盐酸克伦特罗残留酶联免疫吸附(ELISA)检测方法的研究

建立了间接ELISA检测盐酸克伦特罗的方法,并对其参数进行了分析计算。在该检测方法中,抗盐酸克伦特罗(CL)抗体最适稀释度为1∶1000,羊抗兔酶联抗体(HRP-IgG)的最适稀释度为1∶1500。该检测方法的检测灵敏度可达0·1046μg/L,生物检测限为1·452μg/L,线性检测范围为7·26~90·75μg/L。     

盐酸克伦特罗在肉制品中的危害及其检测方法研究进展

盐酸克伦特罗抑制脂肪合成,增加瘦肉率,被非法作为饲料添加剂使用。人食用含有盐酸克伦特罗的肉制品会危害身体健康。介绍盐酸克伦特罗的理化性质及中国、欧盟、日本对其残留量的要求,总结残留在肉及肉制品中的盐酸克伦特罗的检测方法。     

PCX柱净化-气相色谱质谱法测定牲畜瘦肉中的盐酸克伦特罗

<正>盐酸克伦特罗,又名瘦肉精,是一种对动物具有促进生长、减少脂肪含量的兴奋剂类物质,曾广泛应用于生猪的饲养。由于盐酸克伦特罗容易在动物组织内残留,当人们食用含有一定量盐酸克伦特罗的牲畜肉后,可能会引起中毒,所以我国已禁止将盐酸克伦特罗作为生长促进剂使用。但因其具经济效益,故非法使用盐酸克伦特罗的现象依然存在,因此建立快速高效的盐酸克伦特罗检测方法具有重要的意义。气相色谱-质谱检测盐酸克伦特罗的方法较多,如     

盐酸克伦特罗完全抗原的制备及其鉴定

利用重氮化法将小分子半抗原盐酸克伦特罗(CL)偶联到牛血清白蛋白(BSA)上制得盐酸克伦特罗完全抗原(CL-BSA).采用紫外分光光度法快速确定了BSA∶CL偶联比为1∶17,并通过SDS-PAGE法验证了CL与BSA偶联;间接ELISA法检测免疫Balh/c小鼠血清.结果显示1∶1000稀释后的血清在中和掉BSA的抗体后呈阳性;最后,通过细胞融合获得了一株能稳定分泌CL-McAb的细胞株4H5,经鉴定为IgG1.说明合成偶联比为1∶17的完全抗原能刺激小鼠产生针对盐酸克伦特罗的单克隆抗体.     

竞争酶联免疫吸附法测定猪肉中的瘦肉精

介绍了竞争酶联免疫吸附法测定猪肉中的盐酸克伦特罗的方法.利用盐酸克伦特罗试剂盒,对猪肉组织中残留的盐酸克伦特罗经抽提、竞争后,用酶标仪进行检测分析.此法较适用于现场检验,检测速度快、灵敏度高,是保证肉品卫生安全的较好监控方法.     

ELISA检测盐酸克伦特罗残留的方法学评价

以盐酸克伦特罗(clenbuterolhydrochloride)重氮化后分别连接到牛血清蛋白(BSA)和卵清蛋白(OVA)上制得免疫原BSA-CL和包被抗原OVA-CL。通过免疫兔获得含有多克隆抗体的血清,经硫酸铵沉淀、纯化,得到兔源抗CL的抗体,在此基础上建立了间接酶联免疫检测方法。实验结果表明,抗CL抗体最适稀释度为1:1000,羊抗兔酶联抗体(HRP-IgG)的最适稀释度为1:1500。该检测方法的检测灵敏度为1.452μg/L,线性检测范围为7.26～90.75μg/L。     

动物源性食品及生物材料中克伦特罗测定方法的研究进展

克伦特罗是一种非法的饲料药物添加剂，含有其残留的食品可引起食物中毒。为给我国检测食品中盐酸克伦特罗提供借鉴，从样品提取、净化、检测方法及其测定的发展趋势等方面综述了国内外献中动物性食品及生物材料中的克伦特罗残留测定方法。     

