Investigations into the Environmental Conditions Experienced During Ambient Sample Transport: Impact to Dried Blood Spot Sample Shipments

Bowen, Chester L.; Dopson, Wesley; Kemp, Daniel C.; Lewis, Michael M.; Lad, Rakesh; Overvold, Carol

doi:10.4155/bio.11.128

Cited by 35 publications

(27 citation statements)

References 8 publications

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“…Stability-One of the greatest advantages of DBS sampling is improved analyte stability over a wide range of temperatures during storage and transportation (56). In this work, we stored DBS samples at Ϫ20°C, 4°C, RT, and 37°C for 1, 2, 7, 16, 29, 77, and 154 days.…”

Section: Resultsmentioning

confidence: 99%

Multiple Reaction Monitoring Enables Precise Quantification of 97 Proteins in Dried Blood Spots

Chambers

Percy

Yang

et al. 2015

Molecular & Cellular Proteomics

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The dried blood spot (DBS) methodology provides a minimally invasive approach to sample collection and enables room-temperature storage for most analytes. DBS samples have successfully been analyzed by liquid chromatography multiple reaction monitoring mass spectrometry (LC/MRM-MS) to quantify a large range of small molecule biomarkers and drugs; however, this strategy has only recently been explored for MS-based proteomics applications. Here we report the development of a highly multiplexed MRM assay to quantify endogenous proteins in human DBS samples. This assay uses matching stable isotope-labeled standard peptides for precise, relative quantification, and standard curves to characterize the analytical performance. A total of 169 peptides, corresponding to 97 proteins, were quantified in the final assay with an average linear dynamic range of 207-fold and an average R 2 value of 0.987. The total range of this assay spanned almost 5 orders of magnitude from serum albumin (P02768) at 18.0 mg/ml down to cholinesterase (P06276) at 190 ng/ml. The average intra-assay and interassay precision for 6 biological samples ranged from 6.1-7.5% CV and 9.5-11.0% CV, respectively. The majority of peptide targets were stable after 154 days at storage temperatures from ؊20°C to 37°C. Furthermore, protein concentration ratios between matching DBS and whole blood samples were largely constant (<20% CV) across six biological samples. This assay represents the highest multiplexing yet achieved for targeted protein quantification in DBS samples and is suitable for biomedical research applications. Molecular & Cellular Proteomics 14: 10.1074/mcp.O115.049957, 3094-3104, 2015.The dried blood spot (DBS) 1 methodology provides several advantages over traditional plasma or serum samples throughout the entire pre-analytical workflow including sample collection, transportation, and storage (1, 2) These blood samples are typically generated using a small sterile lancet to prick the skin and then spotting a drop onto a collection card. Therefore, DBS sampling is less invasive than venipuncture and does not require a trained phlebotomist. This sampling approach also does not require time-sensitive centrifugation, which is crucial for plasma and serum samples to prevent degradation. Many analytes have been determined to be stable in the DBS format at room temperature, eliminating the cost associated with cold-chain logistics for sample transportation and storage. These considerations are also important for sample collection in remote locations that may be without reliable access to a centrifuge and/or a freezer designated for biohazardous materials. Quantitative bioanalytical methods using the DBS methodology have been developed for genomic, metabolomic, and proteomic applications including newborn screening (3, 4), therapeutic drug monitoring (5, 6), toxicology and drugs of abuse (7,8), viral disease management (9, 10), and many others (2, 11).Targeted MS, in particular selected/multiple reaction monitoring (SRM/MRM) using internal standards...

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Section: Resultsmentioning

confidence: 99%

Multiple Reaction Monitoring Enables Precise Quantification of 97 Proteins in Dried Blood Spots

Chambers

Percy

Yang

et al. 2015

Molecular & Cellular Proteomics

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show abstract

“…We therefore selected peptide targets that lack easily modifiable amino acid residues, such as methionine which may readily undergo oxidation, to improve sample stability. Bowen and coworkers recently monitored the environmental conditions for DBS samples transported without refrigeration on international air cargo shipments (32). Their work demonstrated that samples commonly encounter temperatures between 0°C and 30°C and recommended that future method validation cover Ϫ20°C to 40°C.…”

