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

Abstract

Introduction. "Eltrombopag" is a thrombopoietin receptor agonist (TPO-RA) that is approved for the treatment of immune thrombocytopenic purpura (ITP). According to the literature, very few analytical methods for determining eltrombopag in biological samples have been reported. To study the pharmacokinetics of new formulations of eltrombopag, a sensitive and specific method is required that allows one to accurately determine the concentration of eltrombopag in human blood plasma. Normally, HPLC methods should provide time, accuracy, and sensitivity as a result, it is necessary to develop fast or ultra-fast methods such as LC-MS/MS without any loss in sensitivity or separation efficiency.Aim. We aimed to develop and validate a method for the quantitative determination of eltrombopag levels in human plasma by using HPLC with mass spectrometric detection for performing the analytical part of pharmacokinetic studies.Materials and methods. Eltrombopag levels were determined in human plasma by HPLC with mass spectrometric detection. The samples were prepared using protein deposition.Results and discussion. The method was validated for selectivity, matrix effect, calibration curve, accuracy, precision, the limit of quantification, carry-over effect, and sample stability.Conclusion. The method for the determination of eltrombopag levels in human plasma has been developed and validated by HPLC-MS. The analytical range of eltrombopag levels in human plasma was 10–6750 ng/ml. This method could be used to determine eltrombopag levels in plasma for PK and BE studies.

Keywords

• «элтромбопаг»
• плазма
• вэжх-мс/мс
• валидация
• биоэквивалентность