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LC-MS/MS quantitation of plasma progesterone in cattle

2011; Elsevier BV; Volume: 76; Issue: 7 Linguagem: Inglês

10.1016/j.theriogenology.2011.05.033

1879-3231

Raquel Maria Trindade Fernandes, G.C. Gomes, Andréia M. Porcari, José Rodrigo Valim Pimentel, Patrı́cia Marafon Porciúncula, Helio A. Martins‐Júnior, Patrícia Helena Paiva Miguez, José Luiz Costa, Phellipe Honorio Amaral, Felipe Perecin, Eduardo C. Meurer, Priscila Viau Furtado, Rosineide C. Simas, Marcos N. Eberlin, Christina R. Ferreira, Ed Hoffmann Madureira,

Pharmacological Effects and Assays

Quantitation of progesterone (P(4)) in biological fluids is often performed by radioimmunoassay (RIA), whereas liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) has been used much less often. Due to its autoconfirmatory nature, LC-MS/MS greatly minimizes false positives and interference. Herein we report and compare with RIA an optimized LC-MS/MS method for rapid, efficient, and cost-effective quantitation of P(4) in plasma of cattle with no sample derivatization. The quantitation of plasma P(4) released from three nonbiodegradable, commercial, intravaginal P(4)-releasing devices (IPRD) over 192 h in six ovariectomized cows was compared in a pairwise study as a test case. Both techniques showed similar P(4) kinetics (P > 0.05) whereas results of P(4) quantitation by RIA were consistently higher compared with LC-MS/MS (P < 0.05) due to interference and matrix effects. The LC-MS/MS method was validated according to the recommended analytical standards and displayed P(4) limits of detection (LOD) and quantitation (LOQ) of 0.08 and a 0.25 ng/mL, respectively. The high selective LC-MS/MS method proposed herein for P(4) quantitation eliminates the risks associated with radioactive handling; it also requires no sample derivatization, which is a common requirement for LC-MS/MS quantitation of steroid hormones. Its application to multisteroid assays is also viable, and it is envisaged that it may provide a gold standard technique for hormone quantitation in animal reproductive science studies.