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Preparation of Hydroxypropyl-β-cyclodextrin Cross-linked Multi-walled Carbon Nanotubes and Their Application in Enantioseparation of clenbuterol

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Chinese Journal of Chemistry 2011年第5期29卷 893-897页

作者：于金刚黄笃树黄可龙洪涌 College of Chemistry and Chemical Engineering Central South University Changsha Hunan 410083 China College of Science Honghe University Honghe Yunnan 661100 China

A method of cross-linking multi-walled carbon nanotubes by a nucleophilic substitution of brominated multi-walled carbon nanotubes using hydroxypropyl-β-cyclodextrin anions was studied. The modified multi-walled carbon nanotube samples were characterized using thermogravimetric analysis, energy-dispersive X-ray spectroscopy, transmission electron microscopy, scanning electron microscopy, Raman spectroscopy and Fourier transform infrared spectroscopy. The hydroxypropyl-β-cyclodextrin modified multi-walled carbon nanotubes were used as a chiral stationary phase additive for thin-layer chromatography to separate clenbuterol enantiomers, and the chiral separation factor was increased.

关键词： hydroxypropyl-β-cyclodextrin cross-linking multi-walled carbon nanotubes resolution clenbuterol

Fast Determination of clenbuterol and Salbutamol in Feed and Meat Products Based on Miniaturized Capillary Electrophoresis with Amperometric Detection

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Chinese Journal of Chemistry 2007年第12期25卷 1832-1835页

作者：楚清脆耿成怀周惠叶建农 Department of Chemistry East China Normal University Shanghai 200062 China

The fast separation capability of a novel miniaturized capillary electrophoresis with an amperometric detection （μCE-AD） system was demonstrated by determining clenbuterol and salbutamol in real samples. The effects of several factors such as the acidity and concentration of the running buffer, the separation voltage, the applied potential and the injection time on CE-AD were examined and optimized. Under the optimum conditions, the two β-agonists could be baseline separated within 60 s at a separation voltage of 2 kV in a 90 mmol/L H3BO3-Na2B4O7 running buffer （pH 7.4）, which was not interfered by ascorbic acid and uric acid. Highly linear response was obtained for above compounds over three orders of magnitude with detection limits ranging from 1.20×10^-7 to 6.50×10^-8 mol/L （S/N=3）. This method was successfully used in the analysis of feed and meat products with relatively simple extraction procedures.

关键词： miniaturized capillary electrophoresis-amperometric detection β-agonist clenbuterol salbutamol

Development of a Porcine cDNA Microarray:Analysis of clenbuterol Responding Genes in Pig(Sus scrofa) Internal Organs

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Journal of Integrative Agriculture 2012年第11期11卷 1877-1883页

作者： ZHANG Jin GUO Wei SHEN Liang-cai LILT Qiu-yue DENG Xue-mei HU Xiao-xiang LINing College of Life Science and Biotechnology Hebei Normal University of Science & Technology Qinhuangdao 066600 P.R.China State Key Laboratory for Agrobiotechnology China Agricultural University Beijing 100193 P.R.China College of Animal Science and Technology China Agricultural University Beijing 100193 P.R.China

Pig （Sus scrofa） fat accumulation can be reduced by feeding with high dosages of clenbuterol, but the molecular mechanism has not yet been explained. In our study, a porcine cDNA microarray representing 3 358 pig genes was successfully developed. This microarray is the first porcine DNA microarray in China and its false positive rate is 0.98%, which means the microarray platform is reliable. The microarray can be used to study gene expression profiles in multiple pig tissues because the present genes percentage of adipose, skeletal muscle, heart, liver, lung, kidney, and spleen were all more than 60%. This microarray was used to identify the genes responding to clenbuterol stimulation in pig internal organs, including heart, liver, lung, spleen, and kidney. Many genes were identified including enzymes involved in lipids metabolism （lipoprotein lipase up-regulated in liver, heart and lung, ATP-citrate lyase and carnitine palmitoyltransferase II precursor up-regulated in liver, succinyl-CoA up-regulated in lung, mitochondrial malate dehydrogenase down-regulated in spleen）, and signaling pathway genes （cAMP-protein kinase A signaling pathway was found up-regulated in liver, heart, lung, and kidney as reported previously, while transforming growth factor was found down-regulated in heart and lung）. However, no common gene responding to clenbuterol administration was found in all tissues. The expression levels of 14 genes were analyzed using real-time PCR with 82.1% of them induced to express similar magnitudes as in the microarray analyses. This work offers some understanding of how clenbuterol so effectively reduces pig adipose accumulation on the molecular level.

