Ceramides biomarkers determination in quantitative dried blood spots by UHPLC-MS/MS

Abstract

A method was developed for the analysis of four ceramide species; namely C16:0, C18:0, C24:0 and C24:1 in quantitative Dried Blood Samples (qDBS) by LC-MS/MS and validated with the aim to give prominence to an interesting application of at-home blood microsampling for health monitoring. Ceramides, being key-role metabolites implicated in regulation of diverse cellular processes have been considered as emerging biomarkers for different disease states, such as cardiovascular diseases, type 2 diabetes and others. Here, Capitainer device was utilized to provide accurate and consistent volumes of samples, ideal for accurate determinations. The method requires a 10 μL sample offering duplicate analysis by device, is quick and enables the sample collection by distance as it was proved that ceramides under study were stable at various conditions, including RT. Intra and inter-day accuracy of the determination were estimated between 87.6% - 113% and 90.6% -113%, respectively, while intra- and inter-day precision were calculated from 0.2% to 9.9% %RSD and 0.1% - 8.0% %RSD, respectively. The data acquired by ten healthy individuals indicated that circulating ceramides are at higher levels in whole blood taken from the fingertip in comparison to the reported values in plasma or serum.

Keywords: Blood microsampling; Dried blood spot; LC-MS/MS; qDBS.