Testosterone, Total (LC-MS/MS) Test
The total testosterone LC-MS/MS test quantifies all circulating testosterone using liquid chromatography–tandem mass spectrometry, the reference method that avoids immunoassay cross-reactivity and is essential for women, children, and hypogonadal men.
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Biomarkers
Blood (serum)
Sample
2–4 business days
Turnaround
Preferred but not mandatory
Fasting
What is the Total Testosterone (MS) test?
The Total Testosterone (LC-MS/MS) Test quantifies all circulating testosterone using liquid chromatography–tandem mass spectrometry, the gold-standard method that avoids the cross-reactivity and insensitivity of immunoassays at low concentrations. Practitioners order it to diagnose androgen deficiency in men, androgen excess in women, and pubertal or developmental disorders in children, and to monitor patients receiving testosterone therapy or androgen suppression. Mass spectrometry is particularly important in populations where testosterone concentrations are low or borderline—women, children, and hypogonadal men—because older immunoassays often overestimate low values or fail to detect them reliably. The result is interpreted in the context of the time of collection, symptoms, repeat sampling, and companion tests such as luteinising hormone, sex hormone-binding globulin, and, when appropriate, free or bioavailable testosterone calculated from total testosterone, SHBG, and albumin.
What does the Total Testosterone (MS) test measure?
Testosterone is the principal circulating androgen, synthesised in testicular Leydig cells under luteinising-hormone control in males and in smaller amounts by the ovarian theca and adrenal cortex in females. In the circulation roughly 98% is protein-bound—approximately 60% to sex hormone-binding globulin (SHBG) with high affinity and 38% loosely to albumin—leaving 2% free. Total testosterone measured by liquid chromatography–tandem mass spectrometry captures all circulating forms without the cross-reactivity and poor sensitivity at low concentrations that limit immunoassays, making LC-MS/MS the reference method for women, children, and hypogonadal men.
Total testosterone is the initial test for suspected androgen deficiency or excess. In adult men it is central to the diagnosis of hypogonadism, interpreted alongside clinical features and a confirmatory early-morning repeat if low. In women it helps evaluate hirsutism, virilisation, and polycystic ovary syndrome, though free or bioavailable testosterone and other androgens may add precision. In children testosterone measurement guides investigation of delayed or precocious puberty and differences of sex development. The LC-MS/MS method is essential when concentrations are expected to be low, because immunoassays lack the sensitivity and specificity required in those ranges.
Why is the Total Testosterone (MS) test ordered?
- Evaluation of suspected male hypogonadism in men with low libido, erectile dysfunction, fatigue, reduced muscle mass, or infertility, confirmed by a second early-morning sample if the first is low
- Investigation of hyperandrogenism in women presenting with hirsutism, acne, virilisation, menstrual irregularity, or suspected polycystic ovary syndrome
- Assessment of delayed puberty, precocious puberty, ambiguous genitalia, or differences of sex development in children and adolescents
- Monitoring of testosterone-replacement therapy to confirm that circulating concentrations reach the target range without supraphysiological peaks
- Monitoring of androgen-deprivation therapy in prostate cancer to verify effective suppression
- Work-up of an incidental testosterone result (immunoassay) that is borderline or discordant with the clinical picture, replaced by the more accurate LC-MS/MS measurement
- Suspected androgen-secreting tumour of the ovary, testis, or adrenal, where testosterone may be markedly elevated
- Investigation of unexplained anaemia, osteoporosis, or loss of bone density in men, in whom hypogonadism is a reversible cause
Sample collection and turnaround
Sample type
Blood (serum)
Fasting required
Preferred but not mandatory
Collection method
Venipuncture at patient service centre
Turnaround
2–4 business days
Collection notes
Phlebotomy at the Quest draw site. Collect between 07:00 and 11:00, ideally fasting, particularly in men; early-morning timing is critical because testosterone exhibits marked diurnal variation. Confirm a low male result on a repeat early-morning sample collected on a separate day. Document current testosterone or anabolic-steroid therapy, opioid or glucocorticoid use, and any acute illness, all of which alter interpretation. In women and children mass-spectrometry measurement is important for reliable quantification at low concentrations.
Specimen requirements
Minimum 1 mL serum. Separate from cells and refrigerate or freeze if not analysed promptly; testosterone is stable frozen for months.
