What is the advantage of LC-MS over immunoassay for salivary hormones?

Liquid chromatography-tandem mass spectrometry separates and quantifies steroids without antibody cross-reactivity, providing greater specificity for low-concentration analytes and for patients on exogenous hormones that can interfere with immunoassays.

Can a single salivary cortisol diagnose adrenal insufficiency or Cushing syndrome?

No. A late-night salivary cortisol is validated for Cushing screening, but a single daytime or unspecified-time result does not replace formal dynamic testing. Adrenal insufficiency requires ACTH stimulation or morning serum cortisol; a low single saliva value is not diagnostic.

How should patients time the saliva collection?

Timing is critical. Menstruating women should collect sex-steroid samples in the mid-luteal phase (days 19–21 of a 28-day cycle) unless otherwise instructed. Cortisol timing must be recorded precisely; a bedtime sample is used for Cushing screening, while waking or daytime samples assess different parts of the diurnal curve.

What precautions prevent contamination of the saliva sample?

Avoid food, drink, and tooth-brushing for at least 30 minutes before collection. Patients using topical oestradiol, progesterone, or testosterone must apply it away from the face and mouth and avoid touching the collection device to the lips, as contamination produces spuriously elevated results.

Are salivary hormone reference ranges interchangeable with serum?

No. Salivary concentrations are a small fraction of serum total levels and represent only the free hormone. Reference intervals are assay- and platform-specific and not interchangeable with serum thresholds or with other saliva laboratories.

What medications or conditions interfere with salivary hormone results?

Exogenous hormones, oral contraceptives, spironolactone, and glucocorticoids all alter results. Acute stress or illness transiently raises cortisol. Gum disease or mouth bleeding can contaminate samples. A full medication and supplement history is essential for interpretation.

Is this panel suitable for diagnosing polycystic ovary syndrome or menopause?

It provides supporting data — elevated testosterone in PCOS or low oestradiol in menopause — but diagnosis requires clinical criteria, serum confirmation where appropriate, and often ultrasound or other investigations. Salivary results are an adjunct, not a stand-alone diagnostic.

How long do results take and how are samples returned?

Results are typically available within 5–7 business days of ZRT Laboratory receiving the specimen. Samples are mailed at room temperature in the provided kit; follow the kit instructions for timing to ensure stability.

BML-ADR-005adrenal stressUSA

Saliva LC-MS Hormones 5 Panel

The Saliva LC-MS Hormones 5 Panel quantifies free oestradiol, progesterone, testosterone, DHEA, and a single time-point cortisol by liquid chromatography-tandem mass spectrometry. It is used to assess bioavailable sex-steroid and adrenal-androgen status, monitor transdermal hormone therapy, and investigate symptoms attributed to hormone imbalance, with interpretation against platform-specific…

Biomarkers

Saliva

Sample

5–7 business days

Turnaround

No, but avoid food, drink, and tooth-brushing for at least 30 minutes before collection

Fasting

What is the Saliva LC-MS 5 Panel test?

The Saliva LC-MS Hormones 5 Panel measures oestradiol, progesterone, testosterone, DHEA, and a single time-point cortisol by liquid chromatography-tandem mass spectrometry in saliva. It is ordered by practitioners investigating symptoms attributed to hormone imbalance — fatigue, mood changes, menstrual irregularity, libido disturbance, or signs of androgen excess or deficiency — and by those monitoring transdermal or sublingual hormone replacement, where salivary concentrations may better reflect tissue delivery than serum. The panel sits within functional and integrative medicine rather than mainstream endocrine diagnosis. Salivary steroid measurement offers the advantage of capturing free, bioavailable hormone without venepuncture, and LC-MS avoids the cross-reactivity that limits immunoassay. Yet reference data are platform-specific, single time-point cortisol does not replace validated adrenal function tests, and many interpretive thresholds lack large-scale validation. Practitioners use this panel as one input into a broader clinical assessment rather than as a stand-alone diagnostic.

