Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass Spectrometry

Metabolites

Gopal Peddinti¹  Alberto Pessia²  Meri Kokkonen²  Vidya Velagapudi²  Jatin Nandania² 

[1] Computational Systems Medicine group, University of Helsinki, 00290 Helsinki, Finland;Metabolomics Unit, Institute for Molecular Medicine Finland (FIMM), University of Helsinki, HiLIFE, Tukholmankatu 8, Biomedicum 2U, 00290 Helsinki, Finland;

关键词: high-throughput;    targeted;    semiquantitation;    metabolomics;    LC-MS;    multianalyte method;    validation;    cross-platform comparability;    automation;    biomarkers;   

DOI  :  10.3390/metabo8030044

来源: DOAJ

【 摘 要 】

The use of metabolomics profiling to understand the metabolism under different physiological states has increased in recent years, which created the need for robust analytical platforms. Here, we present a validated method for targeted and semiquantitative analysis of 102 polar metabolites that cover major metabolic pathways from 24 classes in a single 17.5-min assay. The method has been optimized for a wide range of biological matrices from various organisms, and involves automated sample preparation and data processing using an inhouse developed R-package. To ensure reliability, the method was validated for accuracy, precision, selectivity, specificity, linearity, recovery, and stability according to European Medicines Agency guidelines. We demonstrated an excellent repeatability of retention times (CV < 4%), calibration curves (R2 ≥ 0.980) in their respective wide dynamic concentration ranges (CV < 3%), and concentrations (CV < 25%) of quality control samples interspersed within 25 batches analyzed over a period of one year. The robustness was demonstrated through a high correlation between metabolite concentrations measured using our method and the NIST reference values (R2 = 0.967), including cross-platform comparability against the BIOCRATES AbsoluteIDQp180 kit (R2 = 0.975) and NMR analyses (R2 = 0.884). We have shown that our method can be successfully applied in many biomedical research fields and clinical trials, including epidemiological studies for biomarker discovery. In summary, a thorough validation demonstrated that our method is reproducible, robust, reliable, and suitable for metabolomics studies.

【 授权许可】

Unknown