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      Presoaking dried blood spot with water improves efficiency for small-molecule analysis

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          Abstract

          The current method of extracting small molecules from dried blood spots (DBSs) and liquid blood is similar. However, owing to their different physical characteristics, a modification of the extraction process for DBS is required. We propose a modified method involving presoaking in water that results in better extraction efficiency for small-molecule analysis than the conventional protein precipitation method. Using blood and DBSs from eight subjects, the similarities, recovery rates and extraction efficiencies of both methods were compared. Quantitative analysis showed that seven and six out of ten conditions for the modified method group exhibited almost 100% recovery and extraction efficiency rates, respectively, compared with the conventional method group. Taken together, the results suggest that a presoaking step is needed for efficient DBS analysis.

          METHOD SUMMARY

          This study describes the increased efficiency resulting from inclusion of a water addition step before small-molecule extraction from DBS when using the protein precipitation principle.

          Most cited references24

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          What a drop can do: dried blood spots as a minimally invasive method for integrating biomarkers into population-based research.

          Thomas McDade, Sharon Williams, James Snodgrass (2007)
          Logistical constraints associated with the collection and analysis of biological samples in community-based settings have been a significant impediment to integrative, multilevel bio-demographic and biobehavioral research. However recent methodological developments have overcome many of these constraints and have also expanded the options for incorporating biomarkers into population-based health research in international as well as domestic contexts. In particular using dried blood spot (DBS) samples-drops of whole blood collected on filter paper from a simple finger prick-provides a minimally invasive method for collecting blood samples in nonclinical settings. After a brief discussion of biomarkers more generally, we review procedures for collecting, handling, and analyzing DBS samples. Advantages of using DBS samples-compared with venipuncture include the relative ease and low cost of sample collection, transport, and storage. Disadvantages include requirements for assay development and validation as well as the relatively small volumes of sample. We present the results of a comprehensive literature review of published protocols for analysis of DBS samples, and we provide more detailed analysis of protocols for 45 analytes likely to be of particular relevance to population-level health research. Our objective is to provide investigators with the information they need to make informed decisions regarding the appropriateness of blood spot methods for their research interests.
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            Solvent-dependent metabolite distribution, clustering, and protein extraction for serum profiling with mass spectrometry.

            Elizabeth Want, Grace O'Maille, Colin A Smith … (2006)
            The aim of metabolite profiling is to monitor all metabolites within a biological sample for applications in basic biochemical research as well as pharmacokinetic studies and biomarker discovery. Here, novel data analysis software, XCMS, was used to monitor all metabolite features detected from an array of serum extraction methods, with application to metabolite profiling using electrospray liquid chromatography/mass spectrometry (ESI-LC/MS). The XCMS software enabled the comparison of methods with regard to reproducibility, the number and type of metabolite features detected, and the similarity of these features between different extraction methods. Extraction efficiency with regard to metabolite feature hydrophobicity was examined through the generation of unique feature density distribution plots, displaying feature distribution along chromatographic time. Hierarchical clustering was performed to highlight similarities in the metabolite features observed between the extraction methods. Protein extraction efficiency was determined using the Bradford assay, and the residual proteins were identified using nano-LC/MS/MS. Additionally, the identification of four of the most intensely ionized serum metabolites using FTMS and tandem mass spectrometry was reported. The extraction methods, ranging from organic solvents and acids to heat denaturation, varied widely in both protein removal efficiency and the number of mass spectral features detected. Methanol protein precipitation followed by centrifugation was found to be the most effective, straightforward, and reproducible approach, resulting in serum extracts containing over 2000 detected metabolite features and less than 2% residual protein. Interestingly, the combination of all approaches produced over 10,000 unique metabolite features, a number that is indicative of the complexity of the human metabolome and the potential of metabolomics in biomarker discovery.
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              Volumetric absorptive microsampling: a dried sample collection technique for quantitative bioanalysis.

              Philip Denniff, Neil Spooner (2014)
              Volumetric absorptive microsampling (VAMS) is a novel approach to obtaining a dried blood sample for quantitative bioanalysis that overcomes the area bias and homogeneity issues associated with conventional dried blood spot (DBS) sample when a subpunch is taken. The VAMS sampler absorbs a fixed volume of blood (∼10 μL) in 2-4 s with less than 5% volume variation across the hematocrit range of 20-70% with low tip-to-tip variability. There is no evidence of selective absorption by the tip of the plasma component over whole blood. Recommendations for best practice when collecting samples were developed based upon the results of tests examining a number of potential abuse scenarios.
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                Author and article information

                Journal
                Journal ID (publisher-id): BTN
                Title: BioTechniques
                Publisher: Future Science Ltd (London, UK )
                ISSN (Print): 0736-6205
                ISSN (Electronic): 1940-9818
                Publication date (Electronic, pub): 16 October 2019
                Publication date (Print, pub): November 2019
                Volume: 67
                Issue: 5
                Pages: 219-228
                Affiliations
                1Department of Senior Healthcare, BK21 Plus Program, Graduate School, Eulji University, Daejeon 34824, Republic of Korea
                2Department of Biomedical Laboratory Science, College of Health Sciences, Eulji University, Seongnam-si, Gyeonggi-do 13135, Republic of Korea
                Author notes
                [* ]Author for correspondence: kanghg@ 123456eulji.ac.kr
                [‡]

                These authors contributed equally

                Article
                DOI: 10.2144/btn-2019-0043
                SO-VID: aecf20da-044d-436d-8a04-107ea7a32caa
                Copyright © © 2020 Hee-Gyoo Kang
                License:

                This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License

                History
                Date received : 06 April 2019
                Date accepted : 31 May 2019
                Date: 16 October 2019
                Page count
                Pages: 10
                Categories
                Subject: Reports

                ScienceOpen disciplines: General life sciences,Cell biology,Molecular biology,Biotechnology,Genetics,Life sciences
                Keywords: small molecule,extraction method,dried blood spot,recovery,extraction efficiency
                Data availability:
                ScienceOpen disciplines: General life sciences, Cell biology, Molecular biology, Biotechnology, Genetics, Life sciences
                Keywords: small molecule, extraction method, dried blood spot, recovery, extraction efficiency

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