Comprehensive screening of anabolic steroids, corticosteroids, and acidic drugs in horse urine by solid-phase extraction and liquid chromatography–mass spectrometry

Ho, Emmie N.M.; Leung, David K.K.; Wan, Terence S.M.; Yu, Nola H.

doi:10.1016/j.chroma.2006.03.089

Cited by 99 publications

(65 citation statements)

References 16 publications

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“…Although full-scan MS/MS spectra are not acquired in this mode of operation [termed selected reaction monitoring (SRM) and, its extension, multiple reaction monitoring (MRM)], the method provides high selectivity by monitoring chromatographic coelution of multiple transitions for a given peptide. It is important to note that SRM and MRM have already been successfully applied to a variety of biological applications, including quantification of DNA adducts purified from tissue (8) and detection of doping substances in human urine and plasma (9)(10)(11)(12). To date, the use of SRM and MRM in proteomics is less widespread, although MRM has been used to quantify protein expression (13), to find protein biomarkers for disease severity in rheumatoid arthritis (14), and for detection and quantitative analysis of protein phosphorylation (15)(16)(17).…”

mentioning

confidence: 99%

Multiple reaction monitoring for robust quantitative proteomic analysis of cellular signaling networks

Wolf-Yadlin

Hautaniemi

Lauffenburger

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

472

434

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Although recent developments in MS have enabled the identification and quantification of hundreds of phosphorylation sites from a given biological sample, phosphoproteome analysis by MS has been plagued by inconsistent reproducibility arising from automated selection of precursor ions for fragmentation, identification, and quantification. To address this challenge, we have developed a new MS-based strategy, based on multiple reaction monitoring of stable isotope-labeled peptides, that enables highly reproducible quantification of hundreds of nodes (phosphorylation sites) within a signaling network and across multiple conditions simultaneously. We have applied this strategy to quantify temporal phosphorylation profiles of 222 tyrosine phosphorylated peptides across seven time points following EGF treatment, including 31 tyrosine phosphorylation sites not previously known to be regulated by EGF stimulation. With this approach, 88% of the signaling nodes were reproducibly quantified in four analyses, as compared with only 34% by typical information-dependent analysis. As a result of the improved reproducibility, full temporal phosphorylation profiles were generated for an additional 104 signaling nodes with the multiple reaction monitoring strategy, an 88% increase in our coverage of the signaling network. This method is broadly applicable to multiple signaling networks and to a variety of samples, including quantitative analysis of signaling networks in clinical samples. Using this approach, it should now be possible to routinely monitor the phosphorylation status of hundreds of nodes across multiple biological conditions. epidermal growth factor receptor ͉ mass spectrometry ͉ signal transduction ͉ tyrosine phosphorylation L igand binding to cell surface receptors activates multiple protein tyrosine phosphorylation-mediated signaling cascades that regulate many cell biological processes, including proliferation, differentiation, migration, and cell death (1-3). To mechanistically define the relationship between signaling networks and downstream biological responses, it is necessary to quantify the dynamics of protein phosphorylation sites across multiple cell states and to correlate this information to quantitative phenotypic measurements. Until recently, antibody-based assays (e.g., FACS, Western blots, tissue microarrays) have been favored for most signaling network studies. These assays provide excellent quantitative information on the phosphorylation status of selected nodes within the network, but require a priori knowledge of the proteins and phosphorylation sites to be studied and are limited by the need to have high-quality, non-cross-reactive antibodies recognizing specific sites within the network. By comparison, analysis of protein phosphorylation by MS provides the capability of identifying novel phosphorylation sites on novel proteins within the network, requires minimal a priori knowledge, and is therefore compatible with both well and poorly characterized signaling networks. Recent developments in M...

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mentioning

confidence: 99%

Multiple reaction monitoring for robust quantitative proteomic analysis of cellular signaling networks

Wolf-Yadlin

Hautaniemi

Lauffenburger

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

472

434

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“…[104] Moreover, a comprehensive screening methodology has been developed and includes a single deconjugation procedure with a solution of protease and β-glucuronidase, and separate collection of basic and acidic/neutral fractions after SPE with mixed mode C8-cartridge. [105,106] Furthermore, a unified sample preparation procedure has been established that combines different deconjugation, extraction, and derivatization procedures. [107] It begins with two parallel procedures which include (1) enzymatic hydrolysis of sulfate and glucuronide conjugates (β-glucuronidase and arylsulfatase) and (2) methanolysis of the 17β-sulfate steroid conjugates.…”

