presence of CPC and its analytical application.

A novel method with high sensitivity was designed for the determination of trace nucleic acids by using cationic surfactant cetylpyridinium chloride (CPC) and camellia sinensis (CS) as resonance Rayleigh light scattering (RLS) probes. It was found DNA could combine with CS and CPC in Tris-HCl buffer (pH=7.4). Under optimum conditions, the RLS intensity of DNA can be enhanced by CPC-CS obviously at 294nm, and the enhanced RLS intensity was directly proportional to DNA concentration in the range from 0.024 to 3.48μgmL(-1) with a good linear relationship (r=0.9940). The limit of detection (LOD) was 1.49ngmL(-1) (S/N=3). In addition, the effects of some interferences including K(+), Na(+), Mg(2+), Zn(2+), Cu(2+), Ca(2+) and glucose on the determination were studied. The developed RLS assay was successfully applied to three synthetic samples to measure DNA, the recovery was 94.7-106.3% and RSD was 0.58-3.33%.

Study of the interaction of salmon sperm DNA with myricitrin-CPB based on the enhanced resonance light scattering signal and its potential application.

A new assay of salmon sperm DNA at nanogram level was established based on enhanced resonance light scattering (RLS) signals of DNA with myricitrin and cetylpyridinium bromide dihydrate (CPB). The RLS spectral characteristics of DNA with myricitrin-CPB and the optimum conditions for determination of DNA samples have been studied. At pH 7.4, myricitrin-CPB could enhance the intensity of RLS signal of DNA at 468 nm. The enhanced RLS intensity was directly proportional to DNA concentration in the range of 0.076-4.2 μg mL(-1) with a good linear relationship (r=0.9944). The detection limit was 4.1 ng mL(-1). The synthetic samples were analyzed with satisfactory results that the recovery was 100.9-102.6% and RSD was 1.4-2.1%, which proved that the new method was reliable and applicable.