关键词： cDNA microarray pig clenbuterol adipose accumulation internal organ

RISK信号通路在β_2-肾上腺素受体激动剂clenbuterol减轻心肌细胞缺氧/复氧损伤中的作用

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中国药理学通报 2015年第10期31卷 1368-1374页

作者：张秋芳谭艳汪选斌潘龙瑞李洪亮刘慧向继洲付琴华中科技大学同济医学院药理系湖北武汉430071 湖北医药学院药理教研室湖北十堰442000 湖北医药学院武当特色中药湖北省重点实验室湖北十堰442000

目的研究β2-肾上腺素受体激动剂clenbuterol对原代培养的心肌细胞缺氧/复氧损伤的作用及其是否与激活再灌注损伤挽救激酶(reperfusion injury salvage kinase,RISK)信号通路有关。方法将原代培养的新生Wistar大鼠乳鼠心肌细胞分为8组,... 详细信息

目的研究β2-肾上腺素受体激动剂clenbuterol对原代培养的心肌细胞缺氧/复氧损伤的作用及其是否与激活再灌注损伤挽救激酶(reperfusion injury salvage kinase,RISK)信号通路有关。方法将原代培养的新生Wistar大鼠乳鼠心肌细胞分为8组,1正常培养组;2缺氧/复氧(A/R)组;3clenbuterol(1μmol·L-1)+A/R;4ICI118,551(10μmol·L-1)+clenbuterol(1μmol·L-1)+A/R组;5美托洛尔metoprolol(10μmol·L-1)+clenbuterol(1μmol·L-1)+A/R组;6 metoprolol(10μmol·L-1)+A/R组;7 PD98059(20μmol·L-1)+clenbuterol(1μmol·L-1)+A/R组;8LY294002(10μmol·L-1)+clenbuterol(1μmol·L-1)+A/R组。采用MTT法测定各组细胞存活率;比色法检测心肌细胞培养液的乳酸脱氢酶(LDH)含量;Hoechst 33342荧光染色法检测细胞凋亡率;分子探针DCFH-DA检测细胞内活性氧的水平;Western blot检测心肌细胞缺氧/复氧后ERK及p-ERK1/2蛋白的表达水平。结果与A/R组比较,clenbuterol+A/R组明显增高细胞存活率,降低LDH含量,降低细胞凋亡率,ROS产生减少,p-ERK1/2蛋白表达水平增高,而选择性β2受体阻断剂ICI 118,551可取消clenbuterol的上述作用,β1受体阻断剂Metoprolol对clenbuterol的作用无影响,PI3K抑制剂LY294002和ERK1/2抑制剂PD98059可阻断clenbuterol对心肌细胞缺氧/复氧损伤的保护作用。结论clenbuterol能够减轻心肌细胞缺氧/复氧损伤,加入选择性β2受体阻断剂ICI 118,551,PI3K抑制剂LY294002和ERK抑制剂PD98059均使clenbuterol的保护作用取消,表明clenbuterol可通过激动β2肾上腺素受体,激活RISK信号通路发挥抗心肌细胞缺氧/复氧损伤的作用。

