Rapid screening of plasma volume expanders in urine using matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry

Abstract

The use of plasma volume expanders, especially those based on chemically modified polysaccharides such as hydroxyethyl starch, has found its way from the medical field to the athletic community in the everlasting drive for performance enhancement. As such, plasma volume expanders have been placed on the list of banned substances by the International Olympic Committee, and in turn require accurate and sensitive analytical tools for their detection in complex biological matrices. Here we present a relatively straightforward method for the detection of polysaccharide-based plasma volume expanders (PVE) in urine, based on the carefully controlled partial acid hydrolysis of urine (20 microL) in a total volume of 500 microL 4 M trifluoroacetic acid. Following the incubation (30 min at 100 degrees C) an aliquot of the hydrolysate is dried, re-suspended in the analytical matrix (e.g. 2,5-dihydroxybenzoic acid) and examined by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS). The obtained mass spectrometric profile reveals a high number of characteristic peaks in the mass range between 500 and 3000 Da, a region that in urine samples devoid of PVE appears relatively clean, and thus allows the unambiguous identification of the presence of such PVE. This approach is fast (the mass profile can be obtained within 90 min), highly sensitive (the effective sample amount on the MALDI target is equivalent to 100 nL urine), needs little sample handling (four steps), requires no derivatisation and is devoid of interference from other biomolecules. The approach has been worked-out for hydroxy ethyl starch but can be applied to other polymer-derived plasma expanders such as dextran and probably the newly developed acetyl starch.

A rapid analytical method for the detection of plasma volume expanders and mannitol based on the urinary saccharides and polyalcohols profile

“…To speed up the dextran and HES screening analysis, different kinds of colourimetric and electrochemical methods have been proposed by previous investigators. [16][17][18][19][20][21][22][23] Nonetheless, mass spectrometric approaches seem to remain as the methods of choice due to their high specificity. Here we present an analytical screening procedure, involving only one enzymatic hydrolysis step and the direct injection into the LC-MS/MS system, specifically developed to detect, in a single run, the intake of dextran, HESl and mannitol in human urine.…”

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“…HES and dextran oligomers were obtained by hydrolysis with trifluo-roacetic acid (TFA) 3M, adapting a procedure previously described by Gallego and Segura [ 6 ]. Briefly, hydrolyzed HES (10 mg · mL −1 ) was prepared by pipetting 150 µ L of TFA in a tube containing 500 µ L of the commercial solution Voluven ® .…”

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2003–2004

“…Partial acid hydrolysis of urine was carried out with TFA and the product was examined directly by MALDI-TOF using super-DHB (s-DHB). The method was claimed to provide unambiguous identification in only 90 min (Gallego & Segura, 2004).…”