Spectrofluorimetric study on the interaction between antimicrobial drug sulfamethazine and bovine serum albumin

The interaction between the antimicrobial drug sulfamethazine (STM) and bovine serum albumin (BSA) has been studied using steady state and synchronous fluorescence spectroscopy. Fluorescence emission data revealed that BSA (2×10⁻⁶ M) fluorescence was statically quenched by STM at various concentrations, which implies that STM-BSA complex has been formed. The fluorescence emission data was analyzed via applying the Stern-Volmer analysis in combination with thermodynamic investigation, where obtained results revealed that quenching is static with quenching constants of 2.371, 1.658, and 0.916×10⁵ M⁻¹ at 298, 304, and 310 K, respectively. Binding constants and number of binding sites at different temperatures were also determined by applying the Scatchard method, which in turn were used to construct the van't Hoff plot in order to estimate the enthalpy (ΔH) and entropy changes (ΔS) for the complexation process. An average of 1.00±0.17 was estimated for the number of sites of BSA, which indicated that STM binds to BSA with stoichiometric ratio of 1:1. The values that were estimated from the van't Hoff plot for ΔH and (ΔS) were −36.8 kJ mol⁻¹ and −14.9 J mol⁻¹ K⁻¹, respectively, which indicate that the STM-BSA complex is stabilized with hydrogen bonds and van der Waals interactions. Synchronous fluorescence data was obtained at Δλ of 15 and 60 nm, where obtained results confirmed that STM binds to BSA at the tryptophan residue (Trp. 213). In addition, the distance between STM and the Trp. 213 was estimated via employing the Förster's non-radiative energy-transfer theory, and was found to be 2.73 nm, which in turn indicated that STM can bind to BSA with high probability.     

Simultaneous determination of sulfaquinoxaline, sulfamethazine and pyrimethamine by liquid chromatography

A liquid chromatography method is described to determine sulfaquinoxaline (SQX), sulfamethazine (SMT), and pyrimethamine (PMT), by using a Kromasil C₁₈ column and a 40 mM NaH₂PO₄ buffer solution, containing 10 mM NaClO₄ (pH 3.0)–acetonitrile (65:35) as mobile phase. The mobile phase flow-rate and sample volume injected were 1.5 ml/min and 20 μl, respectively and the samples were dissolved in the mobile phase. The limits of quantification were found to be about 180 μg/l (3.6 ng) for each compound. The method was applied in veterinary commercial formulations. Analyses were made by means of the standard addition method, whose results were compared with those obtained by preparing “tests” (from the stock solutions) and with those obtained by a capillary electrophoresis method. Both methods showed similar results, and then it was proved that some commercial claimed levels were not in agreement with the obtained results by using our analytical method, as they were in other cases.     

Preparation and characterization of molecularly imprinted silica monolith for screening sulfamethazine

In this work, a novel approach of preparing molecularly imprinted film‐derivatized silica monolith materials was developed by a two‐step procedure. The silica monolithic support was first prepared by the sol–gel method with tetramethoxysilane as the precursor. Subsequently, vinyl groups were introduced onto the surface of silica monolith by immobilization of γ‐methacryloxypropyltrimethoxysilane. The prepolymerization mixtures, consisting of methacrylic acid as a functional monomer, ethylene dimethacrylate as a crosslinker, sulfamethazine as a template molecule and an ionic liquid as porogen, were injected into the silica monolith immobilized vinyl groups to form the molecularly imprinted films on the surface of the vinyl functionalized silica monolith. The monolithic materials were characterized by SEM, Fourier transform IR and solid‐state reflection UV spectra. The resulted imprinted materials were evaluated under CEC and HPLC mode. The results indicated that there were enough recognition sites on the surface of the imprinted film‐derivatized monolithic materials for selectively recognizing sulfamethazine from the sulfonamide mixture. Ionic liquids, which was utilized as the porogens, could improve the flow‐through property and the imprinting effect of the molecularly imprinted film‐functionalized silica monolithic materials.     

