High-throughput, low-volume, multianalyte quantification of plasma metabolites related to one-carbon metabolism using HPLC-MS/MS
Ø Midttun, G Kvalheim, PM Ueland - Analytical and bioanalytical chemistry, 2013 - Springer
Ø Midttun, G Kvalheim, PM Ueland
Analytical and bioanalytical chemistry, 2013•SpringerRisk of chronic diseases, like cardiovascular disease and cancer, has been associated with
biomarkers related to one-carbon metabolism, which comprises a metabolic network of
cross-talking pathways. To address this complexity in epidemiological studies, we have
established an isotope dilution HPLC-MS/MS method for quantification of 12 biomarkers and
metabolites. All sample handling is performed by a robotic workstation. The assay uses 45
μL of plasma, and sample treatment consists of protein precipitation by trichloroacetic acid …
biomarkers related to one-carbon metabolism, which comprises a metabolic network of
cross-talking pathways. To address this complexity in epidemiological studies, we have
established an isotope dilution HPLC-MS/MS method for quantification of 12 biomarkers and
metabolites. All sample handling is performed by a robotic workstation. The assay uses 45
μL of plasma, and sample treatment consists of protein precipitation by trichloroacetic acid …
Abstract
Risk of chronic diseases, like cardiovascular disease and cancer, has been associated with biomarkers related to one-carbon metabolism, which comprises a metabolic network of cross-talking pathways. To address this complexity in epidemiological studies, we have established an isotope dilution HPLC-MS/MS method for quantification of 12 biomarkers and metabolites. All sample handling is performed by a robotic workstation. The assay uses 45 μL of plasma, and sample treatment consists of protein precipitation by trichloroacetic acid. The analytes were separated on a Fortis Phenyl column using an isocratic mobile phase that contained water, methanol and acetic acid. Methionine, methionine sulfoxide, choline, betaine, dimethylglycine, arginine, asymmetric dimethylarginine, symmetric dimethylarginine, homoarginine, creatinine, cystathionine and trimethyllysine all showed limits of detection well below the 5th percentile of plasma distributions in healthy humans, coefficients of variation were in the range 2.2–12.3 %, and recoveries were 80–131 %. Simple sample processing, low-volume consumption, multiplexing and high capacity/short run time of this method make it suitable for large-scale metabolic profiling of precious biobank samples.
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