Investigation of the 12-Month Stability of Dried Blood and Urine Spots Applying Untargeted UHPLC-MS Metabolomic Assays

Abstract

The use of dried blood spot (DBS) and dried urine spot (DUS) samples represents an attractive opportunity for researchers in biomedical metabolomics to collect whole blood and urine samples in the absence of a processing laboratory and so to allow collection in remote areas or in longitudinal studies away from the clinic. The 12-month stability of the thousands of metabolites present in these biofluids and the applicability of DBS and DUS samples for untargeted metabolomics applications has not previously been investigated in detail and compared to blood and urine samples. Here, the 12-month stability of DBS and DUS at different storage temperatures (−20, +4, and +21 °C) have been compared to plasma and urine biofluids stored at the same storage temperatures and time. Samples were analyzed applying complementary HILIC and C18 reversed-phase UHPLC-MS untargeted metabolomic assays. Results show that metabolites demonstrate increased stability in DBS and DUS compared to whole blood and urine at all storage temperatures and times. DBS and DUS stored at +21 °C are stable for up to 4 weeks but are not stable over a 1 year period. DBS and DUS showed good stability when stored at −20 °C for 1 year. We recommend that DBS and DUS samples are collected and transported within 28 days at room temperature and are stored for longer periods of time at −20 or −80 °C. The metabolomes of DUS samples and urine were very similar but the metabolome of DBS included additional metabolites not detected in plasma and therefore proposed to be released from cells in whole blood.

Metabolomics for personalized medicine: the input of analytical chemistry from biomarker discovery to point-of-care tests

“…The temperature of − 80 °C was considered as the most convenient urine long-term storage condition. In another study, Palmer et al have investigated the 12-month stability of dried blood spots (DBS) and dried urine spots (DUS) at different storage temperatures (− 20, + 4, and + 21 °C) and compared it to plasma and urine biofluids stored at the same storage temperatures and time by using LC/HRMS-based untargeted metabolomics [ 115 ]. Inter-batch normalization was achieved using a pooled QC sample.…”

The rise of the nanomaterial metabolite corona, and emergence of the complete corona

“…A general rule is to store metabolite samples at the lowest possible temperature available as many metabolites such as lipids degrade at room temperature or even in a fridge. 101 While many methods are available for the analysis of metabolites in the literature it is vital that the NM bionano interactions and corona community follow the reporting guidelines published by the metabolomics community 102 alongside those already in place in the NM community [103][104][105] to ensure that high quality data is generated and to enable the data to be re-used by computational modelers to model the formation and interactions of the metabolite and eventually the complete corona.…”

Bridging the Polar and Hydrophobic Metabolome in Single-Run Untargeted Liquid Chromatography-Mass Spectrometry Dried Blood Spot Metabolomics for Clinical Purposes

“…6 − 8 Palmer et al reported detecting more metabolites in DBS than in plasma samples, likely due to red and white blood cell metabolites. 9 DBS are also widely used in newborn screening, 10 , 11 allowing newborn screening even in remote areas, as samples can be gathered anywhere in the world and sent to laboratories elsewhere for analysis.…”