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Área de trabajo: Clínica.

Título: Método LC-MS/MS para la Determinación de Esteroides en Orina usando columnas SPE y columnas HPLC Accucore. 

Título original: LC-MS/MS Method for the Determination of Steroids from Urine Using SOLA and Core Enhanced Technology Accucore HPLC Column.

Autor: Eilidh MacRitchie, Kim Phipps, Thermo Fisher Scientific, Runcorn, Cheshire, UK.

Abstract

This application note demonstrates the use of the Thermo Scientific SOLA well plates for the extraction of steroids from urine and the subsequent fast separation using a Thermo Scientific Accucore RP-MS column.

 

Introduction

SOLA™ products are a revolutionary new Solid Phase Extraction (SPE) product range. This first in class SPE product range introduces next-generation, innovative technological advancements, giving unparalleled performance characteristics compared to conventional SPE, phospholipid and protein precipitation products. This includes:

• Higher levels of reproducibility

• Higher levels of extract cleanliness

• Reduced solvent requirements

• Increased sensitivity SOLA products have significant advantages for the analyst when processing compounds in complex matrices particularly in high throughput bioanalytical and clinical laboratories where reduced failure rate, higher analysis speed and lower sample/solvent requirements are critical.

 

The increased performance from SOLA products provides higher confidence in analytical results and lowers cost without compromising ease of use or requiring complex method development.

Accucore™ HPLC columns use Core Enhanced Technology to facilitate fast and high efficiency separations. The 2.6 µm diameter particles are not totally porous, but rather have a solid core and a porous outer layer. The optimized phase bonding creates a series of high coverage, robust phases. Accucore RP-MS uses an optimized alkyl chain length for more effective coverage of the silica surface. This coverage results in a significant reduction in secondary interactions and thus highly efficient peaks with very low tailing. The tightly controlled 2.6 µm diameter of Accucore particles results in much lower backpressures than typically seen with sub-2 µm materials.

Hydrocortisone, cortisone, corticosterone and 11-α hydroxyprogesterone belong to the progesterone class of steroid horomones. Progesterones are characterized by the 21-carbon backbone and are primarily used to maintain pregnancy. The extraction of these steroids from urine, their separation on an Accucore RP-MS column and their quantification by LC-MS/MS is demonstrated in this application note.

 

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