关键词： clenbuterol 缺氧/复氧心肌细胞磷酸化ERK PI3K reperfusion injury salvage kinase(RISK)

clenbuterol诱导载酯蛋白D在猪脂肪组织中表达加强

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畜牧与兽医 2008年第1期40卷 3-7页

作者：张瑾过伟樊宝良朱文进郑李彬刘秋月邓学梅胡晓湘李宁河北科技师范学院河北秦皇岛066600 中国农业大学农业生物技术国家重点实验室北京100094 河北农业大学动物科技学院河北保定071001

clenbuterol(CLB,盐酸克伦特罗),俗称瘦肉精。由于其在动物体内广泛残留,可能导致食用此类产品的人中毒而被禁止使用,但已有研究表明该制剂可显著减少猪脂肪沉积,因此,对其分子机制进行深入研究将具有重要的理论与实践价值。本研究通过... 详细信息

clenbuterol(CLB,盐酸克伦特罗),俗称瘦肉精。由于其在动物体内广泛残留,可能导致食用此类产品的人中毒而被禁止使用,但已有研究表明该制剂可显著减少猪脂肪沉积,因此,对其分子机制进行深入研究将具有重要的理论与实践价值。本研究通过对家猪饲喂高剂量clenbuterol(≥1.5mg/kg体重),提高瘦肉率10%左右。通过实时荧光定量PCR技术对脂肪组织中与脂类代谢密切相关的4个基因研究发现,饲喂clenbuterol的猪脂肪组织内载酯蛋白D(apolipoprotein D,apoD)mRNA丰度增加超过2倍,而脂蛋白酯酶(lipoprotein lipase,lpl)、硬酯酰辅酶A去饱和酶(stearoyl-CoA desaturase,scd)和激素敏感脂酶(hormone sensitive lipase,hsl)mRNA丰度变化不明显。因此,推断clenbuterol通过改变部分脂代谢关键基因的表达来实现减少脂肪积累。载酯蛋白D可能是CLB应答基因之一。

关键词： clenbuterol 猪脂肪沉积定量PCR 载酯蛋白D

clenbuterol对肥育猪作用效果的研究

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浙江农业大学学报 1995年第3期21卷 227-231页

作者：肖日进许梓荣浙江农业大学饲料科学研究所

７０头“社长大”肥育猪始重（５８±ｌ．０）ｋｇ，按饲养试验要求分成５组，饲喂Ｃｌｅｎｂｕｔｅｒｏｌ含量（ｍｇ／ｋｇ）分别为５，１０，１５，２０的饲粮至猪体重９０ｋｇ左右时结束。结果表明：①饲粮中添加Ｃｌｅｎｂｕｔｅｒ... 详细信息

７０头“社长大”肥育猪始重（５８±ｌ．０）ｋｇ，按饲养试验要求分成５组，饲喂Ｃｌｅｎｂｕｔｅｒｏｌ含量（ｍｇ／ｋｇ）分别为５，１０，１５，２０的饲粮至猪体重９０ｋｇ左右时结束。结果表明：①饲粮中添加Ｃｌｅｎｂｕｔｅｒｏｌ能提高日增重、降低料重比。其中以１５ｍｇ／ｋｇ组效果最佳，日增重提高１５％（Ｐ＜０．０２），料重比下降１０％；②不同添加量的Ｃｌｅｎｂｕｔｅｒｏｌ均提高了猪胴体瘦肉率。其中１５ｍｇ／ｋｇ组瘦肉率升高５％（Ｐ＜０．０５），胴体脂肪比率则降低１５％（Ｐ＜０．０５），而且Ｃｌｅｎｂｕｔｅｒｏｌ对猪胴体组成的。影响存在部位间的差异，对前躯作用最明显；③提高瘦肉率的同时，Ｃｌｅｎｂｕｔｅｒｏｌ还使眼肌中的营养组成发生变化，蛋白质含量升高４．５％（Ｐ＜０．０５），脂肪含量降低３０％（Ｐ＜０．０５）。

关键词： clenbuterol 生产性能胴体特性育肥猪饲料

Studies on the Voltammetric Determination of clenbuterol at One-touch Immunochips

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Journal of Shanghai Jiaotong university(Science) 2005年第S1期10卷 139-145,171页

作者：柴春彦肖红玉刘国艳 School of Agriculture and Biology Shanghai Jiaotong Univ. Shanghai 201101 China