磺胺二甲嘧啶分子印迹聚合物制备与光谱特性

本研究以磺胺二甲嘧啶为模板分子,甲基丙烯酸为功能单体,低温紫外引发聚合制备分子印迹聚合物,用紫外光谱法对磺胺二甲嘧啶与甲基丙烯酸的结合作用及磺胺二甲嘧啶印迹聚合物的静态吸附性能和选择性能进行研究,试验结果表明,溶液中的功能单体与印迹分子之间产生了结合作用;与空白印迹聚合物相比,以甲基丙烯酸为功能单体、磺胺二甲嘧啶为模板的分子印迹聚合物展现了较高的结合容量,与其他结构类似的底物磺胺异唑相比,磺胺二甲嘧啶分子印迹聚合物对磺胺二甲嘧啶模板分子显示出明显的选择性和特异性,静态分配系数K_D可达282.3 mg·mL-1,分离因子α可达3.9,并预测磺胺二甲嘧啶分子印迹聚合物结合机理。分析了甲基丙烯酸、二甲基丙烯酸乙二醇酯和磺胺二甲嘧啶分子印迹聚合物的红外谱图,研究结果表明制备的磺胺二甲嘧啶印迹聚合物中CC双键峰很小,并且功能键羧酸键没有明显变化,经交联聚合得到的聚合物确实存在可以同印迹分子相互作用的化学基团。     

鸡肉中磺胺二甲嘧啶和磺胺吡啶残留的SERS快速鉴别研究

以金溶胶作为活性基底,浓度为1%的NaCl溶液作为活性剂,利用DXRTM显微拉曼光谱仪采集鸡肉的表面增强拉曼光谱（SERS）,实现快速鉴别鸡肉中残留的磺胺二甲基嘧啶（SM-2）和磺胺吡啶（SPD）两种抗生素。用937和1 188 cm-1处是否有拉曼特征峰来判别鸡肉中是否残留SPD和SM-2。采用单因素实验方法,根据937和1 188 cm-1处的特征SERS强度,对试验条件进行优化,得到最佳试验条件：金溶胶加入量为500 μL、NaCl溶液加入量为100 μL和吸附时间为5 min,所选用的金溶胶柠檬酸钠加入量3.7 mL。根据测试集鸡肉中残留的SM-2和SPD的分类精度确定研究用自适应迭代惩罚最小二乘法（air-PLS）、归一化和二阶导数作为原始拉曼光谱的预处理方法,然后用主成分分析（PCA）提取特征向量,最后以前四个PCA得分值作为支持向量机（SVM）分类模型的输入值,建立基于C-SVC类型的SVM分类模型。其中,最优惩罚参数c为0.01、核参数g为0.1。此分类模型对测试集的整体分类精度达到93.23%。对测试集的敏感性和特异性进行计算,敏感性的范围为77.42%~100%,特异性的范围为96%~99.02%,其中,含SM-2+SPD鸡肉的敏感性最高为100%,含SPD鸡肉的特异性最高为99.02%。试验结果表明,该方法具有良好的鉴别效果,可用于实现对鸡肉中SM-2和SPD两种抗生素残留的快速检测和鉴别。     

The performance and degradation mechanism of sulfamethazine from wastewater using IFAS-MBR

Sulfamethazine(SMZ) is an important sulfonamide antibiotic.Although the concentration in the environment is small,it is harmful.The drug residues can be transferred,transformed or accumulated,affecting the growth of animals and plants.In this study,the integrated fixed-film activated sludge membrane bioreactor(IFAS-MB R) were constructed to investigate the performance and degradation mechanism of SMZ.The addition of SMZ had a significant impact on the removal of the chemical oxygen demand(COD) and ammonia nitrogen(NH_4^+-N).The optimal operating conditions were hydraulic retention time(HRT) at 10 h and solid retention time(SRT) at 80 d,respectively.On this basis,the effects of different SMZ concentrations on nutrient removal,degradation,and sludge characteristics were compared.The removal efficiency of SMZ increased with the increase of SMZ concentration.The maximum removal rate was as high as 87%.The SMZ dosage also had an obvious effect on sludge characteristics.As the SMZ concentration increased,the extracellular polymer substances(EPS) concentration and the membrane resistance both decreased,which were beneficial for the reduction of membrane fouling.Finally,seven kinds of SMZ biodegradation intermediates were identified,and the possible degradation pathways were speculated.The microbial community results showed that the microbial diversity and richness in the reactor decreased after adding SMZ to the influent.The relative abundance of Bacteroidetes,Actinobacteria,Saccharibacteria and Nitro spirae increased at the phylum level.Sphingobacteria and Betaproteobacteria became dominant species at the class level.The relative abundance of norankp-Saccharibacteria and Nitrospirae increased significantly,and norank-p-Saccharibacteria may be the dominant bacteria for SMZ degradation.     

