Abstract
BackgroundReal-time and dynamic assessment of an individual’s lipid homeostatic state in blood is complicated due to the need to collect samples in a clinical environment. In the context of precision medicine and population health, tools that facilitate sample collection and empower the individual to participate in the process are necessary to complement advanced bioanalytical analysis. The dried blood spot (DBS) methodology via finger prick or heel prick is a minimally invasive sample collection method that allows the relative ease and low cost of sample collection as well as transport. However, it has yet to be integrated into broad scale personalized lipidomic analysis. Therefore, in this study we report the development of a novel DBS high resolution MS/MSALL lipidomics workflow.MethodsIn this report we compared lipidomic analysis of four types of blood sample collection methods (DBS, venous whole blood, serum, and plasma) across several parameters, which include lipidomics coverage of each matrix and the effects of temperature and time on the coverage and stability of different lipid classes and molecular species. The novel DBS-MS/MSALL lipidomics platform developed in this report was then applied to examine postprandial effects on the blood lipidome and further to explore the temporal fluctuation of the lipidome across hours and days.ResultsMore than 1,200 lipid molecular species from a single DBS sample were identified and quantified. The lipidomics profile of the DBS samples is comparable to whole blood matrix. DBS-MS/MSALL lipidomic analysis in postprandial experiments revealed significant alterations in triacylglyceride species. Temporal analysis of the lipidome at various times in the day and across days identified several lipid species that fluctuate as a function of time, and a subset of lipid species were identified to be significantly altered across hours within a day and within successive days of the week.ConclusionsA novel DBS-MS/MSALL lipidomics method has been established for human blood. The feasibility and application of this method demonstrate the potential utility for lipidomics analysis in both healthy and diverse diseases states. This DBS MS-based lipidomics analysis represents a formidable approach for empowering patients and individuals in the era of precision medicine to uncover novel biomarkers and to monitor lipid homeostasis.
Highlights
Real-time and dynamic assessment of an individual’s lipid homeostatic state in blood is complicated due to the need to collect samples in a clinical environment
We report a high-throughput dried blood spot (DBS)-MS/MSALL lipidomics platform to analyze the blood lipidome via the direct infusion of lipids extracted from DBS samples
Adaption of DBS-MS/MSALL shotgun lipidomics to different blood matrix samples The choice of negative mode or positive mode analysis for different lipid classes is based on the differential propensity of each lipid class to acquire either positive or negative charges under the source of high voltage [15, 22, 33]
Summary
Real-time and dynamic assessment of an individual’s lipid homeostatic state in blood is complicated due to the need to collect samples in a clinical environment. The dried blood spot (DBS) methodology via finger prick or heel prick is a minimally invasive sample collection method that allows the relative ease and low cost of sample collection as well as transport. For over two decades the dried blood spot (DBS) methodology has commonly been used in the diagnosis of inborn errors of metabolism [1,2,3]. More than 95% of newborns are screened for inherited metabolic disorders using DBS in the United States [2, 4]. Using this methodology blood samples are typically obtained from heel or finger pricks and spotted onto filter paper for collection and analysis. A low volume of blood (less than 20 μL) is needed to spot onto filter paper compared to a minimum of 0.5 mL of blood in
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