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Related Concept Videos

Dialysis01:15

Dialysis

Dialysis is a diffusion-based purification process that separates analyte molecules from a complex matrix. This is accomplished by allowing molecules in the solution to pass through a semipermeable membrane into a liquid on the other side. The membrane is usually made of cellulose acetate or cellulose nitrate, and the second liquid must be miscible with the solution. Ions (e.g., chloride or sodium) or organic molecules (e.g., glucose) can pass through the membrane pores, which generally have...
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In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
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Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...

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On-line microdialysis coupled solid-phase extraction to decrease matrix interference in the HPLC analysis of urinary

Shing-Jung Chen1, Fu-Chou Cheng, Jen-Fon Jen

  • 1Department of Chemistry, National Chung Hsing University, Taichung, Taiwan.

Journal of Separation Science
|May 28, 2010
PubMed
Summary

This study introduces a novel microdialysis sampling on-line solid-phase extraction (MDS/SPE) method for analyzing ketamine (K) and its metabolites in urine. The technique effectively reduces matrix interference, enabling accurate detection of K, norketamine (NK), and dehydronorketamine (DHNK).

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Area of Science:

  • Analytical Chemistry
  • Forensic Toxicology
  • Biomedical Analysis

Background:

  • Ketamine (K) and its metabolites, norketamine (NK) and dehydronorketamine (DHNK), are crucial analytes in urine drug testing.
  • Matrix interference in urine samples often complicates the accurate detection of these compounds using High-Performance Liquid Chromatography (HPLC).
  • Dilution techniques can reduce interference but may also decrease analyte concentrations, posing detection challenges.

Purpose of the Study:

  • To develop and optimize a microdialysis sampling on-line solid-phase extraction (MDS/SPE) method for the sensitive and selective determination of K, NK, and DHNK in human urine.
  • To decrease matrix interference and improve detection limits for these analytes.
  • To establish a simple, inexpensive, and eco-friendly analytical procedure.

Main Methods:

  • A combined microdialysis sampling (MDS) and on-line solid-phase extraction (SPE) system was employed for sample preparation prior to HPLC analysis.
  • Urine samples were filtered, diluted (100-fold), and pH adjusted before MDS collection.
  • Optimal conditions for MDS/SPE were determined, including perfusate composition (1 mM sulfuric acid), flow rate (20 µL/min), collection fiber (regenerated cellulose hollow fiber, 8 cm), and SPE trapping time (30 min) on an octadecyl-modified silica phase.

Main Results:

  • The optimized MDS/SPE method successfully reduced matrix interference, allowing for the determination of K, NK, and DHNK concentrations.
  • Recovery rates for K, NK, and DHNK were 41%, 42%, and 28% (m/m), respectively.
  • Excellent detection limits were achieved: 0.38 ng/mL for K, 0.33 ng/mL for NK, and 0.34 ng/mL for DHNK in the 100-fold diluted samples.

Conclusions:

  • The developed MDS/SPE method offers an efficient and effective approach for analyzing ketamine and its primary metabolites in urine.
  • This technique significantly mitigates matrix effects, enhancing the reliability of HPLC-based drug testing.
  • The procedure is characterized by its simplicity, cost-effectiveness, and environmentally friendly nature, making it suitable for routine forensic and clinical analysis.