title: Liquid chromatography-tandem mass spectrometric assay for eltrombopag in 50μL of human plasma: a pharmacokinetic study.
abstract: Eltrombopag is a thrombopoietin receptor agonist, used in the treatment of thrombocytopenia. This paper describes a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay method for the determination of eltrombopag in human plasma samples using eltrombopag 13C4 as internal standard (IS). Analyte and the IS were extracted from 50μL of human plasma using protein precipitation technique with no drying, evaporation and reconstitution steps. The chromatographic separation was achieved on a C18 column by using a mixture of 10mM ammonium formate (pH3) and acetonitrile (10:90, v/v) as the mobile phase at a flow rate of 1.0mL/min. The linearity of the method was established in the concentration range 50.0-10007ng/mL with r(2)≥0.99. The intra-day and inter-day precision and accuracy results in four validation batches across five concentration levels were well within the acceptance limits. The proposed method was found to be applicable to pharmacokinetic studies.
authors.affiliation: Center for Pharmaceutical Sciences, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad 500 085, India; Piramal Clinical Research Laboratories, Ramanthapur, Hyderabad 500 013, India. Electronic address: rambabu1974@gmail.com.
authors.forename: Rambabu
authors.initials: R
authors.lastname: Maddela
authors.affiliation: Piramal Clinical Research Laboratories, Ramanthapur, Hyderabad 500 013, India.
authors.forename: Ramakrishna
authors.initials: R
authors.lastname: Gajula
authors.affiliation: Center for Pharmaceutical Sciences, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad 500 085, India; Piramal Clinical Research Laboratories, Ramanthapur, Hyderabad 500 013, India. Electronic address: nageswarpilli@gmail.com.
authors.forename: Nageswara Rao
authors.initials: NR
authors.lastname: Pilli
authors.affiliation: Department of Pharmaceutical Chemistry, Malla Reddy College of Pharmacy, Maisammaguda, Secunderabad 5000100, India.
authors.forename: Sridhar
authors.initials: S
authors.lastname: Siddiraju
authors.affiliation: Center for Pharmaceutical Sciences, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad 500 085, India.
authors.forename: Srinubabu
authors.initials: S
authors.lastname: Maddela
authors.affiliation: Center for Pharmaceutical Sciences, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad 500 085, India. Electronic address: ajitharaj@hotmail.com.
authors.forename: Ajitha
authors.initials: A
authors.lastname: Makula
mesh_terms: D001565:Benzoates / Q000737:chemistry* / Q000493:pharmacokinetics*; D001681:Biological Assay / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D004355:Drug Stability; D006801:Humans; D006834:Hydrazines / Q000737:chemistry* / Q000493:pharmacokinetics*; D010949:Plasma / Q000737:chemistry*; D011720:Pyrazoles / Q000737:chemistry* / Q000493:pharmacokinetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D001565:Benzoates; D006834:Hydrazines; D011720:Pyrazoles; C520809:eltrombopag
keywords: Eltrombopag; Human plasma; LC–MS/MS; Pharmacokinetics; Protein precipitation (PP)