Saliva LC-MS Hormones 5 with 4-Point Cortisol Panel
The Saliva LC-MS Hormones 5 with 4-Point Cortisol panel measures five salivary steroid hormones — oestradiol, progesterone, testosterone, and DHEA-S — plus a four-point diurnal cortisol curve, analysed by liquid chromatography-mass spectrometry from at-home saliva collections.
5
Biomarkers
Saliva
Sample
5–10 business days
Turnaround
No — however, patients must follow pre-collection instructions (avoiding food, drink, and brushing before sampling) and collect the cortisol samples at the specified times on a representative day
Fasting
What is the Saliva Hormones 5 + 4-Point Cortisol test?
The Saliva LC-MS Hormones 5 with 4-Point Cortisol panel measures five salivary steroid hormones — oestradiol, progesterone, testosterone, and DHEA-S — together with a four-point diurnal cortisol curve, all from at-home saliva collections analysed by liquid chromatography-mass spectrometry. Its purpose is to profile bioavailable sex and adrenal hormones alongside the daily cortisol rhythm in a single integrated assessment. Within hormone assessment, this is a functional and integrative panel rather than a replacement for established endocrine diagnostics. The salivary route is taken to reflect free, bioavailable hormone, and the LC-MS methodology offers analytical specificity over immunoassay. It appeals to practitioners seeking a pattern-based view of sex-hormone balance and the cortisol curve from convenient at-home collection. Its reference intervals are proprietary to the ZRT method and not interchangeable with serum or other salivary assays, and salivary steroid measurement is strongly affected by collection technique and topical hormone contamination. It does not replace serum measurement or dynamic testing — such as the ACTH stimulation or dexamethasone suppression tests, or serum sex-steroid and SHBG measurement — when a defined endocrine disorder is suspected.
What does the Saliva Hormones 5 + 4-Point Cortisol test measure?
Oestradiol is the most potent endogenous oestrogen, produced mainly by the ovaries with peripheral contributions from aromatisation in adipose tissue. The salivary fraction is taken to reflect the free, bioavailable hormone, as only unbound steroid is thought to enter saliva. Measured by liquid chromatography-mass spectrometry, salivary oestradiol is offered as an index of bioavailable oestrogen status across the cycle and life stages.
Salivary oestradiol is used in functional and integrative practice to assess bioavailable oestrogen, particularly in the context of menstrual cycle characterisation, the menopausal transition, and hormone therapy monitoring. It does not replace serum oestradiol for diagnosing defined endocrine disorders, and salivary measurement is influenced by collection technique and any topical hormone use.
Progesterone is the principal luteal-phase hormone, produced by the corpus luteum after ovulation and by the placenta in pregnancy. Salivary progesterone is taken to reflect the free fraction available to tissues. It rises markedly in the luteal phase and is minimal in the follicular phase and in postmenopausal women not on therapy.
Salivary progesterone is used to assess luteal-phase progesterone and the oestrogen-to-progesterone balance in functional hormone assessment, and to monitor progesterone therapy. Because levels are highly cycle-dependent, collection timing is essential, and salivary measurement is sensitive to topical progesterone contamination.
Testosterone is the principal androgen, produced by the gonads and adrenal glands. The salivary fraction is taken to reflect free, bioavailable testosterone, unbound to sex hormone-binding globulin. Measured by LC-MS, salivary testosterone is offered as an index of bioavailable androgen status in both sexes.
Salivary testosterone is used in functional practice to assess bioavailable androgen status, relevant to androgenic symptoms, libido, and hormone therapy monitoring in both sexes. It does not replace serum testosterone with SHBG and calculated free testosterone for diagnosing hypogonadism or androgen excess, where serum measurement is the standard.
DHEA-S is the principal adrenal androgen precursor, produced by the adrenal cortex and declining progressively with age. Salivary DHEA-S is taken to reflect adrenal androgen output. It is interpreted as a marker of adrenal steroidogenic activity alongside cortisol.
Salivary DHEA-S is used to assess adrenal androgen production, often interpreted together with the cortisol curve as part of a broader adrenal and hormonal assessment in functional practice. Serum DHEA-S remains the more established measure for defined adrenal disorders.
Cortisol is the principal glucocorticoid, secreted by the adrenal cortex under hypothalamic-pituitary (CRH-ACTH) control with a marked diurnal rhythm peaking after waking and declining to a nocturnal nadir. Salivary cortisol reflects the free, biologically active fraction. Four samples across the day are collected to construct the diurnal curve — its morning peak, daytime decline, and nighttime nadir.
The four-point salivary cortisol curve is used in functional and integrative practice to characterise the shape of the daily cortisol rhythm rather than a single value, assessing patterns attributed to chronic stress, fatigue, and circadian disruption. Late-night salivary cortisol has an established role in screening for Cushing's syndrome, but the functional curve interpretation is distinct and does not replace dynamic endocrine testing for diagnosing adrenal disease.
