A high-throughput screening (HTS) immunochemical method for the analysis of stanozolol metabolites in cattle urine samples

Salvador, J.‐Pablo; Sánchez‐Baeza, Francisco; Marco, M.‐Pilar

doi:10.1016/j.jchromb.2009.08.027

Cited by 11 publications

(9 citation statements)

References 33 publications

(44 reference statements)

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“…Moreover, to achieve efficient control of AAS, the knowledge of the metabolic pathway is mandatory, allowing us also to recognize metabolites. In particular, for Stz, the 16β-hydroxy-stanozolol ( 16-OH Stz ) has been identified as the major urine metabolite in cattle [ 30 ] and humans [ 31 , 32 ]. In addition, 16-OH Stz is principally secreted as glucuronide or glucuronide of the sulfate conjugate, as often happens for Stz and other steroids.…”

Section: Resultsmentioning

confidence: 99%

Simple Synthesis of 17-β-O-hemisuccinate of Stanozolol for Immunoanalytical Methods

2020

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The use of doping in sports is a global problem that affects athletes around the world. Among the different methods developed to detect doping agents in biological samples, there are antibody-based methods that need an appropriate hapten design. Steroids with a hydroxyl group can be converted to the corresponding hemisuccinates. A novel approach to the synthesis of 17β-O-hemisuccinate of the common doping agent stanozolol is described here. Acylation of stanozolol with methyl 4-chloro-4-oxobutyrate/4-dimethylaminopyridine, followed by mild alkaline hydrolysis with methanolic sodium hydroxide at room temperature, gave the simultaneous protection and deprotection of pyrazole-nitrogen atoms. The proposed new synthetic method allows the desired hemisuccinate derivative to be obtained in only two steps, and with a good total yield starting from stanozolol.

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Section: Resultsmentioning

confidence: 99%

Simple Synthesis of 17-β-O-hemisuccinate of Stanozolol for Immunoanalytical Methods

2020

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“…This is based on the immobilized antigen and the separation of the individual reaction steps. Characteristic features include high sensitivity and the possibility of measurement in biological or food samples of various origins [ 84 ]. In recent years, the use of chemiluminescent enzyme immunoassays (CLEIAs) in clinical diagnostics and analytical tests for food and pharmacological purposes has also become widespread; this is primarily due to their very high sensitivity, broad detection range, and, above all, the speed of their procedure, which is significantly shorter compared to conventional ELISA.…”

Section: Antibody-based Approaches For Aas Determinationmentioning

confidence: 99%

Advances in the Determination of Anabolic-Androgenic Steroids: From Standard Practices to Tailor-Designed Multidisciplinary Approaches

Huml

Tauchen

Rimpelová

et al. 2021

Sensors

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Anabolic-androgenic steroids (AASs), a group of compounds frequently misused by athletes and, unfortunately, also by the general population, have lately attracted global attention; thus, significant demands for more precise, facile, and rapid AAS detection have arisen. The standard methods ordinarily used for AAS determination include liquid and gas chromatography coupled with mass spectrometry. However, good knowledge of steroid metabolism, pretreatment of samples (such as derivatization), and well-trained operators of the instruments are required, making this procedure expensive, complicated, and not routinely applicable. In the drive to meet current AAS detection demands, the scientific focus has shifted to developing novel, tailor-made approaches leading to time- and cost-effective, routine, and field-portable methods for AAS determination in various matrices, such as biological fluids, food supplements, meat, water, or other environmental components. Therefore, herein, we present a comprehensive review article covering recent advances in AAS determination, with a strong emphasis on the increasingly important role of chemically designed artificial sensors, biosensors, and antibody- and fluorescence-based methods.

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“…The preparation and characterization of the immunoreagents for Stanozolol (ST, As147), Tetrahydrogestrinone (THG, As170), Boldenone (B, As138) and Fluoroquinolones (FQ,As172) used in this study have been described before (Calvo et al 2009;Pinacho et al 2012;Salvador et al 2007;Salvador et al 2008Salvador et al , 2009Tort et al 2012b). ST was purchased from Sequoia Research Products, Ltd. (Oxford, UK).…”

Section: Reagents and Immunoreagentsmentioning

confidence: 99%

“…With this scenario, we report here for the first time the development of a multiplexed (multiple target analytes) and multimodal (multiple RPs) LSPR biosensor using biofunctional plasmonic nanostructured chips manufactured on a DNA microarray using DDI to address distinct multifunctional AuNPs to predefined positions of a surface. As a proof of concept, we have used these chips to detect Anabolic Androgenic Steroids (AAS) for which immunoreagents are available in our laboratory (Salvador et al 2007;Salvador et al 2008Salvador et al , 2009Tort et al 2012b;Tort et al 2009).…”

Section: Introductionmentioning

confidence: 99%

Multimodal plasmonic biosensing nanostructures prepared by DNA-directed immobilization of multifunctional DNA-gold nanoparticles

Tort

Salvador

Marco

2017

Biosensors and Bioelectronics

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Biofunctional multimodal plasmonic nanostructures suitable for multiplexed localized surface plasmon resonance (LSPR) biosensing have been created by DNA-directed immobilization (DDI) of two distinct multifunctional biohybrid gold nanoparticles. Gold nanoparticles (AuNP) of distinct sizes, and therefore showing distinct plasmon resonant peaks (RP), have been biofunctionalized and codified with two different single stranded-DNA (ssDNA) chains. One of these oligonucleotide chains has been specifically designed to direct each AuNP to a distinct location of the surface of a DNA microarray chip through specific hybridization with complementary oligonucleotide strands. Scanning Electron Microscopy (SEM) has been used to demonstrate selective immobilization of each AuNP on distinct spots. The second ssDNA chain of the AuNPs provides the possibility to introduce by hybridization distinct types of bioactive molecules or bioreceptors, on a reversible manner. In this work, hapten-oligonucleotide bioconjugate probes, with sequences complementary to the second ssDNA linked to the AuNP, have been synthesized and used to create multiplexed hapten-biofuncionalized plasmonic nanostructures. The oligonucleotide probes consist on anabolic androgenic steroid haptens (AAS) covalently linked to specifically designed oligonucleotide sequences. The biofunctionality of these plasmonic nanostructures has been demonstrated by fluorescent microarray immunoassay and LSPR measurements, recording the shift of the RP produced after the antibody binding to the corresponding hapten-oligonucleotide probes immobilized on the nanostructured surface. Preliminary data show that this approach could allow manufacturing multifunctional multimodal LSPR chips for multiplexed analysis of different substances reaching very good detectability. Thus, small molecular weigh, analytes such as stanozolol (ST,) could be detected at concentrations in the low nM range. The results here presented open the door for an easy way to construct site-encoded multiplexed multimodal LSPR sensor transducers, combining the DDI strategies with multimodal biohybrid nanoparticles showing distinct optical properties.

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A high-throughput screening (HTS) immunochemical method for the analysis of stanozolol metabolites in cattle urine samples

Cited by 11 publications

References 33 publications

Simple Synthesis of 17-β-O-hemisuccinate of Stanozolol for Immunoanalytical Methods

Simple Synthesis of 17-β-O-hemisuccinate of Stanozolol for Immunoanalytical Methods

Advances in the Determination of Anabolic-Androgenic Steroids: From Standard Practices to Tailor-Designed Multidisciplinary Approaches

Multimodal plasmonic biosensing nanostructures prepared by DNA-directed immobilization of multifunctional DNA-gold nanoparticles

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