遗忘因子分光光度法同时测定消尔疴口服液中磺胺噻唑和磺胺二甲嘧啶含量

应用基于递推最小二乘原理的遗忘因子法,本文不经分离直接测定了消尔疴口服液中两主要成分磺胺噻唑(ST)和磺胺二甲嘧啶(SM_2)的含量,平均回收率分别为99.99％和100.4％,相对标准偏差0.23％和0.56％(n=16),与卡尔曼滤波法结果一致。本法保留了卡尔曼滤波法不需成批数据,无须占用大量内存,可以联机应用进行在线测定和控制等优点,同时克服了后者需要测量噪声的先验信息、测量误差加权系数难以确定的不足,通过遗忘因子的引入,提高了收敛速度,改善了递推过程中的适应能力。     

Relevance of a combined process coupling electro-Fenton and biological treatment for the remediation of sulfamethazine solutions – Application to an industrial pharmaceutical effluent

A combined process coupling an electro-Fenton pretreatment and a biological degradation was implemented in order to mineralize synthetic and industrial pharmaceutical effluents, containing a veterinary antibiotic, sulfamethazine (SMT). The electro-Fenton pretreatment of an SMT synthetic solution was first examined and the obtained results showed total SMT degradation after 30 min of electrolysis at pH 3, 18 °C, 500 mA and an initial SMT concentration of 0.2 mM, while the level of mineralization remained low (2.1 and 18.1% for electrolysis times of 30 and 60 min), ensuring significant residual organic content for a subsequent biological treatment. In a second part, biological treatments were performed to complete the mineralization of the electrolyzed solutions of SMT, showing a significant level of mineralization after about 18 days of culture, 61.4% for a 30-min pretreatment. The same electrolysis conditions were then applied to the pretreatment of an industrial pharmaceutical effluent, showing a total SMT removal in the effluent after 100 min of electrolysis, while the mineralization yield remained also low (7.5%), showing the formation of organic intermediate products. Fortunately, the mineralization yield during the subsequent biological treatment increased to almost 80%, namely an overall yield of 81.4%. Consequently, the integrated electro-Fenton biological treatment process proved to be an efficient technology to reprocess industrial pharmaceutical effluents.     

Synthesis,spectroscopic characterization,antimicrobial activity and crystal structure of silver and copper complexes of sulfamethazine

Silver(I) and copper(II) complexes of 4-amino-N-(4,6-dimethyl-2-pyrimidinyl) benzenesulfonamide (smz) have been synthesized and characterized by elemental analysis and infrared (IR), ¹H NMR, and UV–vis spectroscopy. [Ag(smz)(pyridine)] (1) crystallizes in monoclinic system with space group P2₁/c and Z = 4, while [Cu(smz)₂(pyridine)₂]·H₂O (2) crystallizes in triclinic system with space group P-1 and Z = 2. X-ray analysis revealed that silver in 1 is four-coordinate exhibiting distorted tetrahedral geometry, while copper in 2 is coordinated to six nitrogens leading to a highly distorted octahedral geometry. The molecular structures of both 1 and 2 are stabilized by N–H?O and C–H?π intermolecular and C–H?O intramolecular interactions. Water plays a significant role in crystal packing by forming strong N–H?O_water intramolecular as well as O_water–H?N intermolecular interactions in 2. The results of IR, UV–vis, ¹H NMR spectral data and thermal analysis for 1 and 2 suggest that the binding of silver and copper to the sulfonamidic nitrogen is in agreement with the crystal structure determination. Antimicrobial activities of silver (1) and copper (2) complexes of sulfamethazine are studied by the dilution method against Staphylococcus aureus and Escherichia coli strains.     

Ultrasound-Assisted Extraction Combined with HPLC-UV for Fast Determination of Sulfamethazine and Its N4-Acetyl Metabolite in Plasma and Phosphate Buffer

A simple and cost effective HPLC-UV method for fast determination of sulfamethazine (SMZ) and its N₄-acetyl metabolite, N₄-acetyl sulfamethazine (AcSMZ), in plasma and phosphate buffer was developed and validated. The sample treatment was performed by ultrasound-assisted extraction with 1 mL ethyl acetate twice, each time 5 min. Moreover, there was no need for centrifugation and further clean-up, which shortened the measurement time. The total analysis time was less than 30 min. The method proved to be a selective and accurate tool for detection of SMZ and AcSMZ in plasma and phosphate buffer with satisfactory recoveries (85.43–99.29%) and little matrix effects (0.86–0.99). The proposed approach was also applied to the evaluation of pharmacokinetic and drug-metabolite protein binding interaction studies. The results show that SMZ and AcSMZ are non-concentration-dependent, and the metabolite does not bind competitively with the drug to the plasma protein.