Test Catalog

Test Id : ANST

Androstenedione, Serum

Useful For
Suggests clinical disorders or settings where the test may be helpful

Diagnosis and differential diagnosis of hyperandrogenism, in conjunction with measurements of other sex steroids


Diagnosis of congenital adrenal hyperplasia (CAH), in conjunction with measurement of other androgenic precursors, particularly, 17-alpha-hydroxyprogesterone (OHPG), 17 alpha-hydroxypregnenolone, dehydroepiandrosterone sulfate (DHEA-S), and cortisol


Monitoring CAH treatment, in conjunction with testosterone, OHPG, DHEA-S, and DHEA


Diagnosis of premature adrenarche, in conjunction with measurement of follicle-stimulating hormone and luteinizing hormone as well as other adrenal and gonadal sex-steroids and their precursors

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

For information see Steroid Pathways.

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Method Name
A short description of the method used to perform the test

Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS)

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.


Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

Androstenedione, S

Lists additional common names for a test, as an aid in searching




Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

For information see Steroid Pathways.

Specimen Type
Describes the specimen type validated for testing

Serum Red

Additional Testing Requirements

For diagnosis and differential diagnosis of hyperandrogenism, an initial workup in adults should also include total and bioavailable testosterone (TTBS / Testosterone, Total and Bioavailable, Serum) measurements. Depending on results, this may be supplemented with measurements of sex hormone-binding globulin (SHBG1 / Sex Hormone-Binding Globulin, Serum) and other androgenic steroids (eg, dehydroepiandrosterone sulfate [DHEA-S]).


For diagnosis of congenital adrenal hyperplasia (CAH), the following assays should also be ordered:

-OHPG / 17-Hydroxyprogesterone, Serum

-DHES1 / Dehydroepiandrosterone Sulfate, Serum

-CORT / Cortisol, Serum


For monitoring CAH treatment, the following assays should also be ordered:

-TTST / Testosterone, Total, Mass Spectrometry, Serum

-OHPG / 17-Hydroxyprogesterone, Serum

-DHES1 / Dehydroepiandrosterone Sulfate, Serum

-DHEA_ / Dehydroepiandrosterone [DHEA], Serum.


For diagnosis of premature adrenarche, the following assays should also be ordered:

-FSH / Follicle-Stimulating Hormone [FSH], Serum

-LH / Luteinizing Hormone [LH], Serum

-TTBS / Testosterone, Total and Bioavailable, Serum or TGRP / Testosterone, Total and Free, Serum

-EEST / Estradiol, Serum

-DHES1 / Dehydroepiandrosterone Sulfate, Serum

-DHEA_ / Dehydroepiandrosterone (DHEA), Serum

-SHBG1 / Sex Hormone-Binding Globulin, Serum

-OHPG / 17-Hydroxyprogesterone, Serum

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Collection Container/Tube: Red top (serum gel/SST are not acceptable)

Submission Container/Tube: Plastic vial

Specimen Volume: 0.6 mL

Collection Instructions: Centrifuge and aliquot serum into a plastic vial

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the testing laboratory. The minimum volume is sufficient for one attempt at testing.

0.25 mL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

Gross hemolysis Reject
Gross lipemia Reject
Gross icterus OK

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Serum Red Refrigerated (preferred) 28 days
Frozen 28 days
Ambient 7 days

Useful For
Suggests clinical disorders or settings where the test may be helpful

Diagnosis and differential diagnosis of hyperandrogenism, in conjunction with measurements of other sex steroids


Diagnosis of congenital adrenal hyperplasia (CAH), in conjunction with measurement of other androgenic precursors, particularly, 17-alpha-hydroxyprogesterone (OHPG), 17 alpha-hydroxypregnenolone, dehydroepiandrosterone sulfate (DHEA-S), and cortisol


Monitoring CAH treatment, in conjunction with testosterone, OHPG, DHEA-S, and DHEA


Diagnosis of premature adrenarche, in conjunction with measurement of follicle-stimulating hormone and luteinizing hormone as well as other adrenal and gonadal sex-steroids and their precursors

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

For information see Steroid Pathways.