What does the Saliva LC-MS 5 Panel test measure?

E2(Oestradiol (Saliva))

Range: Luteal phase approximately 1.3–3.3 pg/mL; ranges vary by sex, menstrual phase, menopausal status, and assay platform.Optimal: Optimal ranges are context-dependent: pre-menopausal women cycle through follicular and luteal levels, while post-menopausal women on replacement aim for mid-follicular equivalents (approximately 1.5–3 pg/mL in saliva). Functional-medicine targets vary and should be individualised.pg/mL / pmol/L

Oestradiol in saliva represents the free, biologically active fraction unbound to sex hormone-binding globulin. Salivary concentrations are roughly one per cent of serum total oestradiol but reflect tissue-available hormone. LC-MS quantifies the steroid directly without cross-reactivity from metabolites or exogenous oestrogens, making it preferable to immunoassay for patients on hormone therapy.

Salivary oestradiol is used to assess ovarian function, investigate symptoms of oestrogen deficiency or excess, and monitor transdermal or sublingual oestradiol therapy, where serum assays can underestimate tissue delivery. It is not interchangeable with serum oestradiol and should be interpreted against saliva-specific reference data.

P4(Progesterone (Saliva))

Range: Luteal phase approximately 75–270 pg/mL; follicular phase and post-menopause substantially lower.Optimal: Mid-luteal saliva progesterone above approximately 100 pg/mL is generally considered evidence of ovulation; higher values (150–300 pg/mL) are associated with robust luteal function. Post-menopausal women not on therapy typically show values below 20 pg/mL.pg/mL / nmol/L

Salivary progesterone measures the free, unbound steroid that diffuses into saliva. The concentration is a small fraction of serum but closely mirrors tissue-level progesterone. Collection timing is critical: luteal-phase sampling (days 19–21 of a 28-day cycle) captures the corpus-luteum peak, while follicular values are near the assay limit of detection.

Practitioners order salivary progesterone to confirm ovulation, assess luteal sufficiency, and investigate symptoms attributed to progesterone deficiency. It is also used to monitor oral micronised or transdermal progesterone, where first-pass metabolism or binding proteins complicate serum interpretation.

T(Testosterone (Saliva))

Range: Adult men approximately 50–100 pg/mL; pre-menopausal women approximately 20–40 pg/mL; ranges are method- and population-specific.Optimal: Optimal salivary testosterone depends on sex, age, and clinical context. Men with symptoms of hypogonadism and saliva values below approximately 40 pg/mL may benefit from further evaluation; women with values above 50 pg/mL warrant investigation for hyperandrogenism. Functional targets are not universally agreed.pg/mL / pmol/L

Testosterone in saliva represents free, albumin-bound, and other loosely bound fractions that equilibrate across the salivary gland. It correlates well with calculated free serum testosterone and is less influenced by sex hormone-binding globulin than total serum testosterone, making it a useful marker of androgen bioavailability.

Salivary testosterone is ordered to evaluate androgen status in men with suspected hypogonadism or in women with hirsutism, androgenic alopecia, or polycystic ovary syndrome. It is also used to monitor testosterone replacement, particularly transdermal formulations. Results should be correlated with clinical signs rather than treated as diagnostic in isolation.

DHEA(Dehydroepiandrosterone (Saliva))

Range: Adult reference ranges approximately 60–400 pg/mL, age- and sex-dependent; DHEA declines with age.Optimal: Age-adjusted midpoints are often used as targets, with younger adults showing higher values. Functional-medicine protocols sometimes aim for the upper half of the reference range, but evidence for this approach is limited and the marker should be interpreted in the context of symptoms and other adrenal measures.pg/mL / nmol/L

DHEA is an adrenal androgen precursor secreted in response to ACTH. Salivary DHEA measures the unconjugated steroid, which is distinct from the serum sulphate conjugate (DHEA-S) measured in most blood assays. The free form in saliva reflects short-term adrenal output and tissue availability, and its concentration is markedly lower than serum DHEA-S.

