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Journal article
Dried Blood Spot Microsampling: A Semi-Quantitative 4D-Lipidomics Approach Using Ultra High Performance Liquid Chromatography - High Resolution Mass Spectrometry (UHPLC-HRMS)
Talanta (Oxford), Vol.287, 127677
2025
Abstract
Dried blood spot (DBS) sample collections can offer a minimally invasive, cost-effective alternative to traditional venepuncture for remote sampling and high-frequency metabolic profiling. We present an optimised protocol for DBS-based extraction and comprehensive untargeted 4D lipid profiling using ultrahigh-performance liquid chromatography coupled with high-resolution mass spectrometry, designed to support large-scale applications in population-wide lipidomics research. Inclusion of stable isotopically labelled internal standards allowed for semi-quantitative subclass-level correction for 10 μL DBS samples, enhancing the number of reproducible lipids within our curated target list (focussed on 432 unique rule-based lipid annotations out of 6845 features) across positive and negative heated electrospray ionisation modes. The reproducibility of unique lipid features detected in replicate DBS (n = 6) was assessed on both peak areas (351 lipids < 25 % CV) and calculated concentrations relative to internal standards (432 lipids < 25 % CV), underscoring the benefit of internal standard addition. Storage conditions for DBS were also evaluated to determine short-term lipid stability at different temperatures (-20 ˚C, 4 ˚C, room temperature, and 45 ˚C). The majority of lipid subclasses, excluding a minority of glycerophospholipids and oxylipins, were stable up to 1 week at -20 ˚C and 4 ˚C (log2-fold change < 30 % difference), which supports the short-term storage capacity for DBS in field and clinical settings. Similar stability was observed within a week at room temperature, excluding phosphatidylethanolamines and phosphatidylglycerols (log2-fold change > 30 % difference). Application of the optimised workflow to a microsampling device (n = 6) identified 432 lipid features (CV < 25 %) with three repeated samplings over an hour showing minimal impact on lipid profiles by principal component analysis, showing promise for high-frequency, longitudinal DBS monitoring in population health. This work represents a significant advance, highlighting the potential for reliable lipid analysis from DBS samples with short-term stability under various storage conditions, an important logistical benefit for remote or resource-limited settings.
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•Dried blood spots enable minimally invasive, cost-effective sampling in lipidomics•The developed untargeted 4D-lipidomic method annotates 432 lipids in 10 μL DBS•Majority of lipid subclasses are stable on DBS up to 1 week, ideal at -20°C and 4°C•Commercial microsampling devices suit remote, high-frequency lipid profiling
Details
- Title
- Dried Blood Spot Microsampling: A Semi-Quantitative 4D-Lipidomics Approach Using Ultra High Performance Liquid Chromatography - High Resolution Mass Spectrometry (UHPLC-HRMS)
- Authors/Creators
- Jayden Lee Roberts - Australian National Phenome Centre, Health Futures Institute, Harry Perkins Institute, Murdoch University, 5 Robin Warren Drive, Murdoch, WA 6150, AustraliaMonique J. Ryan - Murdoch University, Australian National Phenome CentreLuke Whiley - Murdoch University, Centre for Computational and Systems MedicineMelvin Gay - Bruker Pty Ltd., Preston, VIC 3072, AustraliaVimalnath Nambiar - Murdoch UniversityElaine Holmes - Murdoch University, Centre for Computational and Systems MedicineJeremy K. Nicholson - Murdoch University, Health Futures InstituteJulien Wist - Murdoch University, Centre for Computational and Systems MedicineNicola Gray - Murdoch University, Centre for Computational and Systems MedicineNathan G. Lawler - Murdoch University, Health Futures Institute
- Publication Details
- Talanta (Oxford), Vol.287, 127677
- Publisher
- Elsevier B.V.
- Number of pages
- 12
- Grant note
- Phenome CentreWestern Australian Department of Jobs, Tourism, Science and InnovationAustralian Research Council Laureate FellowshipGovernment of Western Australian Premier's Fellowship
This research is supported by a joint strategic research training program scholarship funded by Murdoch University and Bruker Daltonics to higher degree research students for author J.L.R. We would like to thank the Medical Research Future Fund and Western Australian State Government for funding and supporting the Australian National Phenome Centre. Additionally, we thank the Western Australian Department of Jobs, Tourism, Science and Innovation, Australian Research Council Laureate Fellowship and Government of Western Australian Premier's Fellowship for author E.H. J.L.R. would like to acknowledge N.S.M.
- Identifiers
- 991005741586907891
- Copyright
- © 2025 The Authors.
- Murdoch Affiliation
- Australian National Phenome Centre; Centre for Computational and Systems Medicine; Health Futures Institute
- Language
- English
- Resource Type
- Journal article
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- Citation topics
- 2 Chemistry
- 2.211 Mass Spectrometry
- 2.211.2349 Dried Blood Spot Sampling
- Web Of Science research areas
- Chemistry, Analytical
- ESI research areas
- Chemistry