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

Androstenedione is secreted predominately by the adrenal gland and production is at least partly controlled by adrenocorticotropic hormone (ACTH). It is also produced independent of ACTH in the testes and ovaries from adrenal-secreted dehydroepiandrosterone sulfate (DHEA-S). Androstenedione is a crucial sex-steroid precursor. It lies at the convergence of the 2 biosynthetic pathways that lead from the progestins to the sex steroids, being derived either via:

-C3-dehydrogenation of DHEA

-Catalyzed by 3-beta-hydroxysteroid dehydrogenase-2 (adrenals and gonads)

-17,20-lyase (CYP17A1)-mediated side-chain cleavage of 17-alpha-hydroxyprogesterone (OHPG)


Androstenedione production during life mimics the pattern of other androgen precursors. Fetal serum concentrations increase throughout embryonal development and peak near birth at approximately young adult levels. Levels then fall rapidly during the first year of life to low prepubertal values. With the onset of adrenarche, androstenedione rises gradually, a process that accelerates with the onset of puberty, reaching adult levels around age 18. Adrenarche is a poorly understood phenomenon peculiar to higher primates that is characterized by a gradual rise in adrenal androgen production. It precedes puberty but is not causally linked to it. Early adrenarche is not associated with early puberty, or with any reduction in final height, or overt androgenization, and is generally regarded as a benign condition not requiring intervention. However, girls with early adrenarche may be at increased risk of polycystic ovarian syndrome as adults, and some boys may develop early penile enlargement.


Elevated androstenedione levels can cause symptoms or signs of hyperandrogenism in women. Men are usually asymptomatic but through peripheral conversion of androgens to estrogens, can occasionally experience mild symptoms of estrogen excess, such as gynecomastia.


Most mild-to-moderate elevations in androstenedione are idiopathic. However, pronounced elevations of androstenedione may be indicative of androgen-producing adrenal or gonadal tumors.


In children, adrenal and gonadal tumors are uncommon, but many forms of congenital adrenal hyperplasia can increase serum androstenedione concentrations. Diagnosis always requires measurement of other androgen precursors (eg, OHPG, 17-alpha-hydroxypregnenolone, and DHEA-S) and cortisol, in addition to androstenedione.


For more information see Steroid Pathways.

Reference Values
Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.


Premature infants

26-28 weeks, day 4: 92-282 ng/dL

31-35 weeks, day 4: 80-446 ng/dL

Full-term infants

1-7 days: 20-290 ng/dL

1 month-1 year: <69 ng/dL



Tanner stages

Age (Years)

Reference range (ng/dL)

Stage I (prepubertal)



Stage II



Stage III



Stage IV



Stage V





Tanner stages

Age (Years)

Reference range (ng/dL)

Stage I (prepubertal)



Stage II



Stage III



Stage IV



Stage V



*Soldin SJ, Brugnara C, Wong EC, eds. Androstenedione. In: Pediatric Reference Ranges. 4th ed. AACC Press; 2003: 32-34



Males: 40-150 ng/dL

Females: 30-200 ng/dL


For SI unit Reference Values, see www.mayocliniclabs.com/order-tests/si-unit-conversion.html

Provides information to assist in interpretation of the test results

Elevated androstenedione levels indicate increased adrenal or gonadal androgen production. Mild elevations in adults are usually idiopathic or related to conditions, such as polycystic ovarian syndrome (PCOS) in women or use of androstenedione supplements in men and women. However, levels greater than or equal to 500 ng/dL can suggest the presence of an androgen-secreting adrenal or, less commonly, a gonadal tumor. Androstenedione levels are elevated in more than 90% of patients with benign androgen-producing adrenal tumors, usually well above 500 ng/dL. Most androgen-secreting adrenal carcinomas also exhibit elevated androstenedione levels but more typically show relatively larger elevations in 17-alpha-hydroxyprogesterone (OHPG) and dehydroepiandrosterone sulfate (DHEA-S) than in androstenedione, as they have often lost the ability to produce downstream androgens.


Most androgen-secreting gonadal tumors overproduce androstenedione, often to lesser degrees than adrenal tumors. They also overproduce testosterone. In men and in women with high baseline androgen levels (eg, PCOS), the respective elevations of androstenedione and testosterone may not be high enough to allow unequivocal diagnosis of androgen-producing gonadal tumors. In these cases, an elevation of the usual ratio of testosterone to androstenedione of 1, to a ratio of greater than 1.5, is a strong indicator of neoplastic androgen production.


Diagnosis and differential diagnosis of congenital adrenal hyperplasia (CAH) always requires the measurement of several steroids. Patients with CAH due to 21-hydroxylase gene (CYP21A2) variants, the most common cause of CAH (>90% of cases), usually have very high levels of androstenedione, often 5- to 10-fold elevations. OHPG levels are usually even higher, while cortisol levels are low or undetectable. All 3 analytes should be tested.


