MSACL 2019 EU Abstract

Confident Quantification of Steroids: Analysis in Human Plasma or Serum by LC-MS/MS for Clinical Research

Claudio De Nardi (1), Sergio Indelicato (2), Maura Brambilla (3), Chiara Fania (4)
(1) Thermo Fisher Scientific GmbH, Dreieich, Germany (2) Thermo Fisher Scientific, Les Ulis, France (3) Ospedale di Desio, Desio, Italy (4) Università degli Studi Milano-Bicocca, Milano, Italy

Warning: Undefined variable $headshot in /var/www/html/view_abstract/view_abstract_in_program.php on line 704

Claudio De Nardi (Presenter) Thermo Fisher Scientific >> POSTER (PDF)

Relevant Financial Disclosures (within past 24 months) Honorarium/Expenses Thermo Fisher Scientific Salary Thermo Fisher Scientific Conflict mitigation NOT REQUIRED.

INTRODUCTION
In this report, a robust, reliable LC-MS/MS method for clinical research is developed for the quantification of eight steroids in human plasma or serum. The analytical method includes 11- and 21-deoxycortisol, 17- and 21-hydroxyprogesterone, androstenedione, cortisol, dehydroepiandrosterone sulphate (DHEAS), and testosterone.
OBJECTIVES
Development and implementation of a robust, reliable, sensitive analytical method for quantification of eight steroids in human plasma or serum using the Thermo Scientific™ TSQ Quantis™ triple quadrupole mass spectrometer.
METHODS
Method performance was evaluated using the ClinMass® LC-MS/MS Complete Kit from RECIPE Chemicals + Instruments GmbH (Munich, Germany) in terms of linearity of response within the calibration ranges, carry over, accuracy and intra-assay precision for each analyte. Samples were extracted by protein precipitation followed by LC separation on a Thermo Scientific™ Transcend™ II LX-2 system using mobile phases and analytical column provided by RECIPE. Total runtime was 7.0 minutes. Analytes and internal standards were detected by selected reaction monitoring (SRM) on a TSQ Quantis triple quadrupole mass spectrometer with heated electrospray ionization operated in positive mode. Data were acquired and processed using Thermo Scientific™ TraceFinder™ 4.1 software.
RESULTS
The method proved to be linear in the calibration ranges covered by the calibrators. It was not possible to detect the lowest calibrator for 17-hydroxyprogesterone but the method was linear in the range calibrator 2 – calibrator 6. The maximum registered carry over was 0.08% with the exception of androstenedione which showed a value of 0.36%. The data demonstrated outstanding accuracy of the method with the percentage bias between nominal and average back-calculated concentration for the used control samples ranging between -4.4% and 5.3%. The %CV for intra-assay precision was always below 5.7% for all the analytes.
CONCLUSIONS
A robust, reliable LC-MS/MS method for clinical research for the quantification of eight steroids in human plasma or serum was developed and implemented. The ClinMass LC-MS/MS Complete Kit from RECIPE ensured increased confidence in the result that was obtained. The method was analytically validated on a Transcend II system connected to a TSQ Quantis triple quadrupole mass spectrometer. The sample preparation procedure utilized in this method uses quick and simple offline protein precipitation with concomitant internal standard addition. The described method meets research laboratory requirements in terms of sensitivity, linearity of response, accuracy, and precision.