Extraction of Urinary Hormone Metabolites using Supported Liquid Extraction prior to HPLC-MS/MS Analysis.

Introduction

Estrogen, androgen, and glucocorticoid metabolism can be used to assess overall hormonal balance during hormone therapy. Urine is the recommended testing matrix for quantification of primary estrogen levels as well as secondary estrogen metabolites when monitoring overall hormone balance, therapy, and detoxification. Non-invasive collection allows for sampling over a 24-hour period, providing insight into a patient’s circadian rhythm. Since many samples are generated for a single patient in any one day, a fast and robust testing protocol is needed for extraction and analysis during clinical testing.

Here, we demonstrate a rapid and reliable sample preparation method using Support Liquid Extraction (SLE+) to extract a suite of 30 hormone analytes from a hydrolyzed urine matrix. Single injection analysis by LC-MS/MS shows that matrix effects are eliminated by the SLE+ protocol and that analyte recovery and sensitivity have excellent clinical utility.