Immunoaffinity purification of peptide hormones prior to liquid chromatography–mass spectrometry in doping controls

Thomas, Andreas; Schänzer, Wilhelm; Delahaut, Philippe; Thevis, Mario

doi:10.1016/j.ymeth.2011.08.009

Cited by 84 publications

(77 citation statements)

References 33 publications

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“…Finally, novel, previously unknown macromolecules (i.e., TB500 17-23 fragment), with no current approval for human therapeutic use (i.e., agents under preclinical or clinical development, designer drugs, or compounds approved only for veterinary use), but illegally marketed (e.g., via Internet websites) [5] are included in section S0 ''Non-approved substances''. Different analytical approaches have already been developed and published to detect these compounds in nutritional supplements [6][7][8][9][10] and in doping control samples (either blood [11][12][13][14][15][16][17][18] or urine [19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37]), employing immunological [26,[35][36][37], electrophoretic [16][17][18][19]27], or liquid chromatography-mass spectrometry techniques [6-15, 20-25, 28-34] or a combination of different analytical technologies. In general, the most effective analytical approach (combined sample pretreatment and the instrumental method) has to take into ...…”

Section: Introductionmentioning

confidence: 99%

Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

et al. 2015

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A liquid chromatography-tandem mass spectrometry method was developed and used to simultaneously detect 19 small peptide hormones prohibited in sport and their main metabolites in urine after solid-phase extraction. Detection was achieved using a triple-quadrupole mass spectrometric detector coupled with an electrospray ionization interface after chromatographic separation with an octadecyl column based on fused-core particle technology. Sample pretreatment was performed by solid-phase extraction. The extraction procedure was optimized by comparison of different sorbents and washing/elution protocols. The best results were obtained using a mixed-mode weak cation exchange sorbent, two washing steps (ultrapurified water and methanol), and elution using 300 mM ammonium formate in 25 % ammonia/methanol (10/90) or 25 % ammonia/10 % formic acid/methanol (8/12/80). The procedure was validated in terms of sensitivity (lower limits of detection: 0.05-2.0 ng/ml depending on the target analyte), specificity, recovery [[60 %, coefficient of variation (CV) \15 % except for TB500 17-23 fragment, AOD9604, and ARA290 for which recovery was \50 %), ion suppression/enhancement (\35 %), robustness, carryover, stability of the target analytes [stable for at least for 2 days (25°C, 2 weeks (4°C), 2 months (-20°C)], and repeatability of retention times (CV \0.1 %) and relative abundances of the selected ion transitions (CV \15 %).The suitability of the method was confirmed by analyzing spiked and excreted urines, the latter collected after intravenous injection of 0.1 mg of GHRP-2.

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Section: Introductionmentioning

confidence: 99%

Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

et al. 2015

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“…IC sample cleanup is well suited for nanoLC and Review | Zheng, Mehl, Zhu, Xin & Olah Bioanalysis (2014) 6 (6) had been applied for the quantification of protein and peptide biomarkers in serum [73,[92][93][94].IC sample preparation is compatible with nanoLC for two main reasons: IC removes the bulk of sample matrix, which comprises the majority of the sample mass, the matrix could overload the column capacity, leading to poor chromatography and high background; and IC process can concentrate analytes, thereby lead to low LOQs or increase the assay sensitivity.…”

Section: Low-flow (Nano and Micro) Chromatographymentioning

confidence: 99%

Application and Challenges in Using LC–MS Assays for Absolute Quantitative Analysis of Therapeutic Proteins in Drug Discovery

et al. 2014

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As more protein therapeutics enter the drug-discovery pipeline, the traditional ligand-binding assay (LBA) faces additional challenges to meet the rapid and diverse bioanalytical needs in the early drug-discovery stage. The high specificity and sensitivity afforded by LC-MS, along with its rapid method development, is proving invaluable for the analysis of protein therapeutics in support of drug discovery. LC-MS not only serves as a quantitative tool to complement LBA in drug discovery, it also provides structural details at a molecular level, which are used to address issues that cannot be resolved using LBA alone. This review will describe the key benefits and applications, as well as the techniques and challenges for applying LC-MS to support protein quantification in drug discovery.

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“…As an alternative, hybrid LC-MS/MS platforms have been employed, that is, by combining an immunoaffinity sample preparation step with the LC-MS/MS quantification [8,9]. n LC-MS/MS approach using a digestion step This approach is more complex and mainly used for proteins or larger peptides.…”

Section: Lc-ms(/ms) Approaches For Bioanalysis Of Large Moleculesmentioning

confidence: 99%

LC–MS/MS of Large Molecules in A Regulated Bioanalytical Environment – Which Acceptance Criteria to Apply?

2013

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Immunoaffinity purification of peptide hormones prior to liquid chromatography–mass spectrometry in doping controls

Cited by 84 publications

References 33 publications

Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

Application and Challenges in Using LC–MS Assays for Absolute Quantitative Analysis of Therapeutic Proteins in Drug Discovery

LC–MS/MS of Large Molecules in A Regulated Bioanalytical Environment – Which Acceptance Criteria to Apply?

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