Concordant inter-laboratory derived concentrations of ceramides in human plasma reference materials via authentic standards
Federico Torta, Nils Hoffmann, Bo Burla, Irina Alecu, Makoto Arita, Takeshi Bamba, Steffany A. L. Bennett, Justine Bertrand‐Michel, Britta Brügger, Mónica P. Cala, Dolores Camacho‐Muñoz, Antonio Checa, Michael Chen, Michaela Chocholoušková, Michelle Cinel, Emeline Chu‐Van, Benoît Colsch, Cristina Coman, Lisa Connell, Bebiana C. Sousa, Alex M. Dickens, Maria Fedorova, Finnur Freyr Eiríksson, Héctor Gallart‐Ayala, Mohan Ghorasaini, Martin Giera, Xue Li Guan, Mark Haid, Thomas Hankemeier, Amy C. Harms, Marcus Höring, Michal Holčapek, Thorsten Hornemann, Chunxiu Hu, Andreas J. Hülsmeier, Kevin Huynh, Christina M. Jones, Julijana Ivanišević, Yoshihiro Izumi, Harald Köfeler, Sin Man Lam, Mike Lange, Jong Cheol Lee, Gerhard Liebisch, Katrice A. Lippa, Andrea F. Lopez‐Clavijo, Malena Manzi, Manuela Martinefski, Raviswamy G H Math, Satyajit Mayor, Peter J. Meikle, Marı́a Eugenia Monge, Myeong Hee Moon, Sneha Muralidharan, Anna Nicolaou, Thao Nguyen-Tran, Valerie B. O’Donnell, Matej Orešič, Arvind Ramanathan, Fabien Riols, Daisuke Saigusa, Tracey B. Schock, Heidi Schwartz, Guanghou Shui, Madhulika Singh, Masatomo Takahashi, Margrét Þorsteinsdóttir, Noriyuki Tomiyasu, Anthony Tournadre, Hiroshi Tsugawa, Victoria J. Tyrrell, Grace van der Gugten, Michael J.O. Wakelam, Craig E. Wheelock, Denise Wolrab, Guowang Xu, Tianrun Xu, John A. Bowden, Kim Ekroos, Robert Ahrends, Markus R. Wenk
Abstract
In this community effort, we compare measurements between 34 laboratories from 19 countries, utilizing mixtures of labelled authentic synthetic standards, to quantify by mass spectrometry four clinically used ceramide species in the NIST (National Institute of Standards and Technology) human blood plasma Standard Reference Material (SRM) 1950, as well as a set of candidate plasma reference materials (RM 8231). Participants either utilized a provided validated method and/or their method of choice. Mean concentration values, and intra- and inter-laboratory coefficients of variation (CV) were calculated using single-point and multi-point calibrations, respectively. These results are the most precise (intra-laboratory CVs ≤ 4.2%) and concordant (inter-laboratory CVs < 14%) community-derived absolute concentration values reported to date for four clinically used ceramides in the commonly analyzed SRM 1950. We demonstrate that calibration using authentic labelled standards dramatically reduces data variability. Furthermore, we show how the use of shared RM can correct systematic quantitative biases and help in harmonizing lipidomics. Collectively, the results from the present study provide a significant knowledge base for translation of lipidomic technologies to future clinical applications that might require the determination of reference intervals (RIs) in various human populations or might need to estimate reference change values (RCV), when analytical variability is a key factor for recall during multiple testing of individuals. Here, the authors compared measurements between 34 laboratories from 19 countries, to quantify by mass spectrometry four ceramides of clinical relevance in human blood plasma Standard Reference Materials. The main goals were to evaluate concordance obtained in a large inter-laboratory trial and to report absolute concentrations of four circulating lipids in a publicly available standard.