Practitioners use salivary DHEA to assess adrenal androgen reserve and as a component of 'adrenal stress' or functional-hormone panels. Low values are interpreted as a sign of diminished adrenal function, though the clinical significance of isolated salivary DHEA reduction is contested and not recognised in mainstream endocrinology guidelines.

Cortisol(Cortisol, single time-point (Saliva))

Range: Morning (waking) approximately 3.7–9.5 ng/mL; afternoon approximately 1.2–3.0 ng/mL; evening/bedtime approximately 0.4–1.0 ng/mL. Interpretation depends on the collection time.Optimal: For a late-night (bedtime) sample, values below 1.0 ng/mL are typically normal; elevation above that threshold warrants further evaluation for Cushing syndrome. For waking or daytime samples, optimal ranges are defined by the time of collection and the clinical question, and single-point interpretation is fraught with biological variability.ng/mL / nmol/L

Cortisol in saliva represents free, unbound glucocorticoid that crosses into saliva by passive diffusion. It follows a diurnal rhythm, peaking shortly after waking and declining through the day. A single time-point sample provides a snapshot of free cortisol at that moment but does not capture the rhythm; diurnal assessment requires multiple timed samples.

A single salivary cortisol is most often ordered as part of a hormone panel rather than for adrenal diagnosis. Late-night salivary cortisol is validated for Cushing syndrome screening, but a daytime or unspecified-time result has limited diagnostic value and should not be used to diagnose adrenal insufficiency or hypercortisolism. Functional practitioners interpret single values in the context of symptoms, but this approach lacks robust evidence.

Why is the Saliva LC-MS 5 Panel test ordered?

Investigation of suspected oestrogen or progesterone deficiency in pre-menopausal women with menstrual irregularity, anovulation, or luteal-phase symptoms
Monitoring of transdermal oestradiol or progesterone therapy, where serum assays may underestimate tissue-level hormone or be confounded by first-pass metabolism
Assessment of androgen status in women with hirsutism, androgenic alopecia, or suspected polycystic ovary syndrome, where free testosterone is more informative than total
Evaluation of suspected androgen deficiency in men considering testosterone replacement, particularly when SHBG is high or serum free testosterone is equivocal
Functional assessment of adrenal androgen reserve (DHEA) and diurnal cortisol output in the context of fatigue or 'adrenal stress,' interpreted with caution given contested clinical utility
Patients who prefer non-invasive collection or for whom venepuncture is problematic, provided the practitioner understands the limitations of saliva-specific reference data

Sample collection and turnaround

Sample type

Saliva

Fasting required

No, but avoid food, drink, and tooth-brushing for at least 30 minutes before collection

Collection method

At-home collection kit

Turnaround

5–7 business days

Collection notes

Collect per kit instructions, timing the sample to the menstrual cycle where relevant and recording the exact collection time for cortisol interpretation. Avoid food, drink, and tooth-brushing before collection, and avoid topical hormone contamination of the mouth and device. Document hormone therapy and route and any relevant medications. Note that this panel provides a single cortisol, not a diurnal curve.

Specimen requirements

Saliva collected into the laboratory-provided vial or swab device, typically 1–2 mL. Samples are stable at room temperature for mailing within the timeframe specified in the kit instructions. Record collection date, time, cycle day (if menstruating), and hormone medications on the requisition.

