Development of a prototype blood fractionation cartridge for plasma analysis by paper spray mass spectrometry

Brandon J Bills,Nicholas E Manicke

doi:10.1016/j.clinms.2016.12.002

Abstract

Drug monitoring of biofluids is often time consuming and prohibitively expensive. Analysis of dried blood spots offers advantages, such as reduced sample volume, but depends on extensive sample preparation and the presence of a trained lab technician. Paper spray mass spectrometry allows rapid analysis of small molecules from blood spots with minimal sample preparation, however, plasma is often the preferred matrix for bioanalysis. Plasma spots can be analyzed by paper spray MS, but a centrifugation step to isolate the plasma is required. We demonstrate here the development of a paper spray cartridge containing a plasma fractionation membrane to perform automatic on-cartridge plasma fractionation from whole blood samples. Three commercially available blood fractionation membranes were evaluated based on: 1) accuracy of drug concentration determination in plasma, and 2) extent of cell lysis and/or penetration. The accuracy of drug concentration determination was quantitatively determined using high performance liquid chromatography–mass spectrometry (HPLC–MS). While the fractionation membranes were capable of yielding plasma samples with low levels of cell lysis, the membranes did exhibit drug binding to varying degrees, as indicated by a decrease in the drug concentration relative to plasma obtained by centrifugation. Using the membrane exhibiting the lowest binding, we developed a composite paper spray cartridge incorporating the selected fractionation membrane. Quantitative analysis of the plasma samples by paper spray MS yielded results similar to those found with HPLC–MS, but without the need for offline extraction or chromatography.

Highlights

Monitoring biofluid drug concentration is important for a number of applications
NoviPlex cards were obtained from Novilytic (West Lafayette, IN), while CytoSep and Vivid blood fractionation membranes were obtained from the Pall Corporation (Port Washington, NY)
Three commercially available plasma separation membranes were evaluated for their ability to obtain reproducible amounts of red blood cell-free plasma (Table 1)

Summary

Introduction

Monitoring biofluid drug concentration is important for a number of applications. Pharmaceuticals, for example, sometimes require that concentrations be maintained within patient-specific ranges to effect desired outcomes and, if possible, avoid toxicity. Current methods that analyze whole blood require time-consuming sample preparation. Whole blood requires special handling and refrigeration during transit. Because of these factors, therapeutic drug monitoring is often prohibitively expensive. Many fields in which small molecules are being monitored, such as forensics and toxicology, are faced with a similar problem. To address this need, there exists a continual push to develop rapid and cost effective analytical techniques that require minimal sample handling and preparation

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