Toward the Use of a Molecularly Imprinted Polymer in Doping Analysis: Selective Preconcentration and Analysis of Testosterone and Epitestosterone in Human Urine

Abstract

A molecularly imprinted polymer (MIP), templated with methyltestosterone, has been synthesized for the cleanup of hydrolyzed urine samples for subsequent testosterone (T) quantification by LC-MS/MS. A concentration of 2 ng/mL testosterone could be quantified after a single step extraction on the MIP. The limit of detection and quantification with the criteria of a signal-to-noise ratio of 3 and 5 were 0.3 and 2 ng/mL, respectively. These values meet the conditions set by the World Anti-Doping Agency for the minimum required performance limits for doping controls, between 2 and 10 ng/mL. Epitestosterone (E) was also separated on this polymer and could be detected at concentrations down to 0.3 ng/mL. The quantification of T and E gives access to the determination of the T/E ratio, essential in doping analysis. Hence, our polymers can offer a more specific extraction procedure, resulting in increased sensitivity with limits of detection 10 times lower than the ones achieved by the standard SPE C(18) sorbents employed in official testing laboratories.

Preparation and evaluation of a molecularly imprinted polymer for the selective recognition of testosterone—application to molecularly imprinted sorbent assays

“…We recently reported a molecularly imprinted polymer, templated with methyltestosterone, for the single step extraction of testosterone in hydrolyzed urine samples. The eluted testosterone was then quantified by LC/MS-MS. A limit of detection of 7 nM, that is, 10 times more sensitive than the one obtained by standard SPE C 18 sorbents used in doping analysis, was achieved (Tse Sum Bui et al, 2010). We therefore wanted to exploit this MIP furthermore as an alternative to antibodies in pseudoimmunoassays for the determination of testosterone.…”

“…The eluted testosterone was quantified by LC/MS-MS. The limit of quantification of testosterone after a single step extraction on the MIP was 7 nM, the sensitivity being 10 times superior to the one obtained with standard SPE C 18 sorbents (Tse Sum Bui et al, 2010). In this article, we study the viability of this MIP as an antibody mimic in a RIA for the detection and quantification of testosterone.…”

Molecularly Imprinted Polymer Nanomaterials and Nanocomposites: Atom‐Transfer Radical Polymerization with Acidic Monomers

“…The drugs testosterone and S‐propranolol were used as model templates. First, bulk polymers were synthesized using protocols adapted from previous publications 19. 20 MAA and ethylene glycol dimethacrylate (EGDMA) were used as monomers.…”

Molecularly Imprinted Polymer Nanomaterials and Nanocomposites: Atom‐Transfer Radical Polymerization with Acidic Monomers

“…The IC 50 value (the concentration of competing ligand required to displace 50 % of the specifically bound radioligand) for testosterone, determined from a nonlinear regression fit was 3.8 mm, which is comparable to that obtained with MIPs synthesized through FRP. [19,21] In the molecular imprinting field, commonly preferred ATRP catalysts are CuBr/N,N,N',N'',N''-pentamethyldiethylenetriamine (PMDETA) [7,8,[22][23][24] and CuBr/Tris[2-(dimethylamino)ethyl] amine (Me 6 TREN). [6,25] It was thus interesting to compare their performance with that of fac-[Ir(ppy) 3 ].…”

“…First, bulk polymers were synthesized using protocols adapted from previous publications. [19,20] MAA and ethylene glycol dimethacrylate (EGDMA) were used as monomers. MAA constituted 24 and 16 mol % of the total monomer amount in the testosterone and S-propranolol, respectively, MIP pre-cursor solutions.…”