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UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction

by 1,2, 1,3, 1, 1 and 1,4,*
1
Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China
2
College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100049, China
3
School of Ocean, Yantai University, Yantai 264005, China
4
Center for Ocean Mega-Science, Chinese Academy of Sciences, Qingdao 266071, China
*
Author to whom correspondence should be addressed.
Academic Editor: Stefano Dall'Acqua
Molecules 2021, 26(20), 6213; https://doi.org/10.3390/molecules26206213
Received: 14 September 2021 / Revised: 3 October 2021 / Accepted: 13 October 2021 / Published: 14 October 2021
The quantification of steroid hormones of individual zebrafish (Danio rerio) provides perspective to understand endogenous hormone function. A UPLC–TOF–MS method was developed to provide a reproducible, sensitive, and efficient assay to determine the concentration of steroid hormones, including cortisol, testosterone, androstenedione, 11-deoxycortisol, 11-deoxycorticosterone, and 17-hydroxyprogesterone in whole-body homogenates of each zebrafish. Solid-phase extraction was used to sample matrix clean-up and acquired a recovery from 89.7% to 107.9%. The analytes were separated on an Aquity BEH C18 column using gradient elution. Mass spectrometric analysis was performed by single reaction monitoring (SRM) using positive electrospray ionization mode. The total running time was 6 min, which was greatly shortened compared with a previously reported method. The developed method exhibited excellent linearity for all the analytes, with regression coefficients higher than 0.99. The limit of detection varied between 0.1 and 0.5 ng/L and the limit of quantification was 0.5–1.7 ng/L for all analytes. The precision of the method was assessed on replicate measurements and was found to be in the ranges of 1.9 % to 6.6% and 4.3% to 8.6%, for intra- and inter-day analysis, respectively. This method was validated according to FDA guidance and applied to determine steroid hormone levels in the tissue homogenate of zebrafish acutely treated with caffeine and ethanol. View Full-Text
Keywords: quantitative analysis; validation; steroid hormone; stressor; behavioral ecology quantitative analysis; validation; steroid hormone; stressor; behavioral ecology
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MDPI and ACS Style

Li, Y.; Yan, Z.; Li, X.; Yin, X.; Li, K. UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction. Molecules 2021, 26, 6213. https://doi.org/10.3390/molecules26206213

AMA Style

Li Y, Yan Z, Li X, Yin X, Li K. UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction. Molecules. 2021; 26(20):6213. https://doi.org/10.3390/molecules26206213

Chicago/Turabian Style

Li, Yaxi, Zhi Yan, Xiaodong Li, Xiuli Yin, and Ke Li. 2021. "UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction" Molecules 26, no. 20: 6213. https://doi.org/10.3390/molecules26206213

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