LC-MS/MS Method for the Determination of Docetaxel in Human Serum for Clinical Research

Significance of the Topic

Docetaxel is a cornerstone chemotherapeutic agent used in various cancer treatments. Accurate and sensitive quantification of docetaxel levels in human serum is critical for pharmacokinetic studies, dose optimization, and safety monitoring in clinical research.

Objectives and Study Overview

This application note describes the development and validation of a rapid, high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for measuring docetaxel concentrations in human serum using solid-phase extraction and a core-shell HPLC column.

Methodology and Used Instrumentation

Sample Preparation

• Solid-phase extraction (SPE) performed on Thermo Scientific SOLA 10 mg/2 mL plates.
• Conditioning with methanol and water, sample application, wash with 70:30 water/acetonitrile, elution with acetonitrile/ammonia.
• Evaporation under nitrogen, reconstitution in water/acetonitrile with 20 µM sodium acetate to stabilize sodium adduct formation.

Liquid Chromatography

• Thermo Scientific Dionex UltiMate 3000 RSLC system.
• Accucore RP-MS column (2.6 µm, 50 × 2.1 mm) at 30 °C.
• Gradient elution from 60 % to 70 % organic phase in 2 minutes, flow rate 0.6 mL/min.

Mass Spectrometry

• Thermo Scientific TSQ Vantage triple quadrupole MS with heated electrospray ionization in positive mode.
• MRM transitions: docetaxel m/z 830.26→549.24; paclitaxel (IS) m/z 876.25→308.25.
• Optimized spray voltage, temperatures, gas pressures, collision energies, and resolution settings.

Main Results and Discussion

• Linearity achieved over 0.25–10 ng/mL (r² = 0.9974).
• Lower limit of quantification (LLOQ) determined at 0.25 ng/mL with clear chromatographic peak shape.
• Accuracy data showed deviations within ±8.8 % across QC levels (0.3, 1.5, 6 ng/mL).
• Precision demonstrated CV ≤ 8.8 % for six replicates at each QC level.
• Extraction recovery averaged 109 %, indicating effective SPE cleanup.

Benefits and Practical Applications

• Short 2-minute analysis cycle time supports high-throughput workflows.
• SOLA SPE cartridges reduce solvent consumption and simplify sample handling.
• High sensitivity enables monitoring of low-dose docetaxel regimens.
• Robust reproducibility and recovery support reliable pharmacokinetic and bioequivalence studies.

Future Trends and Applications

• Extension of the method to other anticancer drugs and endogenous biomarkers.
• Integration with robotic sample preparation for large-scale clinical trials.
• Development of multiplexed LC-MS/MS assays for simultaneous drug monitoring.
• Continued improvements in SPE and column materials to further reduce run times and sample sizes.

Conclusion

The described LC-MS/MS workflow combining SOLA SPE and Accucore RP-MS core-shell columns delivers a rapid, sensitive, and reproducible method for quantifying docetaxel in human serum, meeting the stringent requirements of clinical research environments.