Section: Fig 2 the Precision Of The Measured Concentration Of 44 Pementioning

confidence: 99%

Multiplexed Quantitation of Endogenous Proteins in Dried Blood Spots by Multiple Reaction Monitoring - Mass Spectrometry

Chambers

Percy

Yang

et al. 2013

Molecular & Cellular Proteomics

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Dried blood spot (DBS) sampling, coupled with multiple reaction monitoring mass spectrometry (MRM-MS), is a well-established approach for quantifying a wide range of small molecule biomarkers and drugs. This sampling procedure is simpler and less-invasive than those required for traditional plasma or serum samples enabling collection by minimally trained personnel. Many analytes are stable in the DBS format without refrigeration, which reduces the cost and logistical challenges of sample collection in remote locations. These advantages make DBS sample collection desirable for advancing personalized medicine through population-wide biomarker screening. Dried Blood Spot (DBS)1 samples have many advantages over blood serum or plasma and are the preferred clinical sample for newborn screening for metabolic diseases (1, 2). These samples are collected by pricking a newborn's heel and spotting a drop of blood onto specially designed filter paper collection cards. Samples are then dried under ambient conditions and are usually stored with desiccant at room temperature until analysis. This sampling procedure is simpler and less invasive then intravenous blood draws, which require a trained phlebotomist. Not surprisingly, the majority of adult patients prefer the small lancet used in finger-prick blood sampling methods to the larger needles used in intravenous blood draws (3, 4). Unlike plasma or serum samples, which consume Ն250 l of blood and must be centrifuged within an hour of collection, DBS samples can be prepared using a volume of only 10 l, and do not require any specialized equipment at the collection site (5). The simplicity and reduced safety risks associated with DBS sampling enables collection by minimally trained staff or by the patients themselves. In addition, many analytes are stable in the DBS format at room temperature, reducing sample transportation and storage costs, as well as the impact on the environment. Finally, DBS samples are safer to transport and are considered exempt from dangerous goods regulations (6, 7). These advantages make DBS sampling very attractive for advancing personalized medicine and population-based biomarker research (8).Numerous biomolecular targets covering genomics, metabolomics, and proteomics applications have been quantified in DBS samples using a wide array of analytical techniques (9). The most common clinical application of DBS sampling is screening newborns for metabolomics disorders by targeting small molecule biomarkers. Early screening programs relied on bacterial inhibition assays and later immunoassays, both of which required a different assay for each target of interest (2). However, the time and cost required to perform each assay independently has limited the number of 1 The abbreviations used are: DBS, dried blood spot; IEF, isoelectric focusing; HPLC, high performance liquid chromatography; SIS, stable isotope-labeled internal standards; MRM, multiple reaction monitoring; LDR, linear dynamic range; WB, whole blood.

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“…In contrast, the conventional protocol was more accurate than the DMF protocol for the quantification of benzethonium chloride and ibuprofen, with measured accuracies of 112% and 98.8% (conventional), and 66.9% and 119.5% (DMF), respectively. It is possible that the benzethonium chloride and ibuprofen in the DBS samples that were analyzed by DMF (that were mailed from the UK to Toronto) were subject to temperature and humidity changes, which are known to degrade some analytes [46]. But it is also likely that simple changes in methodology could improve these results, for example, tuning the extraction solvent to favor the solubility of weak acids [47] or quaternary amines [48], or adjusting the solvent composition after extraction for higher ionization efficiency.…”

Section: Ms/ms Analysis and Optimizationmentioning

confidence: 99%

Multiplexed Extraction and Quantitative Analysis of Pharmaceuticals from DBS Samples Using Digital Microfluidics

et al. 2014

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Investigations into the Environmental Conditions Experienced During Ambient Sample Transport: Impact to Dried Blood Spot Sample Shipments

Cited by 35 publications

References 8 publications

Multiple Reaction Monitoring Enables Precise Quantification of 97 Proteins in Dried Blood Spots

Multiple Reaction Monitoring Enables Precise Quantification of 97 Proteins in Dried Blood Spots

Multiplexed Quantitation of Endogenous Proteins in Dried Blood Spots by Multiple Reaction Monitoring - Mass Spectrometry

Multiplexed Extraction and Quantitative Analysis of Pharmaceuticals from DBS Samples Using Digital Microfluidics

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