A novel immunosensor for determination of clenbuterol was described in this paper. Horseradish peroxidase (HRP) and Monoclonal antibody against clenbuterol was covalently immobilized onto the cellulose acetate membrane which was used as one-touch immunochips. After 20 min of competitive reaction of clenbuterol in samples against glucose oxidase-conjugated clenbuterol (CLB-GOD) with anti-clenbuteol antibody immobilized on the membrane, the one-touch transducer was fixed on the Nafion - ferrocene modified glass-carbon electrode (GCE, working electrode) by “O” ring. The increased current changes (ΔI) were recorded for the quantitative determination of clenbuterol by derivative cyclic voltammetry. Experimental results showed that a very effective system for electron transmission had been established using dual-enzyme system and Nafion-ferrocene modified GCE. Quantitative analysis of clenbuterol could be easily performed on this kind of immunosensor, which showed the advantage of time-saving and high sensitivity (The detecting limit of this kind of immunosensor can be down to 100 ng/L). In addition, other conditions affecting the electrochemical response were also discussed in detail.

关键词： clenbuterol immunosensor Nafion-ferrocen modified electrode One-touch immunochips cyclic voltammetry

Illegal Use of clenbuterol in Cattle Production in Mexico

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Health 2014年第8期6卷 673-676页

作者： Benjamin Valladares-Carranza Romulo Banuelos-Valenzuela Silvia D.Pena-Betancourt Valente Velázquez-Ordonez Yadira Velázquez-Armenta Alejandro Nava-Ocampo Centro de Investigacion y Estudios Avanzados en Salud Animal Facultad de Medicina Veterinaria y ZootecniaUniversidad Autonoma del Estado de MéxicoUnidad Académica de Medicina Veterinaria y ZootecniaUniversidad Autonoma de ZacatecasZacatecasMéxico Unidad Académica de Medicina Veterinaria y Zootecnia Universidad Autonoma de ZacatecasZacatecasMéxico Laboratorio de Toxicología Departamento de Produccion Agrícola y AnimalUniversidad Autonoma MetropolitanaMéxicoD.F.México Pharma Reasons TorontoONCanada

Slaughtered cattle (n = 582) from México State were bled for a serological screening of clenbuterol residues, using a commercial enzyme-linked immunosorbent assay (ELISA). clenbuterol residues were found in a total of 153/582 (26.2%) sera analysed. These results reinforced the assumption of the illegal use of clenbuterol in cattle production in México;therefore, routine screening examinations in slaughtered cattle were strongly advised considering the toxic potential for humans.

关键词： clenbuterol ELISA Cattle México

clenbuterol对大鼠脊髓损伤后运动功能恢复作用研究

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中国康复理论与实践 2005年第7期11卷 546-547页

作者：张岩李艺影赵继宗王硕刘相名北京天坛医院神经外科北京市100050 北京市神经外科研究所北京市100050

目的探讨肾上腺素能β2受体激动剂clenbuterol对脊髓损伤后运动功能恢复的作用。方法重力冲击方法建立大鼠胸髓(T10)损伤动物模型,以不同高度确立试验组。根据BassoBeattieBresnahan(BBB)评分标准评测运动功能情况。试验动物存活6周,进... 详细信息

目的探讨肾上腺素能β2受体激动剂clenbuterol对脊髓损伤后运动功能恢复的作用。方法重力冲击方法建立大鼠胸髓(T10)损伤动物模型,以不同高度确立试验组。根据BassoBeattieBresnahan(BBB)评分标准评测运动功能情况。试验动物存活6周,进行挫伤脊髓的组织形态学观察。结果clenbuterol治疗组的平均BBB分值为(10.0±2.1),而对照组分值为(6.3±1.5),Pclenbuterol治疗组周边组织残存区面积明显大于对照组。结论在脊髓损伤后应用肾上腺素能β2受体激动剂clenbuterol可保护脊髓残存的神经组织,帮助改善运动功能的恢复。

关键词： clenbuterol 脊髓损伤运动功能肾上腺素能β2受体激动剂大鼠