Understanding Your Medical Test: What It Is, How It's Done, and How to Get Ready

Published: August 9, 2026Updated: August 15, 2026

This test measures androstenedione levels in blood samples taken at specific intervals after a hormone stimulation challenge to evaluate adrenal gland function.

Quick summary

{"Androstenedione is a hormone produced by adrenal glands and ovaries","The challenge test checks how your adrenal glands respond to stimulation","Multiple blood samples are collected over 2-4 hours after receiving a stimulating medication","The test helps diagnose conditions like adrenal insufficiency or congenital adrenal hyperplasia","No special preparation is needed except following your doctor's instructions about medications"}

Medically reviewed by WellAlly Editorial TeamLast reviewed August 15, 2026Editorial policy

Key facts at a glance

LOINC: 16382-4
Short code: AND6PC
Synonyms: Androst sp6 p chal SerPl-mCnc
Category: chemistry
Sample: serum
Container: Red-top serum tube (no anticoagulant) or serum separator tube (SST); plasma collected in heparinized (green-top) or EDTA (lavender-top) tubes is also acceptable.
Units: N/A
Fasting: false
Population: adult
Sex: all
Age range: 18–65 years
Method: Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)
CPT: 82157
SNOMED: 409622007
Specimen volume: 1.0 mL of serum or plasma (minimum 0.5 mL); draw volume should accommodate the full serial specimen protocol.
Stability: Serum/plasma stable for 7 days at 2-8°C; for longer storage, freeze at -20°C or below for up to 6 months. Avoid repeated freeze-thaw cycles.
Hemolysis max: 100
Lipemia max: 500
Icterus max: 20
TAT: 72–168 hours
Order: routine

What is this test and why might my healthcare provider recommend it for me?

The androstenedione challenge test measures the level of androstenedione in your blood at specific time points after you receive a stimulating substance (usually ACTH - adrenocorticotropic hormone). Androstenedione is a steroid hormone produced by the adrenal glands, ovaries in women, and to a lesser extent by the testes in men. This test evaluates how well your adrenal glands function and respond to stimulation.

How is this test performed, and what can I expect during the procedure?

Before the test, a healthcare provider will insert an intravenous (IV) line or draw blood from a vein in your arm. You will then receive an injection of ACTH (cosyntropin), which stimulates your adrenal glands. Blood samples are collected at baseline (before the injection) and at specific intervals afterward - typically at 30, 60, 90, and 120 minutes. The '6th specimen' refers to one of these timed samples collected after the challenge. The entire procedure usually takes 2-4 hours.

How should I prepare for this test in the days leading up to my appointment?

Follow your doctor's instructions regarding medications. You may need to stop certain medications (such as corticosteroids, hormonal treatments, or supplements that might affect hormone levels) for a period before the test. Tell your doctor about all medications you are taking. You can usually eat and drink normally unless instructed otherwise. Wear comfortable clothing with sleeves that can be easily rolled up.

Are there any risks or side effects I should be aware of before having this test?

This test is generally safe. You may experience mild discomfort when the needle is inserted or when blood is drawn. Some people may experience temporary bruising at the injection site. Rarely, individuals may have an allergic reaction to the stimulating medication. The ACTH injection may cause temporary flushing, increased heart rate, or mild nausea. These symptoms usually subside quickly.

Reference range

Reference ranges for the 6th specimen post-challenge are interpreted in the context of the stimulation/suppression protocol used and should be compared against age- and sex-matched baseline values. Consult the performing laboratory's protocol-specific reference data.N/A

Specimen: serum

No special fasting required. Specimen collection is performed as the 6th timed draw following the administration of a provocative agent (e.g., ACTH stimulation, GnRH stimulation, or hCG stimulation test). Patients should remain at rest during the collection period. Document time of challenge dose administration and exact specimen collection time.

Clinical interpretation

How abnormal results are interpreted and what conditions they suggest.

This test is primarily used to evaluate adrenal gland function and diagnose conditions affecting adrenal hormone production. It helps identify adrenal insufficiency (Addison's disease), where the adrenal glands do not produce enough hormones. The test is also valuable in diagnosing congenital adrenal hyperplasia, a genetic condition affecting cortisol production. Additionally, it can help differentiate between adrenal and ovarian sources of excess androgens in women with symptoms like hirsutism (excessive hair growth) or irregular periods.

In men, this test may be used to evaluate testicular function or investigate early or delayed puberty. The challenge test is particularly useful because it shows how the adrenal glands respond to stress or stimulation, which may not be apparent from a single baseline measurement. Abnormal results may indicate the need for further testing or treatment.

Frequently asked questions

Why do I need multiple blood samples?

Multiple samples are needed because hormone levels change over time after stimulation. The first sample (baseline) shows your resting level, while subsequent samples show how quickly and how much your adrenal glands increase androstenedione production in response to the stimulating hormone. This dynamic response provides more information than a single measurement.

How long does the test take?

The entire test typically takes 2-4 hours, depending on the specific protocol used by your healthcare provider. This includes time for the injection, waiting periods between blood draws, and the actual blood collection procedure.

Will I feel any side effects from the stimulation medication?

Some patients may experience mild side effects such as facial flushing, a warm sensation, mild nausea, increased heart rate, or temporary dizziness. These symptoms are usually short-lived and resolve within a few minutes to an hour. Serious reactions are rare.

When will I get my results?

Results are usually available within a few days to a week, as the samples need to be analyzed in a specialized laboratory. Your healthcare provider will discuss the results with you during a follow-up appointment.

Do I need to fast before this test?

Fasting is typically not required for this test. However, follow the specific instructions given by your healthcare provider, as they may have given you personalized recommendations based on your medical condition.

Sources

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This information is for educational purposes only and is not a substitute for professional medical advice, diagnosis, or treatment.

Medical Disclaimer

This content is for general informational and educational purposes only and is not a substitute for professional medical advice, diagnosis, or treatment. Always seek the guidance of a qualified healthcare provider with any questions you have about a medical condition, and never disregard professional medical advice or delay seeking it because of something you read here.

If you think you may have a medical emergency, call your local emergency number or go to the nearest emergency department immediately.

Medically reviewed by WellAlly Editorial Team Last reviewed August 15, 2026. · Read our editorial policy