Skip to main content

Be a Marine Mammal Hero Year-Round

It’s pupping season and that means pups will be pouring through our hospital doors in need of meals and meds. But did you know you can help not only during pupping season, but year-round?

Yes, as a monthly donor, you will help provide fish meals, meds and care for a marine mammal like Magoo so they can regain their strength and return to their ocean home. Then the next month, you will help another patient. And the next month? You will make it possible again!

Plus, new monthly gifts will be matched up to $1,000 when you sign up by Sunday, February 23.

Be a hero year-round
harbor seal, magoo

Analyzing Stress and Reproductive Steroid Hormones in Gray and Fin Whales

Small Blubber Samples (50 mg) Sufficient for Analyses of 10 Stress and Reproductive Steroid Hormones in Gray and Fin Whales via Liquid Chromatography Mass Spectrometry

Abstract

Information on stress, reproductive fitness, and health is difficult to obtain in wild cetaceans but critical for conservation and management. The goal of this study was to develop a methodology requiring minimal blubber mass for analysis of reproductive and stress steroid hormones and, hence, suitable for cetacean biopsies. Blubber biopsies and samples were collected from free-ranging and stranded gray and fin whales. Steroid hormones were extracted from blubber samples as small as 50 mg using liquid-liquid extraction methodology developed to handle the high fat content of blubber. Samples were analyzed via liquid chromatography with tandem mass spectrometry for 10 hormones: aldosterone, androstenedione, cortisol, cortisone, corticosterone, 17β-estradiol, estrone, 17α-hydroxyprogesterone, progesterone, and testosterone. As part of the optimization, homogenization via bead beating and blade dispersion were compared, and the former found superior. To investigate optimal yet minimal tissue mass required, hormone panels were compared among paired 50, 150, and 400 mg samples, the latter two being commonly reported masses for hormone blubber analysis. Results indicated that 50 mg of blubber was suitable and sometimes superior. Additionally, significant differences in precision values were observed between species, possibly stemming from differences in blubber composition, and relevant to homogenization technique selection and calibration methods that use blubber matrix matches obtained from a species other than the study species. Based on recovery and precision values, our methodology was accurate and precise in the measurement of spiked known quantities for all 10 hormones, confirming the methodology capabilities in 50 mg blubber mass in both species. Altogether, and in our specific sample sets, all endogenous hormones, except corticosterone, were identified above the detection limit in 50 mg gray whale blubber samples while all endogenous hormones, except aldosterone, cortisone, estrone, and progesterone, were detected in 50 mg fin whale blubber samples. We present a robust methodology for the analysis of multiple reproductive and stress steroid hormones in minimal masses of cetacean blubber compatible with small biopsies. Finally, we identified statistically significant differences in corticosteroid concentrations between stranded and free ranging animals. 


Wittmaack, C., Urbán Ramírez, J., Bernot-Simon, D., Martínez-Aguilar, S., Subbiah, S., Surles, J. G., Looney, M., Kumar, N., Halaska, B., Duignan, P. J., Knauss, M., Burns, K., & Godard-Codding, C. A. J., 2022. Small Blubber Samples (50 mg) Sufficient for Analyses of 10 Stress and Reproductive Steroid Hormones in Gray and Fin Whales via Liquid Chromatography Mass Spectrometry. Frontiers in Marine Science, 8(808764).

Pádraig Duignan
Barbie Halaska

Meet The Experts

Related Publications

Recent News