Quantitative LC-MS/MS method for the sensitive and simultaneous determination of natural hormones in bovine serum.
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.
Regal
P
P
Analytical Chemistry, Nutrition and Bromatology, Veterinary Faculty, University of Santiago de Compostela, 27002 Lugo, Spain.
Vázquez
B I
BI
Franco
C M
CM
Cepeda
A
A
Fente
C
C
D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D012739:Gonadal Steroid Hormones / Q000097:blood*; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D012739:Gonadal Steroid Hormones