Presenter Bio: I have been with Thermo Fisher Scientific in a sales support role for Clinical Research & Forensic Toxicology Life Science Mass Spectrometry for over three years. My first experience of mass spectrometry lies in an area of biotoxin research during my PhD, which I completed in Cork Institute of Technology, Republic of Ireland. Later I worked at University College Cork as a Post-doctoral researcher for Vitamin D Mass Spectrometry analysis. In my current role, I specialise in clinical and toxicology applications utilising liquid chromatography coupled to mass spectrometry.
Relevant Financial Disclosures
(within past 24 months)
No relevant financial relationship(s) to disclose.
Abstract
Introduction: Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has been widely adopted as an analytically sensitive and selective technique for measuring estrone and estradiol in complex matrices such as human blood plasma or serum. Quantitation of these steroids down to low-picogram per milliliter levels are required by many clinical researchers. We endeavored to achieve this using a triple-stage quadrupole mass spectrometer with the most efficient production, isolation and transmission of precursor ions and fastest selective reaction monitoring.
Objective: For detailed studies, scientists need to quantitate ever lower concentrations of estrone and estradiol in serum samples. Here we demonstrate a method capable of detecting low pg/mL of both estrone and estradiol in human serum for clinical research.
Methods: Samples were prepared by liquid-liquid extraction (LLE). Following chromatographic separation by a reversed-phase high performance liquid chromatographic (HPLC) gradient, analytes were detected on a triple quadrupole mass spectrometer. Precision was determined by analyzing replicate concentrations over three days. Accuracy was determined by analyzing Center for Disease Control (CDC) Hormone Standardization (HoSt) Program Phase 1 samples.
Results: Both estrone and estradiol were able to be quantitated down to 2 pg/mL. Inter-assay precisions of replicate samples across the calibration range were better than 8.4%. Accuracy of estradiol analysis was demonstrated using the CDC HoSt samples with 90% of the samples agreeing within 15% of reference values.
Conclusions: We have demonstrated a sensitive, precise and accurate method for the quantitation of estrone and estradiol in human matrix for clinical research. A detection limit of 2 pg/mL with ion ratio confirmation was achieved for both estrone and estradiol using a TSQ Altis triple quadrupole mass spectrometer. Accuracy of the method was demonstrated by analysis of CDC HoSt program Phase 1 samples.
= Discovery stage.
= Translation stage.
= Clinically available.
