Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Automated Supported Liquid Extraction for the analysis of a panel of 12 endogenous steroids in human plasma by LC-MS/MS

Version 1 : Received: 19 November 2020 / Approved: 21 November 2020 / Online: 21 November 2020 (08:36:10 CET)

How to cite: Denham, S.G.; Just, G.; Kyle, C.J.; Richardson, J.; Lee, P.; Simpson, J.P.; Gifford, R.M.; Andrew, R.; Reynolds, R.M.; Homer, N.Z. Automated Supported Liquid Extraction for the analysis of a panel of 12 endogenous steroids in human plasma by LC-MS/MS. Preprints 2020, 2020110551 (doi: 10.20944/preprints202011.0551.v1). Denham, S.G.; Just, G.; Kyle, C.J.; Richardson, J.; Lee, P.; Simpson, J.P.; Gifford, R.M.; Andrew, R.; Reynolds, R.M.; Homer, N.Z. Automated Supported Liquid Extraction for the analysis of a panel of 12 endogenous steroids in human plasma by LC-MS/MS. Preprints 2020, 2020110551 (doi: 10.20944/preprints202011.0551.v1).

Abstract

Steroid analysis is important in the clinical assessment of endocrine function in health and disease. Although tandem mass spectrometry methods coupled with chromatographic separation are considered the gold standard analytical technique in this setting, enabling profiling of multiple steroids in a single sample, sample processing can be labour-intensive. Here we present a simple, efficient automated 96-well Supported Liquid Extraction method with dichloromethane/isopropanol as organic solvent, carried out on a Extrahera automated sample handler (Biotage), which completes sample preparation of 80 plasma samples (200µL) in 90 minutes. Compounds were separated on a Kinetex C18 column (150x3mm;2.6um) using a mobile phase of methanol and water (0.1% formic acid). The run time was 16 minutes on a Nexera uHPLC system (Shimadzu) with a QTrap 6500+ linear ion trap mass spectrometer (AB Sciex). Precisions ranged 8.1 to 18.1% RSD, bias -10.1-5.8%, and extraction recoveries 73.5-111.9%. LOQs ranged between 0.025–0.500 ng/mL.

Subject Areas

steroids; steroid panel; clinical mass spectrometry; plasma; sample automation; endocrine; Synacthen

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