The metabolomic study of healthy ageing applying dried blood and urine spot collection

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BACKGROUND AND SCIENTIFIC PROBLEM - The lifespan of the UK population is increasing at a greater rate than healthspan providing longer periods of ill health in later life and the associated economic strain on the NHS. There is a necessity to improve the healthy ageing of the UK population. The role of sedentary behaviour vs. exercise is being shown to have a strong influence on the healthy ageing-metabolism axis; exercise is a positive lifestyle intervention available to all. Metabolic changes can be studied applying nontargeted metabolomic analysis of thousands of metabolites. In these studies, biofluid samples are collected in a specialised centre which requires the subject to travel and expert staff and instruments to collect and process the samples; this process is costly and inefficient. However, sample collection away from specialised centres will reduce sample collection costs and allow longitudional studies to be performed cheaply and at minimal inconvenience to the subjects. The collection of dried blood or urine spots (DBS/DUS) away from specialised centres followed by metabolomic analysis offers a simple and robust solution. These devices are currently applied for targeted metabolite assays and we wish to develop this technology for non-targeted metabolomic studies of healthy ageing.SCIENTIFIC OBJECTIVE - We propose the development and application of dried blood and urine spot collection with non-targeted metabolomics to be applied in the study of healthy ageing.AIM 1 - Develop analytical methodologies for non-targeted metabolomic analysis of dried blood and urine spots. The small sample volumes (typically < 5microL) are analytically challenging in relation to sensitivity and range of metabolite classes detected. Here, two analytical techniques will be developed and assessed for non-targeted metabolomic analysis; (1) Liquid Extraction Surface Analysis (LESA) coupled to mass spectrometry for rapid and automated sample extraction and analysis and (2) capillary liquidchromatography-mass spectrometry for analysis of lipid and hydrophilic classes of metabolites.OUTPUT - analytical methods applicable for the non-targeted metabolomic analysis of dried blood and urine spots.AIM 2 - Study the stability of dried blood and urine spots on currently available collection materials applying methods developed in aim 1; the metabolite stability is a concern in these studies. Here, the stability of dried blood and urine spots collected on a range of currently available papers and cards will be assessed for up to 18 months. At the same time new collection systems coated with enzyme and protease inhibitors will be developed and studied to assess their applicability. Both of these studies will assess stability and diversity of metabolite classes detectable.OUTPUT - protocol and device for dried blood and urine spot collection.AIM 3 - Apply the methods developed in (1) and (2) to study metabolism in sedentary behaviour and exercise in adults. With collaborators in healthy ageing and exercise at the University of Birmingham a longitudional metabolomic study of sedentary vs. exercise behaviour in adults applying DBS/DUS will be performed.OUTPUT - mechanistic information on changes in metabolism in sedentary vs. exercising subjects atdifferent ages.