Sample Preparation, Consumables, LC/MS, LC/MS/MS, LC columns, LC/QQQ
IndustriesClinical Research
ManufacturerThermo Fisher Scientific
Importance of the Topic
Capecitabine is an orally administered prodrug of 5-fluorouracil widely used in oncology. Accurate, rapid quantification in human plasma is essential for pharmacokinetic studies and therapeutic monitoring.
Objectives and Study Overview
This study aimed to develop a simple, fast SPE-LC-MS/MS method using Thermo Scientific SOLA cartridges and plates along with an Accucore PFP HPLC column to quantify capecitabine in small plasma volumes.
Methodology and Instrumentation
Sample preparation:
- Condition SOLA SPE with methanol and water
- Load 100 µL plasma spiked with capecitabine and D8 internal standard
- Wash with water/methanol (80:20)
- Elute with methanol, evaporate under nitrogen, reconstitute in water
Chromatography:
- Accela 600 pump and CTC autosampler
- Accucore PFP column (2.6 µm, 30 × 2.1 mm) with gradient water/acetonitrile
- Flow 1.0 mL/min, column temperature 40 °C, 10 µL injection
Mass spectrometry:
- TSQ Vantage with HESI in negative mode
- Transitions m/z 358.30→154.21 for capecitabine, 366.00→153.75 for D8 IS
- Optimized source and collision settings
Main Results and Discussion
- Retention time: 1.8 minutes for both analytes
- Calibration linear over 10–1000 ng/mL with r² ≥ 0.99
- Optimized wash step at 80:20 water/methanol minimized matrix interferences
- Intra-assay reproducibility: 2.3% RSD at mid-level concentration
- Average recovery: 73.2%
Benefits and Practical Applications
This method provides high‐throughput analysis using low sample and solvent volumes, robust extract cleanliness, and rapid turnaround, suitable for clinical pharmacokinetic and therapeutic drug monitoring laboratories.
Future Trends and Opportunities
Potential advancements include automation using 96‐well SPE formats, miniaturized extraction techniques, expanded application to similar prodrugs, and integration with high‐resolution mass spectrometry for multiplexed assays.
Conclusion
A validated SPE‐LC‐MS/MS workflow using SOLA cartridges/plates and Accucore column delivers a rapid, reproducible, and sensitive assay for capecitabine in human plasma.
References
- Farkouh A. A Rapid and Simple HPLC Assay for Quantification of Capecitabine for Drug Monitoring Purposes; Anticancer Research (2010), 30, 5207-5212
Content was automatically generated from an orignal PDF document using AI and may contain inaccuracies.