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Related Concept Videos

Data Validation01:15

Data Validation

Method validation is a crucial process in analytical chemistry designed to confirm that a given method consistently produces reliable and high-quality results. This process is essential when a method is applied to different sample matrices or when procedural modifications are made, ensuring that the results meet acceptable standards across various applications.
Key parameters for method validation include:
Blank Solutions00:56

Blank Solutions

A blank solution is a solution that does not contain the analyte, or the substance of interest being tested or measured. It is typically prepared using the same reagents and procedure as the sample solution but without adding the analyte. The primary purpose of preparing a blank solution is to account for any background interference or contamination that may affect the accuracy and reliability of the analytical method.
In some experimental cases, the reagents, solvents, or lab equipment used in...
Contaminants and Errors01:16

Contaminants and Errors

Effective sample preparation is crucial for accurate and reliable laboratory analysis. During this process, two significant sources of error can arise: concentration bias from improper sample splitting and contamination caused by methods used to reduce particle size, such as grinding or homogenization. Identifying and minimizing these potential errors is crucial to ensuring the validity of the analysis.
Another key consideration is determining the appropriate number of samples required to...

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Related Experiment Video

Updated: Jul 26, 2026

Application and Methodology of the Non-destructive 19F Time-domain NMR Technique to Measure the Content in Fluorine-containing Drug Products
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Addressing Analytical Challenges to Improve Laboratory Consistency in Unbound Flucloxacillin Measurement.

Cathérine Van Herteryck1, Tim Reyns2, Nynke Jager3

  • 1Department of Diagnostic Sciences, Ghent University, Ghent, Belgium.

Therapeutic Drug Monitoring
|March 10, 2026
PubMed
Summary

Accurate quantification of unbound flucloxacillin (FLU) is crucial for therapeutic drug monitoring. A validated LC-HRMS method using ultrafiltration (UF) was developed, showing good interlaboratory agreement and recommending plasma matrix for sample storage due to ultrafiltrate instability.

Keywords:
beta-lactam antibioticscross-validationflucloxacillinstabilityunbound concentration

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Area of Science:

  • Pharmacology and Pharmaceutical Sciences
  • Analytical Chemistry
  • Clinical Chemistry

Background:

  • Accurate quantification of flucloxacillin (FLU) in plasma is essential for therapeutic drug monitoring (TDM).
  • Only unbound FLU is pharmacologically active, necessitating methods like ultrafiltration (UF) for separation.
  • Lack of standardized UF protocols limits interlaboratory reproducibility.

Purpose of the Study:

  • Develop and validate a robust liquid chromatography-high-resolution mass spectrometric (LC-HRMS) method for quantifying unbound FLU.
  • Assess analyte stability and perform cross-validation with an external laboratory to harmonize UF protocols.

Main Methods:

  • Ultrafiltrate obtained after incubation and UF, analyzed by LC-HRMS.
  • Method validation included calibration curves, selectivity, carryover, accuracy, precision, matrix effect, stability, dilution integrity, nonspecific binding, and membrane protein leakage.
  • Cross-validation performed using patient samples from clinical routine.

Main Results:

  • Successful method validation.
  • Ultrafiltrate stability was significantly lower than plasma stability, requiring immediate postfiltration analysis.
  • Interlaboratory comparison showed strong concordance (>90% of samples differing by <±25.0%).

Conclusions:

  • A validated LC-HRMS method for quantifying unbound FLU using UF was developed.
  • Harmonizing key UF parameters (device type, temperature) enables comparable interlaboratory results.
  • Due to limited ultrafiltrate stability, sample storage in plasma matrix is recommended.