Liquid Chromatography–Electrospray Ionization–Mass Spectrometry Method in Multiple Reaction Monitoring Mode To Determine 17α-Ethynylestradiol Residues in Cattle Hair without Previous Digestion

Abstract

A liquid chromatography-mass spectrometric (LC-MS/MS) method has been developed for determination of ethynylestradiol residues in cattle hair. Hair samples were pulverized with a cryogenic mill followed by a simple extraction with acetonitrile. A dansyl derivatization procedure to improve ethynylestradiol detection was used before the LC-MS/MS analysis in multiple reaction monitoring (MRM) mode using alpha-estradiol as an internal standard. The method was validated following the latest EU guidelines using blank hair samples spiked at 2 ng g(-1). The detection capability (CCbeta) was less than 2 ng g(-1), and the decision limit (CCalpha) was 1 ng g(-1). Incurred samples obtained 56 days after cow treatment with ethynylestradiol were analyzed, and the presence of ethynylestradiol in the hair was confirmed in all cases.

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

“…Deproteinization and extraction with acetonitrile, without later steps, permits the HPLC/MS-MS analysis of fluoroquinolones [38,39] or steroids [40] and this procedure has been used in our laboratory for some years.…”

“…The product ion spectra of protonated steroid-oximes were evaluated in each case. The compound-dependent parameters were optimized for every hormone by means of infusion into the source, leading to declustering potential, entrance potential, collision cell entrance potential, collision cell exit potential and collision energy shown in Table 1, collision cell exit potential (CXP) was 4 Dansyl chloride derivatization of the phenolic group has been the most frequently used derivatization procedure for enhancing sensitivity in clinical, environmental or residue analysis of estrogens [37,40]. Quantification using m/z 171 (loss of the dansyl group) is not specific for estrogens and can be problematic during analysis [42].…”

Natural Hormones in Food-Producing Animals:Legal Measurements and Analytical Implications

“…As hormonal esters do not naturally occur in the animal organism, the detection of these synthetic substances in the body of an animal provides irrefutable evidence of the abuse of these promoters. Although the administration of esters of natural hormones can be detected through hair analysis (Duffy et al, 2008;Grataco s-Cubarsi et al, 2006;Pedreira et al, 2007;Rambaud et al, 2005), it has been very difficult to detect intact steroid esters in body fluids or tissues. It is likely that esters quickly hydrolyze in the body of the animal, releasing the corresponding natural hormone (Stolker et al, 2009).…”

Analysis of 17β‐estradiol, estriol and estrone in American eel (Anguilla rostrata) tissue samples using liquid chromatography coupled to electrospray differential ion mobility tandem mass spectrometry

“…The detection and quantification of estrogenic hormones has been achieved using a variety of methods including immunoassays (ELISA), [9,10] gas chromatography (GC) [11][12][13][14][15] and liquid chromatography (LC) coupled to mass spectrometry (MS). [16][17][18][19][20][21][22][23][24][25][26] GC/MS generally requires chemical derivatization of the estrogens prior to analysis, while LC/MS allows the direct analysis of the estrogens without derivatization. When using LC/MS, both atmospheric pressure chemical ionization (APCI) [22,27] and electrospray ionization (ESI) sources have been used.…”