竞争酶联免疫吸附法测定猪肉中的盐酸克伦特罗的研究

介绍了竞争酶联免疫吸附法测定猪肉中的盐酸克伦特罗的方法。利用盐酸克伦特罗试剂盒，对猪肉组织中的盐酸克伦特罗残留经抽提、竞争后，用酶标仪进行检测分析。此法较适用于现场检验，检测速度快、灵敏度高，是保证肉品卫生安全的较好监控方法。     

应用ELISE法测定盐酸克伦特罗的残留

介绍了酶联免疫吸附法测定猪肉中的盐酸克伦特罗的方法.利用试剂盒对猪肉中的盐酸克伦特罗残留处理后,用酶标仪进行检测分析.此法较适用于现场检验,检测速度快、灵敏度高,是保证肉品卫生安全的较好监控方法.     

食品中残留盐酸克伦特罗免疫原的合成与鉴定

盐酸克伦特罗是一种只具有免疫反应性不具有免疫原性的小分子半抗原。本文采用重氮化法将其与牛血清白蛋白交联,用紫外扫描和SDS变性凝胶电泳方法检验。并用合成的CL_BSA免疫小鼠,经ELISA检验,小鼠产生了针对CL的抗体,表明合成的CL_BSA复合物具有免疫原性,为盐酸克伦特罗单克隆抗体的制备及其检测试剂盒的研制奠定了基础。     

微生物传感器在食品分析中的应用

微生物传感器是以活体微生物细胞为感应元件, 感应需要测定的靶标物质浓度并按照一定规律转换为可识别信号的检测装置。早期的微生物传感器以微生物测定和微生物电极的形式出现, 近年来随着分子生物学和合成生物学技术的进步, 基因工程类微生物传感器迅速发展, 并且由于其成本低、使用便捷、功能扩展性强等优势, 使其在食品安全领域具有良好的应用潜力。本文介绍了微生物传感器的发展历程和基本原理, 总结了微生物传感器在食品安全检测、食品品质分析、食品安全毒性评估、食品污染脱毒等方面的最新研究成果, 分析了微生物传感器发展趋势和当前的技术瓶颈。本综述可为食品安全快速检测技术研究和应用人员提供信息和技术参考, 推动微生物传感器领域的进一步发展。     

Development of a specific radioimmunoassay for the detection of clenbuterol residues in treated cattle

A radioimmunoassay for clenbuterol detection in cattle has been validated and used to monitor treated cattle. The tracer used was 4-amino-3,5-dichloro-alpha(tert-butylamino-methyl) benzyl alcohol (benzyl-3H)(clenbuterol) prepared by catalytic tritiation with tritium gas of 4-amino-3,5-dibromo-alpha-(tert-butylamino)-acetophenone, followed by chlorination at positions 3 and 5 in the aromatic ring. The rabbit antiserum was raised against a diazotized clenbuterol/human serum albumin conjugate. The assay described was sensitive (7.8 pg/tube) and reproducible. The intra- and inter-assay variability, which was assessed by measuring known quantities of clenbuterol in plasma, urine and faeces, was satisfactory for RIA. When this assay was used to monitor treated cattle the concentrations of clenbuterol in plasma, urine and faeces were directly related to the administered dose. The absorption and elimination of clenbuterol in cattle was rapid. Data obtained were consistent with results obtained in other species where a rapid clearance rate was also demonstrated.     

