Quantitative analysis of underivatized 17 β-estradiol using a high-throughput LC-MS/MS assay – application to support a pharmacokinetic study in ovariectomized guinea pigs

Publication date: Available online 28 September 2019Source: Journal of Pharmaceutical and Biomedical AnalysisAuthor(s): Adarsh Gandhi, Murali K. Matta, Sharron Stewart, Ashok Chockalingam, Alan Knapton, Rodney Rouse, Wendy Wu, Vikram PatelAbstractDifference in female sex hormone, β-estradiol (E2), levels can contribute to sex differences in biological processes that underlie target tissue functions (QT interval), vulnerability to diseases (hepatitis or HIV), and response toward therapies. Accurate quantification of plasma E2 level is thus an important aspect in both basic science research examining hormone-regulated physiological mechanisms and in clinical settings to support patient care associated with altered E2 levels. Due to lack of a high-throughput high-sensitivity analytical method, we developed and validated a LC-MS/MS assay for accurate low-level quantification of E2 and demonstrated its application to a guinea pig pharmacokinetic study in which guinea pigs were treated with 10 or 40 µg/kg E2 subcutaneously and blood samples collected at 0 (pre-dose), 0.25, 0.5, 1, 2, 4, 8, 12 and 24 h post-dosing. E2 was extracted using 90 µL ovariectomized guinea pig plasma by liquid-liquid extraction. The method was robust, sensitive with linear range from 3.9-1000 pg/mL, and the assay met acceptance criteria for validation parameters listed in the current FDA Guidance on Bioanalytical Method Validation. Compared to the 10 µg/kg dose, more than dose proportional inc...
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research