HPLC-MS/MS method for the quantitative determination of bortezomib in dried blood microsamples collected by volumetric absorptive microsampling

Sobia Noreen1, Roberto Mandrioli2, Michele Protti1, Laura Mercolini1, Oliver Scherf-Clavel3

 

1 Research group of Pharmaco-Toxicological Analysis (PTA Lab), Department of Pharmacy and Biotechnology (FaBiT), Alma Mater Studiorum - University of Bologna, Via Belmeloro 6, Bologna, 40126 (Italy);

2 Research group of Advanced Pharmaceutical Analysis (APA Lab), Department of Pharmacy and Biotechnology (FaBiT), Alma Mater Studiorum - University of Bologna, Corso D'Augusto 237, Rimini, 47921 (Italy);

3 Department of Analytical Chemistry and Organic Chemistry, Ludwig-Maximilians-University (LMU) Munich, Butenandtstraße, 5, Munich, D-81377 (Germany)

*Email: sobia.noreen3@unibo.it

 

Bortezomib is a proteasome inhibitor widely used for multiple myeloma treatment. Its variable pharmacokinetic profile and toxicity concerns, including peripheral neuropathy and haematological effects [1], underline the need for robust quantitative methods to support therapeutic drug monitoring. This study aimed to develop an HPLC-MS/MS method for the quantitative determination of bortezomib in dried blood microsamples collected using volumetric absorptive microsampling (VAMS), enabling accurate-volume (10 µL) sample collection with minimal invasiveness, simplified handling, increased stability and reduced overall costs [2]. Chromatographic and MS/MS conditions were optimised by evaluating precursor/product ion selection, multiple reaction monitoring (MRM) transitions and collision energies. Analyte extraction from the dried blood microsamples was performed using methanol under ultrasound-assisted conditions. The developed VAMS-HPLC-MS/MS workflow was validated for linearity, accuracy, precision, recovery, matrix effects and stability. The method showed excellent linearity (R² ≥ 0.995), accuracy (90.62-101.90%), precision (≤13.75%) and mean extraction recoveries of 88.25% and 89.95% at low and high QC levels, respectively, supporting VAMS-HPLC-MS/MS as a promising minimally invasive approach for bortezomib therapeutic monitoring and clinical bioanalysis.

 

References:

[1] Y. Yang, B. Zhao, H. Lan, J. Sun, G. Wei, Crit Rev Oncol Hematol., 197 (2024) 104353.

[2] M. Protti, E. Milandri, R. Di Lecce, L. Mercolini, R. Mandrioli, Adv. Sample Prep., 13 (2025) 100161.

Acknowledgements:

This study was supported by the scientific collaboration between the Alma Mater Studiorum - University of Bologna and Ludwig-Maximilians-University Munich (LMU Munich). Fondazione Cassa di Risparmio di Imola is acknowledged for supporting the doctoral fellowship of the presenting author.