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The use of coenzyme Q0 as a template in the development of a molecularly imprinted polymer for the selective recognition of coenzyme Q10
Authors:Mario Contin  Sabrina Flor  Manuela Martinefski  Silvia Lucangioli  Valeria Tripodi
Affiliation:1. Department of Analytical Chemistry and Physicochemistry, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina;2. Consejo Nacional de Investigaciones Científicas y Tecnológicas, CONICET, Buenos Aires, Argentina;3. Department of Pharmaceutical Technology, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina
Abstract:In this work, a novel molecularly imprinted polymer (MIP) for use as a solid phase extraction sorbent was developed for the determination of coenzyme Q10 (CoQ10) in liver extract. CoQ10 is an essential cofactor in mitochondrial oxidative phosphorylation and a powerful antioxidant agent found in low concentrations in biological samples. This fact and its high hydrophobicity make the analysis of CoQ10 technically challenging. Accordingly, a MIP was synthesised using coenzyme Q0 as the template, methacrylic acid as the functional monomer, acetonitrile as the porogen, ethylene glycol dimethacrylate as the crosslinker and benzoyl peroxide as the initiator. Various parameters affecting the polymer preparation and extraction efficiency were evaluated. Morphological characterisation of the MIP and its proper comparison with C18 as a sorbent in solid phase extraction were performed. The optimal conditions for the molecularly imprinted solid phase extraction (MISPE) consisted of 400 μL of sample mixed with 30 mg of MIP and 600 μL of water to reach the optimum solution loading. The loading was followed by a washing step consisting of 1 mL of a 1-propanol solution (1-propanol:water, 30:70,v/v) and elution with 1 mL of 1-propanol. After clean-up, the CoQ10 in the samples was analysed by high performance liquid chromatography. The extraction recoveries were higher than 73.7% with good precision (3.6–8.3%). The limits of detection and quantification were 2.4 and 7.5 μg g−1, respectively, and a linear range between 7.5 and 150 μg g−1 of tissue was achieved. The new MISPE procedure provided a successful clean-up for the determination of CoQ10 in a complex matrix.
Keywords:MIP, molecularly imprinted polymers   CoQ10, coenzyme Q10   MISPE, molecularly imprinted solid phase extraction   SPE, solid phase extraction   ATP, adenosine-triphosphate   HPLC, high performance liquid chromatography   ECD, electrochemical detection   MS, mass spectrometry   CoQ0, coenzyme Q0   UC, ubicromenol   MAA, methacrylic acid   EGDMA, ethylene glycoldimethacrylate   FEG-SEM, field emission gun scanning electron microscopy   NIP, non-imprinted polymer   LOD, limit of detection   LOQ, limit of quantification   RSD, relative standard deviation   SEM, scanning electron microscopy   BET, Brunauer&ndash  Emmett&ndash  Teller   IPB, imprinting-induced promotion of binding   N, theoretical plates
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