Doxorubicin in Human Serum Using SOLA and Hypersil GOLD Column

Importance of the Topic

This application focuses on accurate quantification of doxorubicin, a widely used anthracycline anticancer agent, in human serum. Reliable measurement of drug levels supports therapeutic drug monitoring, pharmacokinetic studies, dose optimization and safety assessments in oncology.

Objectives and Overview of the Study

The primary aim was to establish a fast, sensitive and reproducible liquid chromatography method for doxorubicin in human serum. Key goals included minimal sample preparation time, low solvent consumption, high accuracy and a short chromatographic run time of 4 minutes.

Methodology and Instrumentation

Sample preparation employed Thermo Scientific SOLA 10 mg/1 mL solid phase extraction cartridges:

• Condition and equilibrate with methanol and water
• Load 200 µL spiked serum (with internal standard daunorubicin)
• Wash with water/methanol (90:10, v/v)
• Elute with methanol containing 0.1% formic acid

Chromatography was performed on a Thermo Scientific Accela 600 pump with CTC autosampler and a Hypersil GOLD 3 µm column (50 × 2.1 mm) at ambient temperature. Mobile phase gradient between 25 mM ammonium acetate (pH native) and acetonitrile delivered at 1.5 mL/min achieved separation within 4 min. Fluorescence detection used 480 nm excitation and 560 nm emission. Data integration was managed in ChromQuest 5 software.

Main Results and Discussion

The calibration curve was linear over 1–1000 ng/mL with r2=0.9991. Single-level accuracy checks at 20 ng/mL across six replicates yielded a mean measured concentration of 20.2 ng/mL (101.2% nominal) and RSD of 6.9%. The method demonstrated low matrix interference, high reproducibility and rapid turnaround.

Benefits and Practical Applications

• Reduced solvent usage and shorter drying times lower operational costs
• High extract cleanliness improves sensitivity and reduces background
• Fast run time enhances sample throughput in clinical and bioanalytical labs
• Simple SPE workflow decreases method complexity and failure rates

Future Trends and Applications

Future developments may include coupling with mass spectrometry for enhanced selectivity, automation of SPE for higher throughput, miniaturized formats for limited volumes and extension of the approach to other anthracycline drugs or metabolites in therapeutic monitoring and pharmacokinetic research.

Conclusion

The combination of SOLA SPE cartridges and Hypersil GOLD HPLC columns provides a robust, accurate and efficient method for quantifying doxorubicin in human serum. The approach meets the demands of high-throughput bioanalytical laboratories and supports reliable therapeutic drug monitoring.

References

1. Barpe D.R., Rosa D.D., Froehlich P.E. Pharmacokinetic evaluation of doxorubicin plasma levels in normal and overweight patients with breast cancer and simulation of dose adjustment by different indexes of body mass. Eur J Pharm Sci. 2010;41(3-4):458–463.
2. Current Cancer. Pharmacology and mechanisms of doxorubicin. http://currentcancer.com/doxorubicin.html (accessed 06/09/2011).