目的建立采用快速基质分散净化QuEChERS(quick,easy,cheap,effective,rugged,safety)-气相色谱-串联质谱法(gas chromatography-tandem mass spectrometry,GC-MS/MS)快速测定香肠和火腿肠制品中13种N亚硝胺类化合物的分析方法。方法样...目的建立采用快速基质分散净化QuEChERS(quick,easy,cheap,effective,rugged,safety)-气相色谱-串联质谱法(gas chromatography-tandem mass spectrometry,GC-MS/MS)快速测定香肠和火腿肠制品中13种N亚硝胺类化合物的分析方法。方法样品加入内标,经乙腈提取,QuCEhERS萃取盐包和基质分散净化包净化,采用DB-WAXETR(30 m×0.25 mm,0.25μm)色谱柱分离,在多反应监测模式(mult-reaction monitoring,MRM)下测定,内标法定量。结果13种N-亚硝胺化合物在1~100μg/L范围内线性关系良好,相关系数(r)均大于0.99,定量限为0.03~2.84μg/kg;添加水平为3~30μg/kg,平均回收率为59.1%~119.2%,相对标准偏差为0.6%~15.1%(n=6)。结论该方法简单准确、灵敏度高、重复性好,适用于批量肉制品样品中13种N-亚硝胺类化合物的快速检测。展开更多
Objective To clone the cDNA of rat α-Syn gene, investigate its prokaryotic expression and produce purified recombinant rat α-Syn protein. Methods Rat α-Syn cDNA was amplified from the rat brain total RNA by RT-PCR ...Objective To clone the cDNA of rat α-Syn gene, investigate its prokaryotic expression and produce purified recombinant rat α-Syn protein. Methods Rat α-Syn cDNA was amplified from the rat brain total RNA by RT-PCR and was cloned into pGEX-4T-1, a prokaryotic expressing vector. The recombinant plasmid containing rat α-Syn gene was transformed into E. Coli BL21 to express a fusion protein with rat α-Syn protein tagged by glutathione-S-transferase （GST）. The fusion protein was then cleaved by thrombin during passing through the GST-agarose 4B column to release the recombinant rat α-Syn protein. The recombinant rat a-Syn protein was further purified using Superdex S200 gel filtration. Results DNA sequencing confirmed that the cloned cDNA contained 420 base pairs encoding 140 amino acids, which was identical to the reported amino acid sequence of rat α-Syn. After transformation, the recombinant plasmid pGEX-ra-Syn expressed a soluble protein that was inducible by IPTG. The purified recombinant protein was shown to be single band on SDS-PAGE, with a molecular size of around 18000, which was identical to the reported molecular size of rat α-Syn. Western blot analysis demonstrated that the recombinant protein was recognized by specific antibody against α-Syn. Conclusion The rat α-Syn gene was successfully expressed in prokaryotic expression system and highly purified rat α-Syn recombinant protein was produced.展开更多
The reversed micelles were formed with cationic cetyltrimethylammonium bromide (CTAB) as surfactant and n-hexanol as cosolvent in the CTAB (50mmol.L-1)/hexanol (15% by volume)/hexane system. Cibacron Blue 3GA (CB) as ...The reversed micelles were formed with cationic cetyltrimethylammonium bromide (CTAB) as surfactant and n-hexanol as cosolvent in the CTAB (50mmol.L-1)/hexanol (15% by volume)/hexane system. Cibacron Blue 3GA (CB) as an affinity ligand in the aqueous phase was directly introduced to the reversed micelles with electrostatic interaction between anionic CB and cationic surfactant. High molecular weight (Mr) protein, yeast alcohol dehydrogenase (YADH, Mr = 141000) from baker's yeast, has been purified using the affinity reversed micelles by the phase transfer method. Various parameters, such as CB concentration, pH and ionic strength, on YADH forward and backward transfer were studied. YADH can be transferred into and out from the reversed micelles under mild conditions (only by regulation of solution pH and salt concentration) with the successful recovery of most YADH activity. Both forward and backward extractions occurred when the aqueous phase pH＞pI with electrostatic attraction between YADH and CTAB. The recovery of YADH activity and purification factor have been improved with addition of a small amount of affinity CB. The recovery of YADH activity obtained was ～99% and the purification factor was about 4.0-fold after one cycle of full forward and backward extraction. The low ionic strength in the initial aqueous phase might be responsible for the YADH transfer into the reversed micellar phase.展开更多
[Objective] This study was conducted to investigate the effect of laundry wastewater on the quality of river water and the dilution purification effect of river water on laundry wastewater. [Method] The effects of lau...[Objective] This study was conducted to investigate the effect of laundry wastewater on the quality of river water and the dilution purification effect of river water on laundry wastewater. [Method] The effects of laundry wastewater on the contents of total nitrogen(TN), total phosphorus(TP), suspended solids(SS), chemical oxygen demand(COD) and linear alkylbenzene sulfonic acid(LAS) were studied in 7 rivers of Shaoxing City. [Result](1) The contents of TN, TP, SS, COD and LAS increased by 92%, 99%, 340%, 351% and 923%, respectively, at the discharging moment of laundry wastewater; and(2) the five pollutional indexes significantly decreased over time, and especially 2 h after the discharge of laundry wastewater, compared with former the discharge of laundry wastewater, the contents of TN, TP, COD and LAS increased by 6%, 11%, 9% and13%, respectively,while the contents of SS still increased by 76%, i.e., SS required a longer time to achieve self-purification. [Conclusion] Laundry wastewater has some influence on thequality of river water, and the self-purification function of river water could effectively remove pollutants.展开更多
Hydrogen sulfide in rural biogas was removed with liquid-phase catalytic oxidation.By using rare earth as catalyst,and sulfosalicylic acid as stabilizer,H2S purification efficiency could increase as high as 96%,and su...Hydrogen sulfide in rural biogas was removed with liquid-phase catalytic oxidation.By using rare earth as catalyst,and sulfosalicylic acid as stabilizer,H2S purification efficiency could increase as high as 96%,and sulfur capacity of the composite solution was about 3 g/L.The results show that purification efficiency was affected by catalyst addition,pH,experimental temperature,and sulfur capacity.The parameters effects on catalytic oxidation were studied,and the optimized conditions were that Fe3+ concentration 0.08 mg/L,reaction temperature 70°C,pH 9.0,with a absorption solution volume of 50 mL,a gas flow rate 200 mL/min,and H2S mass concentration of 1.58-2.02 mg/m3.展开更多
基金This work was supported by Key Project of National Natural Science Foundation of China （30430280） National Natural Science Foundation of China（ 30271437,30270482 ） Natural Science Foundation of Beijing（ 7022011 ）.
