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Repisalud author Vialas, Vital author Colomé-Calls, Núria author Abian, Joaquín author Aloria, Kerman author Alvarez-Llamas, Gloria author Antúnez, Oreto author Arizmendi, Jesus M author Azkargorta, Mikel author Barceló-Batllori, Silvia author Barderas, Maria G author Blanco, Francisco author Casal, José Ignacio author Casas, Vanessa author de la Torre, Carolina author Chicano-Gálvez, Eduardo author Elortza, Felix author Espadas, Guadalupe author Estanyol, Josep M author Fernandez-Irigoyen, Joaquín author Fernandez-Puente, Patricia author Fidalgo, María José author Fuentes, Manuel author Gay, Marina author Gil, Concha author Hainard, Alexandre author Hernaez, Maria Luisa author Ibarrola, Nieves author Kopylov, Arthur T author Lario, Antonio author Lopez, Juan Antonio author López-Lucendo, María author Marcilla, Miguel author Marina-Ramírez, Anabel author Marko-Varga, Gyorgy author Martín, Luna author Mora, Maria I author Morato-López, Esperanza author Muñoz, Javier author Odena, Maria Antonia author de Oliveira, Eliandre author Orera, Irene author Ortea, Ignacio author Pasquarello, Carla author Ray, Kevin B author Rezeli, Melinda author Ruppen, Isabel author Sabidó, Eduard author Del Pino, Manuel M Sanchez author Sancho, Jaime author Santamaría, Enrique author Vazquez, Jesus author Vilaseca, Marta author Vivanco, Fernando author Walters, James J author Zgoda, Victor G author Corrales, Fernando J author Canals, Francesc author Paradela, Alberto funder Instituto de Salud Carlos III funder Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF) 2018-11-20T14:21:22Z 2018-11-20T14:21:22Z 2017 J Proteomics. 2017; 152:138-149 10.1016/j.jprot.2016.10.014 1876-7737 1874-3919 Journal of proteomics 27989941 http://hdl.handle.net/20.500.12105/6647 Despite the maturity reached by targeted proteomic strategies, reliable and standardized protocols are urgently needed to enhance reproducibility among different laboratories and analytical platforms, facilitating a more widespread use in biomedical research. To achieve this goal, the use of dimensionless relative retention times (iRT), defined on the basis of peptide standard retention times (RT), has lately emerged as a powerful tool. The robustness, reproducibility and utility of this strategy were examined for the first time in a multicentric setting, involving 28 laboratories that included 24 of the Spanish network of proteomics laboratories (ProteoRed-ISCIII). According to the results obtained in this study, dimensionless retention time values (iRTs) demonstrated to be a useful tool for transferring and sharing peptide retention times across different chromatographic set-ups both intra- and inter-laboratories. iRT values also showed very low variability over long time periods. Furthermore, parallel quantitative analyses showed a high reproducibility despite the variety of experimental strategies used, either MRM (multiple reaction monitoring) or pseudoMRM, and the diversity of analytical platforms employed. BIOLOGICAL SIGNIFICANCE: From the very beginning of proteomics as an analytical science there has been a growing interest in developing standardized methods and experimental procedures in order to ensure the highest quality and reproducibility of the results. In this regard, the recent (2012) introduction of the dimensionless retention time concept has been a significant advance. In our multicentric (28 laboratories) study we explore the usefulness of this concept in the context of a targeted proteomics experiment, demonstrating that dimensionless retention time values is a useful tool for transferring and sharing peptide retention times across different chromatographic set-ups. eng Inter-laboratory validation Liquid chromatography Multiple reaction monitoring Proteomics Reproducibility Standardization Targeted proteomics A multicentric study to evaluate the use of relative retention times in targeted proteomics research article 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 URL https://repisalud.isciii.es/bitstreams/c6b5b6ab-bd36-47aa-a76c-8434c1376d38/download File MD5 86cd0ca4079782a1369a3c8793b3af92 2497673 application/pdf PME10_previo__.pdf URL https://repisalud.isciii.es/bitstreams/3954c666-5b61-4ed2-b6a8-e4b7b35a3fd3/download File MD5 240be0894d840f85c6e0988f0e123c93 49697 text/plain PME10_previo__.pdf.txt