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Dulal C Kabiraz, Kinichi Morita, Kazuhira Sakamoto, Masashi Takahashi, Toshikazu Kawaguchi
This study proposed the filtration method for removal of inhibitors from real urine samples for immunoassay without centrifuge. Although the inhibitors could not be removed by the physical filtration, the carboxyl group terminated silica effectively removed the inhibitors. In a low pH, antibody formed aggregation due to the protonation. We propose to adjust pH of the sample solution by adding a phosphate buffer solution (pH 7.5). As a result of pretreatment, the SPR immunosensing achieved the SPR signal of 45 mdeg and a low limit of detection with 100 ppq (100 fg mL-1 )...
August 15, 2018: Talanta
Warren Lee, Syed A Ali, Leow Chiuan Yee, Tan Soo Choon, Leow Chiuan Herng
Anti-salbutamol antibodies remain as important tools for the detection of salbutamol abuse in athletic doping. This study evaluated the feasibility and efficiency of the chicken (Gallus gallus domesticus) as an immunization host to generate anti-salbutamol scFv antibodies by phage display. A phage display antibody library was constructed from a single chicken immunized against salbutamol-KLH conjugate. After a stringent biopanning strategy, a novel scFv clone which was inhibited by free salbutamol recorded the highest affinity...
May 15, 2018: Analytical Biochemistry
Turibius Simon, Muthaiah Shellaiah, Perpectual Steffi, Kien Wen Sun, Fu-Hsiang Ko
New glutamic acid (Glu) and polyethylenimine (PE) functionalized ultra-stable gold nanoparticles (PE-Glu-AuNPs) were developed via a simple NaBH4 reduction method. The low toxicity and biocompatibility of PE-Glu-AuNPs were confirmed via an MTT assay in Raw 264.7 cells. Excitingly, PE-Glu-AuNPs were found to be extremely stable at room temperature up to six months and were utilized in an effective colorimetric naked eye assay of clenbuterol (CLB) and ractopamine (RCT) at pH 5. It was found that the selective assay of CLB and RCT is not affected by any other interferences (such as alanine, phenylalanine, NaCl, CaCl2 , threonine, cysteine, glycine, glucose, urea and salbutamol)...
September 6, 2018: Analytica Chimica Acta
Fernando X Moreira, Renata Silva, Maria B André, Paula G de Pinho, Maria L Bastos, João Ruivo, Patrícia Ruivo, Helena Carmo
The use of performance enhancing drugs is not only common in humans, but also in animal sports, including racing of horses, greyhounds and pigeons. The development of accurate analytical procedures to detect doping agents in sports is crucial in order to protect the fair-play of the game, avoid financial fraud in the attribution of eventual awards and, even more important, to protect the animals from harmful drugs and/or dangerous dosage regimens. The present study aimed to develop and validate, a method that enabled the screening and confirmation of the presence of a beta-agonist (clenbuterol) and three corticosteroids (betamethasone, prednisolone and budesonide) in faeces from pigeons...
May 4, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Qiong Huang, Tong Bu, Wentao Zhang, Lingzhi Yan, Mengyue Zhang, Qingfeng Yang, Lunjie Huang, Baowei Yang, Na Hu, Yourui Suo, Jianlong Wang, Daohong Zhang
Immunochromatographic assays (ICAs) are most frequently used for on-site rapid screening of clenbuterol. To improve sensitivity, a novel probe with bacteria as signal carriers was developed. Bacteria can load a great deal of gold nanoparticles (AuNPs) on their surface, meaning much fewer antibodies are needed to produce clearly visible results, although low concentrations of antibody could also trigger fierce competition between free analyte and the immobilized antigen. Thus, a limited number of antibodies was key to significantly improved sensitivity...
October 1, 2018: Food Chemistry
Dongsheng Xu, Huikai Shao, Rongying Luo, Qiqin Wang, Elena Sánchez-López, Salvatore Fanali, Maria Luisa Marina, Zhengjin Jiang
A facile single-step preparation strategy for fabricating vancomycin functionalized organic polymer-based monolith within 100 μm fused-silica capillary was developed. The synthetic chiral functional monomer, i.e 2-isocyanatoethyl methacrylate (ICNEML) derivative of vancomycin, was co-polymerized with the cross-linker ethylene dimethacrylate (EDMA) in the presence of methanol and dimethyl sulfoxide as the selected porogens. The co-polymerization conditions were systematically optimized in order to obtain satisfactory column performance...
May 3, 2018: Journal of Chromatography. A
Fabrizio Schifano, Stefania Chiappini, John M Corkery, Amira Guirguis
Recently, a range of prescription and over-the-counter drugs have been reportedly used as Novel Psychoactive Substances (NPS), due to their potential for abuse resulting from their high dosage/idiosyncratic methods of self-administration. This paper provides a systematic review of the topic, focusing on a range of medications which have emerged as being used recreationally, either on their own or in combination with NPS. Among gabapentinoids, pregabalin may present with higher addictive liability levels than gabapentin, with pregabalin being mostly identified in the context of opioid, polydrug intake...
