动物性食品中残留盐酸克伦特罗的研究进展

介绍了盐酸克伦特罗(瘦肉精)的理化特性,分析了它在动物性食品中残留对人体和国民经济所造成的危害,同时又综述了动物肌肉组织中残留盐酸克伦特罗(瘦肉精)的各种分析检测方法并作了展望。     

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     

猪肉瘦肉精检测方法的比较与运用

20世纪80年代,美国科学家意外发现盐酸克伦特罗(俗称瘦肉精是一种兴奋剂)在提高猪肉瘦肉率上有较大效果,一度就有在饲料中添加一定量的盐酸克伦特罗的方法,并作为科研成果在生猪饲养中推广运用.     

盐酸克伦特罗的残留与食品安全

十年来,食品动物生产使用违禁药物"瘦肉精"--盐酸克仑特罗已受到了社会各个层面的广泛关注,食品安全越来越受到人们的重视.本文阐述了盐酸克伦特罗在机体内的分解代谢、残留和对人畜的危害.     

盐酸克伦特罗的残留与食品安全

十年来,食品动物生产使用违禁药物"瘦肉精"——盐酸克仑特罗已受到了社会各个层面的广泛关注,食品安全越来越受到人们的重视。本文阐述了盐酸克伦特罗在机体内的分解代谢、残留和对人畜的危害。     

出口肉及制品中盐酸克伦特罗的ELISA检测方法的建立

介绍了酶联免疫吸附法测定出口肉及制品中的盐酸克伦特罗的方法。利用盐酸克伦特罗试剂盒对出口肉及制品中残留的盐酸克伦特罗用酶标仪进行测定分析,结果显示,盐酸克伦特罗的交叉反应为100%,猪肉罐头和牛肉中的检测限(LOD)为0.026(μg/kg)和0.028(μg/kg),样品的平均回收率在83.6%～91.3%之间,相对标准偏差为3.5%～5.6%,经HPLC确证假阳性率≤2.0%。表明酶联免疫分析定量法可快速、准确实现对食品中盐酸克伦特罗残留量的快速筛选。     

旋转生物传感器高灵敏检测盐酸克伦特罗方法研究

盐酸克伦特罗(clenbuterol hydrochloride, CL),又名双氯醇胺、氨哮素、克喘素,为白色结晶状粉末, 无味略苦,是一种人工合成的β2-肾上腺素受体激动剂。盐酸克伦特罗又称"瘦肉精",被非法用于肉用动物生产。彻底查禁CL 的非法滥用,必须要有高效、灵敏、特异的残留检测方法。本研究着重应用免疫学和生物化学方法,构建了基于量子点标记的ATP合酶分子马达免疫旋转生物传感器 (immuno-rotary biosensor, IRB),结合荧光技术,利用中国科学院生物物理研究所张仲伦老师研发的BPCL弱光检测器实现了对盐酸克伦特罗快速、高灵敏度的检测。     

在线净化LC-MS/MS系统对动物组织中克伦特罗残留的检测分析

克伦特罗俗称瘦肉精,常被用作饲料添加剂以提高家畜的瘦肉含量,但由于其严重的毒副作用,欧盟于1988年1月1日起禁止盐酸克伦特罗物质当饲料添加剂使用。美国食品药品管理局(FDA)也于1991年被禁止了其作为饲料添加剂的用途。1997年,中华人民共和国农业部下文,严禁β-肾上腺素类激素在饲料和畜牧生产中使用,盐酸克伦特罗被列为第一位。     

荧光分光光度法测定猪肉中的盐酸克伦特罗的含量

利用荧光胺与盐酸克伦特罗（瘦肉精）生成高度荧光衍生物,检测猪肉中盐酸克伦特罗的含量.实验表明,荧光胺与盐酸克伦特罗反应生成的强荧光物质的最大激发波长和最大发射波长分别为500 nm、490 nm,利用L934）正交试验确定最佳衍生条件为衍生温度37℃,衍生时间40 min,衍生剂荧光胺（1μg/mL）加入量0.3 mL.荧光信号与盐酸克伦特罗的质量浓度变化呈良好的线性关系,线性方程为y=82 492x＋957.96,相关系数为0.993 3,此法可用于检测猪肉中瘦肉精的含量.     

