An analytical method using salting-out assisted liquid-liquid extraction to quantify ceftriaxone from micro volumes of human serum

Yuji Mukai ¹, Narushi Sugii ², Kosuke Doki ^1,3

⁰Department of Pharmacy, University of Tsukuba Hospital, Tsukuba, Ibaraki, Japan.

Biomedical Chromatography : Bmc +

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July 8, 2024

View abstract on PubMed

Summary

This study developed a validated method to measure ceftriaxone (CTRX) in human serum. The new analytical technique ensures accurate and precise monitoring of this important antibiotic for safe clinical use.

Area Of Science

Analytical Chemistry
Pharmacokinetics
Clinical Diagnostics

Background

Ceftriaxone (CTRX) is a widely used cephalosporin antibiotic.
Therapeutic drug monitoring of CTRX serum concentrations is crucial for optimizing patient safety and treatment efficacy.
Existing methods may require improvement in terms of efficiency and adherence to regulatory guidelines.

Purpose Of The Study

To develop and validate a novel analytical method for quantifying ceftriaxone (CTRX) in human serum.
To ensure the method complies with the International Conference on Harmonization (ICH) M10 guideline.
To provide a reliable tool for therapeutic drug monitoring of CTRX.

Main Methods

Serum samples (10 μL) underwent salting-out assisted liquid-liquid extraction using magnesium sulfate.
Analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with a 12-minute run time.
Method validation included linearity, accuracy, precision, selectivity, matrix effects, stability, and carryover assessments.

Main Results

A linear calibration curve was established for CTRX concentrations ranging from 5 to 500 μg/mL.
Within-run and between-run accuracy ranged from 0.2% to 6.5% and 0.7% to 5.6%, respectively.
Within-run and between-run precision were ≤8.0% and ≤6.4%, respectively, with acceptable recovery rates (44.4% and 43.4%) and minimal carryover.

Conclusions

The developed LC-MS/MS method is accurate, precise, and reliable for measuring ceftriaxone (CTRX) in human serum.
The method meets ICH M10 guideline requirements, making it suitable for clinical application.
This validated assay facilitates effective therapeutic drug monitoring of ceftriaxone.

Keywords:

ceftriaxone liquid chromatography–tandem mass spectrometry magnesium sulfate salting‐out assisted liquid–liquid extraction