In the much less common CYP11A1 variant, androstenedione levels are elevated to a similar extent as in the CYP21A2 variant, and cortisol is also low, but OHPG is only mildly, if at all, elevated.


Also less common, 3-beta hydroxysteroid dehydrogenase (HSD) type 2 deficiency is characterized by low cortisol and substantial elevations in DHEA-S and 17-alpha hydroxypregnenolone, while androstenedione is either low, normal, or, rarely, very mildly elevated (as a consequence of peripheral tissue androstenedione production by 3-beta HSD-1).


In the very rare STAR (steroidogenic acute regulatory protein) deficiency, all steroid hormone levels are low, and cholesterol is elevated.


In the also very rare 17-alpha-hydroxylase deficiency, androstenedione, all other androgen-precursors (17-alpha-hydroxypregnenolone, OHPG, DHEA-S), androgens (testosterone, estrone, estradiol), and cortisol are low, while production of mineral corticoid and their precursors, in particular progesterone, 11-deoxycorticosterone, corticosterone, and 18-hydroxycorticosterone, are increased.


The goal of CAH treatment is normalization of cortisol levels and, ideally, also of sex-steroid levels. Traditionally, OHPG and urinary pregnanetriol or total ketosteroid excretion are measured to guide treatment, but these tests correlate only modestly with androgen levels. Therefore, androstenedione and testosterone should also be measured and used for treatment modifications. Normal prepubertal levels may be difficult to achieve, but if testosterone levels are within the reference range, androstenedione levels up to 100 ng/dL are usually regarded as acceptable.


Girls younger than 7 to 8 years of age and boys younger than 8 to 9 years of age who present with early development of pubic hair or, in boys, penile enlargement, may be suffering from either premature adrenarche or premature puberty, or both. Measurement of DHEA-S, DHEA, and androstenedione, alongside determination of sensitive estradiol, total and bioavailable or free testosterone, sex hormone binding globulin (SHBG), and luteinizing hormone/follicle-stimulating hormone levels will allow correct diagnosis in most cases. In premature adrenarche, only the adrenal androgens, chiefly DHEA-S, and to a lesser degree, androstenedione, will be above prepubertal levels, whereas early puberty will also show a fall in SHBG levels and variable elevations of gonadotropins and gonadal sex-steroids above the prepuberty reference range.


For more information see Steroid Pathways.

Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

Any condition that can result in partial or complete adrenal or gonadal failure may result in low androstenedione levels, diminishing the diagnostic usefulness of the test in these settings.


Androstenedione and, to a lesser degree, dehydroepiandrosterone sulfate (DHEA-S) supplements can result in elevations of serum androstenedione level. With large androstenedione doses of 300 to 400 mg/day, serum androstenedione levels can almost double in some patients. Testosterone levels and, particularly in men, estrone and estradiol levels, may also increase but to a much lesser degree.


Although compared with DHEA-S, less information has been published with regard to the effects of hormones and drugs on androstenedione levels, it is likely that many drugs and hormones can result in changes in androstenedione levels. In particular, agents that induce hepatic enzymes, drugs that affect lipid metabolism, and other steroid hormones are likely to affect androstenedione levels, more commonly resulting in lowered levels. Whether any of these secondary changes are of clinical significance and how they should be related to the established normal reference ranges is unknown. In most cases, the drug-induced changes are not large enough to cause diagnostic confusion.

Supportive Data

To establish pediatric reference ranges, we compared adult levels obtained with our liquid chromatography tandem mass spectrometry methodology with adult levels obtained in other labs with their respective methodologies. We found excellent correlation (R=0.92, regression-trendline slope 1.07) with an extracted radioimmunoassay (RIA) method with preanalytical Sephradex column chromatography. This is the method used in Pediatric Reference Ranges, 4th edition and is based on reference 1. The ranges were further verified by comparison with another pediatric reference range publication, which used the same extracted RIA method with Sephradex column chromatography.(2)

Clinical Reference
Recommendations for in-depth reading of a clinical nature

1. Bidlingmaier F, Wagner-Barnack M, Butenandt O, Knorr D. Plasma estrogens in childhood and puberty under physiologic and pathologic conditions. Pediatr Res. 1973;7(11):901-907. doi:10.1203/00006450-197311000-00006

2. Von Schnakenburg K, Bidlingmaier F, Knorr D. 17-hydroxyprogesterone, androstenedione, and testosterone in normal children and in prepubertal patients with congenital adrenal hyperplasia. Eur J Pediatr. 1980;133(3):259-267