Why is the Saliva Hormones 5 + 4-Point Cortisol test ordered?
- Functional assessment of sex-hormone balance combining bioavailable oestradiol, progesterone, testosterone, and DHEA-S from a single at-home collection
- Characterisation of the diurnal cortisol rhythm in patients with chronic stress, fatigue, or disrupted sleep using the four-point curve
- Monitoring of hormone therapy where bioavailable salivary hormone levels are tracked over time
- Assessment of suspected oestrogen-progesterone imbalance across the cycle or menopausal transition
- Evaluation of androgen and adrenal steroid status combining testosterone and DHEA-S with the cortisol curve
- Integrative workups requiring sex-steroid and adrenal-rhythm markers from non-invasive saliva collection rather than venous draws
- Functional follow-up of adrenal-support or hormone interventions where a pattern-based salivary profile is preferred
Sample collection and turnaround
Sample type
Saliva
Fasting required
No — however, patients must follow pre-collection instructions (avoiding food, drink, and brushing before sampling) and collect the cortisol samples at the specified times on a representative day
Collection method
At-home collection kit
Turnaround
5–10 business days
Collection notes
Collect the four cortisol samples at the specified times (typically morning, noon, evening, and night) and the sex-steroid sample per kit instructions, timed to the cycle where relevant. Avoid food, drink, and tooth-brushing before collection, and avoid topical hormone contamination of the mouth and device. Document hormone therapy and route, medications, and collect on a typical day.
Specimen requirements
Saliva collection tubes per the ZRT Laboratory kit; minimum volumes as specified. Follow the laboratory's stability, storage, and transport requirements (saliva samples are typically frozen or returned promptly per instructions).
What can affect Saliva Hormones 5 + 4-Point Cortisol results?
The diurnal cortisol curve depends on collecting saliva at the specified times across the day, and sex-steroid interpretation depends on cycle timing. Mistimed or missed collections invalidate the rhythm and cycle interpretation.
Topical hormone preparations can contaminate the mouth and collection device, producing markedly and falsely elevated salivary steroid results. Hormone therapy, route, and application site must be documented, and contamination precautions followed.
Eating, drinking, vigorous brushing, and gingival bleeding before collection can introduce blood or food into saliva and distort steroid results. Patients should follow pre-collection abstinence and rinsing instructions.
Stimulated versus passive-drool collection and salivary flow rate can affect measured concentrations, so a consistent, specified collection method is essential for reliable and comparable results.
Corticosteroids, hormonal contraceptives, and agents altering steroid synthesis or metabolism can change the profile. A full medication and supplement history is required.
Altered sleep-wake timing, intercurrent illness, and acute stress shift the cortisol rhythm, so collections should ideally be made on a representative, typical day.
What do Saliva Hormones 5 + 4-Point Cortisol results mean?
Elevated
Elevated salivary sex steroids are interpreted against sex, cycle phase, menopausal status, and any hormone therapy, with topical contamination a key cause of spuriously high results to exclude first. An elevated or non-declining cortisol curve may reflect stress or, if markedly raised especially at night, prompt consideration of hypercortisolism requiring confirmation by established testing.
Normal
Results within the method's reference intervals, with an expected morning cortisol peak and progressive daytime decline, suggest a typical pattern. Because these intervals are proprietary and pattern-based, a normal profile does not exclude a defined endocrine disorder warranting conventional testing.
Reduced
Low salivary sex steroids are read against age, sex, and reproductive status, and may reflect ovarian or testicular decline or suppression. A flattened or low cortisol curve is interpreted in functional practice as a pattern associated with chronic stress or fatigue, but is not diagnostic of adrenal insufficiency, which requires ACTH stimulation testing.
Note: This is a functional, pattern-based, proprietary salivary panel whose LC-MS reference intervals are method-specific and not interchangeable with serum or other salivary assays. It is highly dependent on collection timing, technique, and avoidance of topical-hormone and oral contamination. It does not replace serum sex-steroid measurement or dynamic adrenal testing for diagnosing defined disorders, and results should be interpreted by a practitioner familiar with the methodology.
Frequently asked questions
It measures five salivary steroid hormones — oestradiol, progesterone, testosterone, and DHEA-S — together with a four-point diurnal cortisol curve, analysed by liquid chromatography-mass spectrometry from at-home saliva samples.
How to access the Saliva Hormones 5 + 4-Point Cortisol test
The Saliva Hormones 5 + 4-Point Cortisol 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.
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Published: June 2, 2026
Last reviewed: June 2, 2026
- Nieman LK, et al. The diagnosis of Cushing's syndrome: an Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. (2008)
- Raff H. Utility of salivary cortisol measurements in Cushing's syndrome and adrenal insufficiency. J Clin Endocrinol Metab. (2009)
- Gröschl M. Current status of salivary hormone analysis. Clin Chem. (2008)
- Keevil BG. LC-MS/MS analysis of steroids in the clinical laboratory. Clin Biochem. (2016)
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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