{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T22:38:11Z","timestamp":1792103891718,"version":"4.4.0"},"reference-count":40,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2020,2,27]],"date-time":"2020-02-27T00:00:00Z","timestamp":1582761600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"UCSD Christini Fund","award":["philanthropic gift"],"award-info":[{"award-number":["philanthropic gift"]}]},{"name":"The Lennox Foundation","award":["philanthropic gift"],"award-info":[{"award-number":["philanthropic gift"]}]},{"name":"UCSD Mitochondrial Disease Research Fund","award":["philanthropic gift"],"award-info":[{"award-number":["philanthropic gift"]}]},{"name":"Jane Botsford-Johnson Foundation","award":["philanthropic gift"],"award-info":[{"award-number":["philanthropic gift"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>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 \u00b5L 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 \u221280 \u00b0C 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.<\/jats:p>","DOI":"10.3390\/metabo10030082","type":"journal-article","created":{"date-parts":[[2020,3,2]],"date-time":"2020-03-02T07:50:53Z","timestamp":1583135453000},"page":"82","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":65,"title":["Improved Dried Blood Spot-Based Metabolomics: A Targeted, Broad-Spectrum, Single-Injection Method"],"prefix":"10.3390","volume":"10","author":[{"given":"Kefeng","family":"Li","sequence":"first","affiliation":[{"name":"The Mitochondrial and Metabolic Disease Center, School of Medicine, University of California, San Diego, CA 92103, USA"},{"name":"Department of Medicine, School of Medicine, University of California, San Diego, CA 92103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jane C.","family":"Naviaux","sequence":"additional","affiliation":[{"name":"The Mitochondrial and Metabolic Disease Center, School of Medicine, University of California, San Diego, CA 92103, USA"},{"name":"Department of Neuroscience, School of Medicine, University of California, San Diego, CA 92103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan M.","family":"Monk","sequence":"additional","affiliation":[{"name":"Department of Medicine, School of Medicine, University of California, San Diego, CA 92103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Wang","sequence":"additional","affiliation":[{"name":"The Mitochondrial and Metabolic Disease Center, School of Medicine, University of California, San Diego, CA 92103, USA"},{"name":"Department of Medicine, School of Medicine, University of California, San Diego, CA 92103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert K.","family":"Naviaux","sequence":"additional","affiliation":[{"name":"The Mitochondrial and Metabolic Disease Center, School of Medicine, University of California, San Diego, CA 92103, USA"},{"name":"Department of Medicine, School of Medicine, University of California, San Diego, CA 92103, USA"},{"name":"Department of Pediatrics, School of Medicine, University of California, San Diego, CA 92103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,2,27]]},"reference":[{"key":"ref_1","first-page":"E5472","article-title":"Metabolic features of chronic fatigue syndrome","volume":"113","author":"Naviaux","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Naviaux, R.K., Naviaux, J.C., Li, K., Wang, L., Monk, J.M., Bright, A.T., Koslik, H.J., Ritchie, J.B., and Golomb, B.A. (2019). Metabolic features of Gulf War illness. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0219531"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1038\/nrm.2016.25","article-title":"Metabolomics: Beyond biomarkers and towards mechanisms","volume":"17","author":"Johnson","year":"2016","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1158","DOI":"10.1373\/clinchem.2018.287045","article-title":"Preanalytical Processing and Biobanking Procedures of Biological Samples for Metabolomics Research: A White Paper, Community Perspective (for \u201cPrecision Medicine and Pharmacometabolomics Task Group\u201d\u2014The Metabolomics Society Initiative)","volume":"64","author":"Kirwan","year":"2018","journal-title":"Clin. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1542\/peds.32.3.338","article-title":"A Simple Phenylalanine Method for Detecting Phenylketonuria in Large Populations of Newborn Infants","volume":"32","author":"Guthrie","year":"1963","journal-title":"Pediatrics"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1945","DOI":"10.1093\/clinchem\/47.11.1945","article-title":"Tandem mass spectrometric analysis for amino, organic, and fatty acid disorders in newborn dried blood spots: A two-year summary from the New England Newborn Screening Program","volume":"47","author":"Zytkovicz","year":"2001","journal-title":"Clin. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.jchromb.2012.05.040","article-title":"Distribution of 25-hydroxyvitamin D3 in dried blood spots and implications for its quantitation by tandem mass spectrometry","volume":"901","author":"Kvaskoff","year":"2012","journal-title":"J. Chromatogr. B"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Martial, L.C., Aarnoutse, R.E., Schreuder, M.F., Henriet, S.S., Bruggemann, R.J., and Joore, M.A. (2016). Cost Evaluation of Dried Blood Spot Home Sampling as Compared to Conventional Sampling for Therapeutic Drug Monitoring in Children. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0167433"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1002\/mnfr.201500483","article-title":"Exploring the association of dairy product intake with the fatty acids C15:0 and C17:0 measured from dried blood spots in a multipopulation cohort: Findings from the Food4Me study","volume":"60","author":"Albani","year":"2016","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.chroma.2012.11.030","article-title":"Dried blood spots: Liquid chromatography-mass spectrometry analysis of Delta9-tetrahydrocannabinol and its main metabolites","volume":"1271","author":"Mercolini","year":"2013","journal-title":"J. Chromatogr. A"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.ijantimicag.2018.04.020","article-title":"Evaluation of dried blood spot sampling for pharmacokinetic research and therapeutic drug monitoring of anti-tuberculosis drugs in children","volume":"52","author":"Martial","year":"2018","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1286","DOI":"10.1074\/mcp.RA117.000015","article-title":"Stability of Proteins in Dried Blood Spot Biobanks","volume":"16","author":"Bjorkesten","year":"2017","journal-title":"Mol. Cell. Proteom."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1186\/s12986-017-0182-6","article-title":"Dynamic and temporal assessment of human dried blood spot MS\/MS(ALL) shotgun lipidomics analysis","volume":"14","author":"Gao","year":"2017","journal-title":"Nutr. Metab. Lond"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1018","DOI":"10.1007\/s11306-014-0628-z","article-title":"The development and validation of a fast and robust dried blood spot based lipid profiling method to study infant metabolism","volume":"10","author":"Koulman","year":"2014","journal-title":"Metabolomics"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Haijes, H.A., Willemsen, M., Van der Ham, M., Gerrits, J., Pras-Raves, M.L., Prinsen, H., Van Hasselt, P.M., De Sain-van der Velden, M.G.M., Verhoeven-Duif, N.M., and Jans, J.J.M. (2019). Direct Infusion Based Metabolomics Identifies Metabolic Disease in Patients\u2019 Dried Blood Spots and Plasma. Metabolites, 9.","DOI":"10.3390\/metabo9010012"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1007\/s11306-016-1153-z","article-title":"An untargeted metabolomics method for archived newborn dried blood spots in epidemiologic studies","volume":"13","author":"Petrick","year":"2017","journal-title":"Metabolomics"},{"key":"ref_17","first-page":"1389","article-title":"A dried blood spot mass spectrometry metabolomic approach for rapid breast cancer detection","volume":"9","author":"Wang","year":"2016","journal-title":"Onco Targets Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1002\/iub.1617","article-title":"Rapid Differentiating Colorectal Cancer and Colorectal Polyp Using Dried Blood Spot Mass Spectrometry Metabolomic Approach","volume":"69","author":"Jing","year":"2017","journal-title":"IUBMB Life"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"16080","DOI":"10.1038\/nrdp.2016.80","article-title":"Mitochondrial diseases","volume":"2","author":"Gorman","year":"2016","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1016\/j.bbagen.2011.09.005","article-title":"Molecular pathology of MELAS and L-arginine effects","volume":"1820","author":"Koga","year":"2012","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1007\/s11306-017-1264-1","article-title":"A robust, single-injection method for targeted, broad-spectrum plasma metabolomics","volume":"13","author":"Li","year":"2017","journal-title":"Metabolomics"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1373\/clinchem.2014.224840","article-title":"From lost in translation to paradise found: Enabling protein biomarker method transfer by mass spectrometry","volume":"60","author":"Grant","year":"2014","journal-title":"Clin. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/j.jpba.2018.12.030","article-title":"Bioanalytical method validation: New FDA guidance vs. EMA guideline. Better or worse?","volume":"165","author":"Kaza","year":"2019","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6457","DOI":"10.1021\/acs.analchem.6b01145","article-title":"Estimation and Correction of the Blood Volume Variations of Dried Blood Spots Using a Postcolumn Infused-Internal Standard Strategy with LC-Electrospray Ionization-MS","volume":"88","author":"Liao","year":"2016","journal-title":"Anal. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/j.jchromb.2015.02.023","article-title":"\u201cCenter punch\u201d and \u201cwhole spot\u201d bioanalysis of apixaban in human dried blood spot samples by UHPLC-MS\/MS","volume":"988","author":"Zheng","year":"2015","journal-title":"J. Chromatogr. B"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6538","DOI":"10.1021\/acs.analchem.6b01321","article-title":"A Novel, Nondestructive, Dried Blood Spot-Based Hematocrit Prediction Method Using Noncontact Diffuse Reflectance Spectroscopy","volume":"88","author":"Capiau","year":"2016","journal-title":"Anal. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1177\/193229680900300118","article-title":"A liquid chromatography\/tandem mass spectrometry method for determination of 25-hydroxy vitamin D2 and 25-hydroxy vitamin D3 in dried blood spots: A potential adjunct to diabetes and cardiometabolic risk screening","volume":"3","author":"Newman","year":"2009","journal-title":"J. Diabetes Sci. Technol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1595","DOI":"10.1016\/j.jchromb.2009.03.024","article-title":"Analysis of cyclosporin A in dried blood spots using liquid chromatography tandem mass spectrometry","volume":"877","author":"Wilhelm","year":"2009","journal-title":"J. Chromatogr. B"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1046\/j.1365-2257.2003.00484.x","article-title":"Significant differences between capillary and venous complete blood counts in the neonatal period","volume":"25","author":"Kayiran","year":"2003","journal-title":"Clin. Lab. Haematol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1373\/clinchem.2017.278457","article-title":"Short-Term Stabilities of 21 Amino Acids in Dried Blood Spots","volume":"64","author":"Han","year":"2018","journal-title":"Clin. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1039\/C8AN01422A","article-title":"Development of a method for multiple vitamin D metabolite measurements by liquid chromatography coupled with tandem mass spectrometry in dried blood spots","volume":"144","author":"Rola","year":"2018","journal-title":"Analyst"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.pcl.2017.11.006","article-title":"Inborn Errors of Metabolism with Myopathy: Defects of Fatty Acid Oxidation and the Carnitine Shuttle System","volume":"65","author":"Vockley","year":"2018","journal-title":"Pediatr. Clin. N. Am."},{"key":"ref_33","first-page":"5968","article-title":"Hydroxylated Long-Chain Acylcarnitines are Biomarkers of Mitochondrial Myopathy","volume":"104","author":"Vissing","year":"2019","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1442","DOI":"10.1016\/j.bbadis.2012.05.008","article-title":"Functions of plasmalogen lipids in health and disease","volume":"1822","author":"Braverman","year":"2012","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/S0163-7827(01)00003-0","article-title":"Plasmalogens: Biosynthesis and functions","volume":"40","author":"Nagan","year":"2001","journal-title":"Prog. Lipid Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1186\/s12944-019-1044-1","article-title":"Plasmalogens and Alzheimer\u2019s disease: A review","volume":"18","author":"Su","year":"2019","journal-title":"Lipids Health Dis."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Calvano, C.D., Ventura, G., Sardanelli, A.M.M., Savino, L., Losito, I., Michele, G., Palmisano, F., and Cataldi, T.R.I. (2019). Searching for Potential Lipid Biomarkers of Parkinson\u2019s Disease in Parkin-Mutant Human Skin Fibroblasts by HILIC-ESI-MS\/MS: Preliminary Findings. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20133341"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"6245","DOI":"10.1074\/jbc.272.10.6245","article-title":"Peroxisomal impairment in Niemann-Pick type C disease","volume":"272","author":"Schedin","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/s12944-018-0685-9","article-title":"Plasmalogen lipids: Functional mechanism and their involvement in gastrointestinal cancer","volume":"17","author":"Messias","year":"2018","journal-title":"Lipids Health Dis."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Enns, G.M., Moore, T., Le, A., Atkuri, K., Shah, M.K., Cusmano-Ozog, K., Niemi, A.K., and Cowan, T.M. (2014). Degree of glutathione deficiency and redox imbalance depend on subtype of mitochondrial disease and clinical status. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0100001"}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/10\/3\/82\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:02:08Z","timestamp":1760173328000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/10\/3\/82"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,2,27]]},"references-count":40,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2020,3]]}},"alternative-id":["metabo10030082"],"URL":"https:\/\/doi.org\/10.3390\/metabo10030082","relation":{},"ISSN":["2218-1989"],"issn-type":[{"value":"2218-1989","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,2,27]]}}}