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22294 Androstenedione, Serum (ANST)

Androstenedione, Serum (ANST)
Test Code: AND-SO
Synonyms/Keywords
4-Androstene-3, 17-dione; 4-Androstenedione; Delta-4-Androstenedione
Useful For

Diagnosis and differential diagnosis of hyperandrogenism (in conjunction with measurements of other sex-steroids). An initial workup in adults might 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 (SHBG / Sex Hormone Binding Globulin [SHBG], Serum) and other androgenic steroids (eg, dehydroepiandrosterone sulfate [DHEA-S]).

Diagnosis of congenital adrenal hyperplasia (CAH), in conjunction with measurement of other androgenic precursors, particularly, 17-alpha-hydroxyprogesterone (OHPG) (OHPG / 17-Hydroxyprogesterone, Serum), 17 alpha-hydroxypregnenolone, DHEA-S (DHES / Dehydroepiandrosterone Sulfate [DHEA-S], Serum), and cortisol (CORT / Cortisol, Serum).

Monitoring CAH treatment, in conjunction with testosterone (TTST / Testosterone, Total, Serum), OHPG (OHPG / 17-Hydroxyprogesterone, Serum), DHEA-S (DHES / Dehydroepiandrosterone Sulfate [DHEA-S], Serum), and DHEA (DHEA_ / Dehydroepiandrosterone [DHEA], Serum).

Diagnosis of premature adrenarche, in conjunction with gonadotropins (FSH / Follicle-Stimulating Hormone [FSH], Serum; LH / Luteinizing Hormone [LH], Serum) and other adrenal and gonadal sex-steroids and their precursors (TTBS / Testosterone, Total and Bioavailable, Serum or TGRP / Testosterone, Total and Free, Serum; EEST / Estradiol, Serum; DHES / Dehydroepiandrosterone Sulfate [DHEA-S], Serum; DHEA_ / Dehydroepiandrosterone [DHEA], Serum; SHBG / Sex Hormone Binding Globulin [SHBG], Serum; OHPG / 17-Hydroxyprogesterone, Serum).

Specimen Requirements
Specimen Type Preferred Container/Tube Acceptable Container/Tube Specimen Volume Specimen Minimum Volume
(allows for 1 repeat)
Pediatric Minimum Volume
(no repeat)
​Serum Red Top Tube (RTT)​ 0.6 mL​ 0.25 mL​
Specimen Stability Information
Specimen Type Temperature Time
Serum​ ​ Refrigerated (preferred)​ 28 days​
Frozen ​ 28 days​
​Ambient​7 days
Rejection Criteria
Gross hemolysis
​Gross lipemia
​Serum gel tube
Interference

​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.

Performing Laboratory Information
Performing Location Day(s) Test Performed Report Available Methodology/Instrumentation
Mayo Clinic Laboratories
Monday through Friday​
2 to 5 days
Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)​
Reference Lab
Reference Range Information
PEDIATRICS
      Premature infants 26-28 weeks, day 4:  92-282 ng/dL
      Premature infants 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

Males
Tanner Stages                  Age (years)         Reference Range (ng/dL)
Stage I (prepubertal)            <9.8                           <51  
Stage II                                9.8-14.5                     31-65    
Stage III                             10.7-15.4                    50-100
Stage IV                             11.8-16.2                    48-140  
Stage V                              12.8-17.3                    65-210 

Females
Tanner Stages                  Age (years)         Reference Range (ng/dL)
Stage I prepubertal)             <9.2                            <51        
Stage II                                9.2-13.7                     42-100
Stage III                             10.0-14.4                     80-190
Stage IV                             10.7-15.6                     77-225
Stage V                              11.8-18.6                     80-240

ADULTS
Males: 40-150 ng/dL
Females: 30-200 ng/dL
Interpretation

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 also overproduce androstenedione, but 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 >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) mutations 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 mutation, androstenedione levels are elevated to a similar extend as in CYP21A2 mutation, and cortisol is also low, but OHPG is only mildly, if at all, elevated.

Also less common, 3 beta HSD-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 below the age of 7 to 8 and boys before age 8 to 9 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. 

