Síntese de um polímero de impressão molecular restrito à ligação com macromoléculas por meio de revestimento com albumina (RAMMIP-BSA), para análise direta de clorpromazina em plasma por sistema de cromatografia líquida bidimensional

Abstract

A restrict access moleculary imprinted polymer coated with albumin (RAM-MIPBSA) has been synthesized and used for direct analysis of chlorpromazine from human plasma by using bidimensional liquid chromatography. The synthesis was carried out in three steps. First, methacrylic acid (functional monomer), ethylene glycol dimethacrylate (cross-linker), 2,2 azoisobutironitrile (initiator) and chlorpromazine were dissolved in chloroform and the mixture was maintained at 60°C during 60 min. In the second step, 2-hydroxy ethyl acrylate and glycidil (both hydrophilic co-monomers) were added into the flask and the synthesis was processed during 24h at 60°C. The obtained polymer was washed several times and coated with bovine serum albumin by using glutaraldehide as cross-linker. The material presented a macromolecules elimination ability larger than 99%. It was packed in a column (4.4 x 10 mm) and used in a column switching system. Deionized water and 45:55 (v/v) acetonitrile:sodium acetate buffer (pH=4.1) were employed as extraction solvent and mobile phase, respectively. The analytical curve was built using a human plasma pool of different volunteers, added with the drug in 8 different concentrations. The method has shown to be linear in the range from 30 to 350 mg L-1, with r >0.99 and quantification limit of 30 mg L-1. Precision and accuracy were evaluated by using relative standard deviation and relative error, respectively, and all the results are in accordance with FDA recommendations. Additionally, good recoveries and selectivity were also obtained.