What can affect Total Testosterone (MS) results?
Testosterone in men peaks in the early morning and declines through the day. Guidelines recommend collection between 07:00 and 11:00 for diagnostic accuracy; afternoon samples may underestimate true morning values by 20–25%.
Acute systemic illness and severe stress suppress the hypothalamic–pituitary–gonadal axis, transiently lowering testosterone; testing should be deferred until recovery to avoid misdiagnosis of permanent hypogonadism.
Chronic opioid use suppresses gonadotrophin secretion, often causing profound hypogonadism; glucocorticoids suppress the axis centrally. Both are common iatrogenic causes of low testosterone.
Total testosterone reflects both bound and free fractions; conditions that raise SHBG (ageing, hyperthyroidism, oestrogen, liver disease) can elevate total testosterone while free testosterone remains low, and vice versa. Free or bioavailable testosterone should be measured or calculated when SHBG is abnormal.
Testosterone therapy and anabolic-steroid use raise measured total testosterone; they also suppress endogenous production, so a high result in the context of a low LH indicates exogenous androgen.
Obesity lowers SHBG and increases peripheral aromatisation of testosterone to oestradiol, commonly producing borderline-low total testosterone with relatively preserved free testosterone; the clinical significance is debated and weight loss often normalises levels.
What do Total Testosterone (MS) results mean?
Elevated
In men: exogenous testosterone or anabolic-steroid use (LH suppressed), androgen-secreting tumour (testicular, adrenal; rare), resistance to androgen (even rarer). In women: polycystic ovary syndrome (modest elevation, usually <150–200 ng/dL), androgen-secreting ovarian or adrenal tumour (marked elevation, often >200 ng/dL), congenital adrenal hyperplasia, Cushing syndrome, or exogenous androgen. Interpret with DHEAS, androstenedione, 17-hydroxyprogesterone, and LH/FSH.
Normal
A normal total testosterone in a symptomatic man does not exclude androgen deficiency if SHBG is elevated, because free testosterone may be low; calculate or measure free testosterone in that setting. In women a normal total testosterone does not exclude milder hyperandrogenism if clinical features are present; free testosterone and other androgens should be assessed.
Reduced
In men: primary hypogonadism (testicular failure; LH/FSH elevated), secondary hypogonadism (hypothalamic or pituitary; LH/FSH low or inappropriately normal), chronic opioid or glucocorticoid therapy, obesity, ageing, systemic illness. Two early-morning samples below the lower reference limit with consistent symptoms are required for diagnosis. In women: primary ovarian insufficiency, hypopituitarism, or adrenal insufficiency, though low female testosterone is rarely symptomatic or clinically significant.
Note: Total testosterone must be collected in the early morning in men, confirmed on a second sample if low, and interpreted with LH, FSH, and SHBG. LC-MS/MS is the method of choice in women, children, and men with expected low values. A result is valid only if the patient is clinically stable; defer testing during acute illness. Reference intervals are sex-, age-, and assay-specific, and 'low normal' in an older man may not be pathological.
Frequently asked questions
Liquid chromatography–tandem mass spectrometry is the reference method because it avoids the cross-reactivity and poor sensitivity at low concentrations that limit immunoassays. It is essential for accurate measurement in women, children, and hypogonadal men, where testosterone levels are low and immunoassays often overestimate or fail to detect them reliably.
How to access the Total Testosterone (MS) test
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Published: October 7, 2026
Last reviewed: October 7, 2026
- Bhasin S, et al. Testosterone therapy in men with hypogonadism: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. (2018)
- Rosner W, et al. Position statement: utility, limitations, and pitfalls in measuring testosterone — an Endocrine Society position statement. J Clin Endocrinol Metab. (2007)
- Legro RS, et al. Diagnosis and treatment of polycystic ovary syndrome: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. (2013)
- Stanworth RD, Jones TH. Testosterone for the aging male; current evidence and recommended practice. Clin Interv Aging. (2008)
- Wang C, et al. Investigation, treatment, and monitoring of late-onset hypogonadism in males: ISA, ISSAM, EAU, EAA, and ASA recommendations. J Androl. (2009)
This content is for licensed healthcare practitioners only and does not constitute medical advice. Clinical decisions should be based on the full clinical picture and not on any single test result.