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Quantification by nano liquid chromatography parallel reaction monitoring mass spectrometry of human apolipoprotein A‐I, apolipoprotein B, and hemoglobin A1c in dried blood spots
Author(s) -
Henderson Clark M.,
Bollinger James G.,
Becker Jessica O.,
Wallace Jennifer M.,
Laha Thomas J.,
MacCoss Michael J.,
Hoofnagle Andrew N.
Publication year - 2017
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201600103
Subject(s) - venipuncture , apolipoprotein b , chromatography , glycation , chemistry , hemoglobin , biomarker , medicine , biochemistry , receptor , surgery , cholesterol
Purpose Proteomic analysis of blood proteins in dried blood spots (DBS) is gaining attention as a possible replacement for measurements in plasma/serum collected by venipuncture. We aimed to develop and provisionally validate a nanoflow LC‐PRM‐MS method for clinical use. Experimental design We used Skyline to develop a nanoflow LC‐PRM‐MS method to quantify glycated hemoglobin‐β, apolipoprotein A‐I, and apolipoprotein B in DBS. Precision, linearity, interferences, and stability were determined and the method was used to analyze samples from 36 human volunteers. The method was compared with clinically validated measurements in paired blood collected via venipuncture. Results The method was relatively precise for these proteins (10–11% CV) and linear across the normal concentration ranges of these proteins. Interference from high total serum protein concentration (>8 g/dL) was noted for apolipoprotein A‐I and apolipoprotein B. Proteins in DBS were stable for 14 days at temperatures below 25°C and trypsinized samples were stable for 48 h at 7°C. There was moderate correlation with clinical methods ( r = 0.783–0.858) and significant bias in individual samples. Conclusions and clinical relevance Although the method had adequate precision and linearity for a biomarker, the accuracy compared with clinically validated assays raises concerns regarding the use of DBS compared with venipuncture for clinical use.