Neil Loftus (Presenter)
SHIMADZU MSO
Authorship: Neil Loftus (1), Stéphane Moreau (2), Mikaël Levi (3), Anja Grüning (2)
(1) SHIMADZU MSO, Manchester, UK, (2) SHIMADZU Europa, Duisburg, Germany, (3) SHIMADZU France, Noisiel, France
| Short Abstract Steroid hormones play an important role as modulators of the autoimmune disease onset/perpetuation. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is considered to be the method of choice for quantification of steroids as a consequence of the selective detection, precision and high sensitivity. To enhance the scope of steroid research a UHPLC-MS/MS was developed to support higher sample throughput and extended steroid panels. |
Long Abstract
Steroid hormones play a crucial role in controlling metabolism, inflammation and immune functions. Changes in steroid profiling reflect disease status and help research into a number of disorders including congenital adrenal hyperplasia (CAH), Cushing’s disease and polycystic ovarian disease. To overcome many limitations of immunoassays, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has the potential to find its place in the clinical laboratory medicine for quantification of steroid hormones to profile steroids in a disease process. To enhance steroid profiling a highly sensitive analysis method for simultaneous determination of multiple steroids by UHPLC-MS/MS was developed to support clinical research.
A method was developed using an autosampler capable of overlapping sample injections to deliver a higher sample throughput with an extended steroid panel. The Shimadzu Nexera MX multiplex LCMS system has a Dual Stream Technology (MX-DST) using two separate analysis systems to perform overlapping sample injections. This design supports either an injection cycle of two batches of samples at the same time or to inject one batch of samples using the two separate streams of the Nexera MX system. This study also considers the influence on the results when using the two separate streams for one batch compared to the use of only one stream for one batch.
A synthetic surrogate serum (SigMatrix) was spiked with the steroids panel to prepare calibration and quality control samples. After addition of the respective internal standards and dilution with water sample preparation using supported liquid extraction (SLE) was performed. The extracts were evaporated to dryness and reconstituted in water/methanol. Analysis was performed using a binary gradient on the Nexera MX multiplex UHPLC system equipped with an LCMS-8060 triple quadrupole mass spectrometry detector. Multi reaction monitoring (MRM) was used for the MS analysis.
The method was successfully applied for the quantification of an extended steroid panel in biological fluids. This approach resulted in a highly sensitive and selective detection with a higher sample throughput using Nexera MX multiplexing system.
References & Acknowledgements:
| Description | Y/N | Source |
| Grants | no | |
| Salary | yes | SHIMADZU |
| Board Member | no | |
| Stock | no | |
| Expenses | no |
IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | yes |