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Abstract: In this work, a novel method is described for the determination of bromhexine in biological fluids using molecularly imprinted solid-phase extraction as the sample cleanup technique combined with high performance liquid chromatography (HPLC). The water-compatible molecularly imprinted polymers (MIPs) were prepared using methacrylic acid as functional monomer, ethylene glycol dimethacrylate as cross-linker, chloroform as porogen and bromhexine as the template molecule. The novel imprinted polymer was used as a solid-phase extraction sorbent for the extraction of bromhexine from human serum and urine. Various parameters affecting the extraction efficiency of the polymer have been evaluated. The optimal conditions for molecularly imprinted solid-phase extraction (MISPE) consisted of conditioning 1ámL methanol and 1ámL of deionized water at neutral pH, loading of 5ámL of the water sample (25áμgáL-1) at pH 6.0, washing using 2ámL acetonitrile/acetone (1/4, v/v) and elution with 3+ 1ámL methanol/acetic acid (10/1, v/v). The MIP selectivity was evaluated by checking several substances with similar molecular structures to that of bromhexine. Results from the HPLC analyses showed that the calibration curve of bromhexine using MIP from human serum and urine is linear in the ranges of 0.5-100 and 1.5-100áμgáL-1 with good precisions (3.3% and 2.8% for 5.0áμgáL-1), respectively. The recoveries for serum and urine samples were higher than 92%
Template and target information: bromhexine
Author keywords: molecularly imprinted polymer, Solid-phase extraction, Bromhexine, human serum, urine