Outreach CPTs
CPT Modifier
(if needed)
Quantity Description Comments
​82157
Synonyms/Keywords
4-Androstene-3, 17-dione; 4-Androstenedione; Delta-4-Androstenedione
Ordering Applications
Ordering Application Description
​Centricity ​None
​Cerner ​Androstenedione (9709)
If the ordering application you are looking for is not listed, contact your local laboratory for assistance.
Specimen Requirements
Specimen Type Preferred Container/Tube Acceptable Container/Tube Specimen Volume Specimen Minimum Volume
(allows for 1 repeat)
Pediatric Minimum Volume
(no repeat)
​Serum Red Top Tube (RTT)​ 0.6 mL​ 0.25 mL​
Specimen Stability Information
Specimen Type Temperature Time
Serum​ ​ Refrigerated (preferred)​ 28 days​
Frozen ​ 28 days​
​Ambient​7 days
Rejection Criteria
Gross hemolysis
​Gross lipemia
​Serum gel tube
Interference

​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.

Useful For

Diagnosis and differential diagnosis of hyperandrogenism (in conjunction with measurements of other sex-steroids). An initial workup in adults might 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 (SHBG / Sex Hormone Binding Globulin [SHBG], Serum) and other androgenic steroids (eg, dehydroepiandrosterone sulfate [DHEA-S]).

Diagnosis of congenital adrenal hyperplasia (CAH), in conjunction with measurement of other androgenic precursors, particularly, 17-alpha-hydroxyprogesterone (OHPG) (OHPG / 17-Hydroxyprogesterone, Serum), 17 alpha-hydroxypregnenolone, DHEA-S (DHES / Dehydroepiandrosterone Sulfate [DHEA-S], Serum), and cortisol (CORT / Cortisol, Serum).

Monitoring CAH treatment, in conjunction with testosterone (TTST / Testosterone, Total, Serum), OHPG (OHPG / 17-Hydroxyprogesterone, Serum), DHEA-S (DHES / Dehydroepiandrosterone Sulfate [DHEA-S], Serum), and DHEA (DHEA_ / Dehydroepiandrosterone [DHEA], Serum).

Diagnosis of premature adrenarche, in conjunction with gonadotropins (FSH / Follicle-Stimulating Hormone [FSH], Serum; LH / Luteinizing Hormone [LH], Serum) and other adrenal and gonadal sex-steroids and their precursors (TTBS / Testosterone, Total and Bioavailable, Serum or TGRP / Testosterone, Total and Free, Serum; EEST / Estradiol, Serum; DHES / Dehydroepiandrosterone Sulfate [DHEA-S], Serum; DHEA_ / Dehydroepiandrosterone [DHEA], Serum; SHBG / Sex Hormone Binding Globulin [SHBG], Serum; OHPG / 17-Hydroxyprogesterone, Serum).

Reference Range Information
PEDIATRICS
      Premature infants 26-28 weeks, day 4:  92-282 ng/dL
      Premature infants 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

Males
Tanner Stages                  Age (years)         Reference Range (ng/dL)
Stage I (prepubertal)            <9.8                           <51  
Stage II                                9.8-14.5                     31-65    
Stage III                             10.7-15.4                    50-100
Stage IV                             11.8-16.2                    48-140  
Stage V                              12.8-17.3                    65-210 

Females
Tanner Stages                  Age (years)         Reference Range (ng/dL)
Stage I prepubertal)             <9.2                            <51        
Stage II                                9.2-13.7                     42-100
Stage III                             10.0-14.4                     80-190
Stage IV                             10.7-15.6                     77-225
Stage V                              11.8-18.6                     80-240

ADULTS
Males: 40-150 ng/dL
Females: 30-200 ng/dL
Interpretation

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 also overproduce androstenedione, but 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 >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) mutations 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 mutation, androstenedione levels are elevated to a similar extend as in CYP21A2 mutation, and cortisol is also low, but OHPG is only mildly, if at all, elevated.

Also less common, 3 beta HSD-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 below the age of 7 to 8 and boys before age 8 to 9 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 visit:
Performing Laboratory Information
Performing Location Day(s) Test Performed Report Available Methodology/Instrumentation
Mayo Clinic Laboratories
Monday through Friday​
2 to 5 days
Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)​
Reference Lab
For billing questions, see Contacts
Outreach CPTs
CPT Modifier
(if needed)
Quantity Description Comments
​82157
For most current information refer to the Marshfield Laboratory online reference manual.