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

Importance of the topic

The precise measurement of docetaxel concentration in human serum is critical in clinical research to support pharmacokinetic studies, optimize dosing regimens, and ensure patient safety. High-throughput, sensitive, and reproducible assays enable researchers to generate reliable data for drug development and therapeutic monitoring.

Aims and overview

This study presents a rapid LC-MS/MS method for quantifying docetaxel in human serum. The approach combines Thermo Scientific SOLA solid phase extraction (SPE) technology with Accucore RP-MS columns for fast chromatographic separation and tandem mass spectrometric detection, targeting a lower limit of quantification (LLOQ) of 0.25 ng/mL.

Methodology and sample preparation

Sample processing was performed on 96-well SOLA SPE plates with the following steps:

• Condition with methanol and water
• Apply 200 µL spiked serum (10 µL docetaxel spike + 10 µL paclitaxel internal standard)
• Wash with water/acetonitrile (70:30, v/v)
• Elute with acetonitrile containing 1% ammonia
• Dry extracts under nitrogen and reconstitute in 50:50 water/acetonitrile with 0.5% 20 µM sodium acetate

Used instrumentation

The analytical setup included:

• Thermo Scientific UltiMate 3000 RSLC HPLC
• Accucore RP-MS, 2.6 µm, 50 × 2.1 mm column
• Thermo Scientific TSQ Vantage triple quadrupole MS with HESI source
• Thermo Scientific LC Quan software for data processing

Main results and discussion

Calibration over 0.25–10 ng/mL was linear (r2 = 0.9974). Key performance metrics included:

• Lower limit of quantification: 0.25 ng/mL
• Precision (%CV) for QC levels (0.3, 1.5, 6 ng/mL): ≤ 8.8%
• Accuracy deviations: within ±8.2% across standards
• Mean extraction recovery: 109%

The method delivered sharp chromatographic peaks in a 2-minute cycle, low background, and consistent sodium adduct formation for quantification.

Benefits and practical applications

• High sensitivity enabling low-level detection in serum
• Fast throughput with 2-minute runs and 96-well processing
• Robust reproducibility and recovery across sample batches
• Minimal solvent consumption and simplified sample cleanup

Future trends and opportunities

• Integration with automated SPE platforms for higher throughput
• Expansion to multi-analyte panels in clinical trials
• Development of novel core-shell phases for further speed gains
• Application in therapeutic drug monitoring and personalized medicine

Conclusion

The described LC-MS/MS assay offers a rapid, reliable, and sensitive solution for quantifying docetaxel in human serum. By leveraging advanced SPE and core-shell chromatography, the method meets stringent clinical research requirements while maintaining high efficiency and low operating costs.