3. Sciarra F, Tosti-Croce C, Toscano V. Androgen-secreting adrenal tumors. Minerva Endocrinol. 1995;20(1):63-68

4. Collett-Solberg P. Congenital adrenal hyperplasia: from genetics and biochemistry to clinical practice, part I. Clin Pediatr. 2001;40(1):1-16

5. Speiser PW, Azziz R, Baskin LS, et al. Congenital adrenal hyperplasia due to steroid 21-hydroxylase deficiency: an Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab 2010;95(9):4133-4160

6. Nordenstrom A, Falhammar H. Management of endocrine disease: Diagnosis and management of the patient with non-classic CAH due to 21-hydroxylase deficiency. Eur J Endocrinol. 2019;180(3):R127-R145

7. Young WF Jr. Primary aldosteronism: A common and curable form of hypertension. Cardiol Rev. 1999;7(4):207-214

8. Young WF Jr. Pheochromocytoma and primary aldosteronism: diagnostic approaches. Endocrinol Metab Clin North Am. 1997;26(4):801-827

9. Wudy SA, Hartmann M, Svoboda M. Determination of 17-hydroxypregnenolone in plasma by stabile isotope dilution/benchtop liquid chromatography-tandem mass spectrometry. Horm Res. 2000;53(2):68-71

10. Therrell BL. Newborn screening for congenital adrenal hyperplasia. Endocrinol Metab Clin North Am. 2001;30(1):15-30

11. Bachega TA, Billerbeck AE, Marcondes JA, Madureira G, Arnhold IJ, Mendonca BB. Influence of different genotypes on 17-hydroxyprogesterone levels in patients with nonclassical congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Clin Endocrinol. 2000;52(5):601-607

12. Kao PC, Machacek DA, Magera MJ, Lacey JM, Rinaldo P. Diagnosis of adrenal cortical dysfunction by liquid chromatography-tandem mass spectrometry. Ann Clin Lab Sci. 2001;31(2):199-204

13. Young WF Jr. Management approaches to adrenal incidentalomas. A view from Rochester, Minnesota. Endocrinol Metab Clin North Am. 2000;29(1):159-185

14. Ibanez L, DiMartino-Nardi J, Potau N, Saenger P. Premature adrenarche-normal variant or forerunner of adult disease? Endocr Rev. 2000;21(6):671-696

15. Allolio B, Arlt W. DHEA treatment: myth or reality? Trends Endocrinol Metab. 2002;13(7):288-294

16. Lin CL, Wu TJ, Machacek DA, Jiang NS, Kao PC. Urinary free cortisol and cortisone determined by high performance liquid chromatography in the diagnosis of Cushing's syndrome. J Clin Endocrinol Metab. 1997;82:151-155

17. Findling JW, Raff H. Diagnosis and differential diagnosis of Cushing's syndrome. Endocrinol Metab Clin North Am. 2001;30(3):729-747

18. Buchman Al. Side effects of corticosteroid therapy. J Clin Gastroenterol. 2001;33(4):289-297

18. Dodds HM, Taylor PJ, Cannell GR, Pond SM. A high-performance liquid chromatography-electrospray-tandem mass spectrometry analysis of cortisol and metabolites in placental perfusate. Anal Biochem. 1997;247(2):342-347

20. Cengiz H, Demirci T, Varim C, Cetin S. Establishing a new screening 17 hydroxyprogesterone cut-off value and evaluation of the reliability of the long intramuscular ACTH stimulation test in the diagnosis of nonclassical congenital adrenal hyperplasia. Eur Rev Med Pharmacol Sci. 2021;25(16):5235-5240. doi:10.26355/eurrev_202108_26537

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Method Description
Describes how the test is performed and provides a method-specific reference

Deuterated stable isotopes (d4-cortisol, d7-androstenedione, d8 17-hydroxyprogesterone) are added to the serum sample as internal standards. Cortisol, androstenedione, 17-hydroxyprogesterone, and the internal standards are extracted from specimens online using a guard cartridge. The analytes are transferred online to an analytical column and are analyzed by liquid chromatography tandem mass spectrometry.(Unpublished Mayo method)

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information


Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Monday through Friday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

2 to 5 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

14 days

Performing Laboratory Location
Indicates the location of the laboratory that performs the test


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Test Classification
Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

This test was developed, and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information
Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.


LOINC® Information
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.

Test Id Test Order Name Order LOINC Value
ANST Androstenedione, S 1854-9
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
7730 Androstenedione, S 1854-9

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Pdf

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports