The measurement of urinary free cortisol (UFC) is used in the investigation of Cushing’s Syndrome, a hormonal disorder caused by prolonged exposure to high levels of this hormone. Only a small percentage of circulating cortisol is biologically active (free), with the majority of cortisol inactive (protein bound). UFC represents excretion of the circulating free cortisol. The manual procedure is time consuming and repetitive and consequently has significant potential for human error. This application note presents a fully automated method for UFC featuring automated internal standard addition, SPE clean-up and extraction. Sample preparation is coupled directly to LC-MS/MS. The system is capable of running 98 samples in 12 hours and 21 minutes. Sample preparation time is under 5 minutes fitting within the LC runtime ensuring highest possible throughput.
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Automating the Extraction and Analysis of Urinary Free Cortisol Using ITSP, GERSTEL MPS and Agilent LC-MS/MS