Is the hematocrit still an issue in quantitative dried blood spot analysis?

Velghe, Sofie; Delahaye, Lisa; Stove, Christophe

doi:10.1016/j.jpba.2018.10.010

Cited by 158 publications

(116 citation statements)

References 69 publications

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“…To investigate the influence of blood viscosity on different AHD concentrations, blood with different hematocrit levels (0.3, 0.35, 0.4, and 0.45 L/L) was administered to increasing AHD concentrations on DBS. 22,23 Samples were measured using the UHPLC-MS/MS method, as previously described. 7…”

Section: Hematocritmentioning

confidence: 99%

Clinical Validation of a Dried Blood Spot Assay for 8 Antihypertensive Drugs and 4 Active Metabolites

Peeters¹,

Feyz

Hameli³

et al. 2020

Therapeutic Drug Monitoring

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Background: Drug nonadherence is one of the major challenges faced by resistant hypertension patients, and identification of this problem is needed for optimizing pharmacotherapy. Dried blood spot (DBS) sampling is a minimally invasive method designed to detect and determine the degree of nonadherence. In this study, a DBS method for qualifying 8 antihypertensive drugs (AHDs) and 4 active metabolites was developed and validated using ultra highperformance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Method: The DBS assay was validated analytically and clinically, in accordance with FDA requirements. Analytical validation was accomplished using UHPLC-MS/MS. For clinical validation, paired peak and trough levels of DBS and plasma samples were simultaneously collected and comparatively analyzed using Deming regression and Bland-Altman analyses. All concentrations below the set lower limit were excluded. Deming regression analysis was used to predict comparison bias between the collected plasma and DBS samples, with DBS concentrations corrected accordingly. Results: The UHPLC-MS/MS method for simultaneously measuring 8 AHDs and their metabolites in DBS, was successfully validated. With Deming regression no bias was observed in N = 1; constant bias was seen in N = 6 and proportional bias in N = 11 of the AHDs and metabolites. After correction for bias, only one metabolite (canrenone) met the 20% acceptance limit for quantification, after Bland-Altman analyses. In addition, amlodipine, valsartan, and [enalaprilate] met the 25% acceptance limit. Conclusions: A novel DBS assay for simultaneously qualifying and quantifying 8 AHDs and their metabolites, has been successfully developed and validated. The DBS assay is therefore a suitable method to detect drug nonadherence. However, with the exception of canrenone, the interchangeable use of plasma and DBS sampling to interpret drug quantities should be avoided.

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Section: Hematocritmentioning

confidence: 99%

Clinical Validation of a Dried Blood Spot Assay for 8 Antihypertensive Drugs and 4 Active Metabolites

Peeters¹,

Feyz

Hameli³

et al. 2020

Therapeutic Drug Monitoring

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show abstract

“…20 μL) into a porous substrate, independent of hematocrit, and potentially improve the ease of sampling for the patient [17][18][19]. Although the effects of the hematocrit on the sample volume can be overcome by VAMS, this does not necessarily apply for the effect of hematocrit on extraction recovery from VAMS tips [20][21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Volumetric absorptive microsampling and dried blood spot microsampling vs. conventional venous sampling for tacrolimus trough concentration monitoring

Veenhof

Koster

Junier

et al. 2020

Clinical Chemistry and Laboratory Medicine (CCLM)

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ObjectivesMonitoring tacrolimus blood concentrations is important for preventing allograft rejection in transplant patients. Our hospital offers dried blood spot (DBS) sampling, giving patients the opportunity to sample a drop of blood from a fingerprick at home, which can be sent to the laboratory by mail. In this study, both a volumetric absorptive microsampling (VAMS) device and DBS sampling were compared to venous whole blood (WB) sampling.MethodsA total of 130 matched fingerprick VAMS, fingerprick DBS and venous WB samples were obtained from 107 different kidney transplant patients by trained phlebotomists for method comparison using Passing-Bablok regression. Bias was assessed using Bland-Altman. A multidisciplinary team pre-defined an acceptance limit requiring >80% of all matched samples within 15% of the mean of both samples. Sampling quality was evaluated for both VAMS and DBS samples.Results32.3% of the VAMS samples and 6.2% of the DBS samples were of insufficient quality, leading to 88 matched samples fit for analysis. Passing-Bablok regression showed a significant difference between VAMS and WB, with a slope of 0.88 (95% CI 0.81–0.97) but not for DBS (slope 1.00; 95% CI 0.95–1.04). Both VAMS (after correction for the slope) and DBS showed no significant bias in Bland-Altman analysis. For VAMS and DBS, the acceptance limit was met for 83.0% and 96.6% of the samples, respectively.ConclusionsVAMS sampling can replace WB sampling for tacrolimus trough concentration monitoring, but VAMS sampling is currently inferior to DBS sampling, both regarding sample quality and agreement with WB tacrolimus concentrations.

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“…Peptide Standard Synthesis-Peptide synthesis was performed as previously described (30). Briefly, SIS peptides were synthesized by incorporating 13 C/ 15 N heavy isotope L-arginine or L-lysine on the C terminus. Peptides were purified using HPLC and the parent mass to charge ratio confirmed using an Ultraflex III-MALDI TOF/TOF Mass Spectrometer (Bruker LTD., Milton, ON).…”

Section: Methodsmentioning

confidence: 99%

Concentration Determination of >200 Proteins in Dried Blood Spots for Biomarker Discovery and Validation

Eshghi

Pistawka

et al. 2020

Molecular & Cellular Proteomics

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The use of protein biomarkers as surrogates for clinical endpoints requires extensive multilevel validation including development of robust and sensitive assays for precise measurement of protein concentration. Multiple reaction monitoring (MRM) is a well-established mass-spectrometric method that can be used for reproducible protein-concentration measurements in biological specimens collected via microsampling. The dried blood spot (DBS) microsampling technique can be performed non-invasively without the expertise of a phlebotomist, and can enhance analyte stability which facilitate the application of this technique in retrospective studies while providing lower storage and shipping costs, because cold-chain logistics can be eliminated. Thus, precise, sensitive, and multiplexed methods for measuring protein concentrations in DBSs can be used for de novo biomarker discovery and for biomarker quantification or verification experiments. To achieve this goal, MRM assays were developed for multiplexed concentration measurement of proteins in DBSs.The lower limit of quantification (LLOQ) was found to have a median total coefficient of variation (CV) of 18% for 245 proteins, whereas the median LLOQ was 5 fmol of peptide injected on column, and the median inter-day CV over 4 days for measuring endogenous protein concentration was 8%. The majority (88%) of the assays displayed parallelism, whereas the peptide standards remained stable throughout the assay workflow and after exposure to multiple freeze-thaw cycles. For 190 proteins, the measured protein concentrations remained stable in DBS stored at ambient laboratory temperature for up to 2 months. Finally, the developed assays were used to measure the concentration ranges for 200 proteins in twenty same sex, same race and age matched individuals.

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Is the hematocrit still an issue in quantitative dried blood spot analysis?

Cited by 158 publications

References 69 publications

Clinical Validation of a Dried Blood Spot Assay for 8 Antihypertensive Drugs and 4 Active Metabolites

Clinical Validation of a Dried Blood Spot Assay for 8 Antihypertensive Drugs and 4 Active Metabolites

Volumetric absorptive microsampling and dried blood spot microsampling vs. conventional venous sampling for tacrolimus trough concentration monitoring

Concentration Determination of >200 Proteins in Dried Blood Spots for Biomarker Discovery and Validation

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