Quantitative LC–MS/MS method for the sensitive and simultaneous determination of natural hormones in bovine serum

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2009-08, Vol.877 (24), p.2457-2464
• Main Authors: Regal, P., Vázquez, B.I., Franco, C.M., Cepeda, A., Fente, C.
• Format: Article
• Language: English
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Bovine serum; Cattle; Chromatography, Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; Gonadal Steroid Hormones - blood; Hormone; LC–MS/MS; Medical sciences; Pharmacology. Drug treatments; Sensitivity and Specificity; Tandem Mass Spectrometry - methods
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2009.06.025

We have developed a liquid chromatography tandem mass spectrometric (LC–MS/MS) method for the simultaneous quantitative analysis of several free forms of steroid hormones in bovine serum [pregnenolone (P 5), progesterone (P 4), 17hydroxyP 5, 17hydroxyP 4, testosterone (T), dehydroepiandrosterone (DHEA), androstenedione (A), estrone (E 1), 2, 4 and 16 hydroxyE 1, 2 and 4 methoxyE 1]. Deuterated analogs were used as internal standards. Serum proteins were eliminated with acetonitrile. Oxime derivatives of steroids were extracted with tert-butylmethylether and analyzed in positive MRM mode. Methodology was validated in accordance with the European Commission Decision 2002/657/EC. Performance characteristics permit the use of this methodology for steroid determination in animal serum samples.