April 22, 2018: Brain Sciences
L Herpin, E Bichon, L Rambaud, F Monteau, B Le Bizec
β-agonistic drugs have been forbidden as growth promoters in rearing animals in Europe since the late 1980s (Dir 96/22/EC). Specific and sensitive analytical methods based on UHPLC-MS/MS allow to monitor a large set of these substances. However, optimal performances are not observed for all the target analytes, especially for those exhibiting the highest polarities. We developed an SFC-MS/MS approach to cover the huge elution window of β-agonists, from the most polar which are usually eluted in the void volume when using reversed phase chromatography in conventional HPLC to the most apolar ones...
April 11, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Weilin L Shelver, David J Smith
Zilpaterol is a β-adrenergic agonist feed additive approved in the United States to increase weight gain and improve feed efficiency of cattle. A zilpaterol immunochromatographic assay was developed as an economical and user friendly rapid detection method for zilpaterol and validated using urine and tissue samples derived from animal studies. The assay sensitivity was 1.7 - 23.2 ng g-1 or mL-1 across a variety of feed and animal matrices and did not cross react with clenbuterol or ractopamine. No sample pre-treatment of cattle and sheep urine was needed, but horse urine and feed required dilution; skeletal muscle required solvent extraction prior to testing...
April 12, 2018: Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment
Nurul Ain A Talib, Faridah Salam, Yusran Sulaiman
Development of an immunoassay for clenbuterol (CLB) detection required an anti-CLB antibody as an important bioreceptor. In this study, we report our work on production and purification of a rabbit-derived polyclonal anti-CLB antibody. The antibody was then purified by nProtein A Sepharose affinity column and the antibody purity was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The activities of purified antibody were evaluated based on high antibody titer determined from enzyme-linked immunosorbent assay (ELISA)...
March 29, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Renyue Ji, Shuai Chen, Wei Xu, Zhen Qin, Jing Fu Qiu, Chao Rui Li
An ultrasensitive immunosensor for the direct detection of the illegally used livestock feed clebuterol (CLB) is described. It is based on the use of a glassy carbon electrode modified with an MoS2 -AuPt nanocomposite and on biotin-streptavidin interaction. The use of MoS2 -AuPt accelerates electron transfer, and this leads to a sharp increase in the electrochemical signal for the electrochemical probe hydrogen peroxide. Differential pulse voltammetry was used to record the current signal at a peak potential of -0...
March 6, 2018: Mikrochimica Acta
Muthaiah Shellaiah, Turibius Simon, Parthiban Venkatesan, Kien Wen Sun, Fu-Hsiang Ko, Shu-Pao Wu
Nanodiamonds were modified such that they carry thiol groups (ND-thiol). Gold nanoparticles were reacted with ND-thiol to obtain a highly stable conjugate of the type ND@AuNPs. Both ND-thiol and the ND@AuNPs were characterized by SEM, TEM, AFM, DLS, zeta potential, XPS, XRD, UV-Vis, Raman, FTIR and cytotoxicity studies. Their biocompatibility was confirmed via an MTT assay with HeLa cells. At a pH value of 6, the ND@AuNPs represent a colorimetric probe that can be used to selectively detect the illegally used β-adrenergic drug clenbuterol (CLB) and the pollutant chromium(III)...
December 20, 2017: Mikrochimica Acta
Kaylee R Mastrianni, Kalmorat Metavarayuth, William E Brewer, Qian Wang
An analytical procedure for the analysis of 10 β-adrenergic agonists (cimaterol, terbutaline, salbutamol, isoxsuprine, ractopamine, cimbuterol, clenbuterol, brombuterol, mabuterol and mapenterol) in pork meat was developed and validated using LC-MS/MS. An automated dispersive pipette extraction (DPX) was employed on a Hamilton Microlab® NIMBUS96® platform to extract the analytes of interest prior to LC-MS/MS analysis. The extraction time was <20 min with a total LC-MS/MS run time of 9.6 min. The method was fully validated in accordance with the international guidelines (European Commission Decision 2002/657/EC and National Standards of People's Republic of China, GB/T 22286-2008) for limit of detection, limit of quantitation, carryover, extraction efficiency, matrix effects, linearity, and within and between-run precision...