基于酶联免疫法检测盐酸克伦特罗的微流控系统

基于微流控技术和酶联免疫法的原理建立动物性食品中盐酸克伦特罗残留的检测技术,确保食品安全。设计制作的微流控芯片结合搭建的微流控酶联免疫实验平台,可以实现对动物性食品中盐酸克伦特罗的现场、简便、快速的免疫反应和检测。结果表明:通过在芯片上包被盐酸克伦特罗抗原,由实验平台自动完成进样、清洗过程,30 min内即可实现对盐酸克伦特罗的快速检测。标准品检测范围为(0.10~8.10)ng/m L,加标回收率在95.5%~107.3%之间。使用微流控盐酸克伦特罗残留检测芯片系统,具有快速、自动化程度高、灵敏度高、成本低、方法学性能良好等优势,特别适用于非专业人士进行盐酸克伦特罗残留的现场、快速检测。     

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.     

Evaluation of commercial immunoassays for cross-reactivity to clenbuterol stereoisomers and bovine metabolites

Several commercially available immunoassay kits have been developed to detect the beta-adrenergic agonist clenbuterol HCl. Technical materials supplied with the kits do not generally report cross-reactivity with clenbuterol metabolites. Use of such kits to quantitate clenbuterol might lead to an overestimation of parent drug if metabolites were present. The objective of this study was to measure the cross-reactivity of clenbuterol metabolites with several commercially available clenbuterol immunoassays. Three clenbuterol-glucuronide conjugates, clenbuterol-sulphamate, 4-amino-3,5-dichloro-hippuric acid (clenbuterol-hippurate), and purified clenbuterol-stereoisomers were tested for cross-reactivity. The clenbuterol-sulphamate metabolite showed significant cross-reactivity (42-77%), but clenbuterol-hippurate showed very little competition (< 0.2%) towards clenbuterol. Clenbuterol-glucuronides had little (0.1-1.6%) cross-reactivity. In addition, (R)-, (S)-, and racemic clenbuterol were used to determine the stereospecificity of the kits. Both (R)- and (S)-clenbuterol competed for binding in two of the kits, however, in one kit the (S)-clenbuterol stereoisomer had an affinity 100 times greater than the (R)-stereoisomer. The presence of significant quantities of the sulphamate metabolite of clenbuterol in a biological matrix would cause an overestimation of the amount of parent clenbuterol. This study illustrates the inherent problems of using unvalidated immunoassays for quantitation purposes.     

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.     

氨基化聚苯乙烯微球的制备及在克伦特罗抗体纯化中的应用

进行氨基聚苯乙烯微球制备方法及其应用于克伦特罗抗体纯化的研究。通过分散聚合法制备聚苯乙烯微球,硝基反应成硝基化聚苯乙烯微球,再还原成氨基化微球。并利用电子显微镜及红外光谱仪对微球形态及结构进行表征。成功制备氨基聚苯乙烯微球后,将微球与盐酸克伦特罗分子发生重氮化反应,得到连接克伦特罗分子的微球。将这种微球免疫亲和吸附分离盐酸克伦特罗的抗体。最后将纯化后的抗体通过紫外检测,电泳检测等方法证明所制备的微球用于抗体的纯化可行。     

Evaluation of the illegal use of clenbuterol in Portuguese cattle farms from drinking water,urine, hair and feed samples

The recent discovery of clenbuterol contamination in Portuguese food led to the specific inspection of 16 cattle farms for β-agonists, involving the analysis of a total of 486 samples (78 feed, 106 drinking water, 168 urine and 134 hair). The samples were screened for the β-agonists: bromobuterol, cimaterol, clenbuterol, clenpenterol, clenproperol, hydroxymethylclenbuterol, mapenterol, salbutamol and terbutaline. Only clenbuterol was found in all analyzed matrices and the most likely method of illegal administration to animals was through drinking water. Of all samples analysed, 14.15% of drinking water were found positive in the range 0.03–3.80 mg l^?1 clenbuterol. Inclusion of hair samples in the Portuguese plan for clenbuterol residue control in live animals is discussed.     

