Generation of anti-azoxystrobin monoclonal antibodies from regioisomeric haptens functionalized at selected sites and development of indirect competitive immunoassays

Azoxystrobin is a modern strobilurin fungicide used around the world to combat prime diseases affecting highly valuable crops. Accordingly, residues of this chemical are frequently found in food, even though mostly under maximum tolerated levels. We herein describe the development of an indirect competitive immunoassay for the determination of azoxystrobin residues. A panel of monoclonal antibodies displaying subnanomolar affinity to azoxystrobin was generated using, as immunizing haptens in mice, four functionalized derivatives carrying the same spacer arm located at different rationally chosen positions. This collection of antibodies was thoroughly characterized with homologous and heterologous antigens, and the immunoassay consisting of monoclonal antibody AZo6#49 and the coating conjugate OVA–AZb6, which displayed an IC₅₀ value of 0.102 μg L⁻¹ and a LOD of 0.017 μg L⁻¹, was eventually optimized. The response to different pH and ionic strength conditions of the specific assay was studied using a biparametric approach. In addition, the influence of Tween 20 and organic solvents over the assay parameters was also evaluated. After optimization, the developed immunochemical assay was applied to the analysis of azoxystrobin in spiked juices of relevant fruits and vegetables, showing excellent recoveries between 2 and 500 μg L⁻¹.     

电化学间接测量法研究谷胱甘肽在多壁碳纳米管和活性炭上的吸附

选取谷胱甘肽(GSH)作为小分子代表物,利用Cr(VI)与GSH的相互作用,即K2Cr2O7/H2SO4溶液加入GSH前后Cr(VI)还原峰电流值的差异,通过差分脉冲伏安法(DPV)的测定,可以间接测出GSH的含量.尝试将这种电化学间接测量法应用于研究多壁碳纳米管(MWCNTs)与商业活性炭(AC)对GSH的吸附行为.利用该方法可以确立GSH在二者的吸附量(Q)与吸附平衡浓度(Ce)的关系,绘制吸附等温曲线.根据Langmuir方程和Freundlich方程的拟合分析,证明了与商业活性炭相比,GSH在MWCNTs上的吸附更倾向Freundlich模型,即多分子层吸附.利用扫描电子显微镜(SEM)和透射电子显微镜(TEM)进行形貌表征,发现MWCNTs具有发达的堆积孔结构,有利于GSH小分子内扩散和吸附,与实验结果一致.     

离子色谱法-间接紫外检测离子液体中三种含氟酸根阴离子

建立了离子色谱-间接紫外检测法同时分析离子液体中的三氟乙酸根(CF3COO-)、四氟硼酸根(BF4-)和三氟甲烷磺酸根(CF3SO3-)的方法。用Shodex IC NI-424阴离子交换柱作为分离柱,探讨了影响上述三种离子保留和检测的因素及其保留规律。最终选用色谱条件为:以1.75 mmol·L-1邻苯二甲酸-1.68 mmol·L-1三(羟甲基)氨基甲烷为淋洗液,柱温45℃,流速1.0 m L·min-1,UV检测波长设定在265 nm。在此条件下,可实现上述三种阴离子的同时分析,且色谱峰形良好。所测阴离子的检出限(S/N=3)为0.39~1.57 mg·L-1,保留时间和峰面积的相对标准偏差(n=5)均在1%以下。将此方法应用于测定离子液体中的三氟乙酸根、四氟硼酸根及三氟甲烷磺酸根,加标回收率为99%~102%。该方法简单、准确、可靠,具有较好的实用性。     

谷胱甘肽在高岭土修饰碳糊电极上的电化学研究

制备了高岭土修饰碳糊电极(KCPE),并以氢醌(HQ)为电催化介质测定还原型谷胱甘肽(GSH)。实验结果表明,在该电极上谷胱甘肽(GSH)的加入对氢醌的电化学响应有明显提高,且电催化氧化峰电流与GSH浓度呈良好的线性关系,由此可采用差分脉冲伏安法(DPV)对GSH进行定量测定。对修饰电极的电化学性能进行了表征,测得电极有效表面积为0.118 6 cm2、电荷传递系数为0.68、表观速率常数为1.08×104cm3·mol-1·s-1。在优化条件下,测得GSH在KCPE上的线性范围为0.1~1,2.5~25μmol/L,检出限为0.073μmol/L。用该方法对含GSH的市售药进行测定,结果满意。     

What is the form of muscimol from fly agaric mushroom (Amanita muscaria) in water? An insight from NMR experiment supported by molecular modeling

The biologically active alkaloid muscimol is present in fly agaric mushroom (Amanita muscaria), and its structure and action is related to human neurotransmitter _γ-aminobutyric acid (GABA). The current study reports on determination of muscimol form present in water solution using multinuclear ¹H and ¹³C nuclear magnetic resonance (NMR) experiments supported by density functional theory molecular modeling. The structures of three forms of free muscimol molecule both in the gas phase and in the presence of water solvent, modeled by polarized continuous model, and nuclear magnetic isotropic shieldings, the corresponding chemical shifts, and indirect spin–spin coupling constants were calculated. Several J-couplings observed in proton and carbon NMR spectra, not available before, are reported. The obtained experimental spectra, supported by theoretical calculations, favor the zwitterion form of muscimol in water. This structure differs from NH isomer, previously determined in dimethyl sulfoxide (DMSO) solution. In addition, positions of signals C3 and C5 are reversed in both solvents.     

