Application of a Small Protein-Coated Column to Trap, Extract and Enrich Carbamazepine Directly from Human Serum for Direct Chromatographic Analysis

SEPARATIONS(2023)

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摘要
An automated solid phase extraction (SPE) protocol to determine carbamazepine in human serum has been developed and validated using a simple, rabid and sensitive liquid chromatography-based bio-analytical method. Extraction of carbamazepine was carried out using an on-line SPE tool of a short protein-coated (PC) ODS silica pre-column (PC-ODS-pre-column) and phosphate buffer saline (PBS) with a pH of 7.4 as an extraction solvent. There are two distinct chromatographic modes used by PC-ODS-pre-column. While carbamazepine trapping required reversed-phase liquid chromatography, proteins were extracted from serum samples using PBS by size-exclusion liquid chromatography. Then, carbamazepine was eluted from the PC-ODS-pre-column onto the quantification position using a mixture of methanol-distilled deionized water (50:50, v/v) as an eluent and ODS analytical column. At room temperature (22 +/- 1 degrees C), carbamazepine was completely separated from the co-eluted matrix components and detected at 230 nm. Carbamazepine's linearity was obtained at concentrations ranging from 50 to 10,000 ng/mL. With good accuracy and precision, carbamazepine recoveries in serum samples ranged from 86.14 to 97.82%. The extraction step was conducted using PBS as a safe and green extraction solvent, making this protocol both cost-effective and ecologically safe.
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关键词
carbamazepine,protein-coated ODS silica pre-column,human serum,on-line solid phase extraction,UV detection
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