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QC 20190805

Obtaining plasma from a blood sample and preparing it for subsequent analysis is currently a laborious process involving experienced health-care professionals and centrifugation. We circumvent this by utilizing capillary forces and microfluidic engineering to develop an autonomous plasma sampling device that filters and stores an exact amount of plasma as a dried plasma spot (DPS) from a whole blood sample in less than 6 min. We tested 24 prototype devices with whole blood from 10 volunteers, various input volumes (40-80 mu L), and different hematocrit levels (39-45%). The resulting mean plasma volume, assessed gravimetrically, was 11.6 mu L with a relative standard deviation similar to manual pipetting (3.0% vs 1.4%). LC-MS/MS analysis of caffeine concentrations in the generated DPS (12 duplicates) showed a strong correlation (R-2 = 0.99) to, but no equivalence with, concentrations prepared from corresponding plasma obtained by centrifugation. The presented autonomous DPS device may enable patient-centric plasma sampling through minimally invasive finger-pricking and allow generatation of volume-defined DPS for quantitative blood analysis., QC 20190625

Dried blood spot (DBS) sampling is a promising method for collection of microliter blood samples. However, hematocrit-related bias in combination with subpunch analysis can result in inaccurate quantification of analytes in DBS samples. In this study we use a microfluidic DBS card, designed to automatically collect fixed volume DBS samples irrespective of the blood hematocrit, to measure caffeine concentration in normal finger prick samples obtained from 44 human individuals. Caffeine levels originating from blood drops of unknown volume collected on the volumetric microfluidic DBS card were compared to volume-controlled pipetted DBS samples from the same finger prick. Hematocrit independence and volumetric sampling performances were also verified on caffeine-spiked blood samples in vitro, using both LC-MS/MS and gravimetric methods, on hematocrits from 26 to 62%. The gravimetric measurements show an excellent metering performance of the microfluidic DBS card, with a mean blood sample volume of 14.25 μL ± 3.0% (n = 51). A measured mean bias below 2.9% compared to normal hematocrit (47%) demonstrates that there is no significant hematocrit-induced bias. LC-MS/MS measurements confirm low CV and hematocrit independence of the sampling system and exhibit no substantial mean bias compared to pipetted DBS. Tests with 44 individuals demonstrated applicability of the microfluidic DBS card for direct finger prick blood sampling, and measured caffeine concentrations show a good agreement with measurements of pipetted DBS. The presented concept demonstrates a good volumetric performance which can help to improve the accuracy of DBS analysis by analyzing a whole spot, equivalent to a defined volume of liquid blood., QC 20190528

Phosphatidylethanol (PEth) is a group of phospholipids formed in cell membranes following alcohol consumption. PEth measurement in whole blood samples is established as a specific alcohol biomarker with clinical and medico-legal applications. This study further evaluated the usefulness of dried blood spot (DBS) samples collected on filter paper for PEth measurement. Specimens used were surplus volumes of venous whole blood sent for routine LC–MS/MS quantification of PEth 16:0/18:1, the major PEth homolog. DBS samples were prepared by pipetting blood on Whatman 903 Protein Saver Cards and onto a volumetric DBS device (Capitainer). The imprecision (CV) of the DBS sample amount based on area and weight measurements of spot punches were 23–28%. Investigation of the relationship between blood hematocrit and PEth concentration yielded a linear, positive correlation, and at around 1.0–1.5 μmol/L PEth 16:0/18:1, the PEth concentration increased by ~ 0.1 μmol/L for every 5% increase in hematocrit. There was a close agreement between the PEth concentrations obtained with whole blood samples and the corresponding results using Whatman 903 (PEthDBS = 1.026 PEthWB + 0.013) and volumetric device (PEthDBS = 1.045 PEthWB + 0.016) DBS samples. The CV of PEth quantification in DBS samples at concentrations ≥ 0.05 μmol/L were ≤ 15%. The present results further confirmed the usefulness of DBS samples for PEth measurement., QC 20180115