Development of a label-free electrochemical immunosensor based on carbon nanotube for rapid determination of clenbuterol

We have fabricated a label-free electrochemical immunosensor for the detection of clenbuterol, a kind of β-agonist. Clenbuterol was covalently linked to multi-wall carbon nanotubes (MWCNTs) through a two-step process using 1-(3-(dimethylamino)-propyl)-3-ethylcarbodiimide and N-hydroxysulfo-succinimide as crosslinkers. The clenbuterol-MWCNT conjugates were cast on a glassy carbon electrode. Cyclic voltammetry and differential pulse voltammetry were employed to monitor the fabrication steps of immunoreaction system using the redox probe of K₃Fe(CN)₆. In the presence of monoclonal antibody against clenbuterol, the redox peak current of [Fe(CN)₆]^3−/4− was decreased, presumably due to that antibody in solution could adsorb on the electrode surface modified clenbuterol-MWCNT conjugates. The selected monoclonal antibody showed very high sensitivity and specificity for clenbuterol, and was used for the detection and quantitative determination of clenbuterol in solution with a competitive mechanism. This approach provided a detection limit of 0.32 ng mL⁻¹. Accurate detection of clenbuterol in spiked animal feeds was demonstrated by comparison with conventional ELISA assays and LC–MS method.     

高效液相色谱内标法测定动物饲料中盐酸克伦特罗

以茶碱为内标物,建立高效液相色谱内标法测定禽畜饲料中盐酸克伦特罗的检测方法。样品经三氯乙酸溶液提取,经过Oasia HLB固相萃取小柱（3 mL/60 mg）处理,再用体积分数为5%的甲醇溶液冲洗后,采用Nova-Pak C18色谱柱（3.9 mm×150 mm×5 μm）分离,以乙腈-28 g/L磷酸二氢钠缓冲溶液（25:75,V/V）为流动相进行洗脱,流速0.8 mL/min,检测波长210 nm。结果表明,茶碱在该条件下能够与盐酸克伦特罗分离完全,可以作为测定盐酸克伦特罗的内标物。盐酸克伦特罗的加标回收率为97.8%～99.2%,精密度试验结果的相对标准偏差（RSD）为0.13%,表明该方法操作简单、准确度高、精密度良好,可用于饲料中盐酸克伦特罗含量分析。     

Cimaterol and clenbuterol residue analysis by HPLC-HPTLC in liver.

A method has been developed for the analysis of cimaterol and clenbuterol residues in liver, with detection limits of 0.25 micrograms/kg and 0.5 micrograms/kg, respectively. The recovery varied from 55% to 60%. After extraction, a clean-up procedure with Baker-spe C-18 columns was performed. The two chemical compounds of interest were eluted with methanol. Cimaterol and clenbuterol were quantitatively determined by high-performance liquid chromatography (HPLC) using an RP-Select B (5 microns) column and a post-column reaction procedure. The positive results were confirmed by high-performance thin-layer chromatography (HPTLC) as this technique reaches the same level of sensitivity as the HPLC method.     

Clenbuterol residues in pig muscle after repeat administration in a growth-promoting dose

The aim of this study was to determine the level of clenbuterol residues in muscle tissue of pigs after repeat administration in a growth-promoting dose. An anabolic dose of clenbuterol (20 μg/kg body mass per day) was administered orally to experimental group (n = 12) for 28 days, whereas control animals (n = 3) were left untreated. Clenbuterol treated pigs were randomly sacrificed (n = 3) on days 0, 3, 7 and 14 of treatment discontinuation and clenbuterol residues determined in muscle tissue. Determination of residual clenbuterol was by enzyme-linked immunosorbent assay (ELISA) as a screening method and liquid chromatography tandem mass spectrometry (LC-MS/MS) as a confirmation method. The highest clenbuterol content in the muscle of treated animals was recorded on day 0 of treatment cessation (4.40 ± 0.37 ng/g) and significantly (p < 0.05) exceeded the maximum residue limit (MRL) of 0.1 ng/g. On day 3 of withdrawal, it was 0.49 ± 0.22 ng/g and on day 7 0.10 ± 0.02 ng/g (at MRL); on day 14 of treatment discontinuation, clenbuterol content was below the limit of detection (< 0.1 ng/g) in all samples. Administration of clenbuterol as a growth promoter in pig production could lead to residues in meat for human consumption up to 7 days after treatment discontinuation.     