Section: Horse Urine Samplesmentioning

confidence: 99%

“…Recently, hydrophilic interaction chromatography (HILIC) has become the technique preferred for the analysis of polar and ionizable compounds, [114][115][116] as occurred in the detection of myo-inositol trispyrophosphate (ITPP) in equine urine and plasma which is a new drug, capable of increasing the amount of oxygen in hypoxic tissues and is an ideal candidate as doping agent in racehorses. [117,118] LC-MS instruments equipped with mass analyzers with MS/MS capabilities, such as triple quadrupole (QqQ) [104][105][106]108] and triple quadrupole/linear ion trap (QTrap), [101,106,110,112,119] have been implemented successfully in common screening methods in equine doping control. These LC-MS/MS systems provide analyses with high sensitivity and specificity which are critical prerequisites for the detection of banned compounds in trace concentration levels in complex biological samples.…”

Section: Mass Spectrometric Techniques In the Equine Doping Controlmentioning

confidence: 99%

“…[105] • Forty anabolic steroids and corticosteroids using MRM in positive ESI mode and 52 acidic dugs using MRM in negative ESI mode, through fast-scanning MS with only 10 min overall turnaround times for both methods because of the high efficiency RP-column utilized. [106] • One hundred forty multiclass drugs in a single 13 min run through a UPLC column coupled with a fast scanning and fast ESI polarity switching tandem MS in SRM mode. [104] • Nineteen multiclass drugs using automated on-line solid phase extraction system coupled with a fast scanning QqQ-MS in positive ESI mode with SRM scan function in a 9.5 min run.…”

Section: Mass Spectrometric Techniques In the Equine Doping Controlmentioning

confidence: 99%

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Challenges in detecting substances for equine anti‐doping

Fragkaki

Kioukia‐Fougia

Kiousi

et al. 2017

Drug Testing and Analysis

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The artificial increase of the physical capability of horses using drugs is well known in racing and other equine sports. Both illicit and therapeutic substances are regarded as prohibited substances in competition in most countries. Some countries make distinctions for a few, specific drugs which are, however, allowed for use in other countries. The primary objective in the case of doping control is the detection of any trace of drug exposure, either parent drug or any of its metabolites, using the most powerful analytical methods which are generally based on chromatographic/mass spectrometric techniques. Of major concern in horseracing is the absence of a single organization regulating the anti-doping framework; instead of this, individual racing authorities provide rules and regulations often resulting in variations in the applied doping control programmes of different countries. The aim of this paper is to review the recent literature (approximately from 2012 to mid-2016) to highlight the numerous and diverse challenges faced in doping control of racing and equestrian sports, including the detection of designer drugs (anabolic steroids or stimulants) and of other emerging prohibited substances, such as peptides and noble gases in horse urine and plasma. Moreover, the application of 'omics' techniques (especially of metabolomics) deserves attention for establishing possible fingerprints of drug abuse as well as the evolution of instrumental analysis resulting a powerful ally in the fight against doping in equine sports.

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“…In recent years there have been a small number of multiresidue methods which detect quite a wide range of NSA IDs. Daeseleire et al analysing NSAIDs in equine plasma and urine [404,405]. These multi-class methods are discussed in greater detail in a later section (5.16 multi-class multi-residue analysis).…”

Section: Non Steroidal Anti-inflammatory Drugsmentioning

confidence: 99%

Current trends in sample preparation for growth promoter and veterinary drug residue analysis

Kinsella

O’Mahony

Malone

et al. 2009

Journal of Chromatography A

179

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A comprehensive review is presented on the current trends in sample preparation for isolation of veterinary drugs and growth promotors from foods. The objective of the review is to firstly give an overview of the sample preparation techniques that are applied in field. The review will focus on new techniques and technologies, which improve efficiency and coverage of residues. The underlying theme to the paper is the developments that have been made in multi-residue methods and particularly multi-class methods for residues of licensed animal health products, which have been developed in the last couple of years. The role of multi-class methods is discussed and how they can be accommodated in future residue surveillance.

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Comprehensive screening of anabolic steroids, corticosteroids, and acidic drugs in horse urine by solid-phase extraction and liquid chromatography–mass spectrometry

Cited by 99 publications

References 16 publications

Multiple reaction monitoring for robust quantitative proteomic analysis of cellular signaling networks

Multiple reaction monitoring for robust quantitative proteomic analysis of cellular signaling networks

Challenges in detecting substances for equine anti‐doping

Current trends in sample preparation for growth promoter and veterinary drug residue analysis

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