What can affect Saliva LC-MS 5 Panel results?

elevatesTopical hormone contamination

Transdermal or sublingual oestradiol, progesterone, or testosterone can contaminate the mouth and saliva collection device, producing spuriously high results. Practitioners should instruct patients to apply hormones well away from the face and mouth and to avoid touching the applicator to the lips.

variableRecent food, drink, or tooth-brushing

Blood contamination from gum irritation or food residue can alter steroid concentrations. Collection should occur after at least 30 minutes without food, drink, or oral hygiene, and patients with active gingivitis may not be suitable candidates.

variableTime of day and menstrual cycle phase

Cortisol follows a steep diurnal rhythm and sex steroids cycle with the menstrual phase. Without documenting exact collection time and cycle day, interpretation is severely limited. A single cortisol without time-stamp is nearly uninterpretable.

variableStress, illness, and medications

Acute stress or illness can transiently raise cortisol; exogenous glucocorticoids suppress endogenous cortisol and DHEA. Spironolactone, oral contraceptives, and other hormone-modulating drugs alter sex-steroid concentrations and binding.

variableAssay platform and reference interval

Salivary steroid ranges are method-specific and not interchangeable with serum or with other saliva assays. Results must be interpreted against the performing laboratory's validated intervals, which for specialty labs may be proprietary or based on smaller reference populations.

What do Saliva LC-MS 5 Panel results mean?

Elevated

Elevated salivary oestradiol or testosterone may indicate excess endogenous production (PCOS, ovarian or adrenal tumour) or over-replacement with exogenous therapy. High progesterone outside the luteal phase suggests exogenous supplementation or contamination. Elevated DHEA can reflect adrenal hyperplasia or a DHEA supplement. A single elevated cortisol, if collected late at night, raises suspicion for Cushing syndrome and warrants further testing; a daytime elevation is less specific.

Normal

Normal results suggest that free, bioavailable hormone concentrations fall within the laboratory's reference population at the time and cycle phase sampled. This does not exclude all hormone imbalance, as symptoms may arise from receptor sensitivity, rhythm disturbances, or metabolite patterns not captured by these five analytes. Clinical correlation is essential.

Reduced

Low oestradiol or progesterone in a pre-menopausal woman may indicate anovulation, luteal insufficiency, or approaching menopause; in post-menopausal women low values are expected unless on replacement. Low testosterone in men suggests hypogonadism; in women it is common and of uncertain significance. Low DHEA is interpreted as diminished adrenal reserve in functional models but lacks validated diagnostic thresholds. A single low cortisol does not diagnose adrenal insufficiency — formal ACTH stimulation or serum morning cortisol is required.

Note: Salivary steroid measurement is a tool, not a diagnosis. Reference ranges are assay- and population-specific and may not match serum-based diagnostic thresholds. A single cortisol time-point does not assess the diurnal curve or replace validated adrenal function tests. Contamination from topical hormones is common and must be ruled out. Results should be interpreted by a practitioner familiar with salivary LC-MS data and integrated with history, examination, and where appropriate, confirmatory serum or dynamic testing.

Frequently asked questions

Saliva captures free, unbound steroid hormones that reflect tissue-available concentrations. It is particularly useful for monitoring transdermal or sublingual hormone therapy, where serum total levels may not accurately represent bioavailability, and for patients who prefer non-invasive collection.

How to access the Saliva LC-MS 5 Panel test

The Saliva LC-MS 5 Panel reference page is free and open to all practitioners. Create a free practitioner account to see pricing, save tests to your list, and generate requisition forms. Verification takes 1–2 business days.

Create Free Account

About this page

Published: October 7, 2026

Last reviewed: October 7, 2026

References:

Kirschbaum C, Hellhammer DH. Salivary cortisol in psychoneuroendocrine research: recent developments and applications. Psychoneuroendocrinology. (1994)
Stanczyk FZ, et al. Standardization of steroid hormone assays: why, how, and when? Cancer Epidemiol Biomarkers Prev. (2007)
Nieman LK, et al. The diagnosis of Cushing's syndrome: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. (2008)
Taylor AE, et al. Determination of steroid hormones in saliva: applications and recommendations. Bioanalysis. (2015)
Wood P. Salivary steroid assays — research or routine? Ann Clin Biochem. (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.

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