Abstract
Dried blood spots (DBS) have proven to be a powerful sampling and storage method for newborn screening and many other applications. However, DBS methods have not yet been optimized for broad-spectrum targeted metabolomic analysis. In this study, we developed a robust, DBS-based, broad-spectrum, targeted metabolomic method that was able to measure over 400 metabolites from a 6.3 mm punch from standard Whatman 903TM filter paper cards. The effects of blood spot volumes, hematocrit, vacutainer chemistry, extraction methods, carryover, and comparability with plasma and fingerstick capillary blood samples were analyzed. The stability of over 400 metabolites stored under varying conditions over one year was also tested. No significant impacts of blood volume and hematocrit variations were observed when the spotted blood volume was over 60 µL and the hematocrit was between 31% and 50%. The median area under the curve (AUC) of metabolites in the DBS metabolome declined by 40% in the first 3 months and then did not decline further for at least 1 year. All originally detectable metabolites remained within detectable limits. The optimal storage conditions for metabolomic analysis were −80 °C with desiccants and without an O2 scavenger. The method was clinically validated for its potential utility in the diagnosis of the mitochondrial disease mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS). Our method provides a convenient alternative to freezing, storing, and shipping liquid blood samples for comparative metabolomic studies.
Highlights
Metabolomics is emerging as an important tool in clinical chemistry to identify biomarkers, find new roles of metabolism in disease, and discover novel metabolic drug targets [1,2,3]
We explore the utility of dried blood spot (DBS) analysis in metabolomic studies
Our analysis showed that samples collected by fingerstick lancet or as Li-heparin venous blood and stored as dried blood spots were superimposable by Principal component analysis (PCA) (Figure 3E)
Summary
Metabolomics is emerging as an important tool in clinical chemistry to identify biomarkers, find new roles of metabolism in disease, and discover novel metabolic drug targets [1,2,3]. Blood-based samples, including serum and plasma, are probably the best studied biofluids for metabolomics [4]. Serum and plasma preparation requires laboratory healthcare professionals and stringent storage conditions to avoid metabolite degradation. Dried blood spots have been used for newborn screening in millions of children since the 1960s [5]. We explore the utility of dried blood spot (DBS) analysis in metabolomic studies. Samples prepared and stored as dried blood spots on standardized filter paper cards may be an attractive alternative to storage, shipment, and analysis of conventional liquid venous blood samples for many studies. One advantage of dried blood spots is that quantitative results for many targeted
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