Simultaneous Determination of Cortisol and Cortisone in Human Sweat | SYED N ALVI

Summary :
⁣A simple reliable ultra-performance liquid chromatography tandem mass spectrometric assay for measurement of cortisol and cortisone in human sweat was developed and validated. Sweat samples 50 μL spiked with cortisol d4 as internal standard was used to measure the levels of cortisol and cortisone simultaneously. Analysis was performed using a reversed phase Atlantis dC18 column and a mobile phase of 2.0 mM of ammonium acetate: acetonitrile (50:50, v:v). The components of interest were detected and quantified in the positive ion mode of electrospray ionization using multiple reaction monitoring transitions set at mass to charge (m/z): 363.11 → 121.00, 361.18 → 163.11, and 367.19 → 121.24 for cortisol, cortisone and IS, respectively. The relationship between cortisol concentration and peak area ratio (cortisol /IS) was linear over the range 0.5 - 50 ng/ml in phosphate buffered saline. Quality control samples were prepared by spiking cortisol and cortisone solution on sweat at concentration 1.5, 25, and 45 ng/ml. The intra- and interday coefficient of variations and bias were within the acceptable limits. The method was successfully used to monitor the levels of cortisol land cortisone levels in sweat samples obtained from healthy volunteers.