Comparative steroid profiling in large animals

Ruth Morgan; Scott Denham; Patricia Lee; Natalie Homer

doi:10.1530/endoabs.77.P8

Large animals are increasingly used a models for endocrine research. Unlike rodents they are cortisol-dominant and thus more similar to humans. In this study we use two novel analytical methods to compare the steroid profile of pigs, sheep and humans; 1. Simultaneous quantification of primary glucocorticoids, glucocorticoid precursors and mineralocorticoids in plasma (>20 steroids); 2.Quantification of glucocorticoids metabolites (>10 steroids) in plasma. Steroids were extracted from plasma (100μL) from healthy adult female pigs(n = 12), sheep(n = 12) and humans(n = 6) by automated supported liquid extraction and analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). We have shown that components of the glucocorticoid/mineralocorticoid synthesis pathways can be simultaneously quantified and that glucocortocoid metabolites, normally only measured in urine, are present at high concentrations in plasma. Application of these methods to clinical samples could offer valuable insight into endocrine disease pathophysiology. We demonstrated that pigs and sheep have similar glucocorticoid/mineralocorticoid profiles to humans but subtle differences, particularly in glucocorticoid metabolism, should be considered when using these animals in disease models.

**Table 1** Plasma concentrations of primary glucocorticoids cortisol and corticosterone (B), glucocorticoid precursor 17-hydroxyprogesterone (17OH-P), glucocorticoid metabolites cortisone (E), 11-dehydrocorticosterone (A), 5α-tetrahydrocortisol (5α-THF), 5β-tetrahydrocortisol (5β-THF) and 20β-dihydrocortisol (20β-DHF) and mineralocorticoids 11-deoxycorticosterone (DOC) and aldosterone.
	Human	Pig	Sheep
Primary Glucocorticoids	Cortisol 210.2 ± 37.86^a	Cortisol 167.8 ± 15.41^a	Cortisol 141.8 ± 10.29^b
	B 6.65 ± 1.71	B 5.34 ± 0.57	B 4.01 ± 0.39
Glucocorticoid precursors	17OH-P 1.08 ± 0.23^a	17OH-P 0.36 ± 0.10^b	17OH-P 0.13 ± 0.34^b
Glucocorticoid Metabolites	E 34.54 ± 2.70^a	E 26.19 ± 2.46^b	E 14.36 ± 0.6419^c
	A 1.64 ± 0.23	A 1.11 ± 0.11	A 1.09 ± 0.12
	5α-THF 10.02 ± 1.49^a	5α-THF 0.64 ± 0.05^b	5α-THF 1.72 ± 0.60^b
	5β-THF 61.35 ± 19.49^a	5β-THF 11.70 ± 1.45^b	5β-THF 70.03 ± 16.34^a
	20β-DHF 3.57 ± 0.56^a	20β-DHF 2.34 ± 0.28^a	20β-DHF 11.73 ± 0.94^b
Mineralocorticoids	DOC 0.07 ± 0.02	DOC 0.08 ± 0.02	DOC 0.18 ± 0.04
	Aldosterone 0.09 ± 0.01	Aldosterone 0.14 ± 0.06	Aldosterone 0.10 ± 0.01

**Table 1** Plasma concentrations of primary glucocorticoids cortisol and corticosterone (B), glucocorticoid precursor 17-hydroxyprogesterone (17OH-P), glucocorticoid metabolites cortisone (E), 11-dehydrocorticosterone (A), 5α-tetrahydrocortisol (5α-THF), 5β-tetrahydrocortisol (5β-THF) and 20β-dihydrocortisol (20β-DHF) and mineralocorticoids 11-deoxycorticosterone (DOC) and aldosterone.
	Human	Pig	Sheep
Primary Glucocorticoids	Cortisol 210.2 ± 37.86^a	Cortisol 167.8 ± 15.41^a	Cortisol 141.8 ± 10.29^b
	B 6.65 ± 1.71	B 5.34 ± 0.57	B 4.01 ± 0.39
Glucocorticoid precursors	17OH-P 1.08 ± 0.23^a	17OH-P 0.36 ± 0.10^b	17OH-P 0.13 ± 0.34^b
Glucocorticoid Metabolites	E 34.54 ± 2.70^a	E 26.19 ± 2.46^b	E 14.36 ± 0.6419^c
	A 1.64 ± 0.23	A 1.11 ± 0.11	A 1.09 ± 0.12
	5α-THF 10.02 ± 1.49^a	5α-THF 0.64 ± 0.05^b	5α-THF 1.72 ± 0.60^b
	5β-THF 61.35 ± 19.49^a	5β-THF 11.70 ± 1.45^b	5β-THF 70.03 ± 16.34^a
	20β-DHF 3.57 ± 0.56^a	20β-DHF 2.34 ± 0.28^a	20β-DHF 11.73 ± 0.94^b
Mineralocorticoids	DOC 0.07 ± 0.02	DOC 0.08 ± 0.02	DOC 0.18 ± 0.04
	Aldosterone 0.09 ± 0.01	Aldosterone 0.14 ± 0.06	Aldosterone 0.10 ± 0.01

Endocrine Abstracts

P8