Steroid Hormones Analysis in Serum
Steroid hormones are defined as effective biological messengers which take part in crucial body processes even at low concentrations. Thus, dependable and simultaneous analysis of a broad panel of steroid hormones plays vital role for the investigation of the hormone profile, which could be useful in detection endocrine disorders resulting from defects in steroid biosynthesis such as congenital adrenal hyperplasia (CAH). Due to the restrictions and shortcomings of immunoassay techniques for measurement of steroid hormones in body fluids, liquid chromatography tandem mass spectrometry (LC-MS/MS) is increasingly becoming the method for multi-class steroid hormone detection.
Jasem’s approach is offering simultaneously quantification of clinically prominent 16 steroid hormones by LC-MS/MS in 16 minutes.
Features
- Using a simple liquid–liquid-extraction (LLE) protocol for preparation of specimen and no derivatization is needed.
- Detection of multiple target analytes in a single run.
- Measurement of multi-class steroids without sacrificing analysis time.
- Ensure accurate quantitation, high sensitivity and selectivity comparing to immunoassays.Parámetros
17-α-hidroxiprogesterona, aldosterona, testosterona, estradiol, progesterona, cortisol, dihidrotestosterona, androsteno-3,17-diona, corticosterona, 11-desoxicortisol, pregnenolona, 11-desoxicorticosterona, DHEA, DHEAS, 17-α-hidroxipregnenolona, androsterona.
MATRIZ: Suero
Preparación de muestra para suero
- Pipetear 225 μl de muestra de suero en un tubo de centrífuga de vidrio.
- Añadir 25 μl de mezcla etiquetada como estándar interno (IS) y agitar en vórtex durante 5 segundos. Añadir 450 μl de Reactivo-1 en la muestra de suero y adicionalmente agitar durante 15 segundos. Centrifugar a 4000 rpm durante 5 minutos.
- Decantar el sobrenadante transparente en un nuevo tubo de centrífuga de vidrio y lavar el precipitado del tubo con 1,5 ml de Reactivo-2. Después combinar las fases líquidas y agitar durante 3 minutos. Luego centrifugar a 4000 rpm durante 5 minutos.
- La fase superior se evapora bajo corriente de nitrógeno y se disuelve con 100 μl de Reactivo-3. Transferir la solución final al vial de HPLC.
