Capecitabine in Human Plasma Using SOLA and Accucore Core Enhanced Technology HPLC Column

Significance of the Topic

Capecitabine is an orally administered prodrug of 5-fluorouracil widely used in oncology. Accurate quantification in human plasma supports pharmacokinetic studies, therapeutic drug monitoring, and drug development. Combining efficient sample preparation with high-resolution separation addresses challenges of low sample volume, complex matrix effects, and regulatory requirements.

Objectives and Study Overview

This study aimed to develop and validate a rapid and sensitive method for detecting capecitabine in 100 µL human plasma. Extraction performance was evaluated using Thermo Scientific SOLA SPE cartridges and 96-well plates. Quantitative analysis employed HPLC–MS/MS with a core–shell Accucore PFP column under gradient elution.

Methodology and Instrumentation

• Solid Phase Extraction: SOLA 10 mg cartridges and 96-well plates conditioned with methanol and water, loading 100 µL plasma, washing with 80:20 water/methanol, eluting with methanol, drying under nitrogen, and reconstitution in water.
• Chromatography: Accucore PFP 2.6 µm, 30×2.1 mm column; mobile phases water (A) and acetonitrile (B); gradient from 100% A to 100% B in 5 min, returning to initial conditions by 5.1 min; flow rate 1.0 mL/min; column temperature 40 °C; injection volume 10 µL.
• Mass Spectrometry: Thermo Scientific TSQ Vantage with HESI source in negative ion mode; transitions m/z 358.30→154.21 for capecitabine and 366.00→153.75 for capecitabine-D8; collision energy 21 eV; full scan widths Q1 and Q3 at 0.7 FWHM.

Key Results and Discussion

• Retention: Capecitabine and its deuterated analogue eluted in 1.8 min, achieving a rapid cycle time under 6.5 min.
• Linearity: Calibration range 10–1000 ng/mL displayed excellent linearity (r² > 0.99).
• Recovery: Mean recovery of 73.2% was consistent across the range, with efficient removal of matrix interferences.
• Reproducibility: Intra-day precision at mid concentration showed %RSD of 2.3% (n=3).
• Matrix Effects: Minimal ion suppression/enhancement observed after optimized wash step.

Benefits and Practical Applications

The method uses only 100 µL plasma, reducing patient sample requirements. SOLA SPE offers high reproducibility and clean extracts, lowering solvent consumption and processing time. The rapid Accucore PFP separation and sensitive MS/MS detection enable high throughput suitable for clinical pharmacokinetic and therapeutic monitoring studies.

Future Trends and Applications

• Expansion of core–shell column chemistry to other bioanalytical targets for faster separations.
• Automation of SPE workflows in 96-well format to enhance throughput and consistency.
• Application to multi-analyte panels, including metabolites, in precision medicine.
• Integration with microflow LC–MS for further sensitivity gains and reduced solvent use.

Conclusion

A robust SPE–LC–MS/MS workflow was established for quantifying capecitabine in small volumes of human plasma. SOLA cartridges/plates and Accucore PFP core–shell columns delivered rapid, sensitive, and reproducible analysis, meeting the demands of bioanalytical and clinical laboratories.

Instrumentation Used

• Thermo Scientific SOLA SPE cartridges (10 mg/1 mL) and 96-well plates.
• Thermo Scientific Accela 600 HPLC system with CTC autosampler.
• Accucore PFP 2.6 µm, 30×2.1 mm column and Defender Guard Column.
• Thermo Scientific TSQ Vantage triple quadrupole mass spectrometer with HESI source.

References

1. Farkouh A. A Rapid and Simple HPLC Assay for Quantification of Capecitabine for Drug Monitoring Purposes. Anticancer Research. 2010;30:5207–5212.