Cortisol, Free, Equilibrium Dialysis and LC/MS-MS

CPT: 82530
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Expected Turnaround Time

6 - 10 days


Specimen Requirements


Specimen

Serum, frozen


Volume

1.6 mL


Minimum Volume

0.8 mL (Note: This volume does not allow for repeat testing.)


Container

Red-top tube or gel-barrier tube


Collection

Separate within two hours of venipuncture. Transfer specimen to a plastic transport tube before freezing.


Storage Instructions

Freeze. Stable at room temperature or refrigerated for seven days. Stable frozen for six months. Freeze/thaw cycles stable x6.


Causes for Rejection

Hemolysis; icteric or lipemic sample


Test Details


Use

Cortisol is the primary glucocorticoid in humans, affecting glucose control, inflammation, and response to stress.


Limitations

Results of this test are for investigational purposes only. The performance characteristics of this assay have been determined by LabCorp. The result should not be used as a diagnostic procedure without confirmation of the diagnosis by another medically established diagnostic product or procedure.


Methodology

Equilibrium dialysis and high-pressure liquid chromatography (HPLC) with tandem mass spectrometry.


Additional Information

Cortisol stimulates gluconeogenesis in the liver and reduces insulin secretion while increasing glucagon release by the pancreas. This increases blood glucose levels. Cortisol is also involved in inhibiting inflammatory responses and maintaining blood pressure by potentiating effects on norepinephrine.

Most of circulating cortisol is bound to protein, primarily transcortin (corticosteroid-binding globulin [CBG]) and albumin. The free hormone hypothesis suggests that the unbound, or free, cortisol is the active fraction, and that this fraction is the most important clinically. Total serum cortisol may be an adequate measure of cortisol activity except when the levels of the binding proteins are abnormal such as in liver disease or acute illness.

A study published in the New England Journal of Medicine demonstrates the utility of measurements of free cortisol in critically ill patients. Patients with critical illness increase cortisol secretion; however, this is best observed when free cortisol levels are measured. In the study, 40% of patients with hypoproteinemia had low levels of total cortisol even though their adrenal function was adequate as demonstrated by robust response to ACTH. Similar results were obtained when salivary cortisol was used as a marker for adrenal sufficiency during illness.

A number of tests to determine free cortisol have been devised. The free fraction depends on the concentrations of the binding proteins and cortisol, and, thus, may be calculated based on these factors. Free cortisol is best measured by equilibrium dialysis. Structure-function observations favor a direct measure of free cortisol. There are polymorphic forms of transcortin that affect cortisol binding, and glycosylation affects cortisol binding to transcortin. The Cortisol, Free, Equilibrium Dialysis and LC/MS-MS assay provides a specific direct test.


References

Arafah BM, Nishiyama FJ, Tlaygeh H, Hejal RJ. Measurement of salivary cortisol concentration in the assessment of adrenal function in critically ill subjects: A surrogate marker of the circulating free cortisol. Clin Endocrinol Metab. 2007 Aug; 92(8):2965-2971. 17535998
Avvakumov GV, Warmels-Rodenhiser S, Hammond GL. Glycosylation of human corticosteroid-binding globulin at asparagine 238 is necessary for steroid binding. J Biol Chem. 1993 Jan 15; 268(2):862-866. 8419363
Bartanusz V, Corneille MG, Sordo S, Gildea M, Michalek JE, Nair PV, Stewart RM, Jezova DJ. Diurnal salivary cortisol measurement in the neurosurgical-surgical intensive care unit in critically ill acute trauma patients. Clin Neurosci. 2014 Jul 22; .pii:S0967-5868(14)00309-9. 25065844
Bendel S, Karlsson S, Pettil V, Loisa P, Varpula M, Ruokonen E. Free cortisol in sepsis and septic shock. Finnsepsis Study Group. Anesth Analg. 2008 Jun; 106(6):1813-1819. 18499615
Hamrahian AH, Oseni TS, Arafah BM. Measurements of serum free cortisol in critically ill patients. N Engl J Med. 2004 Apr 15; 350(16):1629-1638. 15084695
Hill LA, Vassiliadi DA, Simard M, Pavlaki A, Perogamvros I, Hadjidakis D, Hammond GL. Two different corticosteroid-binding globulin variants that lack cortisol-binding activity in a Greek woman. J Clin Endocrinol Metab. 2012 Nov; 97(11):4260-4267. 22948765
Lentjes EG, M, Romijn F, Maassen RJ, de Graaf I, Gautier P, Moolenaar AJ. Free cortisol in serum assayed by temperature-controlled ultrafiltration before fluorescence polarization immunoassay. Clin Chem. 1993 Dec; 39(12):2518-2521. 8252724
le Roux CW, Sivakumaran S, Alaghband-Zadeh J, Dhillo W, Kong WM, Wheeler M. Free cortisol index as a surrogate marker for serum free cortisol. Ann Clin Biochem. 2002; 39(Pt 4):406-408. 12117446
Lin HY, Underhill C, Lei JH, Helander-Claesson A, Lee HY, Gardill BR, Muller YA, Wang H, Hammond GL. High frequency of SERPINA6 polymorphisms that reduce plasma corticosteroid-binding globulin activity in Chinese subjects. J Clin Endocrinol Metab. 2012 Apr; 97(4): E678-686. 22337907

LOINC® Map

Order Code Order Code Name Order Loinc Result Code Result Code Name UofM Result LOINC
504020 Cortisol, Free Dialysis, LCMS 504021 Cortisol, Free Dialysis, LCMS ug/dL 2145-1

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