Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass Spectrometry
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 cove...
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| Format: | Article |
| Language: | English |
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MDPI AG
2018-08-01
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| Series: | Metabolites |
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| Online Access: | http://www.mdpi.com/2218-1989/8/3/44 |
| _version_ | 1828426922759028736 |
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| author | Jatin Nandania Gopal Peddinti Alberto Pessia Meri Kokkonen Vidya Velagapudi |
| author_facet | Jatin Nandania Gopal Peddinti Alberto Pessia Meri Kokkonen Vidya Velagapudi |
| author_sort | Jatin Nandania |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-10T16:53:44Z |
| format | Article |
| id | doaj.art-a9ba63378dbc42619e6ba78ae5a408df |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-12-10T16:53:44Z |
| publishDate | 2018-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-a9ba63378dbc42619e6ba78ae5a408df2022-12-22T01:40:49ZengMDPI AGMetabolites2218-19892018-08-01834410.3390/metabo8030044metabo8030044Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass SpectrometryJatin Nandania0Gopal Peddinti1Alberto Pessia2Meri Kokkonen3Vidya Velagapudi4Metabolomics Unit, Institute for Molecular Medicine Finland (FIMM), University of Helsinki, HiLIFE, Tukholmankatu 8, Biomedicum 2U, 00290 Helsinki, FinlandComputational Systems Medicine group, University of Helsinki, 00290 Helsinki, FinlandMetabolomics Unit, Institute for Molecular Medicine Finland (FIMM), University of Helsinki, HiLIFE, Tukholmankatu 8, Biomedicum 2U, 00290 Helsinki, FinlandMetabolomics Unit, Institute for Molecular Medicine Finland (FIMM), University of Helsinki, HiLIFE, Tukholmankatu 8, Biomedicum 2U, 00290 Helsinki, FinlandMetabolomics Unit, Institute for Molecular Medicine Finland (FIMM), University of Helsinki, HiLIFE, Tukholmankatu 8, Biomedicum 2U, 00290 Helsinki, FinlandThe 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.http://www.mdpi.com/2218-1989/8/3/44high-throughputtargetedsemiquantitationmetabolomicsLC-MSmultianalyte methodvalidationcross-platform comparabilityautomationbiomarkers |
| spellingShingle | Jatin Nandania Gopal Peddinti Alberto Pessia Meri Kokkonen Vidya Velagapudi Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass Spectrometry Metabolites high-throughput targeted semiquantitation metabolomics LC-MS multianalyte method validation cross-platform comparability automation biomarkers |
| title | Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass Spectrometry |
| title_full | Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass Spectrometry |
| title_fullStr | Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass Spectrometry |
| title_full_unstemmed | Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass Spectrometry |
| title_short | Validation and Automation of a High-Throughput Multitargeted Method for Semiquantification of Endogenous Metabolites from Different Biological Matrices Using Tandem Mass Spectrometry |
| title_sort | validation and automation of a high throughput multitargeted method for semiquantification of endogenous metabolites from different biological matrices using tandem mass spectrometry |
| topic | high-throughput targeted semiquantitation metabolomics LC-MS multianalyte method validation cross-platform comparability automation biomarkers |
| url | http://www.mdpi.com/2218-1989/8/3/44 |
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