March 14, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Ruiguo Wang, Wei Zhang, Peilong Wang, Xiaoou Su
An ultrasensitive paper based lateral flow assay is described for rapid and simultaneous fluorometric detection of several β-agonists including clenbuterol and its chemical analogues (mabuterol, brombuterol, cimaterol, cimbuterol, bromchlorbuterol and banbuterol). A nonspecific monoclonal antibody (mAb) against clenbuterol and its analogues was prepared and employed in a competitive immunoassay where mAb conjugated to fluorescent nanoparticles and free β-agonists compete for the binding sites. This enables rapid screening for the 7 β-agonists in a single run that takes about 8 min...
2018: Mikrochimica Acta
Giulia Milano, Stefania Chiappini, Francesca Mattioli, Antonietta Martelli, Fabrizio Schifano
A recent years' increase in misusing levels of image- and performance- enhancing drugs (IPEDs) has been observed. Out of these drugs, beta-2 agonists have recently emerged for their potential of misuse, especially for slimming and bodybuilding purposes. To this perspective, clenbuterol ('the size zero pill') has been reported as being both popular and widely available from the illegal market. All clenbuterol and salbutamol misuse/abuse/dependence/withdrawal/overdose/off-label spontaneous reports (2006-2016) from the European Medicines Agency (EMA) EudraVigilance (EV) database were collected and analysed by age range, gender, concomitant therapies and source of information...
March 2, 2018: Basic & Clinical Pharmacology & Toxicology
Gerard J Marek, Brian P Ramos
5-Hydroxytryptamine2A (5-HT2A ) receptors are enriched in layers I and Va of the rat prefrontal cortex and neocortex and their activation increases the frequency of glutamatergic excitatory post-synaptic potentials/currents (EPSP/Cs) onto layer V pyramidal cells. A number of other G-protein coupled receptors (GPCRs) are also enriched in cortical layers I and Va and either induce (α1 -adrenergic and orexin2 ) or suppress (metabotropic glutamate2 [mGlu2 ], adenosine A1 , μ-opioid) both 5-HT-induced EPSCs and head twitches or head shakes induced by the phenethylamine hallucinogen 2,5-dimethoxy-4-iodoamphetamine (DOI)...
2018: Frontiers in Pharmacology
Paulo Vinicius F da Costa, Rejane Maria P da Silva, Hugo B Suffredini, Wendel A Alves
This work describes the electro polymerization of a poly-L-arginine film onto boron-doped diamond (BDD) electrode and glassy carbon electrode (GCE) surfaces. The morphological and electrochemical properties of the modified electrodes were studied by atomic force microscopy and electrochemical methods, and their potential for terbutaline sulfate (TBS) detection was determined by voltammetry and chronoamperometry techniques. Our results demonstrate that the electrochemical surface area of both uncoated-bare electrodes (GCE e BDD) did not have significant differences in performance...
July 1, 2018: Journal of Nanoscience and Nanotechnology
B Gray, M Viljanto, E Menzies, L Vanhaecke
The detection of drugs in human hair samples has been performed by laboratories around the world for many years and the matrix is popular in disciplines, such as workplace drug testing. To date, however, hair has not become a routinely utilised matrix in sports drug detection. The analysis of hair samples offers several potential advantages to doping control laboratories, not least of which are the greatly extended detection window and the ease of sample collection and storage. This article describes the development, validation, and utilisation of a sensitive ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS) method for the detection of 50 compounds...
February 12, 2018: Drug Testing and Analysis
Pietro Mascagni, Fabio Melandro, Zoe Larghi Laureiro, Gianluca Mennini, Massimo Rossi
This article is the first description of a spontaneous hepatic rupture in a young bodybuilder with a history of clenbuterol and ephedrine alkaloid use. The patient presented with a sudden mid-epigastric pain and vomiting. Hemoglobin levels decreased a few hours later and a computed tomography scan was performed which revealed a rupture of the right liver capsule and hemoperitoneum. Two attempts at transarterial embolization did not control the bleeding and a right hemihepatectomy was performed. The pathological report identified a hepatic adenoma, a capsular tear and diffuse peliosis hepatis...
April 2018: Revista Española de Enfermedades Digestivas
Éadaoin W Griffin, Justin D Yssel, Eoin O'Neill, Katie J Ryan, Noreen Boyle, Peter Harper, Andrew Harkin, Thomas Connor
OBJECTIVES: Clenbuterol is a brain penetrant β2 -adrenoceptor agonist with anti-inflammatory and putative neuroprotective properties. In the present investigation, the effect of clenbuterol was assessed in a rat model of acute brain injury induced by intra-striatal administration of the pro-inflammatory cytokine IL-1β. METHODS: Clenbuterol (0.5 mg/kg; i.p.) was administered one hour prior to stereotactically delivered IL-1β (100 ng) into the striatum. Four hours postinjection, rats were anesthetized, blood samples were collected for circulating cytokine and chemokine analysis, and the ipsilateral striatum and liver tissue were harvested for mRNA expression analysis of target genes...
April 2018: Immunopharmacology and Immunotoxicology
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