An Optimized Whole-Body Corticosteroid Hormones Quantification Method by LC-MS/MS for Assessing Stress Levels in European glass eels (Anguilla anguilla).

The European eel (Anguilla anguilla) juvenile stage exhibits facultative estuarine migration; the causes of this behavior are yet unknown, but it may have an impact on the population's fate by altering the sex ratio. Recent studies have highlighted potential stress-related issues in glass eels settling in estuaries, however studying stress response in small organisms requires sensitive, accurate and precise analytical methods. The aims of the present study are (i) to develop a whole-body Liquid Chromatography-Mass Spectrometry (LC-MS/MS) method for the simultaneous determination of several stress hormones in low-body mass fish; (ii) to apply this method to glass eels to study their responses to acute stress (iii) to test the effect of anxiolytics (diazepam) on these responses. Our results showed that enhanced LC-MS/MS analysis reduced detection limits and improved accuracy and precision for the quantification at the individual level. Following an acute stress, cortisol concentration significantly increased in glass eels and a 15hours diazepam exposure significantly reduced cortisol levels highlighting a marked anxiolytic effect on this species.

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Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.