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Development and Analytical Validation Method of Simultaneous Quantification of Estradiol and Progesterone for LC-MS/MS in Serum/Bovine Blood Plasma

Grant number: 10/15167-0
Support Opportunities:Scholarships in Brazil - Post-Doctorate
Effective date (Start): December 01, 2010
Effective date (End): November 30, 2013
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal Investigator:Guilherme de Paula Nogueira
Grantee:Rosineide Costa Simas
Host Institution: Faculdade de Odontologia (FOA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil


Progesterone and estradiol quantitation in bovine serum is of fundamental importance for studies in the area ofbovine reproductive physiology and for the development of strategies reproductive management in herds. Thequantitation of both hormones is usually performed by the use of immunoassay commercial kits, some of themnot specifically indicated to be used for bovine serum as a biological matrix, which is considered to be aquestionable practice regarding analytical procedures. The present proposal involves a multidisciplinary approachfor the development, validation and application of an optimal analytical methodology for simultaneousquantitation of progesterone and estradiol in bovine serum. The analytical platform will be based on liquidchromatrography and tandem mass spectrometry (LC-MS/MS), which will be performed at the ThoMSon MassSpectrometry Laboratory, at the Institute of Chemistry in the University of Campinas-SP Brazil. The LC-MS/MSmethodology will be applied in two biological models: (i) progesterone and estradiol quantification pubertyevaluation in heifers submitted to protected fat after weaning, and (ii) comparison of progesterone and estradiollevels during the estrous cycle of Holstein cows of high and low milk production. The LC-MS/MS technique willintroduce optimal analytical confidence of the quantitation results of progesterone and estradiol in bovine serum,introducing in the veterinary reproductive medicine a modern, valuable and high-throughput analytical techniquefor steroid hormones quantitation.

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MARTINS-JUNIOR, HELIO A.; PINAFFI, FABIO L. V.; SIMAS, ROSINEIDE C.; TAROUCO, ADRIANA K.; FERREIRA, CHRISTINA R.; SILVA, LUCIANO A.; NOGUEIRA, GUILHERME P.; MEIRELLES, FLAVIO V.; EBERLIN, MARCOS N.; PERECIN, FELIPE. Plasma Steroid Dynamics in Late- and Near-term Naturally and Artificially Conceived Bovine Pregnancies as Elucidated by Multihormone High-resolution LC-MS/MS. Endocrinology, v. 155, n. 12, p. 5011-5023, . (10/05806-5, 10/15167-0, 10/51677-2)
MARTINS-JUNIOR, HELIO A.; SIMAS, ROSINEIDE C.; BROLIO, MARINA P.; FERREIRA, CHRISTINA R.; PERECIN, FELIPE; NOGUEIRA, GUILHERME DE P.; MIGLINO, MARIA A.; MARTINS, DANIELE S.; EBERLIN, MARCOS N.; AMBROSIO, CARLOS E.. Profiles of Steroid Hormones in Canine X-Linked Muscular Dystrophy via Stable Isotope Dilution LC-MS/MS. PLoS One, v. 10, n. 5, . (10/51677-2, 07/51222-2, 10/15167-0)
GOBBO FERRARI, SARITA P.; BONASSA, KARINA P. D.; COELHO, MIRELA B.; FERREIRA, CHRISTINA R.; DA COSTA, HENI FALCAO; JARA, JOSE LUIS P.; MIGUEL, MARIA CAROLINA V.; REYES, FELIX GUILLERMO R.; EBERLIN, MARCOS N.; NOGUEIRA, GUILHERME P.; et al. High precision and selectivity for quantitation of enrofloxacin and ciprofloxacin in five chicken tissues using solid phase extraction and ESI LC-MS/MS for application in monitoring residues. ANALYTICAL METHODS, v. 7, n. 7, p. 3291-3297, . (10/51677-2, 10/15167-0)

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