Development and validation of a liquid chromatography coupled to mass spectrometer (LC-MS) method for the simultaneous quantification of estrone-3-sulfate, progesterone, estrone and estradiol in serum of mares and American bisons
Development and validation of a liquid chromatography coupled to mass spectrometer (LC-MS) method for the simultaneous quantification of estrone-3-sulfate, progesterone, estrone and estradiol in serum of mares and American bisons.pdf
[en] Steroid concentrations in serum are fluctuating during pregnancy of many mammal species. The current
knowledge about endocrinology of gestation is mainly based on immunoassays. However, the lack of specificity
of these assays hampers the reliability of the results. In the present work, we developed and validated a methodology associating liquid chromatography (LC) and mass spectrometry (MS) to simultaneously quantify, with
high specificity and accuracy, estrone-3-sulfate (E3S), progesterone (PRO), estrone (E1) and estradiol (E2) in
serum of two different mammal species.
The sample preparation procedure is based on a simple protein precipitation and a derivatization with dansyl
chloride. After the chromatographical separation, compounds were analyzed with a triple-quadrupole mass
spectrometer operating in multiple reaction monitoring. Mare and American bison serum samples were analyzed
with the validated method and results were compared with concentrations measured with commercial radioimmunoassay (RIA), enzyme linked immunosorbent assay (ELISA) and chemiluminescent microparticle immunoassay (CMIA).
Following these criterions: relative standard deviation <15% and relative bias <15%, lower limits of quantification of 0.5 ng/mL (E3S), 0.1 ng/mL (PRO) and 2 pg/mL (E1 and E2) were achieved. Most of the comparison
between immunoassays and LC-MS showed poor correlation and proportional differences.
Our LC-MS method is able to simultaneously quantify several steroid hormones with high specificity, accuracy
and sensitivity in serum of two different mammal species. Our method constitutes a useful and performant tool
for veterinary clinicians and LC-MS should thus be used to update and refine the current knowledge about the
endocrinology of pregnancy in mammals.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Dufour, Patrice ; Centre Hospitalier Universitaire de Liège - CHU > Unilab > Pool assistant - biologie clinique
SEYNAEVE, Ysaline ; Centre Hospitalier Universitaire de Liège - CHU > Unilab > Pool assistant - biologie clinique
PEETERS, Stéphanie ; Centre Hospitalier Universitaire de Liège - CHU > Unilab > Laboratoire techniques séparatives et stress oxydant
LE GOFF, Caroline ; Centre Hospitalier Universitaire de Liège - CHU > Unilab > Laboratoire techniques séparatives et stress oxydant
CAVALIER, Etienne ; Centre Hospitalier Universitaire de Liège - CHU > Unilab > Service de chimie clinique
Ponthier, Jérôme ; Université de Liège - ULiège > Dép. clinique des animaux de compagnie et des équidés (DCA) > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Language :
English
Title :
Development and validation of a liquid chromatography coupled to mass spectrometer (LC-MS) method for the simultaneous quantification of estrone-3-sulfate, progesterone, estrone and estradiol in serum of mares and American bisons
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