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Simultaneous quantitation of testosterone, estradiol, ethinyl estradiol, and 11‐ketotestosterone in fathead minnow fish plasma by liquid chromatography/positive atmospheric pressure photoionization tandem mass spectrometry
Author(s) -
Zhang Fagen,
Bartels Michael J.,
Geter David R.,
Carr Melissa S.,
McClymount Lynn E.,
Marino Troy A.,
Klecka Gary M.
Publication year - 2009
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.4298
Subject(s) - chemistry , chromatography , dansyl chloride , mass spectrometry , liquid chromatography–mass spectrometry , testosterone (patch) , atmospheric pressure chemical ionization , tandem mass spectrometry , high performance liquid chromatography , chemical ionization , derivatization , endocrinology , ion , ionization , medicine , organic chemistry
In the present work, for the first time, a rapid and sensitive liquid chromatography/positive atmospheric pressure photoionization tandem mass spectrometry (LC/APPI‐MS/MS) method has been developed and validated for the simultaneous quantitation of testosterone, estradiol, ethinyl estradiol, and 11‐ketotestosterone in fathead minnow fish plasma using no more than 10 µL of plasma. Compounds present in plasma were directly derivatized with dansyl chloride and 25 µL of the derivatized mixture was injected into the LC/APPI‐MS/MS system. The gradient chromatographic elution was achieved on an Agilent Zorbax SB‐C18 analytical column (2.1 mm × 50 mm, 1.8 µm particle size) with mobile phases consisting of acetonitrile, water and acetic acid. The flow rate was 0.5 to 0.7 mL/min and the total run time was 11.5 min. The lower limits of quantitation for testosterone, estradiol, ethinyl estradiol, and 11‐ketotestosterone and were 1, 1, 1, and 2.5 ng/mL, respectively. Intra‐batch precision was less than 19.4% and inter‐batch precision was less than 11.7% for all four analytes. Accuracy was within 83.5–115.4% of nominal concentrations. This method is used for quantitation of sex steroid levels in fathead minnow tested in endocrine disruptor screening experiments. Copyright © 2009 John Wiley & Sons, Ltd.