Cr2O72-/Cr3+间接氧化环己醇合成己二酸

以Cr2O72-/Cr3+作为间接氧化剂电氧化环己醇制备己二酸.应用正交实验优化工艺条件,得出在原料比n(环己醇)∶n(Cr2O72-)=0.4∶1,t=35℃和CH2SO4=5mol.L-1条件下己二酸的收率可达70.29%.同时研究了Ag2SO4、(NH4)2SO4、H2SO4浓度、电流密度对Cr3+电氧化为Cr2O72-的影响,Cr3+的转化率可达82.52%.     

离子液体为背景电解质的毛细管电泳-间接紫外检测法同时测定葡萄酒中无机阴离子和阳离子

以离子液体1-乙基-3-甲基咪唑对甲苯磺酸盐([EMIm]TS)为背景电解质,采用双端进样方式,实现了毛细管电泳-间接紫外检测法同时分析测定葡萄酒中无机阳离子(K⁺、Ca²⁺、Na⁺、Mg²⁺和Li⁺)和阴离子(Cl^-、SO■和ClO^-₃)。[EMIm]TS作为电泳缓冲溶液的同时,其阳离子和阴离子分别作为样品中阳离子和阴离子组分的间接紫外检测的背景电解质。在最佳分析条件下,可在6.5 min内完成8种无机离子的同时分离检测,其线性范围为0.005～0.7 g/L,相关系数为0.963～0.995,检出限(S/N=3)为1.2～12.5 mg/L。该方法成功测定了3种不同品牌的市售葡萄酒中8种无机离子。在3个加标水平下,8种无机离子的回收率为90.1%～110.5%,相对标准偏差(RSD)≤4.8%。结果表明,该方法可应用于葡萄酒中无机阴、阳离子的同时分离检测,且方法简单、快速且结果可靠。     

Modelling of a tubular solid oxide fuel cell with different designs of indirect internal reformer

The cell performance and temperature gradient of a tubular solid oxide fuel cell with indirect internal reformer(IIR-SOFC) fuelled by natural gas, containing a typical catalytic packed-bed reformer, a catalytic coated wall reformer, a catalytic annular reformer, and a novel catalytic annular-coated wall reformer were investigated with an aim to determine the most efficient internal reformer system. Among the four reformer designs, IIR-SOFC containing an annular-coated wall reformer exhibited the highest performance in terms of cell power density(0.67 W cm-2)and electrical efficiency(68%) with an acceptable temperature gradient and a moderate pressure drop across the reformer(3.53×10-5kPa).IIR-SOFC with an annular-coated wall reformer was then studied over a range of operating conditions: inlet fuel temperature, operating pressure, steam to carbon(S : C) ratio, gas flow pattern(co-flow and counter-flow pattern), and natural gas compositions. The simulation results showed that the temperature gradient across the reformer could not be decreased using a lower fuel inlet temperature(1223 K–1173 K)and both the power density and electrical efficiency of the cell also decreased by lowering fuel inlet temperature. Operating in higher pressure mode(1-10 bar) improved the temperature gradient and cell performance. Increasing the S : C ratio from 2 : 1 to 4 : 1 could decrease the temperature drop across the reformer but also decrease the cell performance. The average temperature gradient was higher and smoother in IIR-SOFC under a co-flow pattern than that under a counter-flow pattern, leading to lower overpotential and higher cell performance. Natural gas compositions significantly affected the cell performance and temperature gradient. Natural gas containing lower methane content provided smoother temperature gradient in the system but showed lower power density and electrical efficiency.     

毛细管区带电泳技术快速分离检测爆炸残留物中的无机离子

利用间接紫外毛细管区带电泳方法完成了对爆炸残留物中7种无机离子(K+,NH+4,NO-2,NO-3,SO2-4,ClO-3,ClO-4)的分离检测。阳离子测定采用的缓冲体系为10 mmol/L吡啶（pH 4.5）-3 mmol/L冠醚,K+和NH+4在2.6 min内达到基线分离,检出限分别为0.25 mg/L和0.10 mg/L(S/N=3)。阴离子测定采用的缓冲体系为40 mmol/L硼酸-1.8 mmol/L重铬酸钾-2 mmol/L硼酸钠（pH 8.6）,氢氧化四甲铵为电渗流改性剂,5种阴离子在4.6 min内达到基线分离,检出限为0.10～1.85 mg/L。该方法已成功地应用于实际爆炸物样品种类的判定分析,取得了很好的结果。     

毛细管电泳法测定阿霉素脂质体药物中的微量硫酸根离子

建立了以硝酸钾作为背景电解质测定阿霉素脂质体药物中微量硫酸根离子的毛细管电泳分析法。考察了分离电压、背景电解质、电渗流改性剂浓度、pH值对分离测定的影响。结果表明,当毛细管长度为60 cm(有效长度51.5 cm)、分离电压为~15 kV、缓冲溶液采用20 mmol/L硝酸钾(pH 7.0)、电渗流改性剂采用0.4 mmol/L十六烷基三甲基氯化铵(CTAC)、检测波长为202 nm时,阿霉素脂质体破乳液中硫酸根离子和氯离子在3 min内得到了基线分离,硫酸根离子迁移时间和峰面积的相对标准偏差分别小于0.01%和1.0%,检出限为5 μg/L。用该方法对阿霉素脂质体样品中的微量硫酸根离子进行了分析测定,结果令人满意。