Whole-blood microsampling provides many benefits such as remote, patient-centric, and minimally invasive sampling. However, blood plasma, and not whole blood, is the prevailing matrix in clinical laboratory investigations. The challenge with plasma microsampling is to extract plasma volumes large enough to reliably detect low-concentration analytes from a small finger prick sample. Here we introduce a passive plasma filtration device that provides a high extraction yield of 65%, filtering 18 mu L of plasma from 50 mu L of undiluted human whole blood (hematocrit 45%) within less than 10 min. The enabling design element is a wedge-shaped connection between the blood filter and the hydrophilic bottom surface of a capillary channel. Using finger prick and venous blood samples from more than 10 healthy volunteers, we examined the filtration kinetics of the device over a hematocrit range of 35-55% and showed that 73 +/- 8% of the total protein content was successfully recovered after filtration. The presented plasma filtration device tackles a major challenge toward patient-centric blood microsampling by providing high-yield plasma filtration, potentially allowing reliable detection of low-concentration analytes from a blood microsample., QC 20181211

Blood plasma samples are widely used in clinical analysis but easy-to-use sampling methods for defined volumes are lacking. We introduce the first capillary driven microfluidic device that separates a specific volume of plasma from a blood sample of unknown volume. The input to the device is a small amount of whole blood in the range of 30-60 μl which results in a 4 μl isolated plasma sample within 3 minutes, available for subsequent processing and/or analysis, as demonstrated by collecting the sample in a paper substrate., QC 20160122

Background: Inhomogeneous sample distribution in DBS is a problem for accurate quantitative analysis of DBS, and has often been explained by chromatographic effects. Results: We present a model describing formation of inhomogeneous DBS during drying of the spot caused by higher evaporation rates of water at the edge as compared with the center. Color intensity analysis shows that the relative humidity and DBS card position affect the homogeneity of DBS. Conclusion: The so-called coffee-stain effect' explains the typical distribution pattern of analytes with higher concentrations measured along the edge of DBS as compared with the center. The driving mechanism and potential influencing factors should be considered when addressing the inhomogeneity of DBS in the future., QC 20151001

Background: DBS samples collected from a fingerprick typically vary in volume and homogeneity and hence make an accurate quantitative analysis of DBS samples difficult. Results: We report a prototype which first defines a precise liquid volume and subsequently stores it to a conventional DBS matrix. Liquid volumes of 2.2 mu l +/- 7.1% (n = 21) for deionized water and 6.1 mu l +/- 8.8% (n = 15) for whole blood have been successfully metered and stored in DBS paper. Conclusion: The new method of collecting a defined volume of blood by DBS sampling has the potential to reduce assay bias for the quantitative evaluation of DBS samples while maintaining the simplicity of conventional DBS sampling., QC 20151001

Multi-step lab-on-chip assays typically require multiple manual pipetting steps of different reagents and a sophisticated design to run the assay, which is generally unfavorable for point-of-care applications. This work presents the use of dissolvable polyvinylalcohol films for (1) reagent storage and release together with (2) timed valving in capillary driven microfluidics. This allows four different volumes to be split up from a single liquid which is applied to the inlet of the chip. PVA captured reagents are released to each volume forming four different solutions which are separately released in a timed sequence to a common target zone. The presented chip thereby enables a single liquid-triggered multi-reagent sequence with the potential to be used for advanced point of care diagnostics., QC 20150701

This work presents passive time delay valves for micro channels in two different working modes for microfluidic on chip timing. The delay valve designs are compatible with conventional lamination techniques for microfluidics and allow to pre-program advanced sequential operations independent from the geometries of the microfluidic system. The time delay of the dissolvable valves ranges from 1.2 s up to 36 s per valve resulting in a time range from 1.2 seconds up to 11 minutes for 19 serial valves., QC 20151208

This work presents a disposable chip for metering and transferring an exactly defined liquid volume into a paper matrix by capillary filling and emptying of a microchannel together with self-actuated dissolvable valves. Once a liquid droplet of 20-50 μl is applied to the chip, a volume of 1 μl is automatically metered, separated from the applied volume and subsequently transferred into conventional Whatman 903 paper used in Dried Blood Spot (DBS) sampling. The presented concept allows accurate volume metering for lateral flow devices and is here designed to the specific purpose of metering blood spots for DBS analysis. The material costs for each chip are below 0.04 €., QC 20140613