Detection of clenbuterol in beef meat,liver and kidney by mid‐infrared spectroscopy (FT‐Mid IR) and multivariate analysis

Mid‐infrared spectroscopy (FT‐Mid IR) coupled with multivariate analysis was used to predict clenbuterol in beef meat, liver and kidney. A SIMCA model was also developed to discriminate between pure (beef meat, liver and kidney) and spiked with clenbuterol samples (beef meat‐clenbuterol, liver‐clenbuterol and kidney‐clenbuterol). The best models to predict clenbuterol concentrations were obtained using the partial least squares algorithm (PLS) with a R² > 0.9 and SEC and standard error of prediction <0.296 and 0.324, respectively. The SIMCA model used to discriminate pure and spiked with clenbuterol samples showed 100% correct classification rate. Methods detection limit was 2 μg kg^?1. FT‐Mid IR coupled with chemometrics could be a simple and rapid screening tool for monitoring clenbuterol in beef meat, liver and kidney implicated in food poisoning. This method could be use for screening purposes.     

Detection of Cr⁶⁺ by the sulfur oxidizing bacteria biosensor: effect of different physical factors

A biosensor based on sulfur-oxidizing bacteria (SOB) for detection of toxic chemicals in water was developed. SOB are acidophilic microorganisms that get their energy through the oxidation of reduced sulfur compounds in the presence of oxygen to produce sulfuric acid. The reaction results in an increase in electrical conductivity (EC) and a decrease in pH. The bioassay is based on the inhibition of SOB in the presence of toxic chemicals by measuring changes in EC and pH. The effect of different physical factors such as HRT, inorganic sulfur (S°) particle size, and temperature on detection of Cr(6+) was studied. The detection of Cr(6+) (50 ppb) was improved by decreasing the hydraulic retention time (HRT) from 30 to 10 min and increasing S° particle size from 1 to 4.75 mm. Detection time was shorter at 30 °C compared to 45 °C and the SOB were active over a wide range of temperatures with a maximum temperature for growth at 45 °C. This novel biosensor is simple, highly sensitive to low Cr(6+) concentrations (50 ppb), and also minimizes detection time. The present findings can be applied to the proper continuous screening of water ecosystem toxicity.     

基于纳米粒子吸附的电化学生物传感器快速检测茶汤中的有机磷

利用ZrO2纳米粒子与有机磷的强吸附作用,制成ZrO2纳米粒子/金电极(ZrO2/Au)有机磷传感器,可实现茶汤中有机磷农残的快速检测。研究结果表明,该有机磷传感器在10～550ng/mL浓度范围内对目标分析物对硫磷有线性响应,检出限为2.0ng/mL。传感器制备简单、操作方便,灵敏度高,再生性好,非特异性吸附小,在食品安全监测领域具有较大的应用潜力。     

Clenbuterol residue analysis by HPLC-HPTLC in urine and animal tissues

A method for clenbuterol residue analysis in urine and animal tissues has been developed. The detection limits are 0.25 micrograms/l and 0.5 micrograms/kg, respectively. The recovery in urine varies from 85% to 90% and in animal tissues from 70% to 74%. The beta 2-agonist was liberated from the tissues by an enzymatic digestion, purified on Chem Elut columns using alkaline conditions and extracted with 0.01 mol/l HCl. Clenbuterol was quantified by high-performance liquid chromatography (HPLC) on a RP-8 column and a post-column reaction procedure. High-performance thin-layer chromatography (HPTLC) was performed on silica gel 60 plates and clenbuterol visualized by means of the modified Ehrlich's TLC spray reagent. Since this method is sensitive, as is HPLC, it was used to obtain a confirmation and to exclude false positive results.