Micro-Spe in Pipette Tips as A Tool for Analysis of Small-Molecule Drugs in Serum
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Abstract:
Micro-SPE in pipette tips (μ-SPE-PT) with particle sorbent has never been used in small-molecule drug analysis. Methodology & results: μ-SPE-PT was used for the extraction of statins from biological materials followed by UHPLC-MS/MS. The commercial and homemade μ-SPE-PT tips filled with particle sorbent were compared. While the homemade tips enabled direct serum sample loading into the sorbent, protein precipitation (PP) had to be implemented before μ-SPE-PT procedure using commercial tips. Three μ-SPE-PT methods were developed and validated: method A: μ-SPE-PT with homemade tips; method B: PP + μ-SPE-PT with homemade tips; and method C: PP + μ-SPE-PT with commercial tips. Method A enabled a simple high-throughput approach (48 samples in 90 min) compared with methods B and C that required three-times longer time. However, PP increased the recoveries of protein-bound analytes and extracts purity in methods B and C. The matrix effects without internal standards correction for method C were significantly higher than those for the methods A and B.Compared with commercial tips, homemade tips filled with particles were found to be more suitable for drug analysis. Commercial tips tested in this study were found challenging but the conditions under which they could be applicable were also defined.Keywords:
Pipette
Solid phase extraction
Sample Preparation
Matrix (chemical analysis)
Protein precipitation
Low levels of biomarkers and the complexity of bio sample make the analytical assay of several biomarkers a challenging issue. Suitable sample preparation run remain a vital part of the puzzle of diagnostic level. Enhancing the detection limit of bioanalytical methods start during the sample preparation procedure. A robust sample preparation method is needed to evaluate the number of biomarkers. As worldwide environmental issues attract expanding consideration, all the more harmless to the ecosystem investigations are liked. Solid-phase extraction (SPE) is an appealing strategy among the sample treatment methods due to the versatility of sorbent materials, less solvent consumption, and compatibility with analytical devices. Miniaturization of the SPE gives the chance to integrate the other analytical steps in a single run, known as an easy-to-use and effective method. SPE utilizes various SPE sorbent beds such as packed beads, porous polymer monoliths, molecularly imprinted polymers, membranes, or other magnetic form microstructures to achieve high surface-to-volume ratio and appropriate chemical properties effective extraction. Also, SPE is the methodology of interest to fulfill high recovery and efficiency demands. In this review, we intend to explain more recent methods for the rational design of SPE and miniaturized SPE to determine biomarkers from biological media. The headlines are subdivided into (1) packing materials in SPE, (2) setups for sample preparation by magnetic SPE, and (3) and future perspective for the application of SPE in sample preparation for analysis of biomarkers.
Solid phase extraction
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Solid phase extraction
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Solid-phase extraction (SPE) is a common sample preparation method that involves the interaction between an aqueous sample and a solid phase (or sorbent). An important variable in any SPE system is the choice of sorbent. Generally, SPE sorbents can be divided into two generic classes: silica-based sorbents and unmodified substrates. Irrespective of SPE format, the method of operation is the same and can be divided into several steps. Each step is characterized by the nature and type of solvent used, which in turn, is dependent upon the characteristics of the sorbent and the sample. The steps are explained in the context of a reversed-phase SPE system using a C18 sorbent, and with a vacuum manifold applied. The general methodology to be followed for off-line SPE is described using generic examples with emphasis on reversed phase, normal phase, ion exchange, and mixed mode systems.
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