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

Introduction. Tadalafil is a drug used to treat erectile dysfunction. For the quantitative determination of tadalafil in human plasma are used methods of high performance liquid chromatography with ultraviolet and tandem mass spectrometric detection, during the analytical part of pharmacokinetic studies. In the majority of the considered methods the method of liquid-liquid extraction and the method of solid-phase extraction are used, these methods are difficult and expensive. Therefore, the method of protein precipitation was considered as sample preparation. This method is simple and there is important to analysis a lot of clinical samples in bioequivalence studies.

Aim. The aim of this study is to develop method for the quantitative determination of tadalafil in human plasma by HPLC-MS for the analytical part of pharmacokinetic studies.

Materials and methods. Quantitative determination of tadalafil in plasma by HPLC-MS. A sample was prepared using acetonitrile protein precipitation.

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

Conclusion. The method of the quantitative determination of tadalafil in human plasma was developed and validated by HPLC-MS. The analytical range of the was 5,00–1000,00 ng/ml tadalafil in plasma. Method could be applied to determination of tadalafil in plasma for PK and BE studies.

1. Register of medicines. Available at: http://www.rlsnet.ru/mnn_index (In Russ.).

2. DrugBank. Available at: https://www.drugbank.ca/drugs/DB00820.

3. Shakya A. K., Abu-Awwad A. N., Arafat T. A., Melhim M. Validated liquid chromatographic–ultraviolet method for the quantitation of tadalafil in human plasma using liquid–liquid extraction. Journal of Chromatography B. 2007; 852(1-2): 403–408. Doi.org/10.1016/j.jchromb.2007.01.049.

4. Ramakrishna N. V. S., Vishwottam K. N., Puran, S., Koteshwara M., Manoj S., Santosh, M., Sumatha B. Quantitation of tadalafil in human plasma by liquid chromatography–tandem mass spectrometry with electrospray ionization. Journal of Chromatography B. 2004; 809(2): 243–249. Doi.org/10.1016/j.jchromb.2004.06.026.

5. Cheng C. L., Chou C. H. Determination of tadalafil in small volumes of plasma by high-performance liquid chromatography with UV detection. Journal of Chromatography B. 2005; 822(1-2): 278–284. Doi.org/10.1016/j.jchromb.2005.06.017.

6. Ma B., Shang X., Zhang Q., Li J., Liu Y., Cao X., Xu Q. Rapid analysis of tadalafil in human blood plasma and seminal plasma by liquid chromatography/tandem mass spectrometry. Journal of pharmaceutical and biomedical analysis. 2013; 77: 149–157. Doi.org/10.1016/j.jpba.2013.01.019.

7. Yokoyama Y., Tomatsuri M., Hayashi H., Hirai K., Ono Y., Yamada Y., Itoh K. Simultaneous microdetermination of bosentan, ambrisentan, sildenafil, and tadalafil in plasma using liquid chromatography/ tandem mass spectrometry for pediatric patients with pulmonary arterial hypertension. Journal of pharmaceutical and biomedical analysis. 2014; 89: 227–232. Doi.org/10.1016/j.jpba.2013.11.007

8. Rust K. Y., Wilkens H., Kaiser R., Bregel D., Wilske J., Kraemer T. Detection and Validated Quantification of the Phosphodiesterase Type 5 Inhibitors Sildenafil, Vardenafil, Tadalafil, and 2 of Their Metabolites in Human Blood Plasma by LC-MS/MS – Application to Forensic and Therapeutic Drug Monitoring Cases. Therapeutic drug monitoring. 2012; 34 (6): 729–735. Doi.org/10.1097/FTD.0b013e31827318b8.

9. Mironov A. N. Guidelines for the examination of medicines. V. II. M.: Grief and K. 2013; 280 p. (In Russ.).

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

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