文摘Objective To clone the cDNA of rat α-Syn gene, investigate its prokaryotic expression and produce purified recombinant rat α-Syn protein. Methods Rat α-Syn cDNA was amplified from the rat brain total RNA by RT-PCR and was cloned into pGEX-4T-1, a prokaryotic expressing vector. The recombinant plasmid containing rat α-Syn gene was transformed into E. Coli BL21 to express a fusion protein with rat α-Syn protein tagged by glutathione-S-transferase （GST）. The fusion protein was then cleaved by thrombin during passing through the GST-agarose 4B column to release the recombinant rat α-Syn protein. The recombinant rat a-Syn protein was further purified using Superdex S200 gel filtration. Results DNA sequencing confirmed that the cloned cDNA contained 420 base pairs encoding 140 amino acids, which was identical to the reported amino acid sequence of rat α-Syn. After transformation, the recombinant plasmid pGEX-ra-Syn expressed a soluble protein that was inducible by IPTG. The purified recombinant protein was shown to be single band on SDS-PAGE, with a molecular size of around 18000, which was identical to the reported molecular size of rat α-Syn. Western blot analysis demonstrated that the recombinant protein was recognized by specific antibody against α-Syn. Conclusion The rat α-Syn gene was successfully expressed in prokaryotic expression system and highly purified rat α-Syn recombinant protein was produced.
基金the National Natural Science Foundation of China (No. 29836130).
文摘The reversed micelles were formed with cationic cetyltrimethylammonium bromide (CTAB) as surfactant and n-hexanol as cosolvent in the CTAB (50mmol.L-1)/hexanol (15% by volume)/hexane system. Cibacron Blue 3GA (CB) as an affinity ligand in the aqueous phase was directly introduced to the reversed micelles with electrostatic interaction between anionic CB and cationic surfactant. High molecular weight (Mr) protein, yeast alcohol dehydrogenase (YADH, Mr = 141000) from baker's yeast, has been purified using the affinity reversed micelles by the phase transfer method. Various parameters, such as CB concentration, pH and ionic strength, on YADH forward and backward transfer were studied. YADH can be transferred into and out from the reversed micelles under mild conditions (only by regulation of solution pH and salt concentration) with the successful recovery of most YADH activity. Both forward and backward extractions occurred when the aqueous phase pH＞pI with electrostatic attraction between YADH and CTAB. The recovery of YADH activity and purification factor have been improved with addition of a small amount of affinity CB. The recovery of YADH activity obtained was ～99% and the purification factor was about 4.0-fold after one cycle of full forward and backward extraction. The low ionic strength in the initial aqueous phase might be responsible for the YADH transfer into the reversed micellar phase.
基金Supported by Natural Science Foundation of China(31500321)Scientific Research Foundation of Shaoxing University(20145024)
文摘[Objective] This study was conducted to investigate the effect of laundry wastewater on the quality of river water and the dilution purification effect of river water on laundry wastewater. [Method] The effects of laundry wastewater on the contents of total nitrogen(TN), total phosphorus(TP), suspended solids(SS), chemical oxygen demand(COD) and linear alkylbenzene sulfonic acid(LAS) were studied in 7 rivers of Shaoxing City. [Result](1) The contents of TN, TP, SS, COD and LAS increased by 92%, 99%, 340%, 351% and 923%, respectively, at the discharging moment of laundry wastewater; and(2) the five pollutional indexes significantly decreased over time, and especially 2 h after the discharge of laundry wastewater, compared with former the discharge of laundry wastewater, the contents of TN, TP, COD and LAS increased by 6%, 11%, 9% and13%, respectively,while the contents of SS still increased by 76%, i.e., SS required a longer time to achieve self-purification. [Conclusion] Laundry wastewater has some influence on thequality of river water, and the self-purification function of river water could effectively remove pollutants.
基金Project(2008ZX07105-002) supported by the Erhai Lake Project of National Science and Technology Major Project in the 11th Five years Plan of China
文摘Hydrogen sulfide in rural biogas was removed with liquid-phase catalytic oxidation.By using rare earth as catalyst,and sulfosalicylic acid as stabilizer,H2S purification efficiency could increase as high as 96%,and sulfur capacity of the composite solution was about 3 g/L.The results show that purification efficiency was affected by catalyst addition,pH,experimental temperature,and sulfur capacity.The parameters effects on catalytic oxidation were studied,and the optimized conditions were that Fe3+ concentration 0.08 mg/L,reaction temperature 70°C,pH 9.0,with a absorption solution volume of 50 mL,a gas flow rate 200 mL/min,and H2S mass concentration of 1.58-2.02 mg/m3.
