Development and Validation of Pomalidomide Determination in Human Plasma by HPLC-MS/MS Method

Introduction. B-cell malignancies of the plasma cell leads to the second most spread hematological malignancy disease, called multiple myeloma. Pomalidomide is used in case of previous multiple myeloma ineffective treatment. Pomalidomide is a thalidomide synthetic derived, approved as immunomodulatory drug by the Food and Drug Administration (FDA). Nowadays, detection of pomalidomide in blood plasma by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) is not spread. Moreover, the detection and the experimental setting accumulated data are varying greatly. This investigation provides development and validation of pomalidomide aiming to determine human blood plasma by HPLC-MS/MS method. The samples were processed by methanol protein precipitation.

Aim. The aim of this study is to develop a method for the pomalidomide in human plasma by HPLC-MS/MS for pharmacokinetic studies.

Materials and methods. Determination of pomalidomide in plasma by HPLC-MS/MS. The samples were processed by methanol protein precipitation.

Results and discussion. This method was validated by next parameters: selectivity, matrix effect, calibration curve, accuracy, precision, spike recovery, lower limit of quantification, detection limit, carry-over and stability.

Conclusion. The method of the determination of pomalidomide in human plasma was developed and validated by HPLC-MS/MS. The linearity in plasma sample was achieved in the concentration range of 1,00 – 500,00 ng/ml. Method could be applied to pomalidomide determination in plasma for PK and BE studies.

1. Neşetoglu N., Kaplan C., Aslan S. S., Ünal D. Ö. A Simple and Rapid LC-MS/MS Method for Therapeutic Drug Monitoring of Lenalidomide. Pakistan Journal of Analytical & Environmental Chemistry. 2020;21(1):19–26. DOI: 10.21743/pjaec/2020.06.03.

2. Aydoğmuş Z., Yılmaz E. M., Yildiz G. A simple and rapid spectrofluorometric determination of pomalidomide in spiked plasma and urine. Application to degradation studies. Luminescence. 2019;35(4):1–12. DOI: 10.1002/bio.3748.

3. Hoffmann M., Kasserra C., Reyes J., Schafer P., Kosek I., Capone L., Parton A., Kim-Kang H., Surapaneni S., Kumar G. Absorption, metabolism and excretion of [14C]pomalidomide in humans following oral administration. Cancer Chemotherapy and Pharmacology. 2013;71(2):489–501. DOI: 10.1007/s00280-012-2040-6.

4. Shahbazi S., Peer C. J., Polizzotto M. N., Uldrick T. S., Roth J., Wyvill K. M., Aleman K., Zeldis J. B., Yarchoan R., Figg W. D. A sensitive and robust HPLC assay with fluorescence detection for the quantification of pomalidomide in human plasma for pharmacokinetic analyses. Journal of Pharmaceuticaland Biomedical Analysis. 2014;92:63–68. DOI: 10.1016/j.jpba.2014.01.001.

5. Jiang Y., Wang J., Rozewski D. M., Kolli S., Wu C., Chen C., Yang X., Hofmeister C. C., Byud J. C., Johnson A. J. Sensitive liquid chromatography/mass spectrometry methods for quantification of pomalidomide in mouse plasma and brain tissue. Journal of Pharmaceuticaland Biomedical Analysis. 2014;88:26–268. DOI: 10.1016/j.jpba.2013.08.036.

6. Iqbal M., Ezzeldin E., Al-Rashood K. A., Shakeel F. A validated UPLC–MS/MS assay using negative ionization mode for highthroughput determination of pomalidomide in rat plasma. Journal of Chromatography B. 2015;983-984:76–82. DOI: 10.1016/j.jchromb.2014.12.036.

7. Vasanth D. A., Rajkamal B. A UPLC-MS/MS method development and validation for the estimation of pomalidomide from human plasma. International Journal of Applied Pharmaceutics. 2017;9(1):37–43. DOI: 10.22159/ijap.2017v9i1.15653.

8. Mironov A. N. Guidelines for the examination of medicines. V. II. Moscow: Grif and K, 2013. 280 p. (In Russ.).

9. Food and Drug Administration. Available at: https://www.fda.gov/regulatory-information/search-fda-guidance-documents/bioanalytical-method-validation-guidance-industry.

10. European Medicines Agency. Available at: https://www.ema.europa.eu/en/bioanalytical-method-validation.