Flow injection chemiluminescent determination of clenbuterol using GoldMag particles as carrier

A novel flow injection chemiluminescent (CL) enzyme immunoassay for clenbuterol analysis based on GoldMag particles is described. GoldMag is a new type of super-paramagnetic Fe₃O₄/Au composite particle used as a carrier in a flow injection CL system. Clenbuterol conjugated with ovalbumin (OVA) was immobilized onto GoldMag particles and the particles fixed in a micro-channel by an external electromagnetic field. The clenbuterol test sample and clenbuterol polyclonal antibody (Ab) were injected into the channel and incubated with GoldMag particles. Clenbuterol, immobilized on the magnetic particle surfaces, competes for polyclonal antibodies with clenbuterol in the test sample. The free Ab or Ab combined with the clenbuterol sample was washed away and the magnetic particles conjugated with Ag-Ab left in the micro-channel. Horseradish peroxidase (HRP)-labelled goat anti-rabbit immunoglobulin G (IgG) was added and reacted with clenbuterol polyclonal antibodies; excess goat anti-rabbit-HRP was then washed off. When chemiluminescent reagents were injected into the channel, emitted light from the magnetic particle surface was measured and recorded using a photomultiplier-based apparatus. The linear range of this novel method was 0.01-0.1 ng g^-1 and recovery of clenbuterol was 85-105% with a RSD of 3.2% (n = 11).     

应用超高效液相色谱-四级杆-飞行时间质谱 鉴定克伦特罗在猪尿中的主要代谢产物

目的 应用超高效液相色谱/四级杆-飞行时间质谱技术分析鉴定了克伦特罗在猪尿中的代谢产物,并推测克伦特罗在猪体内的主要代谢途径。方法 按10 mg/kg bw的剂量,给猪口服灌食克伦特罗,分别采集给药前及给药后的尿液样品。应用超高效液相色谱/四级杆-飞行时间质谱技术对样品进行分析,采用MetaboLynx XS软件进行数据处理,共检测到6 种克伦特罗的代谢产物,并根据碎片离子信息进行了结构鉴定。结果 猪尿中克伦特罗的代谢产物包括4-N-羟基克伦特罗（4-N-OH-CLE）、4-硝基克伦特罗（4-NO2-CLE）、克伦特罗及4-N-OH-CLE的葡萄糖醛酸结合物（GLU-CLE和GLU-OH-CLE）等。结论 根据所检测到的代谢产物,克伦特罗在猪体内的代谢途径包括4-N-氧化和葡萄糖醛酸结合等。     

Effects of frozen storage on the recovery of clenbuterol from urine samples for official control.

The effects of storage under frozen conditions on clenbuterol recovery from spiked calf urine samples were studied. Urine samples contaminated with 10 micro g l(-1) clenbuterol and frozen at -15 degrees C were analysed every 15 days over 6 months. A single frozen contaminated urine sample was also thawed every 15 days over 3 months for the analysis of 10-ml aliquots, after which the remaining portion was frozen again (-15 degrees C). The presence of clenbuterol was determined by GC-MS. A gradual decline in clenbuterol recovery was observed from the first to the 180th day. It was only possible to recover 1.74 +/- 0.06 micro g l(-1) (17%) clenbuterol on the 180th day in samples stored at -15 degrees C. Likewise, clenbuterol totally disappeared from spiked urine samples that had been successively frozen and thawed over 3 months. The results were confirmed in a study performed on 2704 calf urine samples collected on farms and analysed by HPTLC and GC-MS. Of 73 positive samples, 61 had been frozen for <10 days.