Urinary steroid hormone profiles for wild greater spear-nosed bats

The 117 urine profile samples came from 59 different females and 44 males. Four females and seven males were sampled more than once. Steroid hormones and their metabolites were measured from dried urine by ZRT labs using gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) following (Xu et al., 2017). Briefly, samples were extracted from filter paper strips (1/3 of a 903^TM sample collection card supplied by ZRT labs and made by Eastern Business Forms, Greenville, SC) using both water and methanol and cleaned using C18-solid phase extraction cartridges. The eluents were dried down and enzymatically hydrolyzed. The liquid extracts of the hydrolysis mixture were also dried down and derivatized to trimethylsilyl ethers using MSTFA:NH4I:ethanethiol. A 2 µl volume of product was injected into an Agilent 7890 Triple Quadropole Mass Spectrometer and results were analyzed using MassHunter software, v.12. We corrected for potential differences in hormone levels caused by variation in the water content of urine by measuring and then adjusting for specific gravity (SG).