Capsule Phase Microextraction Combined with Chemometrics for the HPLC Determination of Amphotericin B in Human Serum

Abstract

This article discusses the use of a sorbent-based microextraction technique employing a capsule device to isolate amphotericin B (AMB) from human serum before analysis by high performance liquid chromatography (HPLC). AMB is a macrocyclic compound used for the treatment of invasive fungal infections. Before determining AMB in human serum by HPLC, a sample preparation step is required. Capsule phase microextraction (CPME) integrates the stirring and filtration mechanisms in a single unit, simplifying the sample preparation procedure. Moreover, it results in fast extraction kinetics and high extraction efficiency, while it has proved to be a powerful tool for bioanalysis. Different sol–gel sorbent encapsulated microextraction capsules were investigated, and sol–gel Carbowax 20 M was finally chosen as the basis for the microextraction device. Accordingly, the sample preparation protocol was investigated using a face-centered central composite design to achieve good extraction performance. The optimum protocol was validated in terms of linearity, selectivity, limit of detection (LOD), limit of quantitation (LOQ), precision, and accuracy. The linear range of the developed approach was 0.10–10.0 μg mL−1. The LOD value was 0.03 μg mL−1, and the LOQ value was 0.10 μg mL−1. Method accuracy (expressed as relative recovery) was 87–113%, while the relative standard deviation of the repeatability (sr) and within-laboratory reproducibility (sR) were <12.4%. The sol–gel sorbent encapsulated microextraction capsules were reusable for at least 10 extraction cycles. All things considered, the proposed method exhibited good overall performance, and it could be used in bioanalysis for quality control, therapeutic drug monitoring and research purposes.