The document summarizes various adrenal function tests that can be used to diagnose disorders of the adrenal glands and related hormones. It describes tests such as cortisol levels, ACTH levels and stimulation tests, dexamethasone suppression tests, and tests for aldosterone, electrolytes, and sex hormones. These tests help evaluate adrenal function, detect tumors, diagnose conditions like Cushing's syndrome and Addison's disease, and assess the hypothalamic-pituitary-adrenal axis.

1. ADRENAL FUNCTION TESTS
Suprarenal Glands
By
Dr. Santhosh Kumar N
Associate Professor
Department of Biochemistry

2. Aldosterone:
maintains the
water and
electrolyte balance
& controls BP
Cortisol: stress management,
increases blood sugar &
regulates BP , suppresses
inflammation, regulates
immune response
Androgens & estrogens:
necessary for normal
sexual development

3. Adrenal Medulla
Catacholamines (epinephrine, noreinephrine, Dopamine)
– Triggers the body’s fight or flight response,
– Stimulation of lipolysis,
– Increases rate and force of contraction of the heart, muscle & blood
vessels
– Increased metabolic rate,

4. To diagnose adrenaline disorders, pheochromocytoma
To detect underactive/ overactive adrenal gland
To diagnose cortisol disorder, cushing’s syndrome,
Addison’s disoeders
To diagnosis Aldosterone disorder- conn syndrome
To assess dysfunction of hypothalamus /Pituitary/ Adrenal glands
Dehydro-epiandrosterone sufate (DHEA-S) to determine the causes of
hirsutism, infertility in women and to evaluate puberty in children
Significance
of
AFTs

5. Adrenal Function Tests
• Estimation of blood cortisol levels
• Estimation of blood Adrenocorticotropic hormone (ACTH)
• ACTH stimulation test
• Corticotropin releasing hormone stimulation test
• Dexamethasone suppression test
• Aldosterone
• Electrolytes
• Urea, creatinine and glucose

6. Plasma Level Of Cortisol
• It is determined by RIA, ELISA and Chemiluminiscent immunoassay
(CLIA)
• Levels of cortisol in blood peaks early in morning (at 9am, 10–20 μg/dL)
then declines throughout the day (at 4pm, 3-10 μg/dL), reaching its
lowest levels about midnight (2–5 μg/dL at 10 pm)
• It helps to diagnose cushing’s syndrome or primary or secondary adrenal
insufficiency(addison’s disease)
• Low cortisol levels seen in congenital adrenal hyperplasia

7. Plasma Adrenocorticotropic hormone (ACTH) test
• ACTH is a pituitary hormone that stimulates the adrenal glands to
produce cortisol.
• ACTH levels will be at highest in the morning (at 9am, 10-50 pg/ml)
and usually low at 4pm, (less than 20pg/mL).
• To evaluate and differentiate b/w 1O and 2O adrenal insufficiency

8. • Suppressed ACTH levels –secondary adrenal insufficiency and
due to an adrenal tumor, steroid medication or hypopituitarism
• High ACTH levels – primary adrenal insufficiency (Addison's
disease), Hypoadrenalism &Cushing’s disease

9. ACTH stimulation test
• It measuring the level of cortisol in a patient’s blood before and
after an injection of synthetic ACTH (synacthen/ cosyntropin)
• Diagnosis of adrenal insufficiency

10. Baseline serum cortisol and
ACTH levels measured
Inject 250μg cosyntropin
(ACTH) intravenously or
IM
To check 30 and 60min
cortisol levels
• Rise in cortisol levels to more than
18μg/dl within 60min – normal
• Rise in cortisol to less than 18μg/dl
– abnormal
• An elevated baseline ACTH levels
suggests primary adrenal failure

11. Corticotropin-releasing hormone (CRH) stimulation test
• CRH is the stimulation of the pituitary synthesis of ACTH
• Human CRH is useful test for
i) To differentiating b/w hypothalamic and pituitary causes of adrenal
insufficiency
ii) pituitary ACTH deficiency
iii) to differentiate between pituitary from ectopic source of ACTH in Cushing's
iv) to investigate pseudo-Cushings

12. Patient should fast for at least 4hr prior
to the test
Blood samples should be collected at
15min and 1min before the load
CRH at 1μg/kg bWt is injected IV
Blood samples should be collected at
15,30,45,60,90 and 120min
Blood ACTH and cortisol levels are
measured after administering CRH
• With 1O adrenal insufficiency
(addison’s disease) have high
baseline ACTH with low cortisol
levels before and after CRH
• With 2O adrenal insufficiency
(pituitary) exhibit low baseline
ACTH , cortisol levels are not
affected by CRH
• With 3O (hypothalamic) adrenal
disease exhibit low baseline
ACTH , cortisol levels are do not
reach 20μg/dl

13. Dexamethasone Suppression Test (DST)
• Used to assess adrenal gland functions
• Dexamethasone is a synthetic glucocorticoid that binds to cortisol receptors in
the pituitary and suppresses ACTH release.
• Dexamethasone suppresses the secretion of cortisol due to feedback suppression
of ACTH by the adrenal gland
• Normal - overnight suppression with high dose (2 mg) causes a 50% fall in the
original value
• Adrenal hyperactivity - fail to produce suppression

14. 17-hydroxy-cortisol levels
 17-hydroxycortisol produced in excessive in patients with primary
hyper aldosteronism
 Help to determine whether primary hyper aldosteronism is caused
by adrenal adenoma or by hyperplasia of adrenal tissue .
 Men is 3-10μg/24hrs; women is 2 -6 μg/24hrs

15. Aldosterone levels
• Helps in determining whether the adrenal gland is producing
Aldosterone
• Blood and urine levels are measured to diagnose Addison's
disease (low)

16. • Serum Electrolytes: to monitor the effectiveness of treatment
Low Na+, Cl- and bicarbonate levels and
very high K+ levels is observed in addison’s disease.
• Serum Urea and creatinine: To monitor kidney function
• Blood Glucose levels may be very low during an addisonian crisis
• Renin activity is elevated in primary adrenal insufficiency
because a lack of Aldosterone causes increased renal Na+ losses.
Lowers blood Na+ levels, blood volume and Blood pressure

17. Disease Laboratory findings
Primary testicular
dysfunction
Low testosterone, high LH
Secondary testicular
dysfunction
Hypopituitarism with low testosterone and LH
levels
Delayed puberty Deficiency of pituitary or gonadal hormones
Precocious puberty Premature secretion of gonadotropins
Primary ovarain failure Low estrogen, high LH, FSH
Virilization High androgen, low estradiol, normal LH, FSH
Abnormalities In Sex Hormones

18. LAB TESTS FOR INFERTILITY
TESTS FOR WOMEN
• FSH
• LH
• Prolactin
• Estradiol
• Progesterone
• Anti-Mullerian hormone (AMH)
• Androgens
TESTS FOR MEN
• Semen analysis
• Free and total testosterone
• FSH
• LH
• Prolactin
• Sex hormone binding globulin
(SHBG)

19. THANK YOU
The End